International Journal of Food Microbiology 368 (2022) 109610

Contents lists available at ScienceDirect

International Journal of Food Microbiology

journal homepage: www.elsevier.com/locate/ijfoodmicro

Emerging trends of microbial technology for the production of

oligosaccharides from biowaste and their potential application as prebiotic
Mukesh Kumar Awasthi, Ph.D. a, *, 1, Ayon Tarafdar b, 1, Vivek Kumar Gaur c, 1, K. Amulya d, 1,
Vivek Narisetty e, Dheeraj Kumar Yadav f, Raveendran Sindhu g, h, Parameswaran Binod h,
Taru Negi i, Ashok Pandey j, k, l, Zengqiang Zhang a, *, Ranjna Sirohi l, m, **
a
College of Natural Resources and Environment, Northwest A&F University, Yangling 712100, Shaanxi Province, PR China
b
Livestock Production and Management Section, ICAR-Indian Veterinary Research Institute, Izatnagar, Bareilly 243 122, Uttar Pradesh, India
c
Environment Toxicology Division, CSIR-Indian Institute of Toxicology Research, Lucknow 226001, India
d
Bioengineering and Environmental Sciences (BEES), Department of Energy and Environmental Engineering, CSIR-Indian Institute of Chemical Technology, Hyderabad
500047, India
e
Innovation Centre, Gallows Hill, Warwick CV34 6UW, United Kingdom
f
Department of Food Engineering, National Institute of Food Technology Entrepreneurship and Management, Sonipat 131 028, Haryana, India
g
Department of Food Technology, T K M Institute of Technology, Kollam 691 505, Kerala, India
h
Microbial Processes and Technology Division, CSIR-National Institute for Interdisciplinary Science and Technology (CSIR-NIIST), Trivandrum 695 019, India
i
Department of Food Science and Technology, G. B. Pant University of Agriculture and Technology, Pantnagar, Uttarakhand 263 145, India
j
Center for Innovation and Translational Research, CSIR-Indian Institute of Toxicology Research, Lucknow 226 001, India
k
School of Engineering, University of Petroleum and Energy Studies, Dehradun 248007, Uttarakhand, India
l
Centre for Energy and Environmental Sustainability, Lucknow 226 029, Uttar Pradesh, India
m
Department of Chemical and Biological Engineering, Korea University, 145, Anam-ro, 9 Seongbuk-gu, Seoul 02841, South Korea

A R T I C L E I N F O A B S T R A C T

Keywords: Oligosaccharides are the sugars made up of 3–10 saccharides units and one of the classes of prebiotics obtained
Waste utilization from various biowastes. These biowastes could include rice straw, husk, spent coffee grounds, sugarcane bagasse,
Hydrolysis spent tea leaves, fruits and vegetables peel, corn stalk, corn stover, deoiled meals and brewer's spent grains etc.,
Saccharides
which can be used as a resource for oligosaccharides production. This review aims to provide a comprehensive
Prebiotics
overview of the suitability of different biowaste resources for oligosaccharide production followed by critical
Food waste
analysis of the recent updates and production methods. The review also discusses the tremendous prebiotic
potential of oligosaccharides in food applications with prospects for further advancements in the field.

1. Introduction selectively fermented ingredient that results in specific changes in the

Oligosaccharides are carbohydrates comprising a small number (3-
10) of simple sugars units called mono-saccharides. Oligosaccharides
such as fructooligo-saccharides and galacto-oligosaccharides are
commercially available, however, in recent years much attention has
been diverted towards higher performing and lower cost prebiotic in­
gredients such as pectic oligosaccharides (Awasthi et al., 2022, 2021a;
Babbar et al., 2016). The International Scientific Association for Pro­
biotics and Prebiotics (ISAPP) states that, “a dietary prebiotic is a
composition and/or activity of the gastrointestinal microbiota, thus
conferring benefit(s) upon host health”. Pectic oligosaccharides are
typically non-digestible and have a variety of health benefits, including
the prevention of uro-pathogenic microorganism adhesion, anti-obesity
effects, antibacterial, antioxidant properties, protection of colonic cells
against Shiga toxins, and stimulation of apoptosis in human colonic
adenocarcinoma cells (Gullón et al., 2013; Li et al., 2010; Olano-Martin
et al., 2003). The inability of oligosaccharides to be digested is a highly
valuable feature for their application in functional meals or

* Corresponding authors.
** Correspondence to: R. Sirohi, Department of Chemical and Biological Engineering, Korea University, 145, Anam-ro, 9 Seongbuk-gu, Seoul 02841, South Korea.
E-mail addresses: mukesh_awasthi45@yahoo.com, mukeshawasthi85@nwafu.edu.cn (M.K. Awasthi), zhangzq58@126.com (Z. Zhang), ranjanabce@gmail.com
(R. Sirohi).
1
Authors contributed equally to the work and share co-first authorship.

https://doi.org/10.1016/j.ijfoodmicro.2022.109610
Received 16 January 2022; Received in revised form 17 February 2022; Accepted 1 March 2022
Available online 3 March 2022
0168-1605/© 2022 Elsevier B.V. All rights reserved.
M.K. Awasthi et al.
nutraceutical products (Awasthi et al., 2021). As a result, extracting
oligosaccharides from agro-waste or agro-biomass might be a very
effective waste management and environmental improvement approach
(Awasthi et al., 2020a, 2020b; Duan et al., 2021).
Pectin is composed of diverse structural components, including
homogalacturonan (HG), xylogalacturonan (XGA),
rhamnogalacturonan-I (RG-I), rhamnogalacturoanan-II (RG-II), ara­
binan, and arabinogalactan. Waste valorization of pectin-rich agro-in­
dustrial wastes into pectic oligosaccharides is an intriguing method of
using waste and by-product streams. Babbar et al. (2016) investigated
the synthesis of pectic oligosaccharides with a specific degree of poly­
merization from onion peels. Pectic oligosaccharides were produced
using a chelator-based crude pectin extraction followed by controlled
enzymatic hydrolysis techniques. For the hydrolysis of pectin, three
distinct enzymes were tested: pectinase, EPG-M2, and viscozyme.
However, as compared to free galacturonic acid, the usage of EPG-M2
resulted in a higher proportion of the oligomers (particularly DP3).
Various EPG-M2 concentrations result in different POS fractions, with
26 IU/mL preferring DP3 production over longer incubation periods.
Martinez et al. (2010) also investigated the synthesis of pectic oligo­
saccharides from orange peel wastes by hydrothermal treatment.
Depolymerization of appropriate raw materials or pure pectin by
acid hydrolysis, enzymatic hydrolysis, hydrothermal processing, dy­
namic hydrothermal microfluidization, or photochemical reactions in
TiO2-containing media can provide pectic oligosaccharides (Duan et al.,
2020; Gullón et al., 2013). Many other researchers conducted studies on
the production of oligosaccharides from agro-based waste. Dávila et al.
(2016) assessed the suitability of vine shoots for hemicellulosic oligo­
saccharides production by subjecting them to non-isothermal aqueous
processing. It has been reported that about 2 to 4 tons of vine shoots are
generated from each hectare of grape cultivation (Arun et al., 2021;
Nabais et al., 2010). The overall oligosaccharide concentration was
determined to be 28.4 g/L, whereas the remaining non-volatile com­
ponents were 0.08 g/g. They found that the degree of hemicellulose
depolymerization depends on the severity of the hydrothermal pro­
cessing and proposed that autohydrolysis is a suitable pre-treatment of
vine shoots, such as the first step of a biomass refinery. Similarly, Singh
et al. (2015a, 2015b) concentrated on xylan-derived oligosaccharides,
which are plentiful in nature and serve as an excellent source of hemi­
cellulose. Hemicellulose (25–35%) is the second most common polymer
found in plant cell walls, followed by cellulose (45–55%), and is found in
close relationship with lignin. Hemicelluloses contain β -(1-4)-linked
backbone and consist of xyloglucans, xylans, mannans, and gluco­
mannans, and also β-(1-3, 1-4)-glucans (Scheller and Ulvskov, 2010; Xie
et al., 2020).
Several investigations in the 1990s showed the health benefits of
xylooligosaccharides (Cano et al., 2020). As a result, hydrothermal
processing began to gain attention not only as a technique to improve
cellulose hydrolysis, but also as a method to preferentially create this
type of oligosaccharide. This was the first time autohydrolysis (hydro­
thermal processing) was employed in 1999, with the emphasis on
oligosaccharide synthesis. Oligosaccharides derived from cellulose,
hemicellulose, and pectin can be thought of as intermediates in the
synthesis of the C5 (pentoses) and C6 (hexoses) kinds of biorefinery
platform chemicals. The development of novel acid, basic, thermal, and
physicochemical biomass pretreatments have allowed the observation of
operating parameters sufficient for oligosaccharide synthesis from
pectin and hemicellulose, which are more prone to depolymerization
than cellulose.
Every year, about 100,000 MT of biomass is generated, 1300 MT of
which are food wastes (foods not consumed from any section of the food
supply chain or any component of the food that is inedible and so forms a
residue) (Cano et al., 2020). Biofuels, monosaccharides, oligosaccha­
rides, nanocellulose, lignin, and bioactive compounds are among the
value-added goods that may be made from such biomass waste. They
may be produced from a variety of culinary and agricultural wastes,
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International Journal of Food Microbiology 368 (2022) 109610
including vine shoots (Dávila et al., 2016; Jose et al., 2022), banana
peels, sugar beet residues, and wheat chaff (Đorđević and Antov, 2018;
Liu et al., 2021a). Through chemical and enzymatic hydrolysis tech­
niques, agro-biomass can be investigated as a possible source of oligo­
saccharide. The enzymatic technique is favored for the safe usage of
oligosaccharides in functional foods since chemical methods create
numerous additional side chemicals in addition to oligosaccharides that
may occasionally be hazardous for human ingestion. In this regard,
mannooligosaccharides, xylooligosaccharides, and arabinooligo­
saccharides are the most common oligosaccharides that are beneficial as
prebiotics (Table 1).
Recently, attempts have been made to produce functional foods and
nutraceuticals that contain probiotics supplemented with various pre­
biotics such as isomaltooligosaccharides, fructooligosaccharides, arabi­
nooligosaccharides, xylooligosaccharides, galactooligosaccharides, and
so on (Jana and Kango, 2020). Mannooligosaccharides (MOSs) produced
from plants and yeast are novel and developing members of the prebiotic
family. Mannans are homo- or heteropolysaccharides found mostly as
structural components of softwood or storage components in the hull of
certain seeds. They are primarily made of mannose sugar connected by
β-1, 4-glycosidic linkages (Liu et al., 2021b; Von Freiesleben et al.,
2019). The mannans are present in lignocellulosic agro-wastes such as
copra meal, spent coffee ground, corn stover, potato peels, kernel cakes,
etc. (Jana and Kango, 2020). Mannans can be classified into the
following groups (1) glucomannans (2) galactomannans and, (3) gal­
actoglucomannans (Schröder et al., 2009). MOSs can be synthesised in a
single step using enzymatic, chemical, or chemo-enzymatic techniques.
MOSs are classified into two types: α-MOS and β-MOS oligosaccharides.
They are made up of a linear chain of mannose sugar, have prebiotic
characteristics, and are used as a feed addition to inhibit pathogen
colonization in humans and animals' digestive tracts. Due to processing
inconsistency, a considerable amount of fruits and vegetables are wasted
in India, accounting for around 1.4% of total global export (Liu et al.,
2021c; Singh et al., 2018). As a result, using agro-waste for human
benefit is the primary concern for environmental sustainability.
This review aims to investigate the process of oligosaccharide pro­
duction from different agro wastes along with a comprehensive analysis
of different chemical and biological techniques for enhanced oligosac­
charide production. An overview of the production process has been
shown in Fig. 1. Further, the factors affecting oligosaccharide produc­
tion have also been detailed followed by perspectives for further
research.
2. Substrates and microbial strain for the production of
oligosaccharides
Oligosaccharide are soluble in water and possess slight sweetness
which depends upon the degree of polymerization, molecular weight
and chemical structure (Awasthi et al., 2022). On the basis of physio­
logical properties, oligosaccharides can be digestible and non-digestible.
The former can be easily broken down by digestive enzymes but the
latter cannot undergo digestion by humans (Fig. 2). This section detailed
the production of oligosaccharides from different substrates such as
sugars, starch, dairy, and lignocellulosic waste.
2.1. Production of oligosaccharide from starch based substrate
Starch is a heterogenous polysaccharide comprising of glucose
monomers with alpha-1, 4 linkage and is considered as an energy source
for human nutrition. Amylose (linear chain of glucose) and amylopectin
(highly branched chain of glucose) are two different types of poly­
saccharides present in starch. It is insoluble in water and contains car­
bon, hydrogen, and oxygen atom in the ratio of 6:10:5 (C6H10O5)n (Hii
et al., 2012; Madhavan et al., 2021a). Physical, chemical, and enzymatic
methods have been used for the modification of starch. The physical
method involves heat moisture treatment, freezing, ultra-high-pressure
M.K. Awasthi et al. International Journal of Food Microbiology 368 (2022) 109610

