Imporntate - Advances in Present-Day Frozen Dough Technology and Its Improver and Novel Bio-Tech Ingredients Development Trends - A Review.

DR.
WEINING HUANG (Orcid ID : 0000-0003-1411-1353)
Article type : Review

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Title: Advances in present-day frozen dough technology and its improver and novel bio-tech
ingredients development trends_a review
Jacob Ojobi OMEDI1 , and Weining HUANG*
1
State Key Laboratory of Food Science and Technology; Laboratory of Baking and Fermentation Science,
Cereal/Sourdough and Ingredient Functionality Research, School of Food Science and Technology,
Jiangnan University, Wuxi 214122, China
*Corresponding author. Tel.: +86(510)8591 9139; fax: +86(510)8591 9139. E-mail address:
wnhuang@jiangnan.edu.cn (W. Huang).
ABSTRACT
Background and objectives: In response to the need for product flexibility and fast response to consumer
trends, research interest frozen dough technology has continued to increase since its inception in 1970s.
Common categories of these products are pre-fermented, unfermented and par-baked frozen dough
products; with widely known frozen dough products such as refrigerated cookies and brownies, sweet
rolls, biscuits, dinner rolls, and pizza, sold “as if freshly baked” to the consumer. The underlying catalyst
for the development and growth of the frozen dough products in the 1970s to 1990s is closely related the
development of improver technology and increased research efforts to improve frozen dough quality; thus
a steady market growth in frozen dough products in the following. Despite increased popularity of frozen
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dough products, they are still faced with processing challenges such as reduced yeast activity and viability,
damaged dough gluten-starch network and alteration of individual dough components which are induced
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by freezing and frozen storage treatment. Therefore, increased research efforts by both industry and
academic institutions have been vital in understanding and improving processing parameters as well as the
end product quality of frozen bread-dough products.
This review provides an overview of the effect of ice crystal characteristics on water redistribution and
major components (starch and gluten) and ingredient (yeast) during freezing and frozen storage of frozen
bread-dough systems. Additionally, advances in frozen dough improver technology such as use of
hydrocolloids, ice structuring proteins, ice nucleating agents, and novel fermentation end products and
enzymes that alter ice crystal characteristics to improve steamed bread frozen dough quality are suggested
and discussed. In addition, due to the recent increase in popularity of frozen steamed products, they are
used as a case study and some critical conditions for their processing were suggested.
Findings: Freezing and frozen treatment on key components. Gluten: Gluten protein quality is mainly
indicated by GMP depolymerisation during freezing and frozen storage. In frozen steamed dough systems,
SH groups, an indicator of S-S bond disruption, increased in gluten but fluctuated in glutenin fraction
during frozen storage. This invariably increases the molecular weight distribution of the glutenin subunits
(HMW-GS and LMW-GS). Starch: Freezing and frozen storage induces granule structure re-organization;
decline in amorphous state coupled with increase in crystallinity as starch granule materials leach out,
influences the A- and B- type granules differently. Yeast: Freezing and frozen storage treatment decreases
yeast activity and viability in frozen dough systems; with the degree of damage related to the freezing
conditions used (e.g. freezing rates, ice crystal size and location and ice re-crystallization) and ability to
metabolize different molecules (e.g. glycerol, trehalose, proline, arginine). Despite loss in activity and
viability, yeasts are comparatively advantageous to use than chemical leaveners in frozen steamed dough
systems. The degree of damage can be related to the freezing conditions used and ability to metabolize
different molecules.
Hydrated frozen steamed dough, 3 water sources identified as rigid, confined and bulk water are found in
the dough matrix system and are redistributed differently through ice crystallization and recrystallization in
dough; thus affecting component structure and functionality. Bulk followed by confined water crystallizes
to ice during freezing. The freezing rate applied will also affect the ice characteristics, thus altering

structure and functionality. Hence, studies on water redistribution induced by ice crystallization and
recrystallization may give a deeper understanding in the search of solutions to preserve gluten in frozen
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dough.
Improver and cryoprotective technology: Advances in additives improver technology such as the use of
hydrocolloids, ice-structuring proteins or antifreeze proteins, ice nucleating agents and use of novel biotech
ingredients and enzyme technology was able to; i) increase yeast cell freeze-tolerance, ii) enhance gluten
and starch functional (rheological, thermal) properties during freezing and frozen storage treatment.
Subsequently, the yeast cells in frozen dough have better fermentative capacity, resulting in improved end
product quality in terms of higher specific volume, lower hardness and increased shelf life.
Conclusion: This is attributed to the additives ability to alter ice crystallization and recrystallization
characteristics in frozen dough. Knowledge on development of ice crystallization and recrystallization
during freezing and frozen storage, respectively is useful to enable optimization of freezing conditions and
reduction of temperature fluctuations to maintain yeast viability and frozen dough quality.
Significance and novelty: This overview adds new knowledge and useful insights on: i) use of bio-tech
ingredients for clean label technology in frozen dough and food industry, a consumer demand trend for
clean label products, ii) optimization of frozen dough processing and equipment technology for modern
baking and food industry.
KEY WORDS: Advanced frozen dough technology, Freezing and frozen storage, cryoprotective ability,
Ice crystals, Gluten, Yeast, Starch,
INTRODUCTION
Consumer tastes and preferences for bakery products are continuously changing. Despite this, bread
freshness and soft texture are acceptable quality attributes (Bockstaele, Walle, Dewettinck, Gellynck, &
Ku, 2009); deviation from which significantly decreases consumer bread acceptability (Heenan, Hamid,
Dufour, Harvey, & Delahunty, 2009). Consumers in different markets desire variety of freshly baked

products sourced from local traditional functional and health promoting ingredients (Annunziata &
Vecchio, 2013).
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Excluding microbial spoilage; change in bread texture and freshness collectively termed as staling, occurs
in bread crumb and crust after baking process (Gray & Bemiller, 2003). Attempts to delay bread staling
through several empirical studies are continuously being done by the baking industry and researchers. In
this regard, improved processing methods and innovative technologies like sourdough fermentation,
enzyme technology and frozen dough technology are commonly applied in the baking industry (Bárcenas,
Haros, & Rosell, 2003; Bartkiene, Vizbickiene, Bartkevics, Pugajeva, Krungleviciute, Zadeike,
Zavistanaviciute, & Juodeikiene, 2017; He & Hoseney, 1990; Izadi Najafabadi, Le-Bail, Hamdami, Monteau,
& Keramat, 2014; Jiang, Le Bail, & Wu, 2008; Ronda, Caballero, Quilez, & Roos, 2011; Yi, Johnson, & Kerr,
2009).
Brief history of frozen dough products
In the late 1970s, industrial bakeries were reported to supply unfermented frozen dough for bake-off to
supermarket chains, retail bakeries, food service and institutional users (Decock & Cappelle, 2005).
Therefore, since then, the use of frozen dough technology concept, since its inception in 1970s, continues
to receive great interest from researchers and industry thus being converted into a mainstream business
(Rosell & Gómez, 2007). This trend was primarily driven by the need for product flexibility in terms of
freshness and fast response to consumer trends. To address product freshness, semi-finished doughs were
produced and frozen and stored at subzero temperatures followed by transportation to be processed at the
baking unit stations located at the point of retail such as supermarket chains, and retail bakeries among
others (Decock & Cappelle, 2005). This ensured extension of dough shelf life and addressed bread
freshness concerns of the consumers. Additionally, with increased distance between central production
units (factory) and the point of sale; introduction of the frozen dough concept invariably reduced the
distance between production sites and point of sale, thus improved the response to consumer needs.
Therefore, the use of frozen dough technology in bread industry has been growing since the 1970s.
As presented in Fig. 1a, there are four frozen dough products have been developed and produced overtime.
Unfermented frozen dough (U-FD), where the dough is prepared, shaped and frozen, however, baking is
done after fermentation. Fermentation before freezing should be minimized; for instance, zero pre-
fermentation in Danish pastry without yeast but using baking powder is done, while over 30% relaxation

time in yeast raised Croissant without a separate proofing step, but with a 20min resting period prior to
blast freezing (Decock & Cappelle, 2005). U-FD technology has been commonly used in croissant and
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Danish, with recent applications in bread and rolls. U-FD technology saves on transport costs; when the
final fermentation is eliminated especially in croissants, no thawing is required, thus further increasing on
the level of convenience and product response time. However, the baking times are longer and the ovens
used require steam during operation. Additionally, U-FD products are close to the conventional dough
products, baking products made using this technology usually face problems related to final bread volume.
These are mainly attributed to loss of yeast fermentative activity and lower gas retention of dough.
Pre-fermented frozen dough (P-FD), are frozen dough pieces that no longer require additional proofing
prior to baking with the degree of fermentation prior to freezing varying from 20 to 100% (Gabric, Ben-
Aissa, Le-Bail, Monteau, & Curic, 2011; Le-Bail, Nicolitch, & Vuillod, 2010). P-FD were mainly developed
to offer ready-to-bake products that can be prepared faster than U-FD products. The success of P-FD was
attributed to the introduction and development of improver technology in the late 1900s; with most success
reported in layered dough systems such as croissants and Danish, but more challenging in un-layered
systems such as baguettes and variety breads. After frozen storage, P-FD may directly be transferred from
the freezer to the oven; especially in layered dough products, but a defrosting step is recommended for
most un-layered prior to baking. On the market, P-FD technology products are commonly marketed as
freezer-to-oven, turbo, ready-to-bake, and frozen-to-bake. Compared to other frozen dough products, P-FD
products have been reported to give fast responses to market consumer demands. Compared to pre-baked
goods, P-FD are considered to be less costly to transport, but costlier than un-proofed baked products.
However, most P-FD are prone to thawing due to temperature fluctuations during storage and
transportation, which result in collapse of the pre-fermented structure attributed to pressure drop caused by
freezing, humidity condensation in the cells and carbon dioxide gas transfer towards the dough due to
increased gas solubility with decline in temperature.
Par-baked frozen dough (PB-FD), has become more popular in several shops and supermarkets, and it
involves prebaking the dough product at lower oven temperatures such as 180°C instead of 230°C for
French baguette, while using lots of steam. This enables complete formation of a crust-less baked product,
followed by freezing and frozen storage, and finally a second and final baking which is done at the bake
off point to create a crust. The second baking gives the par-baked product its distinct flavor, taste, and a

crisp crust, as expected by the consumer (Rosell & Gómez, 2007). The concept of PB-FD is reported to
have gained popularity in the mid to late 1980s, but due to product quality inconsistencies such as crust
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flakiness between the first and second bake, led to a decline in its popularity. However, with the
introduction of improver technology in the early 1900s, the shelf life after the second bake and minimized
crust flakiness were achieved thus increased trend in the application of the technology. PB-FD technology
has been mainly used for crusty baked goods such as baguettes and a variety of rolls, but almost non-
existent in laminated items such as croissants or Danish (Decock & Cappelle, 2005; Rosell & Gómez,
2007). This technology has been known to offer high convenience, easy to handle products with little
skilled labor needed at the bake off point, no need for steam in the oven. However, the products have a
limited shelf life after the second baking time and high transport costs are involved.
The underlying catalyst for the development and growth of the frozen dough products in the 1970s to
1990s is closely related the development of improver technology for both layered and un-layered dough
systems and increased research efforts to improve frozen dough quality; thus a steady market growth in
frozen dough products. Therefore, using the frozen dough concept, bakery products such as refrigerated
cookies and brownies, sweet rolls, biscuits, dinner rolls, and pizza have been sold “as if freshly baked” to
the consumer at their convenient time in the market.
Freezing and frozen storage treatment negatively impacts dough structure and composition. For instance,
disrupted gluten network integrity (Wang, Xu, Nikoo, Ocen, Wu, Yang, Jin, & Xu, 2014b), decreased loaf
volume and increased bread textural hardness (Meziani, Kaci, Jacquot, Jasniewski, Ribotta, Muller, Ghoul, &
Desobry, 2012b) are common effects observed in frozen dough products. The changes have been attributed
to ice crystal characteristics among others, during freezing and frozen storage treatment (Baier-Schenk,
Handschin, & Conde-Petit, 2005a; Chen, Jansson, Lustrup, & Swenson, 2012; Chen, Öhgren, Langton, Lustrup,
Nydén, & Swenson, 2013; Huen, Weikusat, Bayer-Giraldi, Weikusat, Ringer, & Lösche, 2014). Despite
growth in global presence of frozen bread-dough products, processing challenges induced by freezing and
frozen storage treatment are still prevalent. The understanding of ice crystal characteristics during freezing
and frozen storage treatment is vital in improving processing parameters as well as end product quality.
Therefore, this review intends to provide an overview of the effect of ice crystal characteristics on water
redistribution and major components (starch and gluten) and ingredient (yeast) during freezing and frozen
storage of frozen bread-dough systems. Additionally, advances in frozen dough improver and

