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Resistant Starch Preparation Methods

Chapter · January 2019

DOI: 10.1016/B978-0-08-100596-5.22491-8

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 Resistant Starch Preparation Methods
Amir Amini Khoozani, John Birch, and Alaa El-Din Ahmed Bekhit, Department of Food Science, University of Otago, Dunedin,
New Zealand
© 2018 Elsevier Inc. All rights reserved.

Introduction 1
Resistant Starch 1
Resistant Starch Health Benefits 2
Preparation Methods 2
Conclusion 4
References 4

Glossary
Prebiotic A carbohydrate-based substance which acts as a metabolite for the growth of probiotics.
Probiotic Microorganisms living in the large intestine with capability of fermentation and producing short chain acids that
exert health benefits from the gastrointestinal system.
Resistant starch A part of starch that is resistant to digestion in the small intestine and can be fermented by colon microbiota.

Introduction

Due to the importance of prebiotics in prevention of certain diseases, they have increasingly attracted the attention of food
technologists (Verma and Banerjee, 2010). Prebiotics are non-digestible oligosaccharides that offer many beneficial effects on
the gastrointestinal system. Typical prebiotics are dietary fibers (DF) where the most well-known are inulin, oligosaccharides
and resistant starch (RS) (Buttriss and Stokes, 2008). The functional features of RS, together with its potential physiological benefits,
provide an opportunity to increase the level of DF in the diet through common foods. However, processing procedures, especially
thermal processing, may decrease the RS content of food products, whether naturally containing RS or RS-enriched products
(Sullivan et al., 2017). In this respect, researchers started to look for ways to either increase the amount of RS or making it more
resistant to processing conditions. In this chapter, in addition to review the preparation methods of RS, health benefits of this
unique prebiotic will also be discussed.

Resistant Starch

Resistant starch is mainly composed of the linear part of the starch molecule (amylose) which is fermented by probiotics, including
Bifidobacterium species in the colon. This gives rise to the production of short chain fatty acids, mainly butyric acid, which plays
a significant role in prevention of colorectal cancer. The effective dosage of RS which exerts beneficial health effects was reported
as 6–12 g/d, whereas the recommendation for daily intake of another DF was 38 g/d (Fuentes-Zaragoza et al., 2011; Behall
et al., 2006). RS naturally occurs in starch-based seeds, cereal grains and cooled heat-treated starchy foods. The highest amount
of RS can be found in raw potato with 75 g/100 g (dry basis). Also, green banana is remarkable due to its high RS percentage of
about 70 g/100 g of peeled fruit (Wang et al., 2017).
There are five types of RS introduced to date. In table 1, types, sources and description of each type are summarized.

Table 1 Resistant starch types, description and sources (Milasinovic-Seremesic et al., 2012; Birt et al.,
2013; Khalili and Amini, 2015)

Type Description Source

RS1 Physically inaccessible in cell walls Grain, legumes, seeds

RS2 Granular native starch with high Raw starchy foods (potato, pasta, high-amylose corn,
crystalline structure unripe banana)
RS3 Retrograded starch Retrograded starchy foods
RS4 Chemically modified starch Esterified, Etherified or phosphorylated cross-linked
starch
RS5 Amylose-lipid complex Fatty acid treatment of debranched starch