Table 1
Different substrates for oligosaccharide production.
S. Substrate Category Mode of production Oligosaccharide produced Concentration References
no

1. Sweet potato Starch Enzymatic saccharification Isomalto-oligosaccharide 68.85 ± 1.82 g/L (Duong Hong et al., 2021; Yusree
and liquefaction et al., 2022)
2. Lactose Dairy Bacterial enzyme Galacto-oligosaccharides 91 g/L (Awasthi et al., 2019;Ji et al.,
2005)
3. Lactose Dairy Bacterial enzyme Galacto-oligosaccharides 52.5% (w/w) (Ji et al., 2005)
4. Lactose Dairy Bacterial enzyme Galacto-oligosaccharides 13 g/L (Awasthi et al., 2021a; Yu and
O’Sullivan, 2014)
5. L-fucose and Dairy Metabolic engineered Human milk oligosaccharide 92 and 503 mg/L (Awasthi et al., 2022; Yu and
Lactose bacteria 2’fucosyllactose O’Sullivan, 2014)
6. Vine shoot Lignocellulosic Non-isothermal aqueous Xylo-oligosaccharide and galacto- 12.2 g/L and 8.64 Dávila et al., 2016(Dávila et al.,
biomass autohydrolysis oligosccharides g/L 2016)
7. Peanut shells Lignocellulosic Non-isothermal aqueous Xylo-oligosaccharide 0.061 g/g (Wang et al., 2014)
biomass autohydrolysis
8. Corn cobs Lignocellulosic Bacterial enzyme Feruloyl xylo-oligosaccharides 100 g/L (Katapodis and Christakopoulos,
biomass 2008; Reshmy et al., 2021b)
9. Orange peel Lignocellulosic Fungal enzyme Pectic oligosaccharides – Qin et al., 2021a; Zhong et al.,
wastes biomass 2020)
10. Beet sugar syrup Sugar Fungal enzyme Fructo-oligosaccharide 388 mg/mL and (Ghazi et al., 2006)
and molasses 235 mg/mL
11. Sugarcane bagasse Sugar Hydrolysis/Catalyst Xylooligosaccharides 53.2% (Wang et al., 2021; Zhou et al.,
2019)
12. Dry coconut meal Lignocellulosic Autohydrolysis Mannooligosaccharides 0.23 g/L (Cano et al., 2020)
biomass
13. Sucrose Sugar Bacterial enzyme Cello-oligosaccharides 39 g/L (Liu et al., 2021b; Zhong et al.,
2020)
14. Wheat husk Lignocellulosic Fungal enzyme Xylooligosaccharides 3.5 g/L (Jagtap et al., 2017)
biomass
15. Sucrose Sugar Fungal enzyme Fructooligosaccharides 6 g/L (Duan et al., 2020; Sánchez et al.,
2008)

Fig. 1. Overview of the oligosaccharide production process.

treatment; the chemical method involves etherification, esterification,
cross-linking and acid treatment while enzymatic modification involves
starch modifying enzymes such as α-amylases. Functional sugars were
reported to be extracted from starchy substrates by employing enzy­
matic approaches (Jaafar et al., 2021). Majority of different factors
affect the production of oligosaccharides such as temperature, enzyme
ratio, enzyme loading, and pH. All these factors affect the generation
and activity of enzymes. It was recorded that the CGTase exhibited
better activity at 7 pH as it yields 1.1 mg of β-CD as compared to 0.8 mg
at 6.0 pH (Jaafar et al., 2021). However, it was also reported that
CGTase showed efficient activity at 6.0 pH when tropical starch rather
than soluble starch. Furthermore the increase in the substrate loading
increases the yield of β-CD. Jaafar and coworkers recorded that increase
in soluble starch loading from 0.1 to 0.2% (w/v) increase the concen­
tration of β-CD from 0.4 mg to 3.0 mg. A further increase to 0.2% (w/v)
significantly increased the concentration. The reason to this may be
attributed to the complete occupancy of active sites of CGTase. In
addition to this, the enzyme CGTase loading from 1 to 10 U increases
25–40% of β-CD from 4.4 mg to 10.2 mg. A further increase upto90 U
does not further affect the production of β-CD (Jaafar et al., 2021). The