cryoprotective technology including; additives, enzymes and novel fermentation end products that alter ice
crystal characteristics are suggested and discussed. Due to their increase in popularity, frozen steamed
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products and some critical conditions for processing high quality frozen steamed dough products were also
suggested as a case study.
Ice crystal characteristics during freezing and frozen storage treatment
To understand the impact of freezing and frozen storage treatment on frozen dough systems, knowledge of
ice crystal characteristics is important (Simmons, Serventi, & Vodovotz, 2012). Based on thermodynamic
principles of aqueous solutions; frozen dough, a heterogeneous mixture of ingredients in water, has a lower
freezing point compared to pure water (Tf=0°C) (Zaritzky, 2010). It is therefore critical to study ice
formation dynamics on frozen dough during freezing and frozen storage. Two distinct ice crystal
characteristics, ice crystallization and ice re-crystallization occurring at during freezing and frozen storage
stages, respectively are discussed.
Ice crystallization during freezing: At the start of freezing, dough undergoes 3 changes (Meziani,
Ioannou, Jasniewski, Belhaj, Muller, Ghoul, & Desobry, 2012a); i) initial cooling phase, characterized by
temperature decrease to the freezing point, ii) pseudo plateau, characterized by progressive crystal
formation due to removal of latent heat of crystallization, and iii) tempering stage, characterized by
temperature reduction in dough to the environmental temperature. In step two, up-to 80% water is
transformed to ice crystals and is key in the freezing process efficiency (Kiani, Zhang, Delgado, & Sun,
2011; Kiani & Sun, 2011). In frozen dough, ice crystallization occurs when water undergoes ice nucleation
and ice growth.
Ice nucleation in dough: Frozen dough ice nucleation is predominantly of the heterogeneous type, and
involves aggregation of water molecules in a crystalline arrangement on the nucleating agent. In frozen
dough, ice nucleation occurs when free energy state of pure water is overcome through initial cooling to
freezing point of dough for phase transition to occur. The initial cooling rate used therefore influences ice
nucleation and location of the crystals in dough; at high initial cooling rate, ice nucleation increases
characterized by increase in nuclei number of small ice crystals formed. This was confirmed in frozen

sweet dough when slow and fast cooling rates resulted in large and small ice crystals formation,
respectively (Meziani et al., 2012a). The ice seeding temperature used influences location of ice nuclei
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formed; as indicated by a study where lowered ice seeding temperature increased intracellular ice crystal
formation, whereas raising it resulted in more extracellular ice crystal formation in frozen dough
component tissue (Nakamura, Takagi, & Shima, 2009).
Additionally, in dough, ice nucleation sites have linked to gas pore interfaces. This was confirmed in a
wheat dough study model with air inclusions by (Baier-Schenk, Handschin, Von Schönau, Bittermann,
Bächi, & Conde-Petit, 2005b); which indicated that ice nucleation was observed to start at the gas pore
interface.
Ice crystal growth in dough: Ice crystal growth occurs readily at temperatures close to freezing point of
dough and is controlled by mass and heat transfer (Delgado & Sun, 2001). For instance, crystal growth is
achieved as ice crystallizes from solution in dough, water molecules diffuse out to the ice crystal surface
and are incorporated into the growing solid phase. Simultaneously, solute molecules are continuously
rejected from the region occupied by pure ice crystals and diffuse away from ice surface. Hence, number
and size of ice crystals formed is a result of both ice nucleation and growth rate; and ice crystal size varies
inversely with number of nuclei formed. Therefore, control of ice nucleation and growth may alleviate
quality shortcomings of frozen dough.
Ice recrystallization during frozen storage of dough:
Many studies on frozen storage effect on dough have cited ice recrystallization as the main detrimental
factor to frozen dough product quality (Eckardt, Öhgren, Alp, Ekman, Åström, Chen, Swenson, Johansson, &
Langton, 2013). Ice recrystallization occurs during frozen storage due to presence of temperature gradient
as temperature fluctuates. Temperature gradient created by fluctuation in temperature during frozen storage
of dough leads to melting of small ice crystals on dough surface and formation of larger ice crystals at this
site. Consequently, larger ice crystals grow at the surface of stored dough. As shown in Fig. 2, this was
confirmed in pre-fermented frozen bread dough using cryoscanning electron microscopy stored at -22°C;
ice recrystallization progressed with an increase in ice crystal size and ice crystal initially embedded in the
pore interface were instead on the surface (Baier-Schenk et al., 2005a). These changes are reported to

affect frozen dough quality, however, several specific studies on ice recrystallization in frozen steamed
bread-dough systems need to be done.

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Thermodynamic properties of frozen dough. Thermodynamic properties of frozen dough have been
used to simulate and model heat transfer, and calculate the size of the ice crystals in the frozen dough.
These properties include; effective thermal conductivity, apparent specific heat, ice melting enthalpy,
freezable water fraction and ice fraction (Matuda, Pessôa Filho, & Tadini, 2011). However, differential
scanning calorimetry (DSC) has been commonly used to directly determine thermal conductivity and ice
melting enthalpy (∆H) in many reported studies.
Freezable water and ice fraction of frozen dough can be calculated according to method by (Kumcuoglu,
Tavman, Nesvadba, & Tavman, 2007); X 100% ; where fw is freezable water of frozen dough
(%), ∆H is ice melting enthalpy in frozen dough (J), ∆Hi is latent heat of fusion for pure water (334 J/g), m
is mass of water in frozen dough (g).
The ice fraction in frozen dough can be calculated as expressed by (Hamdami, Monteau, & Le Bail, 2004);
f= ; where f is ice fraction in frozen dough, ∆Hr is exothermic enthalpy for temperature from T0 to
T1.
However, other methods have been used to simulate and model the determination of thermodynamic
properties of dough during freezing and frozen storage (Cornejo, Cornejo, Ramírez, Almonacid, & Simpson,
2016; Luo, Sun, Zhu, & Wang, 2017).
Freezing rate on ice crystal characteristics in frozen dough.
Studies on impact of freezing rate during freezing and frozen storage treatment of dough has focused
mainly on quality and shelf life attributes. For instance, rapid freezing of cooked rice frozen dough stored
at -18°C for 7 months; starch retrogradation was retarded and textural properties improved (Yu, Ma, & Sun,
2010). However, less focus is placed on effect of ice crystal characteristics on frozen dough.

The freezing rate applied during freezing affects ice crystal characteristics in frozen dough in different
ways. For instance, when fast-rapid freezing rates are used in dough freezing and frozen storage treatment,
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small evenly distributed ice crystals are formed; less damage is done to the dough hence better product
quality reported (Ban, Yoon, Han, Kim, Han, Lim, & Choi, 2016; Yi & Kerr, 2009; Yu et al., 2010). The
reverse is true when slow freezing rates are used (Havet, Mankai, & Le Bail, 2000) which leads to fewer but
large ice crystals formation. This is attributed to the rates’ ability to initiate ice nucleation. This is observed
in slow freezing rate, where formation of initial ice crystals keeps pace with heat removal than in fast-rapid
freezing rate; resulting in heat undercooling and increased frequency of nucleation in fast-rapid freezing
rates, thus more active nucleation sites and an increase in the number of small ice crystals. Additionally,
freezing rates are closely linked to location of ice crystals formed in frozen dough (Delgado & Sun, 2001).
However, an opposite effect of freezing rate is observed on yeast activity and viability in frozen dough.
With a slow freezing rate reported to improve yeast viability while the fast rate is detrimental (Meziani et
al., 2012b). These effects are attributed to water migration dynamics, size and location of ice crystals
formed during the different freezing rate (Acker & Croteau, 2004; Gao & Critser, 2000; Li, Wang, &
Tingrui, 2013; Soveral, Madeira, Loureiro-Dias, & Moura, 2008). As discussed in the next section,
characteristic of the ice crystals formed impacts dough components differently, thus altering their structure,
morphology, and functionality.
EFFECT OF FREEZING AND FROZEN STORAGE ON MAJOR DOUGH COMPONENTS AND
INGREDIENTS
Dough is a complex heterogeneous mixture of constituents like water, starch, gluten and yeast (Goesaert,
Brijs, Veraverbeke, Courtin, Gebruers, & Delcour, 2005). The main ingredients for frozen steamed dough are
mainly flour, water, and yeast among others. Frozen dough quality is impacted by ice crystal
characteristics and freezing conditions used, and is presented in Table I. Frozen steamed bread systems are
mainly composed of yeasts, flour (majorly gluten protein and starch components), water and other
ingredients. In this section, advances on studies on the impact of ice crystal characteristics and freezing
conditions on water redistribution and its effect gluten, starch and yeast activity and viability as they affect
dough quality are discussed.

GLUTEN PROTEIN IN FROZEN DOUGH
Wheat protein, mainly in the form of gluten, is a major component in Chinese steamed bread. Gluten forms
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a viscoelastic network in dough, holding starch and other components inside the product (Zhu, 2014); the
network is stabilized by covalent disulphide bonds (S-S) and other non-covalent interactions (hydrogen,
ionic, and hydrophobic bonds) (Wieser, 2007). Freezing of dough will therefore impact gluten composition
and structure and alter viscoelastic and rheological properties of frozen dough. From studies, frozen
steamed bread dough should be made from flour with protein content of 9.5-11% (Kondakci, Zhang, &
Zhou, 2015; Zhu, 2014), where too high or too low protein content was found to result in rough surface and
bad eating quality.
Changes in gluten protein structure: Gluten macropolymer (GMP), composed of gliadins and glutenins
(high (HMW-GS) and low (LMW-GS) molecular weight glutenin subunits) whose molecular size
distribution in dough influences functionality in bread making (Wieser, 2007). GMP contains same LMW-
GS content irrespective of HMW-GS composition; structurally, LMW-GS is part of GMP through S-S
cross-linking with HMW-GS (Don, Mann, Bekes, & Hamer, 2006). HMW-GS is directly correlated to
amount of GMP. Functionally, GMP fractions contribute to dough rheology in different but synergistic
ways; gliadin is responsible for dough viscosity and extensibility and glutenin is responsible for dough
strength (Delcour, Joye, Pareyt, Wilderjans, Brijs, & Lagrain, 2012; Wieser, 2007). As observed by high
precision techniques, freezing and frozen storage treatment influences gluten functionality.
Macro-molecularly, using an electrophoretic analysis of SDS-soluble protein aggregates extracted from
frozen dough, significant decline in HMW-GS from 129,100 to 88,700 was observed (Ribotta, León, &
5 8
Añón, 2001). A similar decreasing trend in molecular weight (from 3x10 Da to 4x10 Da) and radius of
gyration of gluten after 3 freeze-thaw cycles was observed using size exclusion chromatography in
conjunction with multi-angle laser light scattering (SEC-MALLS) (Zhao, Li, Liu, Chen, Liu, Zhu, & Li,
2013).
On the other hand, decrease in HMW-GS coincides with increase in LMW-GS. This was associated with
weakening S-S cross-link between LMW-GS and HMW-GS resulting in release and increase of LMW-GS
(Xuan, Zhang, Zhao, Zheng, Jiang, & Zhong, 2017). Collectively, the changes observed are due to
depolymerisation of HMW-GS fraction of GMP (Wang, Chen, Mohanad, Xu, Ning, Xu, Wu, Yang, Jin, &

Xu, 2014a; Wang, Tao, Jin, & Xu, 2015b) as gluten structure is weakened with prolonged frozen storage as
illustrated by atomic force microscopy (AFM) (Fig. 3) (Zhao et al., 2013).

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GMP depolymerisation during frozen storage is the main indicator of gluten quality; is linked to S-S bond
disruption and breakage (Wang, Jin, & Xu, 2015a). SH groups, an indicator of disruption of S-S linkages;
was constant in gliadin, but increased in gluten and fluctuated in glutenin fraction during frozen storage
(Wang et al., 2014a). Upon frozen storage, decreased degree of gluten polymerization was observed in
steamed bread made from frozen dough (Wang et al., 2015b). Despite lack of a clear understanding of
GMP depolymerisation mechanism during frozen storage; many recent studies attribute it to interaction
between GMP and water during frozen storage.
Gluten protein interaction with water: In hydrated frozen dough, 3 water sources identified as rigid,
confined and bulk water (Fig. 3) are found in or around gluten (Kontogiorgos, 2011). Water redistribution
through ice crystallization and recrystallization in frozen dough damages gluten structure and functionality.
The gluten-starch matrix network supports gas cells in form of pores which are preferred sites for ice
nucleation and growth; thus water in gas cells crystallize and grow (Esselink, Van Aalst, Maliepaard, & Van
Duynhoven, 2003). This invariably disrupts and weakens the gluten structure and GMP subunits as
confirmed by scanning (SEM) and transmission electron microscopy (TEM) (Esselink et al., 2003; Wang
et al., 2014b).
Conformational changes in gluten have been suggested to originate from the GMP fractions. In gluten-
water and gliadin-water systems, water mobility was induced and increased upon frozen storage; a similar
trend was observed in hydrated gluten (Wang et al., 2014b). Increased water mobility was also observed in
dehydrated gluten after 5 weeks of frozen storage (Jia, Huang, Rayas-Duarte, Zou, Zhang, & Li, 2014b).
Bulk followed by confined water in gluten matrix crystallizes to ice during freezing. The freezing rate
applied affect the ice characteristics, thus alter gluten structure and functionality. During frozen storage,
temperature fluctuations create a temperature gradient leading to ice crystal melting and recrystallization to
form large ice crystals on surface of gluten (Kontogiorgos & Goff, 2006).
This was elaborated when T2 relaxation time distributions of gluten-, glutenin- and gliadin-rich fractions at
different frozen storage times were used. Increased frozen storage time, increased T2relaxation time in
gluten- and gliadin-rich fractions; an indication of weakened association between amino acids and low

mobility of water molecules (Wang et al., 2014b). The rigid and confined water in gluten- and gliadin-rich
fractions were more sensitive to frozen storage than glutenin-rich fractions.