3
M.K. Awasthi et al.
Fig. 2. Beneficial effects of oligosaccharide derived prebiotics in the human body.
reaction time from 0.5–24 h affects the yield of MOS. It was reported
that at the minimum reaction time of 0.5 h gave highest yield of mal­
topentoses. However the increase in the reaction time to 1 h increases
the total MOS production to 146.8 mg/g. Starch modifying enzymes has
been used for the production of branched malto-oligosaccharide with
degree of polymerization between 20 and 100 resulting for the gum­
miness in bread. The conversion of starchy raw material into oligosac­
charide was designed to focus on high quality, low cost, enzyme stability
and increased productivity (Meyer et al., 2015).
To improve the quality of bread, researchers used the de-branching
enzyme known as pullunase that hydrolyses the branched maltooli­
gaosaccharide into smaller maltooligosaccharide (Madhavan et al.,
2021b; Park et al., 2018). The enzymatic hydrolysis of starch, with en­
zymes such as α-amylases or pullunase from Aspergillus niger, β-amylases
from Bacillus species and α-glycosidase from Bacillus subtilis have been
studied for the synthesis of the isomalto-oligosaccharide (IMO). A simple
two step process for the production of IMO from sweet potato starch was
developed by Liu et al. (2021c). Sweet potato starch was simultaneously
transglycosylated and saccharified followed by liquefaction. The reac­
tion mixture was then treated by a cocktail of enzymes namely
α-transglucosidase, pullulanase, and β-amylase. This process yielded
68.85 ± 1.82 g/L of IMO at optimized conditions. The IMO derived from
sweet potato starch was suggested to be utilized in the field of healthcare
for use as prebiotics (Hong et al., 2021). Iso-malto-900 comprising iso­
maltose, isomaltotriose and pannose have been produced from corn-
starch. Starch converting enzyme such as endoamylases present in
bacteria and archaea was well known for the formation of linear and
branched oligosaccharides (Hii et al., 2012; Madhavan et al., 2022). The
use of maltogenic amylases for malto-oligosaccharides (MOS) produc­
tion had several limitations such as low MOS productivity and low
starch specificity. At industrial scale, group of enzymes called the malto-
oligosaccharides forming amylases (de-branching enzymes) are
employed to synthesize MOS from starch (Prapulla et al., 2000).
Therefore, Jaafar et al. (2021) utilized CGTase reactions and maltogenic
amylase in an synchronous and asynchronous conversion strategy to
increase the yield of MOS by utilizing soluble starch. The highest MOS
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International Journal of Food Microbiology 368 (2022) 109610
yield of 307.9 mg/g was obtained by employing the combination of
maltogenic amylase and CGTase from Bacillus lehensis G1. Also, syn­
chronous strategy yields 2.1-fold higher MOS with a catalytic efficiency
of 1746.1 mL/g/h in comparison to 146.8 mg/g MOS production and
18.5 mL/g/h catalytic efficiency by asynchronous approach (Jaafar
et al., 2021).
Furthermore, simultaneous use of cyclodextrin glucosyltransferase
and cyclodextrinase was studied as a novel enzymatic method for con­
version of waxy maize starch to malto-oligosaccharide. Successive and
simultaneous treatment with cyclodextrinase and cyclodextrin gluco­
syltransferase led to a 0.2 × 105 g/mol and 0.3 × 105 g/mol reduction in
molecular weight. Cyclodextrinase hydrolyze cyclodextrins from
cyclodextrin glucosyltransferase and this positively affects the produc­
tion of MOS (Ji et al., 2021). This suggested that combinatorial approach
provides a foundation for industrial MOS production with better effi­
ciency. Zhang et al. (2021) studied the conversion of raw starch by the
action of α-amylase AmyM derived from Corallococcus sp. It was re­
ported that the AmyM did not depend on Ca2+ and exhibits a Vmax of 741
and 1111 μmol/min/mg for corn and wheat raw starch, respectively.
Hydrolysis by AmyM led to the formation of cerviced surfaces and pores
around the periphery of starch granule. Moreover, AmyM at 5 U per mg
of raw starch hydrolyzed 90% of 30% corn and wheat raw starch that
resulted in the formation of 30% dextrin and 70% MOS at 45 ◦ C after 20
h. Interestingly, during the process other di to oligosaccharides namely
maltose, maltotriose and maltotetraose were synchronously produced
(Zhang et al., 2021).
2.2. Production of oligosaccharide from dairy based substrates
A fructose oligosaccharide inulin is a low-calorie oligosaccharide
that has been used in dairy products to replace fat by enhancing flavor
and texture of food. Inulin has applications in low fat cheese, low fat
yogurt, ice-cream, chocolate mousse and custard production (Awasthi
et al., 2022). Fructo-oligosaccharides (FOS) and galacto-
oligosaccharides (GOS) are produced by the trans-glycosylation of
glycosidase enzymes (Zeuner et al., 2019). GOS are prebiotics composed
M.K. Awasthi et al.
of 1 molecule of glucose and 2–20 molecule galactose and are non-
digestible oligosaccharide, that are produced by the trans-
galactosylation activity during hydrolysis of lactose with β-galactosi­
dase enzyme which has great potential for GOS synthesis (Ji et al., 2005;
Narisetty et al., 2021). Lactic acid bacteria namely Lactococcus lactis
employed in the fermentation of dairy products such as cheese and
fermented milk has been used for the production of GOS (Yu and
O’Sullivan, 2014). Several microorganisms such as Aspergillus oryzae,
Sirobasidium magnum, Penicillium simplicissimum, Bacillus circulans, Bifi­
dobacterium infantis, Bullera singular, Escherichia coli, and Kluyveromyces
lactis were reported for GOS production. The solubility of lactose at high
temperature and high initial concentration increases the formation of
GOS. It was suggested that the suitable factors determining the total GOS
yield are initial lactose concentration, enzymatic activity of microor­
ganisms, spectrum of GOS products, optimum pH, and temperature.
Some thermostable microorganisms such as Thermus sp., Pyrococcus
furiosus, Sterigmato myceselviae, Rhodotorula minuta and Sulfolobus sol­
fataricus have been studied for the production of GOS at high tempera­
ture (Ji et al., 2005; Narisetty et al., 2022; Yu and O’Sullivan, 2014). Ji
et al. (2005) studied the production of GOS by a thermostable recom­
binant β-galactosidase from Thermotoga maritima and found that under
optimal conditions: pH 6.0, 80 ◦ C, 500 g lactose/L, and 1.5 units
enzyme/mL, the rate of production of GOS was 91 g/L for 300 min with
the GOS productivity rate of 18.2 g/L/h. Similarly, a hyperthermostable
β-galactosidase enzyme from Sulfolobus solfataricus resulted in the
highest yield of 315 g/L trisaccharide or longer chain of GOS from 60%
initial lactose at high temperature and higher reaction velocity. The
optimum temperature for the production of GOS using β-galactosidase
was 37 ◦ C and 90 ◦ C for E. coli and Thermotoga maritime, respectively.
Bifidobacteria longum (colon bacteria) has an important role in control­
ling the gut microbiota with several health benefits and the production
of 13 g/L GOS was reported by utilizing lactose at an initial concen­
tration of 40% (Yu and O’Sullivan, 2014). According to Otieno (2010),
dairy products containing lactose such as yogurt have been used as a
substrate for the synthesis of GOS. It has been reported that yogurt with
Bifidobacterial infantis exhibited 0.72% GOS production at the end of the
fermentation process than the yogurt with the other bifidobacterial
species. The reason to this was attributed to the higher trans-galactosyl
reaction by β-galactosidase from Bifidobacterial infantis (Otieno, 2010).
Human milk oligosaccharide (HMO) is a group of oligosaccharides
with component of human milk that promotes the infant health and
development. One of the most common HMO is Fucosylated oligosac­
charide that enhances the growth of the bifido-bacteria and prevents the
binding of pathogens and toxins to human gut (Patel et al., 2021; Zeuner
et al., 2019). Yu and O’Sullivan (2014) studied the production of human
milk oligosaccharide 2’fucosyllactose by metabolically engineered
Saccharomyces cerevisiae via salvage pathway using L-fucose and lactose
as a substrate and obtained 92 and 503 mg/L of milk from a 48 h batch
and 120 h fed batch fermentation, respectively using ethanol as a carbon
source.
2.3. Production of oligosaccharides from lignocellulosic substrates
Lignocellulosic biomass such as agricultural residues, grasses, woody
raw material, and forestry wastes are the renewable and sustainable
feedstock for biofuel and biochemical production. Cellulose, hemicel­
lulose and lignin are the major polymers of lignocelluloses (Gaur et al.,
2020). Depolymerization of the biomass is essential for lignocellulosic
utilization, either as feedstock for catalytic process or as a carbon source
for microbial utilization (Jin et al., 2020; Qin et al., 2021a). Lignocel­
lulosic biomass serves as a rich source for oligosaccharide production.
To enhance the availability of polymers and biodegradability through
enzymatic and chemical activity, the pre-treatment of the lignocellulosic
biomass is a vital process. The pre-treatment strategy involves biological
(enzymatic and whole cell pre-treatment), physical (milling, microwave,
ultrasonication, and extrusion), chemical (acid and alkaline hydrolysis,
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International Journal of Food Microbiology 368 (2022) 109610
and organosolv process), and physio-chemical (steam explosion, carbon-
dioxide explosion, and liquid hot water) methods (Baruah et al., 2018;
Isikgor and Becer, 2015). Temperature, residence time, acid load, and
biomass loading are the vital conditions for oligosaccharide production
(refer to Section 4). The oligosaccharide derived from agricultural
wastes such as oat bran, wheat bran, and rice straw were reported to
exhibit wide applications in food, cosmetics, pharmaceuticals, and
nutraceuticals industry (Bhatia et al., 2019). The lignocellulosic sources
such as forest farming residues and other value-added products have
been studied for the production of cello-oligosaccharide (COS) by using
acid based and enzyme-based hydrolysis of the residual cellulose. COS
are linear oligosaccharides having β-1,4-linked glucopyranose unit with
applications in food industry as a prebiotic (Karnaouri et al., 2019a,
2019b). The hemicellulose fraction of the lignocellulosic biomass acts as
a good source for xylo-oligosaccharides (XOS) and manno-
oligosaccharides (MOS) production. Orange peels, banana peels, sug­
arcane bagasse and straw are rich in hemicellulose content that results in
the production of oligosaccharides. It has been reported that the utili­
zation of vine shoot as a feedstock the concentration of galacto-
oligosccharides (GOS) and XOS was 8.64 g/L and 12.2 g/L of galacto-
oligosccharides (GOS) and XOS, respectively. The autohydrolysis of
chestnut shell and peanut shell produced 0.057 g/g and 0.061 g/g of
XOS, respectively. Similarly, 0.23 g of oligosaccharides was produced
during autohydrolysis of 1 g of dry coconut meal (Cano et al., 2020;
Dávila et al., 2016; Qin et al., 2021b). The autohydrolysis of almond
shell led to the production of 3.3 g per 100 g dry solid of XOS with low
degree of polymerization and upon addition of enzymatic hydrolysis and
purification, the yield was further increased to 8.2 g/100 g dry solid.
Furthermore, the autohydrolysis of sugar beet pulp, orange peels and
lemons peels resulted in 13 g/L of arabino-oligosaccharide, 17 g/L and
21 g/L of pecto-oligosaccharide, respectively (Gomez et al., 2015).
The acid hydrolysis method resulted in the hydrolysis of rigid crys­
talline structure of celluloses and produced 0.02 g of oligosaccharide
using concentrated hydrochloric acid and sulfuric acid (4:1) at 22 ◦ C for
4–6 h (Zhang and Lynd, 2003). Auto-hydrolysis and acid pre-treatment
had some harmful effects as it led to the production of inhibitors which
may cause serious health issues such as respiratory, kidney damage, and
inflammation in the human body. The purification process is effective
for the removal of these inhibitory compound and to produce food grade
XOS.
Microbial fermentation, size exclusion and cation exchange chro­
matography, nanofiltration and activated charcoal method etc. are the
techniques for the purification and extraction of oligosaccharide derived
from lignocellulosic feedstock (Bhatia et al., 2019). Several studies were
also reported on enzymatic hydrolysis of agricultural wastes for the
production of XOS and pecto-oligosaccharide (POS) (Baldassarre et al.,
2018; Cano et al., 2020; Mazlan et al., 2019b; Qin et al., 2021c). Mazlan
et al. (2019a) reported the production of XOS from oil palm frond
bagasse by optimizing the important operational factors such as sub­
strate loading (1% w/w) and enzyme activity (50 U/mL of xylanase).
They recorded a yield of 0.175 g of xylo-oligosaccharides (XOS) per
gram of dry substrate (Mazlan et al., 2019a). Baldassarre et al. (2018)
studied the production of POS using 50 g/L onion skin subjected to
enzymatic hydrolysis with an enzyme viscozyme for 15–30 min at 45 ◦ C
and obtained 22 g/L POS with 2–8 degree of polymerization (Baldas­
sarre et al., 2018; Qu et al., 2021). The bio-valorization of lignocellulosic
residues to yield oligosaccharides reduces the difficulties of biomass
management. The production of oligosaccharide in biorefineries inte­
grating lignocellulose energy matrix suggests an appropriate alternative
for the production of high commercial value-added products ultimately
enriching renewable economy under a sustainability regime (Bhatia
et al., 2019; Ravindran et al., 2021).
2.4. Oligosaccharide production from sugar based substrate
Sugar based products are widely known substrates used for the
M.K. Awasthi et al.
production of oligosaccharides. Their simplest chemical structure and
ease of breakdown enables them an efficient candidate to exploit
boundlessly. Ghazi et al. (2006) studied the production of fructo-
oligosaccharide (FOS) from beet sugar syrup and molasses as sugar
feedstock containing 620 and 570 mg/mL of sucrose, respectively. FOS
obtained from these substrates reached a maximum of 388 mg/mL in 30
h and 235 mg/mL in 65 h. Owing to growing demand of the oligosac­
charides, studies have delineated towards microbial production of these
molecules using sugary biomass. Using mixed enzyme system, Sir­
isansaneeyakul et al. (2000) evaluated the production of fructo-
oligosaccharides (kestose and nystose) from sucrose. Using 400 g/L of
sucrose concentration, 0.44 and 0.49 g/g of kestose and nestose with
volumetric productivity of 4.97 and 5.44 g/L/h, respectively were
recorded.
Furthermore, Singh et al. (2018) reported the production of nystose
by B. subtilis natto cct 7712. The strain utilized low cost substrates like
sugarcane molasses, sugarcane juice and commercial sucrose. Maximum
production of 179.77 g/L nystose with 71.73% yield was obtained with
400 g/L of commercial sucrose. Also, during fermentation of sugarcane
juice and molasses, results were found satisfactory as 42.58 and 97.93 g/
L of nystose were produced, respectively. This suggested that the use of
sugar derived biomass offers a promising strategy for synthesizing nys­
tose at a commercial level. Valorization of sugar based substrates not
only minimizes the pollution load from the environment but also aids in
the economic production of value added products from cheapest sources
(Kothari et al., 2021). Sugar beet pectin was used to produce pectic ol­
igosaccharides using cross flow enzyme membrane reactor incorporated
with the low-cost enzyme, viscozyme, for continuous production.
Optimized and continuous production was performed in order to
maximize the yield and minimize the formation of monosaccharides. An
average 82.9 ± 9.9% (w/w) yield of pectic-oligosaccharide was ach­
ieved in 28.5 h (Elst et al., 2018). An enzymatic approach for production
of pectic oligosaccharides from sugar beet pulp was studied by Babbar
et al. (2017). Statistical model of response surface methodology was
applied to investigate the effect of different parameters responsible for
the production. Based on optimization, 135 g/L of sugar beet pulp was
found to correspond for the production of pectic oligosaccharides rich
hydrolysate with a notable yield of 26.5% (w/w) (Babbar et al., 2017).
Furthermore, previously Roy et al. (2002) optimized the production of
galacto-oligossaccharides from lactose. The parameter optimization was
carried out using response surface methodology in which, nearly 40% of
lactose concentration appeared to be the most optimum concentration
for the production of oligosaccharide. Additionally, a green integrated
approach was performed to co-produce xylo-oligosaccharides with glu­
conic acids using sugarcane bagasse as sugary substrate. Highest pro­
duction of XOS and gluconic acid was 39.1% and 10%, respectively
whereas after enzyme hydrolysis 105 g XOS and 340 g gluconic acid
were co-produced from the consumption of 1 kg dried sugarcane bagasse
as a starting feedstock (Zhou and Xu, 2019).
Recently in 2020, Zhong et al. investigated the production of cello-
oligosaccharides from glucose and sucrose. Cello-oligosaccharides are
water soluble dietary fibers produced by employing glycoside phos­
phorylases such as cellodextrin phosphorylase, cellobiose phosphory­
lase, and sucrose phosphorylase. A cello-oligosaccharide yield of 39 g/L
was recorded by employing 0.065 M glucose and 0.2 M sucrose,
exhibiting sucrose conversion of ~ 95%. More importantly, 93 g/L of
COS with 82 mol% yield was recorded from the utilization of 0.5 M
sucrose (Reshmy et al., 2022a; Zhong et al., 2020). This also suggested
that the feedstock concentration had a promising effect on the produc­
tion of oligosaccharides. All these findings from different studies sug­
gested that sugars may serve as an efficient substrate for the production
of different oligosaccharides in the presence of bacteria or enzymes.
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International Journal of Food Microbiology 368 (2022) 109610
3. Processing strategies for the production of oligosaccharides
from biomass
3.1. Pretreatment
Pretreatment strategies are significant to efficiently deconstruct
lignocellulosic biomass to economically produce oligosaccharides. Pre­
treatment is deployed as an essential step to convert the recalcitrant
lignocellulosic fraction for effective biomass conversion. Different types
of pretreatment strategies such as chemical (acid, alkali and ionic
liquid), physical (milling, grinding), physicochemical (liquid hot water,
steam explosion and wet oxidation) and biological (biomass-degrading
microorganisms) have been widely employed (Bhatia et al., 2021;
Reshmy et al., 2022b). These pretreatments depict varied effects on the
fragmentation and physicochemical modification of lignocelluloses and
hence choosing an appropriate pretreatment method is crucial to
determine the environmental and economic feasibility of commercial
production of oligosaccharides (Bhatia et al., 2021).
3.2. Hydrolysis
Simple hydrolysis, physico-chemical pretreatments with or without
the addition of acid catalysts is considered to be effective for hydro­
lyzing the hemicellulose and hydrolysis of cellulose fractions for the
production of xylo-oligomers, cello-oligomers and sugar monomers.
Nevertheless, the operating conditions such as biomass loading, acid
load, temperature and residence time are crucial to obtain optimal yields
of the oligosaccharides from biomass feedstocks (Ávila et al., 2021;
Reshmy et al., 2022c; Wainaina et al., 2019).
3.2.1. Chemical
3.2.1.1. Acidic. Prior to enzymatic pretreatment, acid hydrolysis has
been primarily used for the hydrolysis of lignocellulosic biomass.
Chemical pretreatments under acidic conditions have been exclusively
carried out to obtain glucose, xylose, furan derivatives or levulinic acid
(Ávila et al., 2021; Cano et al., 2020). The β-1, 4 glycosidic bonds are
cleaved during acid hydrolysis and the polysaccharides are broken down
to oligo or monosaccharides. The commonly used acids are sulfuric and
hydrochloric acids and various organic and inorganic acids such as
acetic acid phosphoric acid, oxalic acid, nitric acid or maleic acid. The
type of acid and the severity of its application are of prime importance as
they can produce inhibitors such as furfural and HMF (Reshmy et al.,
2022d; Sirohi et al., 2020). The formation of inhibitors and unwanted
compound can be reduced by the use of low acid concentrations
(<10%). Use of low acidic levels at steaming temperatures of
135–140 ◦ C for 30 min can recover hemicellulose from xylan-lignin
complex efficiently as reported by Yang and co-workers (Yang et al.,
2005).
Metallic salts such as potassium chloride (KCl), iron chloride (FeCl3),
zinc chloride (ZnCl2) and cupric chloride (CuCl2), have also been re­
ported to be agents alternate pretreatment (Li et al., 2020). They are
widely used due to their higher catalytic activity, inexpensive nature
and less toxicity and corrosivity, and the possibility of recycling. Among
these metallic salts, ZnCl2 has been considered as an effective salt due to
its Lewis acid and Brønsted acid characters, as well as the autoionisation
of water in aqueous solution. It was reported by You et al. (2020) that
excellent removal of xylan and release of XOS (DP 2–5) from COS was
achieved with the use of ZnCl2. In order to avoid the hydrolysis of
hemicellulose into monomers, such as xylose, the acid reagent concen­
tration should be low. To increase the production of xylooligo­
saccharides (XOS), random attack on the xylan chain should occur
(Brienzo et al., 2016). The cellulose fraction can be very difficult to
hydrolyze due to the rigid crystalline structure it possesses that makes
the penetration of acid into the dense network very difficult (Bhatia
M.K. Awasthi et al.
et al., 2019; Cano et al., 2020; Reshmy et al., 2022e). In such cases,
hydrolysis can be carried out with the help concentrated or supported
catalyst acids. Various studies have been reported for the production of
oligosaccharides using acid pretreatments. Cello oligosaccharide (COS)
production was carried out via acid hydrolysis using sulfuric acid under
different conditions by Bouchard et al. (2016) from bleached kraft pulp.
In another study, a 4:1 mixture of concentrated hydrochloric acid and
concentrated sulfuric acid was used for the treatment of microcrystalline
cellulose at room temperature (22 ◦ C; 4–6 h) for producing COS. Ion
exchange column chromatography was used for the purification of sol­
uble oligosaccharides, which were precipitated using acetone (Reshmy
et al., 2021b; Zhang and Lynd, 2003).
3.2.1.2. Alkaline. Alkaline pretreatment has also been widely studied as
an effective pretreatments strategy. NaOH is frequently employed as it
promotes effective delignification, has high reaction rate, and limits the
formation of inhibitory compounds. Other compounds like Ca(OH)2 or
Na2SO3 have also been used. The hydrogen bonds between celluloses
and hemicelluloses are broken down by the action of the hydroxide ions
present in the alkaline reagents, solubilizing the hemicellulose portion.
Furthermore, the ester bonds can be broken without degrading other
compounds using alkaline extraction (Brienzo et al., 2016; Sarsaiya
et al., 2019). These pretreatments are carried out in several conditions,
such as at high temperatures, pressures, and short residence times (Singh
et al., 2015a, 2015b). Due to the aforementioned reasons, alkaline
pretreatment is considered as the first option and also selectively sepa­
rate high purity lignin from the biomass (Pinales-Márquez et al., 2021;
Wainaina et al., 2020). Another advantage of this process is that it
selectively removes lignin. Besides, with this pre-treatment, the porosity
and surface area can be enhanced thus improving enzymatic hydrolysis
yields. Various alkaline pre-treatment alternatives such as high and low
alkali concentrations in the presence or absence of high temperatures
have been studied. Studies have reported that NaOH was more efficient
than KOH for the recovery of hemicellulose and that the use of high
levels of alkali leads to higher yield under overnight incubation or by
steam application (Santibáñez et al., 2012). Nevertheless, the disad­
vantages of alkaline pretreatment are the corrosivity and contamination
of water effluents and the formation of toxic compounds at high pres­
sures and temperatures (Pinales-Márquez et al., 2021).
XOS production from lignocellulosic material can also be achieved