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Secondary structural changes like reduction of α-helix structure and conversion to specific β-sheet and β-
turn structures of gluten- and gliadin-rich fractions have also been observed using Fourier transform-
infrared spectroscopy (FTIR) (Jia et al., 2014b). The structural changes increased with longer frozen
storage (Wang et al., 2014b). Presented in Fig. 3, evidence by atomic force microscopy (AFM) images
showing the weakening of a fibril-like branched network formed from gluten chain with increased frozen
storage time (Zhao, Liu, Hu, Li, & Li, 2016). Hence, studies on water redistribution induced by ice
crystallization and recrystallization may give a deeper understanding in the search of solutions to preserve
gluten in frozen dough.
STARCH IN FROZEN DOUGH
Starch has a unique structure and physicochemical properties which determine its functionality in bread
making (Goesaert et al., 2005). Starch granules are intracellularly water insoluble and have different
diameter sizes, morphology and structure depending on botanical source (Vamadevan & Bertoft, 2015). In
conventional bread making, starch granule hydration follow similar pattern of water absorption,
gelatinization and retrogradation upon cooling; leading to loss of birefringence and crystallinity, and re-
association to a more ordered state. The effect of freezing and frozen storage impact on starch composition
and structure are discussed.
Changes in structure of starch during freezing and frozen storage: Changes in starch granule in frozen
dough is important for technological and sensory quality improvement of starch based frozen foods
(Vamadevan & Bertoft, 2015). Structurally, freezing and frozen storage damages starch granules and
changes its morphological make-up. This was observed with increased number of freeze-thaw cycles
greatly impacting wheat starch granular structure (Tao, Yan, Zhao, Tian, Jin, & Xu, 2015). Granule
structure changes are attributed to freezing pressure created due to exposure to phase transformation as
water expands on freezing to form ice crystals in dough. Starch granule is preferentially compressed
resulting in deformation and disruption (Perry & Donald, 2000) (Fig. 4).

Freezing induces re-organization in freeze-thawed starch compared to native starch. This was evidenced by
decline in amorphous state coupled with crystallinity increase in starch granule as materials leach out (Tao,
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Wang, Wu, Jin, & Xu, 2016a). After 10 freeze-thaw cycles, relative crystallinity and short-range order of
native wheat starch increased by 4.3% and 0.251% respectively (Tao et al., 2015). Similar irreversible and
disruptive changes in crystallinity were observed in waxy rice starch (Tao et al., 2015) and potato starch
after multiple freeze-thaw (Szymonska, Krok, Komorowska-Czepirska, & Rebilas, 2003).
Starch granules of common cereals of baking relevance like wheat, barley, and rye exhibit a bimodal size
distribution. Freeze-thaw influences the A- and B- type granules differently. As indicated in Fig. 4, (Tao et
al., 2016a) observed no apparent damage on A-type granular surface, but a cracked structure on B-type
wheat granules; indicative of sensitivity of B-type granules to freeze-thaw treatment. In a reconstituted
bread model made from wheat starch B-granules; smaller specific volume and increased crumb firmness
(p>0.05) was observed. But, no difference was observed in A-granules after freeze treatment (Tao et al.,
2016a). In wheat flour frozen dough bread; increasing A-type granule may improve product quality.
Similar studies using other starch sources will further enrich our understanding of this phenomena. Despite
difference in botanical starches; impact of freezing and frozen treatment on starch granule generally
follows similar trends. This was observed in gel structures were freezing rates and both freezing rates and
thawing rates impacted potato starch and wheat starch, respectively (Freschi, Doran, Malumba, & Blecker,
2014).
Starch interaction with water during frozen storage: Starch granule functionality is closely related to its
hydration properties. To understand starch granule hydration in native wheat, maize and potato, 1H HR-
MAS spectroscopy shows that water mobility exists mainly in minor part of starch polymer, while majority
of chains were densely packed and remained isolated from bulk water (Larsen, Blennow, & Engelsen,
2008). However, chains in the amorphous starch micro-spheres were more accessible for water. Therefore,
the structure adopted a hydrophilic three dimensional network held together by chemical linkages. For
cross-linked starch microspheres, X-ray scattering showed hydration induced evolution in the polymer
structure; a fact linked to swelling as observed with optical microscopy (Wojtasz, Carlstedt, Fyhr, &
Kocherbitov, 2016).

Starch granule interaction with water during freezing and frozen storage impacts starch composition and
functionality (Alcázar-Alay & Meireles, 2015; Przetaczek-Rożnowska, 2017; Zhu, 2017). Pretreatment of
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starch negatively influences functionality in frozen dough. This was observed in sodium dodecyl sulfate
(SDS) pretreated wheat starch were freezing induced irreversible changes on granular structure, but barely
affected the native starch (Fig. 4) (Tao, Wang, Zhang, Wu, Jin, & Xu, 2016b). SDS washing of native starch
could have removed non-starch components like proteins and lipids; thus increasing granule water
absorption and sensitivity to freezing treatment. Functionally, higher gelatinization temperatures, melting
enthalpy and pasting viscosities were observed in pretreated starches. In other words, freezing delayed
thermal properties in native wheat starch samples. In In-situ study by (Molina, Leiva, & Bouchon, 2016),
freezing delayed the gelatinization degree of frozen native starch granules in excess water. However, in
limited water accessibility, difference with the unfrozen samples decreased (Molina et al., 2016). This
indicates the indispensable role water migration plays during freezing in relation to starch functionality.
YEAST ACTIVITY AND VIABILITY IN FROZEN DOUGH
In frozen dough systems, Saccharomyces cerevisiae yeast strains have pre-dominantly been used as an
ingredient during fermentation. The effect of freezing and frozen storage treatment on yeast activity and
viability and its impact in frozen dough quality is discussed.
Changes in yeast activity and viability: Generally, freezing and frozen storage treatment decreases yeast
activity and viability in frozen dough systems. Despite loss in activity and viability, yeasts are
comparatively advantageous to use than chemical leaveners in frozen steamed dough systems (Wang,
Yang, Gu, Xu, & Jin, 2017a). The degree of damage can be related to the freezing conditions used and
ability to metabolize different molecules.
Freezing conditions affect yeast activity and viability in several ways, thereby influence sensory and
physical properties of frozen steamed dough products. Generally, freezing and frozen storage treatment
may affect yeast activity and viability through factors such as yeast cell water permeability, freezing rates,
ice crystal size and location and ice re-crystallization. During freezing, yeast cells can be damaged by the

ice crystals formed resulting in the release of reducing agents such as glutathione which weaken dough
through reduction of gluten S-S bonds (Öztürk, Cerit, Mutlu, & Demirkol, 2017).
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As shown in Fig. 5, damage to yeast due to freezing conditions can be categorized in two parts; firstly,
slow freezing rate, characterized by slow osmotic migration of intracellular water and extracellular
formation of ice crystals (Gao & Critser, 2000). The cells shrink as solute concentration increases in
extracellular environment (Fig. 5); thus cells become dehydrated and may denature (Soveral et al., 2008).
Secondly, when freezing rate is fast; intracellular water has no time to flow out through membrane and ice
crystals are formed intracellularly (Fig 5); this impairs membrane structure and function as ice crystals
form in the cells (Acker & Croteau, 2004; Li et al., 2013). Therefore, high freezing rates correlate with an
extended damage to yeast activity in frozen steamed dough products (Meziani et al., 2012b). The resultant
bread had a lower specific volume and higher hardness. In this regard, in frozen steamed dough, a slow
freezing rate are preferable to maintain yeast cell integrity. Also, in practice, to compensate loss in yeast
activity and viability; high levels of yeast is used.
In a recent study by (Huang, Zhao, Zhang, Xu, Toth, & He, 2017), combined treatment of pre-dehydration
using extracellular trehalose and ice seeding at high subzero temperature resulted in high cell viability of
fibroblasts, adult stem cells and red blood cells. Ice seeding minimized free energy that drives ice re-
crystallization induced cell injury during thawing of cryopreserved cells (Huang et al., 2017). In yeast (S.
cerevisiae), ice-seeding temperatures enhance growth and survival in the log phase of growth during
freezing process; in prolonged storage, this was more important than presence of trehalose (Nakamura et
al., 2009). Hence, the absence of ice nucleation capable sites reduces intracellular ice formation; and the
super-cooling should be eliminated by dehydration before the cell cools to its ice nucleation temperature
(Lindow, Arny, & Upper, 1982). This is confirmed in predictive models by (Kasner, Hunter, & Kariko,
2013); were at nucleation temperature of -25°C, no intracellular ice formation was predicted to occur in
yeasts cooled at a rate of <20°C/min, but would occur with near certainty when cooled at ≥30°C/min. The
predictions were closely confirmed in experimental observations by DSC. Therefore, attempts to control
freezing and frozen storage conditions that affect ice crystallization and recrystallization may offer
solutions to improve yeast activity and viability, resulting in improved fermentation ability of frozen
dough.

In response to freezing and frozen storage, yeast cells have been reported to intracellularly accumulate high
levels of protective molecules such as proline, arginine, and glycerol; which confers high resistance to
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freezing stress (Morita, Nakamori, & Takagi, 2002). Metabolically, non-proline utilizing S. cerevisiae yeast
strains release ∆1-pyroline-5-carboxylate reductase to convert ∆1-pyroline-5-carboxylate into proline,
which accumulates to confer freeze-tolerance. In freeze-tolerant yeast cell mutants, co-overexpression of
mutant γ-glutamyl kinase and wild type γ-glutamyl phosphate reductase, but not ∆1-pyroline-5-carboxylate
reductase were effective for proline accumulation in yeast cells (Terao, Nakamori, & Takagi, 2003). The
two enzymes are considered rate limiting enzymes in yeast (Tsolmonbaatar, Hashida, Sugimoto, Watanabe,
Furukawa, & Takagi, 2016).
The accumulated intracellular molecules impact yeast functionality differently. Trehalose prevents protein
denaturation by slowing rate of polyglutamine mediated protein aggregation and resultant pathogenesis by
stabilizing an aggregation prone model protein; during freeze stress, helping retain yeast cellular integrity
(Mizunoe, Watanabe, Sudo, Kobayashi, Yasukawa, Natori, Hoshino, Negishi, Okita, Komatsu, & Higami ,
2017; Stefanello, Machado, Pasqualin Cavalheiro, Bartholomei Santos, Nabeshima, Copetti, & Fries, 2018;
Zhang, Oldenhof, Sieme, & Wolkers, 2016). Proline, on the other hand combines with intracellular free
water to form strong hydrogen bonds; and at high levels, are associated with high level of superoxide
dismutase which lowers levels of reactive oxygen species; preventing intracellular substances from
oxidation (Sasano, Haitani, Hashida, Ohtsu, Shima, & Takagi, 2012). Glycerol prevents dehydration by
balancing intracellular osmolarity with that of environment (Ballester-Tomás, L., Pérez-Torrado, Rodríguez-
Vargas, Prieto, & Randez-Gil, 2016); as observed when glycerol (2%, flour basis) added to pre-fermented
frozen steamed bread reduced (P<0.05) freezable water proportion by 14-16% compared to control sample.
This subsequently prevented formation of ice crystals in dough during freezing and frozen storage (Huang,
Wan, Huang, Rayas-Duarte, & Liu, 2011); thus improved yeast activity and viability resulting in improved
dough leavening capacity, and reduced proof time after initial freeze-thawing.
Despite suggested mechanisms that allow yeast cells to survive during freezing and frozen storage, there is
still a lot to be studied (Ballester-Tomás et al., 2016). However, accumulation of intracellular molecules is
by far the most important component in enhancing baker’s yeast strains in frozen dough.

Based on this, through genetic engineering techniques, specialized yeast strains have been developed for
commercial frozen steamed dough product production. Common examples predominantly used in frozen
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steamed dough formulations are compressed or active dried; however, instant dry yeast are not
recommended.
FROZEN BREAD DOUGH IMPROVER AND CRYOPROTECTIVE TECHNOLOGY;
ADVANCES IN ADDITIVES, ENZYMES AND FUNCTIONAL MICROBIAL END PRODUCTS.
With the ever increasing popularity of frozen dough products, frozen dough research trends have become
relevant for frozen bread production. Ingredient added and processing techniques used in frozen dough
technology may impact the quality of frozen dough end products. For instance, in an earlier study, salt,
trehalose and dough mixing time interacted and influenced the rheofermentometer parameters and specific
volume of frozen sweet dough in a mixed trend (Huang, Kim, Li, & Rayas-Duarte, 2008).
As discussed already, freezing and frozen storage conditions affect key dough components. In this section,
as highlighted and presented in Table II, recent cryopreservation techniques applied in frozen dough
systems and products are summarized with emphasis placed on techniques that control ice crystallization
and recrystallization. Additionally, developments in enzyme applications and microbial (e.g. lactic acid
bacteria, LAB) fermentation technology aimed at enhancing quality of the frozen dough oriental products
are discussed.
Additives as improvers in frozen dough.
To control ice crystallization and recrystallization during freezing and frozen storage; studies suggest use
of different additives to improve baking quality of frozen dough products. In earlier studies, frozen dough
additives of lipid-related emulsifiers (e.g. DATEM, CSL) and sucrose esters received most attention
(Berton, Ropers, Bertrand, Viau, & Genot, 2012; Matsumiya, Takahashi, Nakanishi, Dotsu, & Matsumura,
2014; Matuda, Parra, Lugão, & Tadini, 2005; Van Steertegem, Pareyt, Brijs, & Delcour, 2013). These
emulsifiers’ mode of action has been linked to i) retardation of retrogradation by interaction with water, ii)
blocking moisture migration between gluten and starch, thus less water uptake by starch, and iii) interact
with added lipids to reduce surface tension in gas bubbles resulting in larger numbers of smaller bubbles
(Selomulyo & Zhou, 2007).

In recent studies, focus has shifted to additives such as ice structuring proteins (ISPs), ice nucleation agents
(INA), and hydrocolloids in cryopreservation of frozen dough.

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Hydrocolloids: Hydrocolloids used as additives capable of controlling both rheology and texture of
aqueous food systems have been extensively studied. During frozen storage, instability of frozen dough
properties is linked to ice recrystallization as temperatures fluctuate (Maity, Saxena, & Raju, 2017).
Hydrocolloid can preserve frozen dough by complexing with gluten and bound water. This increases
dough water holding capacity (WHC), alters moisture migration, and minimizes gluten damage caused by
ice recrystallization in frozen dough (Kontogiorgos & Goff, 2006). Hydrocolloids also decrease water
activity through competition with other flour components. Interaction of hydrocolloids with dough macro-
components (water, starch and protein) varies based on nature of hydrocolloid used (Linlaud, Ferrer,
Puppo, & Ferrero, 2011). For instance, higher and lower molecular mobility of gluten-water matrix were
observed when xanthan gum and pectin respectively were used. Hence, overall cryopreservative
effectiveness of hydrocolloids in frozen dough products may depend on type, solubility, dosage, water
holding capacity, rheological properties and their synergistic effect with other ingredients during freezing
and frozen storage (Ferrero, 2017). Studies to establish clean sources of hydrocolloids are also ongoing.
Ice-structuring proteins: Ice structuring proteins (ISPs) isolated from sources like plants and
microorganisms, improve freeze tolerance of frozen dough. ISPs extracted from oat (Avena sativa L.)
lowered freezable water content of frozen dough; this improved final steamed bread quality (Zhang, Zhang,
Wang, Qian, & Qi, 2015). The improved steamed bread quality was attributed to improved fermentation
capacity of ISP supplemented samples; an indication of improved yeast viability and less gluten matrix
disruption. This is in agreement with an earlier study by (Zhang, Zhang, & Wang, 2007), who reported
decreased yeast mortality during freeze-thaw cycle and improved gas retention capacity of frozen dough
after addition of ISP isolated from carrots (Daucus carota). Ice crystallization was also retarded (Zhang et
al., 2007). In a separate study (Ding, Zhang, Wang, Qian, Qi, & Xiao, 2015), ISP isolated from barley
increased the apparent specific heat of dough after freezing, increased freezing and range of melting and
glass transition temperature when added to frozen dough. As demonstrated by ISPs from oats (Zhang et al.,
2015), barley antifreeze proteins (BaAFP-1) decreased the melting enthalpy and freezable water content of

fresh dough after freeze-thaw cycles (Ding et al., 2015). Presence of ice structuring proteins from white
wheat increased WHC of frozen dough and increased bread specific volume (Xu, Huang, Jia, Kim, & Liu,
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2009). Similar results were observed in water molecular state, microstructure, rheo-fermentation capacity
and baking properties of frozen doughs with added thermostable ice structuring proteins (TSISPs)
extracted from Chinese privet (Ligustrum vulgare) leaves (Jia, Huang, Wu, Zhong, Rayas-duarte, & Guo,
2012).
Ice nucleation agents (INA): During freezing and frozen storage of dough, heterogeneous ice nucleation
is the prevalent pathway for ice formation and it occurs at warmer temperatures (Glatz & Sarupria, 2017).
From studies, different surfaces promote ice nucleation at different temperatures and rates; but surface’
ability to promote ice nucleation are largely unknown. Studies relating surface and water behavior at super-
cooled temperatures using INA attempt to address these questions.
INA mainly minimize the degree of super-cooling and inhibit formation of large ice crystals. Epiphytic
bacteria such as Pseudomonas syringae and Erwinia herbicola widely distributed in leaves of plants have
been extensively studied for their ice nucleation properties through prevention of super-cooling of plants
(Lindow et al., 1982; Vanderveer, Choi, Miao, & Walker, 2014).
In frozen dough, addition of extracellular ice nucleators (ECINs) from Erwinia herbicola alleviated bread
crumb hardness, and increased specific volume by 50% after 3 freeze-thaw cycles compared to controls
(Shi, Yu, & Lee, 2013b). This was associated with increased yeast viability at log- and stationary-phase by
100 and 10 fold respectively, and 17% increase in frozen dough (Shi et al., 2013b). Microbial sourced INA
can be classified by their nucleation temperature and chemical composition. Despite several classification
studies using the nucleation temperature, there are relatively few studies that have characterized bacteria
INA by their chemical composition. In an earlier study, the extracellular ice-nucleating matter produced by
Erwinia uredovora was reported to contain 10%, 43%, 35%, and 12% of lipids, proteins, saccharide and
polyamine, respectively (Kawahara, Maho, & Obata, 1993). This confirms the lipoglycoprotein nature of
ECINs. Interestingly, when zein-based ice nucleation films (INFs) were used to wrap frozen dough; ice
nucleation temperature of water increased from -15°C to -6.7°C, water loss by frozen storage reduced after
5 freeze-thaw cycles (Shi, Yu, Jin, & Lee, 2013a). In bread, specific volume increased to 25% compared to
control dough. This was attributed to improved yeast viability through cryopreservation by INFs.

However, factors such as amount of additives, dough formula, and processing conditions could each affect
cryopreservation effectiveness of additives in frozen dough. Besides, the discussed additives, other
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techniques such as optimization of the freezing process and use of novel technologies have been suggested
in a review by (Luo et al., 2017).
Bio-tech ingredients in frozen dough systems
Many food fermentations especially natural are dominated by lactic acid bacteria (LAB) and yeast
microorganisms (Brandam, Fahimi, & Taillandier, 2016; Nout, 2009; Wang, Mao, Meng, Li, Liu, & Feng,
2014); which have been reported to bio-transform food physicochemical and functional attributes. The
changes are greatly linked to the enzymes synthesized in-situ by specific microorganisms. As processing
aids, enzymes are involved in a series of catalytic activities; which when optimized is beneficial to product
quality (Archer, Connerton, & MacKenzie, 2008).
Microbial end products: In frozen dough systems, the low freezing and frozen storage temperatures will
or may damage and alter metabolic capabilities of microorganisms used as starters in dough fermentation.
To overcome this, studies have attempted to use functional end products synthesized in-situ by several
microbial fermentations or added the enzymes separately. For instance, a low branched high molecular
weight microbial exopolysaccharides (EPS) produced in-situ by LAB Weissella confusa QS813
fermentation significantly improved the freeze-thaw stability of wheat starch gel (Tang, Liu, Huang, Cheng,
Wang, Zhang, Chen, Jiang, Omedi, & Li, 2018). In this instance, the EPS acted as a hydrocolloid by
reducing the rate of syneresis in wheat starch after frozen storage. This invariably enhanced the
technological properties of frozen goods. Additionally, functional end products like organic acids (Couto
& Sanromán, 2006; Erbaş, Kemal Uslu, Ozgun Erbaş, & Certel, 2006; Rhee, Lee, & Lee, 2011), GABA
(Huang, Hu, Tsai, & Chang, 2013), ACE-I inhibiting peptides with emulsifying properties (Omedi, Huang,
Su, Liu, Tang, Xu, & Rayas-Duarte, 2016) among others have been successfully studied in sourdough
fermentation systems, but not in frozen dough systems.

Enzymes: Enzymes have been added singly or in combination to minimize dough freezing and frozen
storage damage on gluten structure, and yeast. For instance, transglutaminase enzyme principally used for
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its protein cross-linking ability, could be applied to frozen dough systems to reduce gluten cross-linking
caused by ice crystallization. In an earlier study (Huang, Yuan, Kim, & Chung, 2008), the use of
transglutaminase improved frozen dough quality through its protein polymerization ability which stabilized
the gluten structure embedded in the starch granules. This notion was confirmed in a study by (Steffolani,
Ribotta, Perez, Puppo, & León, 2012), where transglutaminase added at intermediate levels resulted in the
largest bread volume compared to control (without added enzymes) and other two enzyme (pentosanase
and glucose oxidase) samples of frozen dough stored at -18°C for 0 to 9 weeks. In another study (Tang,
Wang, Huang, Zou, Jia, Jin, Omedi, & Li, 2016), transglutaminase treatment significantly decreased the ratio
of HMW-GS: LMW-GS and GMP content in fresh dough and GMP particle size increased. However,
Lipase known to catalyse hydrolysis and or synthesize the ester bond (s) of glycerol (phospho- or glycol)
lipids (Gerits, Pareyt, Decamps, & Delcour, 2014); had no effect on GMP particle size (Tang et al. 2016),
but its specificity to lipid fractions in frozen dough may exert major roles in gas incorporation and
stabilization after frozen storage (Pareyt, Finnie, Putseys, & Delcour, 2011). Interestingly, the combination
effect of Rhizopus Chinensis lipase and transglutaminase was observed to dramatically increase the
proportion of the larger particles and weighted average volume (D4.3) in GMP (Tang et al. 2016). This
could attributed to the synergistic benefit of combining both enzymes in the treatment which inhibited
GMP depolymerisation during frozen storage. Similar synergistic findings were reported in doughs
containing recombinant lipase (Rhizopus chinensis lipase) and transglutaminase enzymes stored at -18°C
for 7 to 35 days; rheofermentative and water holding capacity of the frozen dough were improved (Li,
Tang, Huang, Liu, Tilley, & Yao, 2011). Additionally, the samples had a more open network and uniform
crumbs, higher specific volume with higher sensory scores for the product (Li et al., 2011).
Glucose oxidase and pentosanase responsible for removal of oxygen and breakdown of pentosane for
reduction of viscosity, respectively; both showed promising results in improving bread volume and
lowering firmness of frozen dough systems (Steffolani et al., 2012). Studies involving use of other
enzymes to enhance key quality parameters of frozen dough systems are being undertaken.

FROZEN STEAMED PRODUCTS: Despite being unpopular in the past decades, frozen steamed
products such as Chinese frozen steamed bread, buns and rolls are becoming readily available in countless
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supermarkets in China and East and Southeast Asia. Additionally, due to their high quality, frozen steamed
products are increasingly being exported to markets in United States, Europe and Australia; a trend that is
increasing the product’ global presence (Huang & Miskelly, 2016). Due to this, increased research efforts
to improve understanding of the freezing and frozen storage of the product has been done by both industry
and academic institutions. Steamed bread, originally from northern China, has increasingly become a
staple food for many Asian countries and the western world as well. Characteristically, this bread has no
browned crust, has a softer crumb and is mainly produced using low protein flour (7.5-11%) depending on
the cultural and geographic location of the local production and consumers (Liu, Li, Hao, Zheng, Bian,
Zhang, & Wang, 2015; Zhu, 2014). However, on a larger scale, there are compositional differences in the
recipes used in its production (Keeratipibul, Luangsakul, Otsuka, Sakai, Hatano, & Tanasupawat, 2010; Liu,
Chang, Li, & Liu, 2012; Liu, Ruan, & He, 2016; Zhu, 2014). For consumption, these products are reheated
in a steamer or microwave and eaten for breakfast or as a snack. With the ever increasing popularity of
steamed bread, frozen dough research trends have become relevant for steamed bread production. As
shown in Fig. 1b, frozen dough for steamed bread production may be pre-proofed (frozen and ready for
steaming) or frozen un-proofed (requires thawing and proofing by end user before steaming); an indication
of frozen dough technology evolution in principles and practices in western bakery products.
Critical processing conditions for frozen steamed product quality.
Control of critical processing conditions of frozen steamed doughs is important to achieve product
acceptability and freshness of the frozen product on re-steaming or microwaving. In this section, control of
fermentation processes and dough temperature during processing are discussed as the two critical
processing conditions for frozen steamed product quality. As shown in Fig 1b, the dough fermentation
differs depending on the frozen product of interest. The control of fermentation at all stages of the process,
through proper dough temperature management is critical to maintain dough quality. In practice, frozen
steamed products are molded, proofed (or unproofed), cooled, freezing treated and packed followed by
frozen storage. For quality products, dough should be mixed optimally and temperatures maintained at 20
to 24°C. For frozen steamed bread products, control of fermentation and proofing temperatures in the

range of 30 to 35°C and 35°C-40°C for proofed frozen steamed products and 20 to 25°C, rest at 30°C-
45°C, followed by a proofing at 25°C to 35°C, 60-70% RH for 90-120min for un-proofed frozen steamed
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products. The control and optimization of the relative humidity and time is normally used.
In addition, use of the recommended flour for processing at the right temperature should be used. The
dough should be worked immediately from the mixer with little rest as possible between the stages.
Adequate packing through lamination to provide extra protection. The steamed products dough need to be
freezing treated as fast as possible. Freezing of steamed products to bring the temperature as fast as
possible to the range of -8°C to -15°C within 10 to 30 minutes and freezing and frozen storage at -18°C to
-22°C.
CONCLUSIONS AND FUTURE PROSPECTS.
Use of frozen dough enables provision of fresh bread to consumer at all times, while reducing
manufacturing costs. However, freezing and frozen storage reduces yeast viability and damages dough
gluten-starch network. As a result, bread from frozen dough is lower in quality than that of freshly baked
bread.
This review discussed ice crystal development characteristics during freezing and frozen storage of frozen
dough as the main factor affecting quality of gluten, starch and yeast viability as they interact with water
re-distribution. Effective strategies used to lower the impact of ice crystallization and re-crystallization on
quality of frozen dough are suggested. The use of hydrocolloids, ice-structuring proteins and ice nucleating
agents as additives increase the ability of yeast to tolerate freezing and frozen storage treatment. Yeasts’
ability to produce intracellular molecules in stress response to ice crystal characteristics in relation to water
re-distribution in dough are the main influencers of quality of frozen dough components. This improves
yeast freeze tolerance and fermentation capacity of yeast. However, there is need for further research on
the interaction of frozen dough added additives on dough ingredients such as starch, water and gluten
during freezing and extended frozen storage.
Enzymes (e.g. lipase, transglutaminase, glucose oxidase) and functional microbial end products (e.g.
dextrans), as discussed, have enhanced some technological properties of frozen dough systems. However,
additional studies are needed to enrich this body of knowledge.