through alkaline extraction. For XOS production, the acetyl groups are
cleaved under this pre-treatment type and this can be considered as an
advantage since the xylan substituents are eliminated and the use of
acetyl esterase enzymes can be avoided (Pinales-Márquez et al., 2021).
However, the solubility of xylan can be reduced by this method thus
reducing the yields of hemicelluloses extraction. Subsequently biomass
rich in cellulose can be obtained which can be further used for the
production of biofuels and reduce the degradation of XOS.
3.2.2. Biological
Biological pretreatments are usually carried out by direct application
of microorganisms or the enzymes produced by them. Biological pre­
treatment is considered to be selective and eliminates the addition of
chemicals, lower energy uses and do not produce inhibitory compounds
due to their mild reactions. Therefore, biological pretreatment is
considered as a green and sustainable process for the pretreatment of
lignocellulosic materials.
3.2.2.1. Microbial processes. Pretreatment through fermentation has
garnered significant attention since it enhances the availability of
biomass. Various types of bacteria belonging to genus Bacillus, Actino­
mycetes, Streptomyces, and fungal species like Aspergillus or Candida have
been widely used for the production of oligosaccharides (Sindhu et al.,
2016; Xu et al., 2020; Wang et al., 2021). Filamentous fungi capable of
producing endo-1, 4-β-xylanase have been widely used for in situ
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International Journal of Food Microbiology 368 (2022) 109610
fermentation hydrolysis. A wide variety of enzymes such as cellulases,
β-glucosidases, xylanases and amylase are also produced by these mi­
croorganisms. These enzymes are responsible for the degradation of
polysaccharides into sugars.
Solid-state cultivations using fungi have been studied to produce
xylanase and arabinofuranosidase and these enzymes were subsequently
used for the production of XOS (Pereira et al., 2018). Over the past few
years, XOS production in biorefinery route has gained tremendous
popularity. Direct fermentation of brewers' spent grain (BSG) using
Trichoderma species and XOS with DP from 2 to 5 was produced indi­
cating an attractive route for the production of oligosaccharides
(Pinales-Márquez et al., 2021). Use of recombinant enzymes produced in
non-cellulolytic or non-xylanolytic hosts is another alternative than can
be explored (Pinales-Márquez et al., 2021). Wild type B. subtilis and a
clone containing xylanase (Xyn2) gene from T. reesei were used for XOS
production (Amorim et al., 2018). The results indicated that 824 mg/g
and 997 mg/g, were obtained from the wild strain and the clone,
respectively and the yield was increased by 16%. Use of eukaryotic
systems such as Pichia pastoriscan reduces the formation of unwanted
enzymes such as xylosidases and cellobiases, when GRAS microorgan­
isms are used. Nevertheless, in these processes different process pa­
rameters such as temperature, stirring, pH, and biomass growth need to
be controlled. Fermentation using organisms can reduce the cost of using
commercial enzymes for the process and can be easily employed in
biorefineries.
3.2.2.2. Enzymatic processes. The expanding knowledge on the various
types and quantities of lignocellulosic materials available has enabled
the use of enzymatic pretreatments for saccharification. Enzymatic hy­
drolysis in comparison to other chemical/physical methods is more
attractive due to the relatively mild reaction conditions and the for­
mation of fewer by-products (Bhatia et al., 2019; Xu et al., 2021). The
group of enzymes that break down cellulose are the cellulases, which are
mainly of three types namely exoglucanases, endoglucanes and β-glu­
cosidases (Barbosa et al., 2020). Several factors such as loading, tem­
perature, pH, agitation, synergy and the property of biomass play an
important role in enzymatic pretreatment (Barbosa et al., 2020). Keep­
ing these parameters in view, various strategies have been put forth for
enzyme-mediated production of cello-oligosaccharides. Design of tailor-
made enzymatic cocktails that permit controlled polysaccharide
breakage to produce fewer monomers (Karnaouri et al., 2019a, 2019b),
changes to the reaction conditions or fine-tuning the activity of enzyme
cocktails available commercially are some of the strategies that were
carried out (Karnaouri et al., 2019a, 2019b; Yusree et al., 2022). During
cellulose hydrolysis to glucose, COS are the intermediates while during
the production of COS monosaccharides, glucose are the intermediates.
It has been reported that the formation of glucose as a byproduct hinders
the application of oligosaccharides at the commercial level. Use of
β-glucosidase enzymes results in the formation of higher quantities of
glucose as the enzyme hydrolyses the glycosidic bond within cellobiose,
reducing the formation of oligosaccharides. This challenge can be
overcome by eliminating certain enzymes or reducing their activities in
the enzyme cocktail.
Additionally, enzymatic hydrolysis of cellulose is carried out in
various stages and after each stage the products formed are removed.
This strategy has proven to be effective to enhance the efficiency of
hydrolysis and stop product inhibition. Chu et al. (2014) reported that
the selective removing of β-glucosidase activity in a corncob residue
doubled the production of cello-oligosaccharides in subsequent enzy­
matic hydrolysis (Chu et al., 2014). In another work by Zhou et al.
(2020), COS production was integrated with simultaneous production of
high value-added products of sugar monoester, to increase the purity of
resultant COS. In this process, the excess glucose was converted to
glucose mono-decanoate, a biodegradable non-ionic surfactant.
Through this process, the purity of the cellooligosaccharides increased
M.K. Awasthi et al.
from 33.3 to 74.3% and glucose mono-decanoate was produced simul­
taneously with 92.3% purity. The process was further verified with
switchgrass, cornstalk, and reed also a good yield of oligosaccharides
was obtained (Zhou et al., 2020).
Apart from cellulases, cellobiohydrolase (CBH) belonging to the
glycosyl hydrolase family 7 (CBH7) and the endoglucanase of the family
5 (EG5) have also been used for the production of COS (Karnaouri et al.,
2019a, 2019b). However, the amorphous areas need to be acted upon by
other enzymes such as endoglucanases of the family 7 (EG7) and cel­
lobiohydrolase of family 6 (CBH6 (Karnaouri et al., 2019a, 2019b).
When CBHs and endoglucanases are utilized together, drastic
enhancement in hydrolysis can be obtained due to the endo-exo synergy
between these two classes of the enzymes (Barbosa et al., 2020; Kar­
naouri et al., 2019a, 2019b).
Fairly recent approaches for COS production have been employed
using endoglucanases, processive endoglucanases, and auxiliary en­
zymes, such as lytic polysaccharide monooxygenases (LPMOs) (Kar­
naouri et al., 2019a, 2019b). LPMOs and endoglucanases act in synergy
to release both neutral and oxidized sugars (Barbosa et al., 2020) and
LPMO treatment results in new chain breaks in crystalline substrates,
reducing the biomass crystallinity (Hemsworth et al., 2015). With the
use of LPMOs, novel essential cellulolytic cocktails could be generated as
these enzymes create new terminals to the cellulose chains, promote
defibrillation of the fibers, and improve overall degradation of the
process (Hemsworth et al., 2015). For the production of XOS through
enzymatic hydrolysis, two steps are employed. In the first step, the
hemicellulose present in the lignocellulosic material is isolated through
various pre-treatments. In the second step, xylan is enzymatically hy­
drolyzed via endo-β-1, 4-xylanase with little or no activity of the
β-xylosidase enzyme to avoid the formation of monosaccharides
(xylose). In the first step it is important to select the appropriate pre­
treatment based on the final characteristics of the hemicellulose
recovered.
Additionally, since the xylan molecule is a complex structure inter­
connected by covalent and ionic bonds, a direct contact between enzyme
and substrate should be made. Xylanases exhibit different substrate
specificities and hence generate different hydrolysis products thus
making it difficult to control the degree of polymerization. Branched
XOS can also be produced by enzymatic hydrolysis in the absence of
auxiliary enzymes. Nevertheless, challenges associated with the cost and
separation of enzymes needs to be overcome for application on a large
scale (Zhou et al., 2020).
4. Factors affecting the production of oligosaccharides
4.1. Temperature and pH
Oligosaccharides are more challenging prebiotics, which renders
them on a platform for their wide application in market. Although the
chemical and biotechnological production of these molecules provide an
opportunity to improve the yield and offer advantages to human health,
but certain parameters are accountable for efficient production of oli­
gosaccharides (Cano et al., 2020). Variables such as temperature, pH,
and substrate concentration have significant rate controlling effect over
the yield of oligosaccharides. It was reported that higher temperature
led to high yield of pentoses with minor by-products accumulation
(Carvalheiro et al., 2016). Molecular weight distribution of oligosac­
charides mainly depends on the temperature and time. The maximum
XOS were obtained when isothermal temperature was altered from
150 ◦ C to 190 ◦ C for 5 min (Carvalheiro et al., 2004). Oguro et al. (2019)
studied the effect of temperature on oligosaccharides production from
Koji amazake, a sweet beverage. It was observed that the production of
oligosaccharides can be enhanced by increasing the temperature of
saccharification. The optimal saccharification temperature was between
50 ◦ C to 60 ◦ C, whereas a higher production of sophorose was reported
when the temperature was increased to 70 ◦ C. Lignocellulosic residues
8
International Journal of Food Microbiology 368 (2022) 109610
serve as substrates for non-digestible oligosaccharide production (NDO).
The potential of rice straw for the production of NDO was evaluated by
Sophonputtanaphoca et al. (2018). The fructo-oligosaccharides yield of
1.55% to 1.61% (w/w) at 60 ◦ C and 1.44% to 1.67% (w/w) at 70 ◦ C
suggested that their production was not affected by changing tempera­
ture. However, in the same study, maximum yield 6.53% (w/w) of XOS
was obtained at 100 ◦ C. This suggested that temperature has a signifi­
cant effect on the production of different types of NDOs. Waste products
of palm industry such as palm kernel cake consist of 35.2% mannan,
which was utilized as a substrate for the production of manno-
oligosaccharides.
The rate of oligosaccharide production can also be modulated by
altering the pH of the process. The pH controls and improves the
selectivity of monomers for oligosaccharide production and minimizes
the formation of degradation products (Cano et al., 2020). Pecto-
oligosaccharides (POS) production from orange peel by-product was
reported by Manderson et al. (2005). They found that pH 1.5 was op­
timum for efficient recovery of these molecules as prebiotics. Milk oli­
gosaccharides are associated with human health especially in infants.
One of the lactose rich co-product of whey protein permeate (colostrum)
was used to produce bovine milk oligosaccharides by lactose hydrolysis.
To obtain high recovery with minimal loss, initially whey permeate was
adjusted at a pH of 4.5, 7.0 and 8.5. It was found that at pH 8.5, high
purity yield of oligosaccharides was recovered which could be used as a
food supplement (Cohen, 2018). Another study by Martínez-Villaluenga
and coworkers evaluated the effect of pH on galacto-oligosaccharide
synthesis via lactose hydrolysis. Synthesis was carried out at different
pH of 5.5, 6.5 and 7.5. Enzyme inactivation was observed during 6′
galactosyl lactose (GOS) synthesis at pH 5.5. At pH 6.5, 15.9% of GOS
was produced but the maximum yield of GOS (17.1%) was obtained at
pH 7.5 (Martínez-Villaluenga et al., 2008). This suggested that pH is a
significant factor to determine the productivity of desired oligosaccha­
rides. Cyclodextrin glucanotransferase is an enzyme that acts on starch
to produce cyclodextrins (CD). It was reported that at pH 6.0 the pro­