Knowledge on the development of ice crystals during freezing and recrystallization during frozen storage
is useful to enable optimization of freezing conditions and reduction of temperature fluctuations to

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maintain yeast activity in frozen dough.
FUTURE PROSPECTS
The concepts in food nutrition and consumption have remained dynamic in the recent decades. The past
decades were mainly characterized by less knowledge awareness on nutrition and consumption by the
individual consumer. However, with the information age era, this has tremendously changed, as more
consumers are more informed and continuously select more health promoting foods. Therefore, to cope
with such changes, the baking frozen technology industry should continue to innovate and develop better
premium quality products with novel ingredients and technologies.
In this view, the use of new and advanced freezing technologies such as high pressure and ultrasound-
assisted freezing techniques to enable better control of temperature fluctuations during frozen storage and
freezing conditions of frozen dough is inevitable. This would enable a better understanding of the
mechanism of ice recrystallization in frozen dough. Also, during freezing and frozen storage treatments;
mostly single freezing techniques or cryopreservation methods have been applied. Studies to explore the
combination effect of these techniques could improve the quality of frozen dough products.
Further search for novel frozen dough improvers will continue. As discussed, the use of ice-structuring
proteins and ice nucleating agents indicate a growing research trend in tackling ice crystal development
characteristics. Finally, the use of bio-tech ingredients for clean label consideration in frozen dough
systems is a very recent research effort. This trend is in line with the consumer need for non-synthetic or
non-chemical food ingredients to be used in food formulations. Bio-tech ingredients such as
exopolysaccharides (dextran), amino acid supplements (GABA, ACE-I inhibiting peptides), emulsifying
agents (gums like xanthan and guar) and flavor enhancers among others have been successfully
synthesized in microbial fermentation systems, but not entirely in frozen dough systems. Enzymes such as
transglutaminase and lipase have been successfully utilized in the frozen steamed dough bread systems;
thus a significant potential for future related studies.

ACKNOWLEDGEMENT
We are grateful for the financial supports of the research from Grants (31071595, 31571877) from the
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National Natural Science Foundation of China, the National High Technology Research and Development
Program of China (863 Program, 2012AA022200), Fujian “Hundreds of Talents Expert” Program of China
(20172022), Science and Technology “LiaoYuan Plan” Program of Quanzhou, Fujian Province, China
(2015G46), of BaihoBake Biotechnology International, Inc. (Nanjing, China), and MagiBake International,
Inc. (Wuxi, China).

REFERENCE
Acker, J.P., & Croteau, I.M. (2004). Pre- and post-thaw assessment of intracellular ice formation. Journal of
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Microscopy, 215, 131–138.
Alcázar-Alay, S.C., & Meireles, M.A.A. (2015). Physicochemical properties, modifications and applications of
starches from different botanical sources. Food Science and Technology, 35, 215–236.
Annunziata, A., & Vecchio, R. (2013). Consumer perception of functional foods: A conjoint analysis with
probiotics. Food Quality Preference, 28, 348–355.
Archer, D.B., Connerton, I.F., & MacKenzie, D. A. (2008). Filamentous fungi for production of food additives
and processing aids. Advances in Biochemical Engineering/Biotechnology, 111, 99–147.
Aslankoohi, E., Rezaei, M.N., & Vervoort, Y. (2015). Glycerol Production by Fermenting Yeast Cells Is
Essential for Optimal Bread Dough Fermentation. PLoS One. 1–13.
Baier-Schenk, A., Handschin, S., & Conde-Petit, B. (2005a). Ice in prefermented frozen bread dough - An
investigation based on calorimetry and microscopy. Cereal Chemistry, 82, 251–255.
Baier-Schenk, A., Handschin, S., Von Schönau, M., Bittermann, A.G., Bächi, T., & Conde-Petit, B. (2005b). In
situ observation of the freezing process in wheat dough by confocal laser scanning microscopy (CLSM):
Formation of ice and changes in the gluten network. Journal of Cereal Science, 42, 255–260.
Ballester-Tomás, L., Pérez-Torrado, R., Rodríguez-Vargas, S., Prieto, J.A., & Randez-Gil, F. (2016). Near-
freezing effects on the proteome of industrial yeast strains of Saccharomyces cerevisiae. Journal of
Biotechnology, 221, 70–77.
Ban, C., Yoon, S., Han, J., Kim, S.O., Han, J.S., Lim, S., & Choi, Y.J. (2016). Effects of freezing rate and
terminal freezing temperature on frozen croissant dough quality. LWT - Food Science and Technology, 73,
219–225.
Bárcenas, M.E., Haros, M., & Rosell, C.M. (2003). An approach to studying the effect of different bread
improvers on the staling of pre-baked frozen bread. European Food Research and Technology, 218, 56–
61.
Bartkiene, E., Vizbickiene, D., Bartkevics, V., Pugajeva, I., Krungleviciute, V., Zadeike, D., Zavistanaviciute,

P., & Juodeikiene, G. (2017). Application of Pediococcus acidilactici LUHS29 immobilized in apple
pomace matrix for high value wheat-barley sourdough bread. LWT - Food Science and Technology, 83,
Accepted Article
157–164.
Berton, C., Ropers, M.-H., Bertrand, D., Viau, M., & Genot, C. (2012). Oxidative stability of oil-in-water
emulsions stabilised with protein or surfactant emulsifiers in various oxidation conditions. Food
Chemistry, 131, 1360–1369.
Bockstaele, F. Van., Walle, D. Van De., Dewettinck, K., Gellynck, X., & Ku, B. (2009). Consumer perception
of bread quality. Appetite, 53, 16–23.
Brandam, C., Fahimi, N., & Taillandier, P. (2016). Mixed cultures of Oenococcus oeni strains: A mathematical
model to test interaction on malolactic fermentation in winemaking. LWT - Food Science and Technology,
69, 211–216.
Chen, G., Jansson, H., Lustrup, K.F., & Swenson, J. (2012). Formation and distribution of ice upon freezing of
different formulations of wheat bread. Journal of Cereal Science, 55, 279–284.
Chen, G., Öhgren, C., Langton, M., Lustrup, K.F., Nydén, M., & Swenson, J. (2013). Impact of long-term
frozen storage on the dynamics of water and ice in wheat bread. Journal of Cereal Science, 57, 120–124.
Cornejo, I., Cornejo, G., Ramírez, C., Almonacid, S., & Simpson, R. (2016). Inverse method for the
simultaneous estimation of the thermophysical properties of foods at freezing temperatures. Journal of
Food Engineering, 191, 37–47.
Couto, S.R., & Sanromán, M.Á. (2006). Application of solid-state fermentation to food industry—A review.
Journal of Food Engineering, 76, 291–302.
Decock, P., & Cappelle, S. (2005). Bread technology and sourdough technology. Trends in Food Science and
Technology, 16, 113–120.
Delcour, J. a, Joye, I.J., Pareyt, B., Wilderjans, E., Brijs, K., & Lagrain, B. (2012). Wheat gluten functionality as
a quality determinant in cereal-based food products. Annual Review of Food Science and Technology, 3,
469–92.
Delgado, A.E., & Sun, D.-W. (2001). Heat and mass transfer models for predicting freezing processes – a

review. Journal of Food Engineering, 47, 157–174.
Ding, X., Zhang, H., Wang, L., Qian, H., Qi, X., & Xiao, J. (2015). Effect of barley antifreeze protein on
Accepted Article
thermal properties and water state of dough during freezing and freeze-thaw cycles. Food Hydrocolloids,
47, 32–40.
Don, C., Mann, G., Bekes, F., & Hamer, R.J. (2006). HMW-GS affect the properties of glutenin particles in
GMP and thus flour quality. Journal of Cereal Science, 44, 127–136.
Eckardt, J., Öhgren, C., Alp, A., Ekman, S., Åström, A., Chen, G., Swenson, J., Johansson, D., & Langton, M.
(2013). Long-term frozen storage of wheat bread and dough - Effect of time, temperature and fibre on
sensory quality, microstructure and state of water. Journal of Cereal Science, 57, 125–133.
Erbaş, M., Kemal Uslu, M., Ozgun Erbaş, M., & Certel, M. (2006). Effects of fermentation and storage on the
organic and fatty acid contents of tarhana, a Turkish fermented cereal food. Journal of Food Composition
and Analysis, 19, 294–301.
Esselink, E.F.J., Van Aalst, H., Maliepaard, M., & Van Duynhoven, J.P.M. (2003). Long-term storage effect in
frozen dough by spectroscopy and microscopy. Cereal Chemistry, 80, 396–403.
Ferrero, C. (2017). Hydrocolloids in wheat breadmaking: A concise review. Food Hydrocolloids, 68, 15–22.
Freschi, J., Doran, L., Malumba, P., & Blecker, C. (2014). Impact of freezing and thawing processes on wheat
and potato starch gel syneresis. Starch/Staerke, 66, 208–215.
Gabric, D., Ben-Aissa, F., Le-Bail, A., Monteau, J.Y., & Curic, D. (2011). Impact of process conditions on the
structure of pre-fermented frozen dough. Journal of Food Engineering, 105, 361–366.
Gao, D., & Critser, J.K. (2000). Mechanisms of cryoinjury in living cells. ILAR Journal, 41, 187–196.
Gerits, L.R., Pareyt, B., Decamps, K., & Delcour, J.A. (2014). Lipases and their functionality in the production
of wheat-based food systems. Comprehensive Reviews in Food Science and Food Safety, 13, 978–989.
Gharaie, Z., Azizi, M.H., Barzegar, M., & Aghagholizade, R. (2015). Effects of Hydrocolloids on the
Rheological Characteristics of Dough and the Quality of Bread Made From Frozen Dough. Journal of
Texture Studies, 46, 365–373.

Glatz, B., & Sarupria, S. (2017). Heterogeneous ice nucleation: Interplay of surface properties and their impact
on water orientations. Langmuir acs.

Accepted Article
Goesaert, H., Brijs, K., Veraverbeke, W.S., Courtin, C.M., Gebruers, K., & Delcour, J.A. (2005). Wheat flour
constituents: how they impact bread quality, and how to impact their functionality. Trends in Food
Science and Technology, 16, 12–30.
Gray, J.A., & Bemiller, J.N. (2003). Bread Staling: Molecular Basis and Control. Comprehensive Reviews in
Food Science and Food Safety, 2, 1–21.
Hamdami, N., Monteau, J.Y., & Le Bail, A. (2004). Thermophysical properties evolution of French partly baked
bread during freezing. Food Research International, 37, 703–713.
Havet, M., Mankai, M., & Le Bail, A. (2000). Influence of the freezing condition on the baking performances of
French frozen dough. Journal of Food Engineering, 45, 139–145.
He, H., & Hoseney, R.C. (1990). Changes in Bread Firmness and Moisture During Long-Term Storage. Cereal
Chemistry, 67, 603–605.
Heenan, S.P., Hamid, N., Dufour, J., Harvey, W., & Delahunty, C.M. (2009). Consumer freshness perceptions
of breads , biscuits and cakes. Food Quality and Preference, 20, 380–390.
Huang, C.-S., Hu, H.-H., Tsai, Y.-M., & Chang, W.-T. (2013). In vitro effects of Monascus purpureus on
antioxidation activity during fermentation of Kinmen sorghum liquor waste. Journal of Bioscience and
Bioengineering, 115, 418–23.
Huang, H., Zhao, G., Zhang, Y., Xu, J., Toth, T.L., & He, X. (2017). Predehydration and Ice Seeding in the
Presence of Trehalose Enable Cell Cryopreservation. ACS Biomaterial Science and Engineering, 3, 1758–
1768.
Huang, L., Wan, J., Huang, W., Rayas-Duarte, P., & Liu, G. (2011). Effects of glycerol on water properties and
steaming performance of prefermented frozen dough. Journal of Cereal Science, 53, 19–24.
Huang, S., & Miskelly, D. (2016). Nutrition of Steamed Products. Steamed Breads. 161–168.
Huang, W., Kim, Y., Li, X., & Rayas-Duarte, P. (2008). Rheofermentometer parameters and bread specific
volume of frozen sweet dough influenced by ingredients and dough mixing temperature. Journal of

Cereal Science, 48, 639–646.
Huang, W., Yuan, Y., Kim, Y., & Chung, O. (2008). Effects of Transglutaminase on rheology, microstructure,
Accepted Article
and baking properties of frozen dough. Cereal Chemistry, 85, 301-306.
Huen, J., Weikusat, C., Bayer-Giraldi, M., Weikusat, I., Ringer, L., & Lösche, K. (2014). Confocal Raman
microscopy of frozen bread dough. Journal of Cereal Science, 60, 555–560.
Izadi Najafabadi, L., Le-Bail, A., Hamdami, N., Monteau, J.Y., & Keramat, J. (2014). Impact of baking
conditions and storage temperature on staling of fully and part-baked Sangak bread. Journal of Cereal
Science, 60, 151–156.
Jia, C., Huang, W., Ji, L., Zhang, L., Li, N., & Li, Y. (2014a). Improvement of hydrocolloid characteristics
added to angel food cake by modifying the thermal and physical properties of frozen batter. Food
Hydrocolloids, 41, 227–232.
Jia, C., Huang, W., Rayas-Duarte, P., Zou, Q., Zhang, L., & Li, Y. (2014b). Hydration, polymerization and
rheological properties of frozen gluten-water dough as influenced by thermostable ice structuring protein
extract from Chinese privet (Ligustrum vulgare) leaves. Journal of Cereal Science, 59, 132–136.
Jia, C., Huang, W., Wu, C., Zhong, J., Rayas-duarte, P., & Guo, C. (2012). Frozen Bread Dough Properties
Modified by Thermostable Ice Structuring Proteins Extract from Chinese Privet ( Ligustrum vulgare )
Leaves 162–167. Cereal Chemistry, 89, 162-167.
Jiang, Z., Le Bail, A., & Wu, A. (2008). Effect of the thermostable xylanase B (XynB) from Thermotoga
maritima on the quality of frozen partially baked bread. Journal of Cereal Science, 47, 172–179.
Kaino, T., Tateiwa, T., Mizukami-Murata, S., Shima, J., & Takagi, H. (2008). Self-cloning baker’s yeasts that
accumulate proline enhance freeze tolerance in doughs. Appllied and Environmental Microbiology, 74,
5845–5849.
Kasner, E., Hunter, C.A., & Kariko, K. (2013). NIH Public Access, 70, 646–656.
Kawahara, H., Mano, Y., & Obata, H. (1993). Purification and characterization of extracellular ice-nucleating
matter from Erwinia uredovora KUIN-3. Biosci. Biotechnol. Biochem., 57, 1429–1432.
Keeratipibul, S., Luangsakul, N., Otsuka, S., Sakai, S., Hatano, Y., & Tanasupawat, S. (2010). Application of

the Chinese steamed bun starter dough (CSB-SD) in breadmaking. Journal of Food Science, 75, E596-
604.
Accepted Article
Kiani, H., & Sun, D.W. (2011). Water crystallization and its importance to freezing of foods: A review. Trends
in Food Science and Technology, 22, 407–426.
Kiani, H., Zhang, Z., Delgado, A., & Sun, D.W. (2011). Ultrasound assisted nucleation of some liquid and solid
model foods during freezing. Food Research International, 44, 2915–2921.
Kondakci, T., Zhang, J.W., & Zhou, W. (2015). Impact of Flour Protein Content and Freezing Conditions on the
Quality of Frozen Dough and Corresponding Steamed Bread. Food and Bioprocess Technology, 8, 1877–
1889.
Kontogiorgos, V., (2011). Microstructure of hydrated gluten network. Food Research Internationl, 44, 2582–
2586.
Kontogiorgos, V., & Goff, H.D. (2006). Calorimetric and microstructural investigation of frozen hydrated
gluten. Food Biophysics, 1, 202–215.
Kumcuoglu, S., Tavman, S., Nesvadba, P., & Tavman, I.H. (2007). Thermal conductivity measurements of a
traditional fermented dough in the frozen state. Journal of Food Engineering, 78, 1079–1082.
Larsen, F.H., Blennow, A., & Engelsen, S.B. (2008). Starch granule hydration-A MAS NMR investigation.
Food Biophysics, 3, 25–32.
Le-Bail, A., Nicolitch, C., & Vuillod, C. (2010). Fermented frozen dough: Impact of pre-fermentation time and
of freezing rate for a pre-fermented frozen dough on final volume of the bread. Food and Bioprocess
Li, Y., Wang, H., & Tingrui, P. (2013). Intracellular ice formation (IIF) during freeze-thaw repetitions.
International Journal of Heat and Mass Transfer, 64, 436–443.
Li, Z., Tang, X., Huang, W., Liu, J.G., Tilley, M., & Yao, Y. 2011. Rheology, Microstructure, and Baking
Characteristics of Frozen Dough Containing Rhizopus chinensis Lipase and Transglutaminase. Cereal
Chemistry, 88, 596–601.
Lindow, S.E., Arny, D.C., & Upper, C.D. (1982). Bacterial Ice Nucleation: A Factor in Frost Injury to Plants.

Plant Physiology, 70, 1084–1089.
Linlaud, N., Ferrer, E., Puppo, M.C., & Ferrero, C. (2011). Hydrocolloid interaction with water, protein, and
Accepted Article
starch in wheat dough. Journal of Agricultural and Food Chemistry, 59, 713–719.
Liu, C., Chang, Y., Li, Z., & Liu, H. (2012). Effect of ratio of yeast to Jiaozi on quality of Chinese steamed
bread. Procedia Environmental Sciences, 12, 1203–1207.
Liu, C., Li, L., Hao, C., Zheng, X., Bian, K., Zhang, J., & Wang, X. (2015). Effects of different milling
processes on whole wheat flour quality and performance in steamed bread making. LWT - Food Science
and Technology, 62, 310-318.
Liu, T., Ruan, H., & He, G. (2016). A study revealing the key aroma compounds of steamed bread made by
chinese traditional sourdough. Journal of Zhejiang University-Science B (Biomedicine & Biotechnology),
1581, 1–11.
Luo, W., Sun, D.-W., Zhu, Z., & Wang, Q.-J. (2017). Improving freeze tolerance of yeast and dough properties
for enhancing frozen dough quality - A review of effective methods. Trends in Food Science and
Maity, T., Saxena, A., & Raju, P.S. (2017). Use of hydrocolloids as cryoprotectant for frozen foods. Critical
Reviews in Food Science and Nutrition, 0, 1–16.
Matsumiya, K., Takahashi, Y., Nakanishi, K., Dotsu, N., & Matsumura, Y. (2014). Diglycerol esters of fatty
acids promote severe coalescence between protein-stabilized oil droplets by emulsifier-protein
competitive interactions. Food Hydrocolloids, 42, 397–402.
Matuda, T.G., Parra, D.F., Lugão, A.B., & Tadini, C.C. (2005). Influence of vegetable shortening and
emulsifiers on the unfrozen water content and textural properties of frozen French bread dough. LWT -
Food Science and Technology, 38, 275–280.
Matuda, T.G., Pessôa Filho, P.A., & Tadini, C.C. (2011). Experimental data and modeling of the
thermodynamic properties of bread dough at refrigeration and freezing temperatures. Journal of Cereal
Science, 53, 126–132.
Meziani, S., Ioannou, I., Jasniewski, J., Belhaj, N., Muller, J.M., Ghoul, M., & Desobry, S. (2012a). Effects of

freezing treatments on the fermentative activity and gluten network integrity of sweet dough. LWT - Food
Science and Technology, 46, 118–126.

Accepted Article
Meziani, S., Kaci, M., Jacquot, M., Jasniewski, J., Ribotta, P., Muller, J.-M., Ghoul, M., & Desobry, S. (2012b).
Effect of freezing treatments and yeast amount on sensory and physical properties of sweet bakery
products. Journal of Food Engineering, 111, 336–342.
Mizunoe, Y., Watanabe, S., Sudo, Y., Kobayashi, M., Yasukawa, H., Natori, D., Hoshino, A., Negishi, A.,
Okita, N., Komatsu, M., & Higami, Y. (2017). Trehalose protects against oxidative stress by regulating
the Keap1–Nrf2 and autophagy pathways. Redox Biology, 15, 115–124.
Molina, M.T., Leiva, A., & Bouchon, P. (2016). Examining the effect of freezing on starch gelatinization during
heating at high rates using online in situ hot-stage video-microscopy and differential scanning calorimetry.
Food Bioproducts Processing, 100, 488–495.
Morita, Y., Nakamori, S., & Takagi, H. (2002). Effect of proline and arginine metabolism on freezing stress of
Saccharomyces cerevisiae. Journal of Bioscience and Bioengineering, 94, 390–394.
Nakagawa, Y., Ogihara, H., Mochizuki, C., Yamamura, H., Iimura, Y., & Hayakawa, M. (2017). Development
of intra-strain self-cloning procedure for breeding baker’s yeast strains. Journal of Bioscience and
Bioengineering, 123, 319–326.
Nakamura, T., Takagi, H., & Shima, J. (2009). Effects of ice-seeding temperature and intracellular trehalose
contents on survival of frozen Saccharomyces cerevisiae cells. Cryobiology, 58, 170–174.
Nout, M.J.R. (2009). Rich nutrition from the poorest - cereal fermentations in Africa and Asia. Food
Microbiology, 26, 685–92.
Omedi, J.O., Huang, W., Su, X., Liu, R., Tang, X., Xu, Y., & Rayas-Duarte, P. (2016). Effect of five lactic acid
bacteria starter type on angiotensin-I converting enzyme inhibitory activity and emulsifying properties of
soy flour sourdoughs with and without wheat bran supplementation. Journal of Cereal Science, 69, 57-63.
Öztürk, S., Cerit, I., Mutlu, S., & Demirkol, O. (2017). Enrichment of cookies with glutathione by inactive yeast
cells (Saccharomyces cerevisiae): Physicochemical and functional properties. Journal of Cereal Science,
78, 19-24.

Pareyt, B., Finnie, S.M., Putseys, J.A., & Delcour, J.A. (2011). Lipids in bread making : Sources , interactions ,
and impact on bread quality. Journal of Cereal Science, 54, 266–279.

Accepted Article
Perry, P. A, & Donald, A. M. (2000). The effects of low temperatures on starch granule structure. Polymer, 41,
6361–6373.
Przetaczek-Rożnowska, I. (2017). Physicochemical properties of starches isolated from pumpkin compared with
potato and corn starches. International Journal of Biological Macromolecules, 101, 536–542.
Rhee, S.J., Lee, J.-E., & Lee, C.-H. (2011). Importance of lactic acid bacteria in Asian fermented foods.
Microbial Cell Factories.
Ribotta, P.D., León, A.E., & Añón, M.C. 2001. Effect of freezing and frozen storage of doughs on bread quality.
Journal of Agricultural and Food Chemistry, 49, 913–918.
Ronda, F., Caballero, P. A., Quilez, J., & Roos, Y.H. (2011). Staling of frozen partly and fully baked breads.
Study of the combined effect of amylopectin recrystallization and water content on bread firmness.
Journal of Cereal Science, 53, 97–103.
Rosell, C.M., & Gómez, M. (2007). Frozen dough and partially baked bread: An update. Food Reviews
International, 23, 303–319.
Sasano, Y., Haitani, Y., Hashida, K., Ohtsu, I., Shima, J., & Takagi, H. (2012). Simultaneous accumulation of
proline and trehalose in industrial baker’s yeast enhances fermentation ability in frozen dough. Journal of
Bioscience and Bioengineering, 113, 592–5.
Selomulyo, V.O., & Zhou, W. (2007). Frozen bread dough: Effects of freezing storage and dough improvers.
Journal of Cereal Science, 45, 1–17.
Shi, K., Yu, H., Jin, J., & Lee, T.C. (2013a). Improvement to baking quality of frozen bread dough by novel
zein-based ice nucleation films. Journal of Cereal Science, 57, 430–436.
Shi, K., Yu, H., & Lee, T.C. (2013b). A novel approach for improving yeast viability and baking quality of
frozen dough by adding biogenic ice nucleators from Erwinia herbicola. Journal of Cereal Science, 57,
237–243.
Simmons, A.L., Serventi, L., & Vodovotz, Y. (2012). Water dynamics in microwavable par-baked soy dough