duction of β-CD was 0.8 mg whereas at pH 7.0 the production increased
to 1.1 mg. Unlike this, the cyclodextrin glucanotransferase was also
reported to exhibit highest activity at pH 6.0 with soluble tapioca starch
substrates. Thus it was concluded that the enzyme cyclodextrin gluca­
notransferase has an optimum pH between 5.0 and 9.0 pH (Elbaz et al.,
2015; Jaafar et al., 2021). These studies suggest that pH does play a
critical role in enhancing the yield of oligosaccharides. Also, majority of
the oligosaccharide producing strategies involved the use of enzymes or
bacteria and they require specific optimum temperature and pH for their
efficient functioning.
4.2. Substrate loading and particle size
The initial load of biomass and its nature is very crucial to have
satisfactory yield of oligosaccharides (Awasthi et al., 2019). Nabarlatz
et al. (2007) used six different agricultural residues as feedstock for the
production of XOS. The productivity of XOS was mainly associated with
percentage of xylan and acetyl content of the raw material (corncobs,
almond shells, olive stones, rice husks, wheat straw, and barley straw)
used. It was observed that utilization of corncob and almond shell
exhibited high yields as 60% and 50% of XOS, respectively. This further
suggested their use in pure XOS production with low lignin content.
Initial concentration of substrate demonstrates the yield of oligosac­
charides. To determine the effect of substrate concentration, Martínez-
Villaluenga et al. (2008) optimized the initial lactose concentration
during 6′ galactosyl lactose (GOS) synthesis via lactose hydrolysis. The
tested lactose concentration was 150, 250 and 350 mg/mL and it was
found that maximum yield of 6′ galactosyl lactose was obtained at a
concentration of 250 mg/mL. Furthermore, Jaafar et al. (2021) reported
that a lower substrate concentration enhances the production of oligo­
saccharides. They tested the concentration of soluble starch in the range
of 0.5–3.0% (w/v) to evaluate the production of MOS. It was recorded
M.K. Awasthi et al.
that lower starch concentration reduced the MOS production by
reducing the cyclodextrins yielded by cyclodextrin glucanotransferase.
The increase in the starch concentration to 1.5% increased the yield to
297.0 mg/g from an initial of 216 mg/g. A further increase in substrate
loading decreases the concentration of MOS. The reason to this may be
attributed to the low hydrolysis efficiency (Jaafar et al., 2021). Also, the
activity and optimized parameters of enzymes was reported to be
affected by changing the substrates. As shown in Section 4.1, the opti­
mum pH for better efficiency of cyclodextrin glucanotransferase to
produce cyclodextrin changes when substrates were altered.
5. Concept of prebiotics in healthcare and functional foods
Prebiotics are fermentable and non-digestible dietary fiberous foods
that constitute carbohydrates or oligosaccharides. Prebiotics like Inulin
based fructooligosaccharides (FOS), galactooligosaccharides (GOS) and
lactulose are commercially established, soybean oligos, gluco, gentio,
isomalto, xylo (XOS) and mannano oligosaccharides (MOS) are found to
be emerging (lab scale research), whereas other sugar derivatives like
maltodextrin, raffinose, arabinose and arabinoxylan have characteristics
influencing the feature prebiotics. These FOS, GOS and XOS can be
derived from cellulose and hemicellulose fractions of lignocellulosic
biomass (Conde et al., 2009; Martinez et al., 2015; Reshmy et al.,
2021a).
Human intestine is a complex ecosystem with dominant population
of microorganisms. These microorganisms infer symbiotic relation with
the host by providing beneficial effects like inhibition of pathogenic
microorganisms, nutrient fermentation, and adsorption of H2O and
stimulation of human immune system. Many different microorganisms
were identified with probiotic potential and two genera, Lactobacillus
and Bifidobacterium, of microorganisms could prevent the harmful ef­
fects of gastrointestinal disorders (Martinez et al., 2015; Varankovich
et al., 2015). Consequently, these beneficial microorganisms are
recognized as the targets to improve health. Moreover, there are indeed
protective carriers required for the delivery of these viable probiotic
strains to the target site, where the prebiotics plays a dominant role. We
require the raw material that is biocompatible, biodegradable, and
resistant to extreme conditions that mimic the acidic conditions in the
stomach and gastrointestinal tract.
These microorganisms digest the prebiotics or polymeric oligosac­
charides to various secondary metabolites like short chain fatty acids
which increases the acidic environment in the colonic lumen. Upon in­
crease of the acidity, the conditions would not be favourable for
opportunistic pathogens or other harmful microorganisms (LeBlanc
et al., 2017). The short chain fatty acids (SCFAa) such as acetate, pro­
pionate, butyrate, format etc. are small molecules with various biolog­
ical functions and act as energy source to epithelial cells. Acetic acid is
involved in counter acting the pathogen invasion and controls inflam­
mation. Propionic acid and butyric acid protect against diet related
obesity and insulin sensitivity. Butyric acid has an important role in
altering gene transcription by inhibiting histone deacetylase activity
resulting in prevention and treatment of colon cancer (Rivière et al.,
2016). The molar ratio of these SCFAs depends on the colonic micro­
biota and the type of dietary fibre.
6. Conclusions and perspectives
Oligosaccharides (OS) can be efficiently produced using starch,
dairy, lignocellulosic and sugar based substrates. Physico-chemical
pretreatment strategies followed by hydrolysis using suitable catalysts
have shown potential to enhance the production of different categories
of OS. For lignocellulosic substrates, random attack with concentrated
acids has been suggested to facilitate breaking the cell wall for enhanced
production of OS. Among alkalis, NaOH has been found efficient for
recovery of hemicelluloses at low pressure and temperature conditions.
Among biological hydrolysis strategies, microbial and enzymatic
9
International Journal of Food Microbiology 368 (2022) 109610
treatments have been recommended by various researchers. However,
the use of commercial enzymes could be a cost intensive process for
which recombinant enzymes produced in non-cellulolytic or non-
xylanolytic hosts has been proposed as another alternative. Simulta­
neous production of sugar monoesters by enzymatic processes could
increase the purity of oligosaccharides while producing other value
byproducts which could also decrease the cost of the process. However,
the separation of enzymes needs to be carefully considered for its
application on a large scale. Using microorganisms could be more
economical than commercial enzymes and can be effectively adminis­
tered in biorefineries. The factors of OS production should also be taken
care of with special consideration to pH, substrate loading, particle size
and temperature. In this regard, process models, simulation studies and
investigation associated with pilot scale validation of the optimum
process conditions could be beneficial for easy industrial adaptation.
Emerging technologies and combination of technologies should be
investigated with controlled process intensification for higher OS pro­
duction from agro waste residues.
CRediT authorship contribution statement
Mukesh Kumar Awasthi, Ayon Tarafdar, Vivek Kumar Gaur, K.
Amulya: Conceptualization, reviewing, editing and resources, Vivek
Narisetty, Dheeraj Kumar Yadav, Taru Negi: Writing-original draft,
Raveendran Sindhu, Parameswaran Binod, and Ashok Pandey:
reviewing and editing, Ranjna Sirohi: Conceptualization, reviewing
and editing, and Zengqiang Zhang reviewing and editing.
Declaration of competing interest
The authors declare that they have no known competing financial
interests or personal relationships that could have appeared to influence
the work reported in this paper.
Acknowledgments
The authors are grateful for the financial support from the Shaanxi
Introduced Talent Research Funding (A279021901 and F1020221012),
and the Introduction of Talent Research Start-up fund (Z101022001),
College of Natural Resources and Environment, Northwest A&F Uni­
versity, Yangling, Shaanxi Province 712100, China. We are also thankful
to all our laboratory colleagues and research staff members for their
constructive advice and help.
References
1 Amorim, C., Silvério, S.C., Silva, S.P., Coelho, E., Coimbra, M.A., Prather, K.L.J.,
Rodrigues, L.R., 2018. Single-step production of arabino-xylooligosaccharides by
recombinant Bacillus subtilis 3610 cultivated in brewers' spent grain. Carbohydr.
Polym. 199, 546–554.
2 Arun, K.B., Madhavan, A., Sindhu, R., Emmanual, S., Binod, P., Pugazhendhi, A.,
Sirohi, R., Reshmy, R., Awasthi, M.K., Gnansounou, E., Pandey, A., 2021. Probiotics
and gut microbiome - prospects and challenges in remediating heavy metal toxicity.
J. Hazard. Mater. 420, 126676.
3 Ávila, P.F., Silva, M.F., Martins, M., Goldbeck, R., 2021. Cello-oligosaccharides
production from lignocellulosic biomass and their emerging prebiotic applications.
World J. Microbiol. 37 (5), 1–11.
4a Awasthi, M.K., Ferreira, J.A., Sirohi, R., Sarsaiya, S., Khoshnevisan, B., Baladi, S.,
Sindhu, R., Binod, P., Pandey, A., Juneja, A., Kumar, D., Zhang, Z., Taherzadeh, M.
J., 2021. A critical review on the development stage of biorefinery systems
towards the management of apple processing-derived waste. Renew. Sust. Energ.
Rev. 143, 110972.
5 Awasthi, M.K., Paul, A., Kumar, V., Sar, T., Kumar, D., Sarsaiya, S., Liu, H.,
Zhang, Z., Binod, P., Sindhu, R., Kumar, V., Taherzadeh, M.J., 2022. Recent trends
and developments on integrated biochemical conversion process for valorization of
dairy waste to value added bioproducts: a review. Bioresour. Technol. 344, 126193.
6a Awasthi, M.K., Ravindran, B., Sarsaiya, S., Chen, H., Wainaina, S., Singh, E.,
Liu, T., Kumar, S., Pandey, A., Singh, L., Zhang, Z., 2020. Metagenomics for
taxonomy profiling: tools and approaches. Bioengineered 11 (1), 356–374.
7b Awasthi, M.K., Sarsaiya, S., Patel, A., Juneja, A., Singh, R.P., Yan, B., Awasthi, S.
K., Jain, A., Liu, T., Duan, Y., Pandey, A., Zhang, Z., Taherzadeh, M., 2020.
Refining biomass residues for sustainable energy and bio-products: an assessment
M.K. Awasthi et al.
of technology, its importance, and strategic applications in circular bio-economy.
Renew. Sust. Energ. Rev. 127, 109876.
8b Awasthi, M.K., Sarsaiya, S., Wainaina, S., Rajendran, K., Awasthi, S.K., Liu, T.,
Duan, Jian, A., Sindhu, R., Binod, P., Pandey, A., Zhang, Z., Taherzadeh, M., 2021.
Technoeconomics and life-cycle assessment of biological and thermochemical
treatment of bio-waste. Renew. Sustain. Energy. Rev. 144, 110837.
9 Awasthi, M.K., Sarsaiya, S., Wainaina, S., Rajendran, K., Kumar, S., Quan, W.,
Duan, Y., Awasthi, S.K., Chen, H., Pandey, A., 2019. A critical review of organic
manure biorefinery models toward sustainable circular bioeconomy: technological
challenges, advancements, innovations, and future perspectives. Renew. Sust.
Energ. Rev. 111, 115–131.
10 Babbar, N., Baldassarre, S., Maesen, M., Prandi, B., Dejonghe, W., Sforza, S.,
Elst, K., 2016. Enzymatic production of pectic oligosaccharides from onion skins.
Carbohydr. Polym. 146, 245–252.
11 Babbar, N., Dejonghe, W., Sforza, S., Elst, K., 2017. Enzymatic pectic
oligosaccharides (POS) production from sugar beet pulp using response surface
methodology. J. Food Sci. Technol. 54, 3707–3715.
12 Baldassarre, S., Babbar, N., Van Roy, S., Dejonghe, W., Maesen, M., Sforza, S.,
Elst, K., 2018. Continuous production of pectic oligosaccharides from onion skins
with an enzyme membrane reactor. Food Chem. 267, 101–110.
13 Barbosa, F.C., Silvello, M.A., Goldbeck, R., 2020. Cellulase and oxidative enzymes:
new approaches, challenges and perspectives on cellulose degradation for
bioethanol production. Biotechnol. Lett. 42, 875–884.
14 Baruah, J., Nath, B.K., Sharma, R., Kumar, S., Deka, R.C., Baruah, D.C., Kalita, E.,
2018. Recent trends in the pretreatment of lignocellulosic biomass for value-added
products. Front. Energy Res. 6, 141.
15 Bhatia, L., Sharma, A., Bachheti, R.K., Chandel, A.K., 2019. Lignocellulose derived
functional oligosaccharides: production, properties, and health benefits. Prep.
Biochem. Biotechnol. 49, 744–758.
16 Bhatia, R., Lad, J.B., Bosch, M., Bryant, D.N., Leak, D., Hallett, J.P., Franco, T.T.,
Gallagher, J.A., 2021. Production of oligosaccharides and biofuels from Miscanthus
using combinatorial steam explosion and ionic liquid pretreatment. Bioresour.
Technol. 323, 124625.
17 Bouchard, J., Méthot, M., Fraschini, C., Beck, S.M., 2016. Effect of oligosaccharide
deposition on the surface of cellulose nanocrystals as a function of acid hydrolysis
temperature. Cellulose 23, 3555–3567.
18 Brienzo, M., Carvalho, A.F.A., Figueiredo, F.C., Neto, P.O., 2016. Sugarcane bagasse
hemicellulose properties, extraction technologies and xylooligosaccharides
production. In: Riley, G.L. (Ed.), Food Waste, pp. 155–188. Chap. 8.
19 Cano, M.E., García-Martin, A., Comendador Morales, P., Wojtusik, M., Santos, V.E.,
Kovensky, J., Ladero, M., 2020. Production of oligosaccharides from agro-food
wastes. Fermentation 6, 31.
20 Carvalheiro, F., Duarte, L.C., Gírio, F., Moniz, P., 2016. Hydrothermal/liquid hot
water pretreatment (autohydrolysis): a multipurpose process for biomass
upgrading. In: Biomass Fractionation Technologies for a Lignocellulosic Feedstock
Based Biorefinery. Elsevier, pp. 315–347.
21 Carvalheiro, F., Esteves, M.P., Parajó, J.C., Pereira, H., Gırio, F.M., 2004.
Production of oligosaccharides by autohydrolysis of brewery's spent grain.
Bioresour. Technol. 91, 93–100.
22 Chu, Q., Li, X., Xu, Y., Wang, Z., Huang, J., Yu, S., Yong, Q., 2014. Functional cello-
oligosaccharides production from the corncob residues of xylo-oligosaccharides
manufacture. Process Biochem. 49 (8), 1217–1222.
23 Cohen, J.L., 2018. Bioprocessing of Dairy Co-products for Glycan Isolation.
University of California, Davis.
24 Conde, E., Cara, C., Moure, A., Ruiz, E., Castro, E., Domínguez, H., 2009.
Antioxidant activity of the phenolic compounds released by hydrothermal
treatments of olive tree pruning. Food Chem. 114, 806–812.
25 Dávila, I., Gordobil, O., Labidi, J., Gullón, P., 2016. Assessment of suitability of vine
shoots for hemicellulosic oligosaccharides production through aqueous processing.
Bioresour. Technol. 211, 636–644.
26 Đorđević, T., Antov, M., 2018. The influence of hydrothermal extraction conditions
on recovery and properties of hemicellulose from wheat chaff–a modeling
approach. Biomass Bioenergy 119, 246–252.
28 Duan, Y., Mehariya, S., Kumar, A., Singh, E., Yang, J., Kumar, S., Li, H., Awasthi, M.
K., 2021. Apple orchard waste recycling and valorization of valuable product-a
review. Bioengineered 12 (1), 476–495.
29 Duan, Y., Pandey, A., Zhang, Z., Awasthi, M.K., Bhatia, S.K., Taherzadeh, M., 2020.
Organic solid waste biorefinery: sustainable strategy for emerging circular
bioeconomy in China. Ind. Crop. Prod. 153, 112568.
30 Duong Hong, Q., Hoang Minh, T., Nguyen Thi, H.G., Nguyen Thi, T.H., Nguyen, N.
H., Nguyen Thi, T.L., Nguyen Thi, V.A., Vu, T.T., Luong, H.N., 2021. Synthesis of
isomaltooligosaccharides (IMOs) from sweet potato starch by simultaneous
saccharification and transglycosylation using Saccharomyces cerevisiae var.
diastaticus BE 134 to improve purity of IMOs. J. Food Qual. 2021, 12.
31 Elbaz, A.F., Sobhi, A., ElMekawy, A., 2015. Purification and characterization of
cyclodextrin β-glucanotransferase from novel alkalophilic bacilli. Bioprocess
Biosyst. Eng. 38, 767–776.
32 Elst, K., Babbar, N., Van Roy, S., Baldassarre, S., Dejonghe, W., Maesen, M.,
Sforza, S., 2018. Continuous production of pectic oligosaccharides from sugar beet
pulp in a cross flow continuous enzyme membrane reactor. Bioprocess Biosyst. Eng.
41, 1717–1729.
33 Gaur, V.K., Sharma, P., Sirohi, R., Awasthi, M.K., Dussap, C.-G., Pandey, A., 2020.
Assessing the impact of industrial waste on environment and mitigation strategies:
a comprehensive review. J. Hazard. Mater. 398, 123019.
34 Ghazi, I., Fernandez-Arrojo, L., Gomez De Segura, A., Alcalde, M., Plou, F.J.,
Ballesteros, A., 2006. Beet sugar syrup and molasses as low-cost feedstock for the
10
International Journal of Food Microbiology 368 (2022) 109610
enzymatic production of fructo-oligosaccharides. J. Agric. Food Chem. 54,
2964–2968.
35 Gomez, B., Miguez, B., Veiga, A., Parajó, J.C., Alonso, J.L., 2015. Production,
purification, and in vitro evaluation of the prebiotic potential of
arabinoxylooligosaccharides from brewer's spent grain. J. Agric. Food Chem. 63,
8429–8438.
36 Gullón, B., Gómez, B., Martínez-Sabajanes, M., Yáñez, R., Parajó, J.C., Alonso, J.L.,
2013. Pectic oligosaccharides: manufacture and functional properties. Trends Food
Sci. Technol. 30 (2), 153–161.
37 Hemsworth, G.R., Johnston, E.M., Davies, G.J., Walton, P.H., 2015. Lytic
polysaccharide monooxygenases in biomass conversion. Trends Biotechnol. 33
(12), 747–761.
39 Hii, S.L., Tan, J.S., Ling, T.C., Ariff, A.B., 2012. Pullulanase: role in starch
hydrolysis and potential industrial applications. Enzyme Res. 2012.
40 Isikgor, F.H., Becer, C.R., 2015. Lignocellulosic biomass: a sustainable platform for
the production of bio-based chemicals and polymers. Polym. Chem. 6, 4497–4559.
41 Jaafar, N.R., Nawawi, N.N., Abd Rahman, N.H., Annuar, N.A.S., Rahman, R.A.,
Illias, R.M., 2021. Synergistic action of cyclodextrin glucanotransferase and
maltogenic amylase improves the bioconversion of starch to malto-
oligosaccharides. Process Biochem. 103, 9–17.
42 Jagtap, S., Deshmukh, R.A., Menon, S., Das, S., 2017. Xylooligosaccharides
production by crude microbial enzymes from agricultural waste without prior
treatment and their potential application as nutraceuticals. Bioresour. Technol.
245, 283–288.
43 Jana, U.K., Kango, N., 2020. Characteristics and bioactive properties of
mannooligosaccharides derived from agro-waste mannans. Int. J. Biol. Macromol.
149, 931–940.
44 Ji, E.-S., Park, N.-H., Oh, D.-K., 2005. Galacto-oligosaccharide production by a
thermostable recombinant β-galactosidase from Thermotoga maritima. World J.
Microbiol. Biotechnol. 21, 759–764.
45 Ji, H., Li, X., Bai, Y., Shen, Y., Jin, Z., 2021. Synergetic modification of waxy maize
starch by dual-enzyme to lower the in vitro digestibility through modulating
molecular structure and malto-oligosaccharide content. Int. J. Biol. Macromol. 180,
187–193.
46 Jin, T., Käldström, M., Benavides, A., Rechulski, M.D.K., Jarboe, L.R., 2020.
Utilization of mechanocatalytic oligosaccharides by ethanologenic Escherichia coli
as a model microbial cell factory. AMB Express 10, 1–6.
47 Jose, A., Hazeena, S.H., Lakshmi, N.M., Arun, K.B., Madhavan, A., Sirohi, R.,
Tarafdar, A., Sindhu, R., Awasthi, M.K., Pandey, A., Binod, P., 2022. Bacterial
biopolymers: from production to applications in biomedicine. Sustain. Chem.
Pharm. 25, 100582.
48 Karnaouri, A., Matsakas, L., Krikigianni, E., Rova, U., Christakopoulos, P., 2019.
Valorization of waste forest biomass toward the production of cello-
oligosaccharides with potential prebiotic activity by utilizing customized enzyme
cocktails. Biotechnol. Biofuels 12, 1–19.
49 Karnaouri, A., Matsakas, L., Krikigianni, E., Rova, U., Christakopoulos, P., 2019.
Valorization of waste forest biomass toward the production of cello-
oligosaccharides with potential prebiotic activity by utilizing customized enzyme
cocktails. Biotechnol. Biofuels 12 (1), 1–19.
50 Katapodis, P., Christakopoulos, P., 2008. Enzymic production of feruloyl xylo-
oligosaccharides from corn cobs by a family 10 xylanase from Thermoascus
aurantiacus. LWT-Food Sci. Technol. 41, 1239–1243.
51 Kothari, R., Singh, A., Pandey, A.K., Tyagi, V.V., Egamberdieva, D., Bellingrath-
Kimura, S.D., Arora, N.K., 2021. Valorization of bio-waste material: future
dimensions for path towards sustainability. Environ. Sustain. 4, 199–200.
52 LeBlanc, J.G., Chain, F., Martín, R., Bermúdez-Humarán, L.G., Courau, S.,
Langella, P., 2017. Beneficial effects on host energy metabolism of short-chain fatty
acids and vitamins produced by commensal and probiotic bacteria. Microb. Cell
Factories 16, 79.
53 Li, T., Li, S., Du, L., Wang, N., Guo, M., Zhang, J., Yan, F., Zhang, H., 2010. Effects
of haw pectic oligosaccharide on lipid metabolism and oxidativestress in
experimental hyperlipidemia mice induced by high-fatdiet. Food Chem. 121 (4),
1010–1013.
54 Li, Y., Xu, J., Yu, W., Ray, A.K., 2020. Multi-objective optimization of sequential
simulated moving bed for the purification of xylo-oligosaccharides. Chem. Eng. Sci.
211, 115279.
55 Liu, C., Ren, L., Yan, B., Luo, L., Zhang, J., Awasthi, M.K., 2021. Electron transfer
and mechanism of energy production among syntrophic bacteria during acidogenic
fermentation: a review. Bioresour. Technol. 323, 124637.
56 Liu, H., Kumar, V., Jia, L., Sarsaiya, S., Kumar, D., Juneja, A., Zhang, Z., Sindhu, R.,
Binod, P., Bhatia, S.K., Awasthi, M.K., 2021. Biopolymer poly-hydroxyalkanoates
(PHA) production from apple industrial waste residues: a review. Chemosphere
284, 131427.
57 Liu, H., Qin, S., Sirohi, R., Ahulwalia, V., Zhou, Y., Sindhu, R., Binod, R.,
Singhania, R.R., Patel, A.K., Juneja, A., Kumar, D., Zhang, Z., Kumar, J.,
Taherzadeh, M., Awasthi, M.K., 2021. Sustainable blueberry waste recycling
towards biorefinery strategy and circular bioeconomy: a review. Bioresour.
Technol. 332, 125181.
58 Madhavan, A., Arun, K.B., Binod, P., Sirohi, R., Tarafdar, A., Reshmy, R.,
Awasthi, M.K., Sindhu, R., 2021. Design of novel enzyme biocatalysts for industrial
bioprocess: harnessing the power of protein engineering, high throughput screening
and synthetic biology. Bioresour. Technol. 325, 124617.
59 Madhavan, A., Arun, K.B., Sindhu, R., Jose, A.A., Pugazhendhi, A., Binod, P.,
Sirohi, R., Reshmy, R., 2022. Engineering interventions in industrial filamentous
fungal cell factories for biomass valorization. Bioresour. Technol. 344, 126209.
M.K. Awasthi et al.
60 Madhavan, A., Arun, K.B., Sindhu, R., Krishnamoorthy, J., Reshmy, R., Sirohi, R.,
Pugazhendi, A., Awasthi, M.K., Szakac, G., Binod, P., 2021. Customized yeast cell
factories for biopharmaceuticals: from cell engineering to process scale up. Microb.
Cell Factories 20, 124.
61 Manderson, K., Pinart, M., Tuohy, K.M., Grace, W.E., Hotchkiss, A.T., Widmer, W.,
Yadhav, M.P., Gibson, G.R., Rastall, R.A., 2005. In vitro determination of prebiotic
properties of oligosaccharides derived from an orange juice manufacturing by-
product stream. Appl. Environ. Microbiol. 71, 8383–8389.
62 Martinez, M., Yanez, R., Alonsó, J.L., Parajó, J.C., 2010. Chemical production of
pectic oligosaccharides from orange peel wastes. Ind. Eng. Chem. Res. 49 (18),
8470–8476.
63 Martinez, R.C., Bedani, R., Saad, S.M., 2015. Scientific evidence for health effects
attributed to the consumption of probiotics and prebiotics: an update for current
perspectives and future challenges. Br. J. Nutr. 114, 1993–2015.
64 Martínez-Villaluenga, C., Cardelle-Cobas, A., Corzo, N., Olano, A., Villamiel, M.,
2008. Optimization of conditions for galactooligosaccharide synthesis during
lactose hydrolysis by β-galactosidase from Kluyveromyces lactis (Lactozym 3000 L
HP G). Food Chem. 107, 258–264.
65 Mazlan, N.A., Samad, K.A., Yahya, N.D., Samah, R.A., Jahim, J., Yussof, H.W.,
2019. Factorial analysis on nitric acid pretreatment of oil palm frond bagasse for
xylan recovery. Mater. Today Proc. 19, 1189–1198.
66 Mazlan, N.A., Samad, K.A., Yussof, H.W., Saufi, S.M., Jahim, J., 2019.
Xylooligosaccharides from potential agricultural waste: characterization and
screening on the enzymatic hydrolysis factors. Ind. Crop. Prod. 129, 575–584.
67 Meyer, T.S.M., Miguel, A.S.M., Fernández, D.E.R., Ortiz, G.M.D., 2015.
Biotechnological production of oligosaccharides-applications in the food industry.
In: Food Prod. Ind. InTech, pp. 25–78.
68 Nabais, J.V., Laginhas, C., Carrott, P.J.M., Carrott, M.R., 2010. Thermal conversion
of a novel biomass agricultural residue (vine shoots) into activated carbon using
activation with CO2. J. Anal. Appl. Pyrolysis 87 (1), 8–13.
69 Nabarlatz, D., Ebringerová, A., Montané, D., 2007. Autohydrolysis of agricultural
by-products for the production of xylo-oligosaccharides. Carbohydr. Polym. 69,
20–28.
70 Narisetty, V., Castro, E., Durgapal, S., Coulon, F., Jacob, S., Kumar, D., Awasthi, M.
K., Pant, K.K., Binod, P., Kumar, V., 2021. High level xylitol production by Pichia
fermentans using non-detoxified xylose-rich sugarcane bagasse and olive pits