evaluated during frozen storage. Food Research International, 47, 58–63.
Soveral, G., Madeira, A., Loureiro-Dias, M.C., & Moura, T.F. (2008). Membrane tension regulates water
Accepted Article
transport in yeast. Biochimica et Biophysica Acta, 1778, 2573–2579.
Stefanello, R.F., Machado, A.A.R., Pasqualin Cavalheiro, C., Bartholomei Santos, M.L., Nabeshima, E.H.,
Copetti, M.V., & Fries, L.L.M. (2018). Trehalose as a cryoprotectant in freeze-dried wheat sourdough
production. LWT - Food Science and Technology, 89, 510–517.
Steffolani, M.E., Ribotta, P.D., Perez, G.T., Puppo, M.C., & León, A.E. (2012). Use of Enzymes to Minimize
Dough Freezing Damage. Food and Bioprocess Technology, 5, 2242–2255.
Szymonska, J., Krok, F., Komorowska-Czepirska, E., & Rebilas, K. (2003). Modification of granular potato
starch by multiple deep-freezing and thawing. Carbohydrate Polymer, 52, 1–10.
Tan, H., Dong, J., Wang, G., Xu, H., Zhang, C., & Xiao, D. (2014). Enhanced freeze tolerance of baker’s yeast
by overexpressed trehalose-6-phosphate synthase gene (TPS1) and deleted trehalase genes in frozen
dough. Journal of Industrial Microbiology and Biotechnology, 41, 1275–1285.
Tang, X., Liu, N., Huang, W., Cheng, X., Wang, F., Zhang, B., Chen, J., Jiang, H., Omedi, J.O., & Li, Z. (2018).
Syneresis rate, water distribution, and microstructure of wheat starch gel during freeze-thaw process: Role
of a high molecular weight dextran produced by Weissella confusa QS813 from traditional sourdough.
Cereal Chemistry. doi:10.1094/CCHEM-08-17-0174-R
Tang, X., Wang, F., Huang, W., Zou, Q., Jia, C., Jin, D., Omedi, J.O., & Li, Z. (2016). The combination of
Rhizopus chinensis Lipase and Transglutaminase affects the rheology and gluten macropolymer properties
of frozen dough. Cereal Chemistry. doi: 10.1094/CCHEM-08-15-017
Tao, H., Wang, P., Wu, F., Jin, Z., & Xu, X. (2016a). Particle size distribution of wheat starch granules in
relation to baking properties of frozen dough. Carbohydrate Polymer, 137, 147–153.
Tao, H., Wang, P., Zhang, B., Wu, F., Jin, Z., & Xu, X. (2016b). A comparative study of sodium dodecyl sulfate
and freezing/thawing treatment on wheat starch: The role of water absorption. Carbohydrate Polymer,
143, 149–154.
Tao, H., Yan, J., Zhao, J., Tian, Y., Jin, Z., & Xu, X. (2015). Effect of multiple freezing/thawing cycles on the

structural and functional properties of waxy rice starch. PLoS One, 10, 1–11.
Tao, H., Zhang, B., Wu, F., Jin, Z., & Xu, X. (2016c). Effect of multiple freezing / thawing-modified wheat
Accepted Article
starch on dough properties and bread quality using a reconstitution system. Journal of Cereal Science, 69,
132–137.
Terao, Y., Nakamori, S., & Takagi, H. (2003). Gene Dosage Effect of L -Proline Biosynthetic Enzymes on L -
Proline Accumulation and Freeze Tolerance in Saccharomyces cerevisiae. Applied and Environmental
Microbiology, 69, 6527–6532.
Tsolmonbaatar, A., Hashida, K., Sugimoto, Y., Watanabe, D., Furukawa, S., & Takagi, H. (2016). Isolation of
baker’s yeast mutants with proline accumulation that showed enhanced tolerance to baking-associated
stresses. International Journal of Food Microbiology, 238, 233–240.
Vamadevan, V., & Bertoft, E. (2015). Structure-function relationships of starch components. Starch/Staerke 67,
55–68.
Van Steertegem, B., Pareyt, B., Brijs, K., & Delcour, J.A. (2013). Impact of mixing time and sodium stearoyl
lactylate on gluten polymerization during baking of wheat flour dough. Food Chemistry, 141, 4179–4185.
Vanderveer, T.L., Choi, J., Miao, D., & Walker, V.K. (2014). Expression and localization of an ice nucleating
protein from a soil bacterium, Pseudomonas borealis. Cryobiology, 69, 110–118.
Wang, P., Chen, H., Mohanad, B., Xu, L., Ning, Y., Xu, J., Wu, F., Yang, N., Jin, Z., & Xu, X. (2014a). Effect
of frozen storage on physico-chemistry of wheat gluten proteins: Studies on gluten-, glutenin- and gliadin-
rich fractions. Food Hydrocolloids, 39, 187–194.
Wang, P., Jin, Z., & Xu, X. (2015a). Physicochemical alterations of wheat gluten proteins upon dough formation
and frozen storage - A review from gluten, glutenin and gliadin perspectives. Trends in Food Science and
Wang, P., Mao, J., Meng, X., Li, X., Liu, Y., & Feng, H. (2014). Changes in flavour characteristics and bacterial
diversity during the traditional fermentation of Chinese rice wines from Shaoxing region. Food Control,
44, 58–63.
Wang, P., Tao, H., Jin, Z., & Xu, X. (2016). Impact of water extractable arabinoxylan from rye bran on the

frozen steamed bread dough quality. Food Chemistry, 200, 117–124.
Wang, P., Tao, H., Jin, Z., & Xu, X. (2015b). The final established physicochemical properties of steamed bread
Accepted Article
made from frozen dough: Study of the combined effects of gluten polymerization, water content and
starch crystallinity on bread firmness. Journal of Cereal Science, 63, 116–121.
Wang, P., Xu, L., Nikoo, M., Ocen, D., Wu, F., Yang, N., Jin, Z., & Xu, X. (2014b). Effect of frozen storage on
the conformational, thermal and microscopic properties of gluten: Comparative studies on gluten-,
glutenin- and gliadin-rich fractions. Food Hydrocolloids, 35, 238–246.
Wang, P., Yang, R., Gu, Z., Xu, X., & Jin, Z. (2017a). Comparative study of deterioration procedure in
chemical-leavened steamed bread dough under frozen storage and freeze/thaw condition. Food Chemistry,
229, 464–471.
Wang, P., Yang, R., Gu, Z., Xu, X., & Jin, Z. (2017b). Comparative study on the freeze stability of yeast and
chemical leavened steamed bread dough. Food Chemistry, 221, 482–488.
Wieser, H., (2007). Chemistry of gluten proteins. Food Microbiology, 24, 115–9.
Wojtasz, J., Carlstedt, J., Fyhr, P., & Kocherbitov, V. (2016). Hydration and swelling of amorphous cross-linked
starch microspheres. Carbohydrate Polymer, 135, 225–233.
Xu, H.N., Huang, W., Jia, C., Kim, Y., & Liu, H. (2009). Evaluation of water holding capacity and breadmaking
properties for frozen dough containing ice structuring proteins from winter wheat. Journal of Cereal
Science, 49, 250–253.
Xuan, Y.F., Zhang, Y., Zhao, Y.Y., Zheng, Z., Jiang, S.T., & Zhong, X.Y. (2017). Effect of
hydroxypropylmethylcellulose on transition of water status and physicochemical properties of wheat
gluten upon frozen storage. Food Hydrocolloids, 63, 35–42.
Yi, J., Johnson, J.W., & Kerr, W.L. (2009). Properties of bread made from frozen dough containing waxy wheat
flour. Journal of Cereal Science, 50, 364–369.
Yi, J., & Kerr, W.L. (2009). Combined effects of freezing rate, storage temperature and time on bread dough
and baking properties. LWT - Food Science and Technology, 42, 1474–1483.
Yu, S., Ma, Y., & Sun, D.W. (2010). Effects of freezing rates on starch retrogradation and textural properties of

cooked rice during storage. LWT - Food Science and Technology, 43, 1138–1143.
Zaritzky, N.E. (2010). Chemical and physical deterioration of frozen foods, Chemical Deterioration and
Accepted Article
Physical Instability of Food and Beverages. Woodhead Publishing Limited.
Zhang, C., Zhang, H., & Wang, L. (2007). Effect of carrot (Daucus carota) antifreeze proteins on the
fermentation capacity of frozen dough. Food Research International, 40, 763–769.
Zhang, M., Oldenhof, H., Sieme, H., & Wolkers, W.F. (2016). Freezing-induced uptake of trehalose into
mammalian cells facilitates cryopreservation. Biochimica et Biophysica Acta, 1858, 1400–1409.
Zhang, Y., Zhang, H., Wang, L., Qian, H., & Qi, X. (2015). Extraction of Oat (Avena sativa L.) Antifreeze
Proteins and Evaluation of Their Effects on Frozen Dough and Steamed Bread. Food Bioprocess
Zhao, L., Li, L., Liu, G., Chen, L., Liu, X., Zhu, J., & Li, B. (2013). Effect of freeze–thaw cycles on the
molecular weight and size distribution of gluten. Food Research International, 53, 409–416.
Zhao, L., Liu, X., Hu, Z., Li, L., & Li, B. (2016). Molecular Structure Evaluation of Wheat Gluten during
Frozen Storage. Food Biophysics,1, 60–68.
Zhu, F. (2017). Structures, properties, modifications, and uses of oat starch. Food Chemistry, 229, 329–340.
Zhu, F. (2014). Influence of ingredients and chemical components on the quality of Chinese steamed bread.
Food Chemistry, 163, 154–162.

List of tables:
Table I: Effect of freezing and frozen storage on starch, gluten and yeast functionality in frozen dough.
Accepted Article
Table II: Effect of different additives on cryopreservation of frozen dough system.
List of figures:
Figure 1a: Manufacturing process of frozen dough products to the point of retail. P/F-FD; pre/fully
fermented frozen dough, PB-FD; partially baked frozen dough, U-FD; unfermented frozen dough, FB-F;
fully baked frozen. Adapted and modified from (Decock & Cappelle, 2005) and (Wang et al., 2015a).
Figure 1b: Production of pre-proofed ( ) and un-proofed ( ) frozen doughs for frozen steamed
products. Adapted from (Huang & Miskelly, 2016).
Figure 2: Micrographs of fractured dough from dough center illustrating ice recrystallization during frozen
storage of pre-fermented frozen dough. A; fast frozen in liquid N2 after proofing (no storage), B; frozen 1h
in freezer (-28°C) to 11°C (no further storage), C; frozen 2h in freezer (-28°C), stored 1 day at -22°C, and
D; frozen 2h in freezer (-28°C), stored 149 days at 22°C in freezing room. Adopted from (Baier-Schenk et
al., 2005a).
Figure 3: Schematic description of structural changes of hydrated gluten network during frozen storage at
-18°C for 0 (A, C) and 60 (D, E) days. Left to right: Atomic scale of atomic force microscopy (AFM)
images and cross-section of gluten protein network. At nanoscale, gluten sheets arranged in nano-capillary
format filled with confined and rigid water, and surrounded by bulk water; state of water redistribution in
gluten upon frozen storage. At microscale, SEM images of gluten and secondary structural changes in
gluten matrix with the increased frozen storage time. Reproduced images adopted from (Kontogiorgos,
2011; Kontogiorgos & Goff, 2006; Zhao et al., 2016; Xuan et al., 2017)
Figure 4: SEM images: 1Effect sodium dodecyl sulfate (SDS) pretreatment and freeze-thawing treatment
on native wheat starch (adopted from (Tao et al., 2016b)). 2Freezing treatment on particle size distribution
of wheat starch granules (adopted from (Tao et al., 2016a)).

Figure 5: TEM micrographs of dough: Effect of freezing treatment/rates on fermentative activity of yeast
cell structure. A; yeast in unfrozen dough. Yeast in dough after, B; slow freezing rate (-0.06°C/min), C;
Accepted Article
slow freezing rate (-0.23°C/min), D; intermediate freezing rate (-0.36°C/min), and E; rapid freezing rate (-
8.6°C/min) (adopted from (Meziani et al., 2012a)).

ccepted Articl TABLE 1: Effect of freezing and frozen storage on starch, gluten and yeast functionality in frozen dough.
Component Object of study Result of treatment Effect on functionality Reference(s)
Gluten Physico-chemical changes on wheat GMP depolymerisation. Loss of elastic (G’) and viscous Lowered rheological property (Wang et al., 2014a)
gluten-, glutenin- and gliadin- modulus (G’’) of gluten and glutenin. No change in gliadin
fractions fraction
Structural behavior of gluten and Enhanced SDS-extractable proteins upon frozen storage; due to Decreased degree of gluten polymerization (Wang et al., 2015b)
starch during steaming process of less incorporated glutenin in the glutenin-gliadin crosslink of
frozen dough. steamed bread
Starch Native starch in water, carrageenan In excess water, delayed gelatinization; limited water diminished Less firmness/ improved specific volume (Molina et al., 2016)
gel or sucrose solution difference of gelatinization.
Functional properties of SDS- Increased the temperature of onset of gelatinization Lower bread specific volume. Increased bread (Tao et al., 2016b)
pretreated native wheat starch Increased the enthalpy change of gelatinization. hardness
Increased pasting viscosities
Multiple freeze/thaw modified wheat Increase in freeze/thaw cycles, the lower starch gelatinization Increased starch crystallinity in bread crumbs. (Tao et al., 2016c)
starch on dough properties. enthalpy and the higher the retrogradation enthalpy of Increased firmness of bread crumbs.
amylopectin.
Structural behavior of gluten and Decreased moisture content of steamed bread; due to ice Increased bread firmness (Wang et al., 2015b)
starch during steaming process of crystallization.
frozen dough Increased starch crystallinity; due to increase in melting enthalpy
of amylopectin and rate of retrogradation.
Wheat starch granule particle size No damage to A-type surface. Cracked B-type structure Lower specific volume, increased hardness in (Tao et al., 2016a)

ccepted Articl distribution in frozen dough reconstituted bread made from B-type
granules
Yeast Self-cloning of yeast cells mutants Increased intracellular proline, trehalose, and glycerol. Higher fermentation level. (Kaino et al., 2008;
Increased freeze-thaw tolerance. Nakagawa et al., 2017;
Sasano et al., 2012;
Tsolmonbaatar et al.,
2016)
Gene engineering of yeast Gene overexpression of GPD1, MAL62, TSP1, PRO1, SNR84, Increased freeze tolerance and freeze-thaw. (Sasano et al., 2013;
PDE2. Increased synthesis of cryo-preservative Sasano et al., 2010; Tan et
Gene deletion of POG1, both NTH1 and PUT1). molecules al., 2014; Aslankoohi, et
al., 2015)
Comparison of yeast and chemical Chemical leavened dough more freeze-stable than yeast Decreased yeast viability, but better gas (Wang et al., 2017b)
leavened frozen dough leavened dough after 3 FT cycles. retention compared to chemical leavened
dough

ccepted Articl
Category
TABLE II: Effect of different additives, enzymes and functional microbial end products on cryopreservation of frozen dough system.
Study objective Result Ingredients used (flour basis) Reference
Hydrocolloid Gum (tragacanth, salep) influence on Increased water absorption, improved bread sensory, 2% salt, 2% yeast, 0.5-1% gum, 53.8- (Gharaie et al., 2015)
rheological properties flat bread from frozen less firmness and chewiness; especially with 58.2% water depending on gum.
dough Tragacanth gum
Gum (xanthan, locust bean, guar), pectin on Increased water restriction increased freezable water. 2% salt, 1.5% hydrocolloid, 60.2-66% (Linlaud et al., 2011)
interaction with water, protein, starch of Decreased starch final gelatinization temperature in water depending on hydrocolloid.
frozen wheat dough. excess water. Disordered and labile network in
presence of pectin. Good compatibility between
hydrocolloid and gluten network; guar gum
Hydrocolloids (carboxymethyl cellulose, Frozen storage negatively affected batter properties 308% egg white, 130% soft sugar, 2.5% (Jia et al., 2014a)
locust bean gum, xanthan gum) and storage (viscosity, specific gravity, bubble size, hardness); salt, 1.5% tartar powder, 1% for each
conditions on angel food cake frozen batter hydrocolloids improved all affected properties hydrocolloid.
properties. affected by frozen storage and freeze-thaw cycle;
increased specific volume and reduced hardness in
cake.
Antifreeze protein (AFP) Oat extracted AFP on frozen dough and Lowered freezable water content in frozen dough. 50% deionized water, 1.5% instant dry (Zhang et al., 2015)
or Ice structuring protein steamed bread quality Improved fermentation capacity. yeast, 0.1g AFP
(ISP) Protect gluten network matrix.
Improved texture of bread
Barley extracted AFP on thermal properties Increased apparent specific heat, freezing Not reported (Ding et al., 2015)

ccepted Articl and water state of frozen dough temperature, influenced the melting performance and
gelation properties of frozen dough, slowed decrease
in moisture content,
Carrot extracted AFP on fermentation capacity Increased yeast viability, improved gas retention 2% instant yeast, 4% sucrose, 1.5% salt, (Zhang et al., 2007)
of frozen dough capacity, retard ice crystal formation 62% water, 5% butter, 0.62% protein
(carrot concentrate protein or bovine
serum albumin or soy protein isolate)
Ice structuring proteins (ISP) isolated from Increased water holding capacity, improved 60% water, 1.5% yeast, 4% sugar, 1.5 salt, (Xu et al., 2009)
wheat on water holding and bread making breadmaking properties (proofing time, specific 4% shortening, 0.3% or 0.6% ISP
properties of frozen dough volume)
Thermostable ice structuring proteins TSISPs inhibited dehydration of gluten proteins, 25g vital gluten flour, 0.125g TSISPs, (Jia et al., 2014b)
(TSISPs) extracted from Chinese privet decreased α-helix, increased β-sheet and 25ml water.
(Ligustrum vulgare) on hydration, HMW/LMW ratio, improved rheological properties
polymerization and rheological properties of of gluten dough
gluten-water frozen dough
Thermostable ice structuring proteins Decreased freedom of water molecules and the 4% sugar, 1.5% salt, 2% yeast, 60% water, (Jia et al., 2012)
(TSISPs) extracted from Chinese privet melting enthalpy of ice (∆H); improved 4% shortening, 0.5% ISPs
(Ligustrum vulgare) on frozen bread dough microstructure, fermentation and baking properties of
properties frozen dough bread
Ice nucleation agents Biogenic ice nucleators from Improve yeast viability, decreased hardness and 56% water, 1.6% yeast, 6% sucrose, 1.5% (Shi et al., 2013b)
(INA) Erwiniaherbicola on yeast viability and improved specific volume salt, 6% shortening, 0.35mg/g of INA

ccepted Articl baking quality of frozen dough
Zein based ice nucleation films on baking Increase specific volume (due to improved yeast 56% water, 1.6% yeast, 6% sucrose, 1.5% (Shi et al., 2013a)
quality of frozen dough viability), reduced water loss (higher water content in salt, 6% shortening, 0.35mg/g of INA
bread), lower firmness
Ingredient and process Flour protein content and freezing conditions Higher flour protein content led to better resistance to Flour’ protein: 7.5% (low), 9.5% (Kondakci et al., 2015)
conditions on frozen dough and steamed bread quality freezing damage of dough; improved specific (medium), 11% (high). Water: 54% (low),
volume, form ratio and texture of steamed bread. 57% (medium), 59% (high). 1% salt, 1%
instant yeast.
Rye bran Water extractible arabinoxylan (WEAX) from Water extractible arabinoxylan improved the loaf 1.1% active dry yeast, 54.4% water, 0, 1, (Wang et al., 2016)
rye bran on frozen steam bread quality volume and lowered rate of firmness of steamed 2% flour replaced by WEAX
bread.
Functional microbial end EPS produced by Weissella confusa on Reduced syneresis rate, altered water distribution and 0.25, 0.5, 0.75, 0.1g purified EPS, 100mL (Tang et al., 2018)
products syneresis rate, water distribution and mobility, more dense and uniform gel microstructure deionized water, 8g unmodified wheat
microstructure of wheat starch gel during starch.
freeze-thaw process
Enzyme Single effect of glucose oxidase (Gox), Compared to the control (no enzyme), bread loaf was 3% compressed yeast, 2.2% salt, 58.5% (Steffolani et al., 2012)
transglutaminase (TG), and pentosanase (Pn) improved differently by each enzyme: high Gox water, enzymes: 0.001%, 0.005%, 0.01%
on frozen dough concentration required, Pn needed a longer time (9 Gox; 0.01%, 0.1%, 0.5% TG; 0.006%,
weeks), while an intermediate TG concentration was 0.012%, 0.018% Pn.
needed.

ccepted Articl Combined effect of Rhizopuschinensis lipase
and transglutaminase on rheology,

Improved G’ and G’’, rheofermentative
characteristics, specific volume and textural

60% water, 2% instant dry yeast, 1% salt,
8% shortening, 8% sugar.
(Li et al., 2011)
microstructure and baking properties of frozen properties of frozen dough
dough

Accepted Article
P-FD PB-FD
Freezing Freezing
Ferment Pre-bake
Ingredients Mixing Molding Ferment Baking
Freezing Freezing
e e
U-FD FB-F
Figure 1: Manufacturing process of frozen dough products to the point of retail. P-FD; pre/fully fermented
frozen dough, PB-FD; partially baked frozen dough, U-FD; unfermented frozen dough, FB-F; fully baked
frozen, FB-F. Adopted and modified from (Decock & Cappelle, 2005) and (Wang et al., 2015a).

Ingredient formulation
Accepted Article
Mixing
Fermentation
Divide and mold Rest
Proof
Cooling, freezing, packing
Frozen storage
Thaw Proof
Steaming
Figure 1b: Production of pre-proofed ( ) and un-proofed ( ) frozen doughs for frozen steamed
products. Adopted from (Huang & Miskelly, 2016).

A B
Accepted Article
C D
Figure 2: Micrographs of fractured dough from dough center illustrating ice recrystallization during frozen
storage of pre-fermented frozen dough. A; fast frozen in liquid N2 after proofing (no storage), B; frozen 1h
in freezer (-28°C) to 11°C (no further storage), C; frozen 2h in freezer (-28°C), stored 1 day at -22°C, and
D; frozen 2h in freezer (-28°C), stored 149 days at 22°C in freezing room. Adopted from (Baier-Schenk et
al., 2005a).

ccepted Articl
Atomic scale Nano scale Microscale Secondary changes
A B C F
D E

ccepted Articl
Figure 3: Schematic description of structural changes of hydrated gluten network during frozen storage at -18°C for 0 (A, C) and 60 (D, E) days. Left to right:
Atomic scale of AFM images and cross-section of gluten protein network. At nanoscale, gluten sheets arranged in nano-capillary format filled with confined and
rigid water, and surrounded by bulk water; state of water redistribution in gluten upon frozen storage. At microscale, SEM images of gluten and secondary structural
changes in gluten matrix with the increased frozen storage time. Reproduced images adopted from (Kontogiorgos, 2011; Kontogiorgos & Goff, 2006; Zhao et al.,
2016; Xuan et al., 2017)

1 1 1 1
Native wheat starch Freezing-treated wheat SDS treated wheat SDS + freezing-treated
Accepted Article
starch (3FTS) starch (SDS) wheat starch
2 2 2 2
A- type starch granule Freezing treated A- type B- type starch granule Freezing treated B-
starch granule type starch granule
Figure 4: SEM images: 1Effect SDS pretreatment and freeze-thawing treatment on native wheat starch
(adopted from (Tao et al., 2016b)). 2Freezing treatment on particle size distribution of wheat starch
granules (adopted from (Tao et al., 2016a)).

Accepted Article
Figure 5: TEM micrographs of dough: Effect of freezing treatment/rates on fermentative activity of yeast
cell structure. A; yeast in unfrozen dough. Yeast in dough after, B; slow freezing rate (-0.06°C/min), C;
slow freezing rate (-0.23°C/min), D; intermediate freezing rate (-0.36°C/min), and E; rapid freezing rate (-
8.6°C/min) (adopted from (Meziani et al., 2012a)).

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DR.

WEINING HUANG (Orcid ID : 0000-0003-1411-1353)

Article type : Review

ingredients development trends_a review

Jacob Ojobi OMEDI1 , and Weining HUANG*

Jiangnan University, Wuxi 214122, China

wnhuang@jiangnan.edu.cn (W. Huang).

end product quality of frozen bread-dough products.

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characteristics in frozen dough. Knowledge on development of ice crystallization and recrystallization

baking and food industry.

Ice crystals, Gluten, Yeast, Starch,

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Brief history of frozen dough products

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increased gas solubility with decline in temperature.

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the consumer at their convenient time in the market.

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suggested as a case study.

Ice crystal characteristics during freezing and frozen storage treatment

stages, respectively are discussed.

and ice growth.

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component tissue (Nakamura, Takagi, & Shima, 2009).

quality shortcomings of frozen dough.

Ice recrystallization during frozen storage of dough:

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bread-dough systems need to be done.

melting enthalpy (∆H) in many reported studies.

is mass of water in frozen dough (g).

2016; Luo, Sun, Zhu, & Wang, 2017).

Freezing rate on ice crystal characteristics in frozen dough.

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morphology, and functionality.

EFFECT OF FREEZING AND FROZEN STORAGE ON MAJOR DOUGH COMPONENTS AND

dough quality are discussed.

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bad eating quality.

Macro-molecularly, using an electrophoretic analysis of SDS-soluble protein aggregates extracted from

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illustrated by atomic force microscopy (AFM) (Fig. 3) (Zhao et al., 2013).

between GMP and water during frozen storage.

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fractions were more sensitive to frozen storage than glutenin-rich fractions.

gluten in frozen dough.

STARCH IN FROZEN DOUGH

and structure are discussed.

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after multiple freeze-thaw (Szymonska, Krok, Komorowska-Czepirska, & Rebilas, 2003).

wheat granules; indicative of sensitivity of B-type granules to freeze-thaw treatment. In a reconstituted

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YEAST ACTIVITY AND VIABILITY IN FROZEN DOUGH

viability and its impact in frozen dough quality is discussed.

ability to metabolize different molecules.

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activity and viability; high levels of yeast is used.

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strains release ∆1-pyroline-5-carboxylate reductase to convert ∆1-pyroline-5-carboxylate into proline,

which accumulates to confer freeze-tolerance. In freeze-tolerant yeast cell mutants, co-overexpression of

Furukawa, & Takagi, 2016).

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FROZEN BREAD DOUGH IMPROVER AND CRYOPROTECTIVE TECHNOLOGY;