hydrolysates. Bioresour. Technol. 342, 126005.
71 Narisetty, V., Parhi, P., Mohan, B., Hazzena, S.H., Kumar, A.N., Gullon, B.,
Sirvastav, A., Nair, L.M., Alphy, M.P., Sindhu, R., Kumar, V., Castro, E., Awasthi, M.
K., Binod, P., 2022. Valorization of renewable resources to functional
oligosaccharides: recent trends and future prospective. Bioresour. Technol. 346,
126590.
72 Oguro, Y., Nakamura, A., Kurahashi, A., 2019. Effect of temperature on
saccharification and oligosaccharide production efficiency in koji amazake.
J. Biosci. Bioeng. 127, 570–574.
73 Olano-Martin, E., Rimbach, G.H., Gibson, G.R., Rastall, R.A., 2003. Pectin and
pectic-oligosaccharides induce apoptosisinvitrohuman colonic adenocarcinoma
cells. Anticancer Res. 23 (1A), 341–346.
74 Otieno, D.O., 2010. Synthesis of β-galactooligosaccharides from lactose using
microbial β-galactosidases. Compr. Rev. Food Sci. Food Saf. 9, 471–482.
75 Park, S.H., Na, Y., Kim, J., Dal Kang, S., Park, K.-H., 2018. Properties and
applications of starch modifying enzymes for use in the baking industry. Food Sci.
Biotechnol. 27, 299–312.
76 Patel, A., Singhania, R.R., Awasthi, M.K., Varjani, S., Bhatia, S.K., Tsai, M.L.,
Hsiesh, S.L., Chen, C.W., Dong, C.D., 2021. Emerging prospects of macro and
microalgae as prebiotic. Microb. Cell Factories 20, 112.
77 Pereira, G.F., de Bastiani, D., Gabardo, S., Squina, F., Ayub, M.A.Z., 2018. Solid-
state cultivation of recombinant Aspergillus nidulans to co-produce xylanase,
arabinofuranosidase, and xylooligosaccharides from soybean fibre. Biocatal. Agric.
Biotechnol. 15, 78–85.
78 Pinales-Márquez, C.D., Rodríguez-Jasso, R.M., Araújo, R.G., Loredo-Treviño, A.,
Nabarlatz, D., Gullón, B., Ruiz, H.A., 2021. Circular bioeconomy and integrated
biorefinery in the production of xylooligosaccharides from lignocellulosic biomass:
a review. Ind. Crop. Prod. 162, 113274.
79 Prapulla, S.G., Subhaprada, V., Karanth, N.G., 2000. Microbial production of
oligosaccharides: a review. Adv Appl. Microbial. 47, 299–343.
80 Qin, S., Giri, B.S., Patel, A.K., Sar, T., Liu, H., Chen, H., Juneja, A., Kumar, D.,
Zhang, Z., Awasthi, M.K., Taherzadeh, M., 2021. Resource recovery and biorefinery
potential of apple orchard waste in the circular bioeconomy. Bioresour. Technol.
321, 124496.
81 Qin, S., Wainaina, S., Liu, H., Soufiani, A.M., Pandey, A., Zhang, Z., Awasthi, M.K.,
Taherzadeh, M.J., 2021. Microbial dynamics during anaerobic digestion of sewage
sludge combined with food waste at high organic loading rates in immersed
membrane bioreactors. Fuel 303, 121276.
82 Qin, S., Wainaina, W., Awasthi, S.K., Mahboubi, A., Liu, T., Liu, H., Zhou, H.,
Zhang, Z., Taherzadeh, M., 2021. Fungal dynamics during anaerobic digestion of
sewage sludge combined with food waste at high organic loading rates in immersed
membrane bioreactors. Bioresour. Technol. 335, 125296.
83 Qu, J., Sun, Y., Awasthi, M.K., Liu, Y., Xu, X., Meng, X., Zhang, H., 2021. Effect of
different aerobic hydrolysis time on the anaerobic digestion characteristics and
energy consumption analysis. Bioresour. Technol. 320, 124332.
84 Ravindran, B., Karmegam, N., Yuvaraj, A., Thangaraj, R., Chang, S.W., Zhang, Z.,
Awasthi, M.K., 2021. Cleaner production of agriculturally valuable benignant
materials from industry generated bio-wastes: a review. Bioresour. Technol. 320,
124281.
11
International Journal of Food Microbiology 368 (2022) 109610
85 Reshmy, R., Balakumaran, P.A., Divakar, K., Philip, E., Madhavan, A.,
Pugazhendhi, A., Sirohi, R., Binod, P., Awasthi, M.K., Sindhu, R., 2022. Microbial
valorization of lignin: prospects and challenges. Bioresour. Technol. 344, 126240.
86 Reshmy, R., Madhavan, A., Arun, K.B., Philip, E., Sindhu, R., Binod, P.,
Puthiyamadam, A., Awasthi, M.K., Pandey, A., 2022. Chili post-harvest residue-
derived nanocellulose composite as a matrix for in vitro cell culture and
Hemigraphis colorata blended nanocellulose extends antimicrobial potential.
Sustain. Chem. Pharm. 25, 100584.
87 Reshmy, R., Paulose, T.A.P., Philip, E., Thomas, D., Madhavan, A., Sirohi, R.,
Binod, P., Awasthi, M.K., Pandey, A., Sindhu, R., 2022. Updates on high value
products from cellulosic biorefinery. Fuel 308, 122056.
88 Reshmy, R., Philip, E., Madhavan, A., Sindhu, R., Pugazhendhi, A., Binod, P.,
Sirohi, R., Awasthi, M.K., Tarafdar, A., Pandey, A., 2021. Advanced biomaterials for
sustainable applications in the food industry: updates and challenges*. Environ.
Pollut. 283, 117071.
89 Reshmy, R., Philip, E., Madhavan, A., Sirohi, R., Pugazhendhi, A., Binod, P.,
Awasthi, M.K., Vivek, N., Kumar, V., Sindhu, R., 2022. Lignocellulose in future
biorefineries: strategies for cost-effective production of biomaterials and bioenergy.
Bioresour. Technol. 344, 126241.
90 Reshmy, R., Philip, E., Madhavan, A., Tarfdar, A., Sindhu, R., Binod, P., Sirohi, R.,
Awasthi, M.K., Pandey, S., 2022. Biorefinery aspects for cost-effective production of
nanocellulose and high value-added biocomposites. Fuel 311, 122575.
91 Reshmy, R., Philip, E., Thomas, D., Madhavan, A., Sindhu, R., Binod, P., Varjani, S.,
Awasthi, M.K., Pandey, A., 2021. Bacterial nanocellulose: engineering, production,
and applications. Bioengineered 12 (2), 11463–11483.
92 Rivière, A., Selak, M., Lantin, D., Leroy, F., De Vuyst, L., 2016. Bifidobacteria and
butyrate-producing colon bacteria: importance and strategies for their stimulation
in the human gut. Front. Microbiol. 28 (7), 979.
93 Roy, D., Daoudi, L., Azaola, A., 2002. Optimization of galacto-oligosaccharide
production by Bifidobacterium infantis RW-8120 using response surface
methodology. J. Ind. Microbiol. 29 (5), 281–285.
94 Sánchez, O., Guio, F., Garcia, D., Silva, E., Caicedo, L., 2008. Fructo
oligosaccharides production by Aspergillus sp. N74 in a mechanically agitated
airlift reactor. Food Bioprod. Process. 86, 109–115.
95 Santibáñez, L., Henríquez, C., Corro-Tejeda, R., Bernal, S., Armijo, B., Salazar, O.,
2012. Xylooligosaccharides from lignocellulosic biomass: a comprehensive review.
Carbohydr. Polym. 1 (251), 117–118.
96 Sarsaiya, S., Jain, A., Awasthi, S.K., Duan, Y., Awasthi, M.K., Shi, J., 2019.
Microbial dynamics for lignocellulosic waste bioconversion and its importance with
modern circular economy, challenges and future perspectives. Bioresour. Technol.
291, 121905.
97 Scheller, H.V., Ulvskov, P., 2010. Hemicelluloses. Annu. Rev. Plant Biol. 61,
263–289.
98 Schröder, R., Atkinson, R.G., Redgwell, R.J., 2009. Re-interpreting the role of endo-
β-mannanases as mannan endotrans glycosylase/hydrolases in the plant cell wall.
Ann. Bot. 104 (2), 197–204.
99 Sindhu, R., Binod, P., Pandey, A., 2016. Biological pretreatment of lignocellulosic
biomass – an overview. Bioresour. Technol. 199, 76–82.
100 Singh, J., Suhag, M., Dhaka, A., 2015. Augmented digestion of lignocellulose by
steam explosion, acid and alkaline pretreatment methods: a review. Carbohydr.
Polym. 117, 624–631.
101 Singh, R.D., Banerjee, J., Arora, A., 2015. Prebiotic potential of oligosaccharides: a
focus on xylan derived oligosaccharides. Bioact. Carbohydr. Diet. Fibre. 5 (1),
19–30.
102 Singh, S., Ghosh, A., Goyal, A., 2018. Manno-oligosaccharides as prebiotic-valued
products from agro-waste. In: Biosynthetic Technology and Environmental
Challenges. Springer, Singapore, pp. 205–221.
103 Sirisansaneeyakul, S., Lertsiri, S., Tonsagunrathanachai, P., Luangpituksa, P., 2000.
Enzymatic production of fructo-oligosaccharides from sucrose. Agric. Nat. Resour.
34, 262–269.
104 Sirohi, R., Pandey, J.P., Singh, A., Sindhu, R., Lohani, U.C., Goel, R., Kumar, A.,
2020. Acid hydrolysis of damaged wheat grains: modeling the formation of
reducing sugars by a neural network approach. Ind. Crop. Prod. 149, 112351.
105 Sophonputtanaphoca, S., Pridam, C., Chinnak, J., Nathong, M., Juntipwong, P.,
2018. Production of non-digestible oligosaccharides as value-added by-products
from rice straw. Agric. Nat. Resour. 52, 169–175.
106 Varankovich, N.V., Nickerson, M.T., Korber, D.R., 2015. Probiotic-based strategies
for therapeutic and prophylactic use against multiple gastrointestinal diseases.
Front. Microbiol. 14 (6), 685.
107 Von Freiesleben, P., Moroz, O.V., Blagova, E., Wiemann, M., Spodsberg, N.,
Agger, J.W., Krogh, K.B., 2019. Crystal structure and substrate interactions of an
unusual fungal non-CBM carrying GH26 endo-β-mannanase from
Yunnaniapenicillata. Sci. Rep. 9 (1), 1–14.
108 Wainaina, S., Awasthi, M., Sarsaiya, S., Chen, H., Singh, E., Kumar, A.,
Ravindran, B., Awasthi, S., Liu, T., Duan, Y., Kumar, S., Zhang, Z., Taherzadeh, M.
J., 2020. Resource recovery and circular economy from organic solid waste using
aerobic and anaerobic digestion technologies. Bioresour. Technol. 301, 122778.
109 Wainaina, S., Lukitawesa, Awasthi, M.K., Taherzadeh, M.J., 2019. Bioengineering
of anaerobic digestion for volatile fatty acids, hydrogen or methane production: a
critical review. Bioengineered 10, 437–458.
110 Wang, X., Chen, Q., Lü, X., 2014. Pectin extracted from apple pomace and citrus
peel by subcritical water. Food Hydrocoll. 38, 129–137.
111 Wang, Y., Jing, Y., Lu, C., Kongjan, P., Wang, J., Awasthi, M.K., Tahir, N.,
Zhang, Q., 2021. A co-fermentation model for bio-hydrogen production. J. Clean.
Prod. 317, 128288.
M.K. Awasthi et al.
38 Xie, H., Feng, X., Wang, M., Wang, Y., Awasthi, M.K., Xu, P., 2020. Implications of
endophytic microbiota in Camellia sinensis: a review on current understanding and
future insights. Bioengineered 11 (1), 1001–1015.
112 Xu, A., Lai, W., Chen, P., Awasthi, M.K., Chen, X., Wang, Y., Xu, P., 2021.
A comprehensive review on polysaccharide conjugates derived from tea leaves:
composition, structure, function and application. Trends Food Sci. Technol. 114,
83–99.
113 Xu, Y., Awasthi, M.K., Li, P., Meng, X., Wang, Z., 2020. Comparative analysis of
prediction models for methane potential based on spent edible fungus substrate.
Bioresour. Technol. 317, 124052.
114 Yang, R., Xu, S., Wang, Z., Yang, W., 2005. Aqueous extraction of corncob xylan and
production of xylooligosaccharides. LWT-Food Sci. Technol. 38 (6), 677–682.
115 You, Y., Zhang, X., Li, P., Lei, F., Jiang, J., 2020. Co-production of
xylooligosaccharides and activated carbons from Camellia oleiferashell treated by
the catalysis and activation of zinc chloride. Bioresour. Technol. 306, 123131.
116 Yu, L., O’Sullivan, D.J., 2014. Production of galactooligosaccharides using a
hyperthermophilic β-galactosidase in permeabilized whole cells of Lactococcus
lactis. J. Dairy Sci. 97, 694–703.
117 Yusree, F.I.F.M., Peter, A.P., Yusree, N.A.B., Nor, M.Z.M., Basri, M.S.M.,
Mokhtar, M.N., Awasthi, M.K., Show, P.L., 2022. Towards green recovery of
β-amylase from slurry of sweet potato (Ipomoea batatas) of VitAto variety via liquid
biphasic system. Sustain. Chem. Pharm. 25, 100579.
12
International Journal of Food Microbiology 368 (2022) 109610
118 Zeuner, B., Teze, D., Muschiol, J., Meyer, A.S., 2019. Synthesis of human milk
oligosaccharides: protein engineering strategies for improved enzymatic
transglycosylation. Molecules 24, 2033.
119 Zhang, L., Zhong, L., Wang, J., Zhao, Y., Zhang, Y., Zheng, Y., Dong, W., Ye, X.,
Huang, Y., Li, Z., 2021. Efficient hydrolysis of raw starch by a maltohexaose-
forming α-amylase from Corallococcus sp.EGB. LWT 152, 112361.
120 Zhang, Y.-H.P., Lynd, L.R., 2003. Cellodextrin preparation by mixed-acid hydrolysis
and chromatographic separation. Anal. Biochem. 322, 225–232.
121 Zhong, C., Ukowitz, C., Domig, K.J., Nidetzky, B., 2020. Short-chain cello-
oligosaccharides: intensification and scale-up of their enzymatic production and
selective growth promotion among probiotic bacteria. J. Agric. Food Chem. 68,
8557–8567.
122 Zhou, P., Liu, C., Wang, W., Wang, F., Nie, K., Deng, L., 2020. The effectively
simultaneous production of cello-oligosaccharide and glucose mono-decanoate
from lignocellulose by enzymatic esterification. Appl. Biochem. Biotechnol. 192 (2),
600–615.
123 Zhou, X., Xu, Y., 2019. Integrative process for sugarcane bagasse biorefinery to co-
produce xylooligosaccharides and gluconic acid. Bioresour. Technol. 282, 81–87.
124 Zhou, X., Zhao, J., Zhang, X., Xu, Y., 2019. An eco-friendly biorefinery strategy for
xylooligosaccharides production from sugarcane bagasse using cellulosic derived
gluconic acid as efficient catalyst. Bioresour. Technol. 289, 121755.