1
2 Resistant Starch Preparation Methods

Resistant Starch Health Benefits

Considering that several digestive diseases are triggered by inadequate or inappropriate diet, increased consumption of indigestible
carbohydrates is important. Among numerous DF, RS has approximately half of the calorific value (8 kJ/g) compared with digestible
starch (15 kJ/g) (Chen et al., 2017). Numerous health benefits have been shown for RS. In addition to its direct impact on the
reduction of the glycemic index, most health effects have been ascribed to the prebiotic nature of RS (Boll et al., 2016).
Colonic microbiota, specifically Bifidobacterium bifidum sp., starts to ferment RS into short chain fatty acids. Therefore, with
reducing pH level of the environment by the presence of acetic, propionic and butyric acid, the proliferation of carcinomatous cells
begins to be prohibited. Some studies have shown a positive effect of RS intake on prevention of colorectal cancer (Homayouni
et al., 2014; Malcomson et al., 2015; Amini et al., 2015; Singh et al., 2016; Hung et al., 2016; Yuan et al., 2017; Yin and Zhao,
2017; Panebianco et al., 2017; Cray et al., 2017). As a result, the indirect effects of RS consumption are mostly due to the activity
of probiotic microbiota which is illustrated in Fig. 1.

Preparation Methods

There are three main strategies to produce resistant starch-rich powdered ingredients: physical, chemical and enzymatic processes.
In the physical method, the main procedure is repeated heating–cooling cycles which lead to reorganization of linear chains of
starch into a new structure which is resistant to hydrolysis by digestive enzymes (Sarawong et al., 2014). When heating is applied
to a high-amylose starch dispersion, disaggregation and breaking of both amylose and amylopectin occurs and leads to formation of
shorter linear chains. In the next process, cooling down of these chains brings about the formation of double helical aggregates
which are denser and more resistant to be hydrolyzed. Consequently, RS produced by this method is more resistant to heat
treatment and can be used as a functional ingredient for fortification of food products (Abioye et al., 2017).
Chemical methods promote modification of starch granules by lintnerization, acetylation, phosphorylation, oxidation, hydrox-
ypropylation, esterification and combinations of these treatments (Nagahata et al., 2013). In each of these treatments, a chemical
reagent prevents enzymes from binding properly to the starch. For instance, acetyl groups can be esterified to starch using free
hydroxyl groups present on the glucose units with acetic anhydride commonly being used (Sha et al., 2012). In oxidation, by
the utilization of reagents such as oxygen, ozone, sodium hypochlorite, periodate and hydrogen peroxide, carbonyl and carboxylic
groups are produced that impedes the starch digestive enzymes (Chung et al., 2008). For hydroxypropylation treatment, propylene
oxide is commonly used. Under high alkaline conditions, the esterified propylene is utilized to free hydroxy groups from the starch
structure resulting in a bulky structure which cannot be digested by carbohydrase enzymes (Juansang et al., 2012). Lintnerization is
the term applied to mineral acid-treated starch. By disruption of amorphous sections of starch, acid treatment (mainly by

Increase Decrease Prevention

excretion rates of postprandial insulin

colon cancer
cholesterol response

postprandial glucose
mineral absorption obesity
response

T-lymphocyte cells pathogenic growth hypertention

duration of rotavirus inflammatory bowel

immune system
diarrhea diseases

HDL-Cholesterol LDL-Cholesterol type one diabetes

excretion rates of bile

appetite type two diabetes
acids

Figure 1 Health effects of resistant starch (Homayouni et al., 2014; Amini et al., 2015).
 Resistant Starch Preparation Methods 3

Table 2 Chemical treatments for the production of resistant starch

Chemical method Starch type Main chemical process Remarks Reference

Lintnerization Banana HCL (heating for 75
C and • Time consuming and less Aparicio-Saguilán et al., 2005,
Corn storage up to 78 h) productive Brumovsky and Thompson,
Waxy corn 2001, Ozturk et al., 2011,
Nagahata et al., 2013
Phosphorylation Pea Using sodium • Legal limitations by FDA at Dupuis et al., 2014,
Normal corn trimetaphosphate and sodium excess amounts (0.4%) Thompson, 2001, Yeo and
Canna tripolyphosphate in alkaline • Destructive effects on Seib, 2009
Rice conditions (around 3 h), bread’s rheology at high level
Wheat following gelatinization at of substitution
100
C
Carboxymethylation Potato Addition of monochloroacetic • Reducing process time by Liu et al., 2012,
Rice acid in alkaline conditions microwave heating Kittipongpatana and
with subsequent microwave • Low RS yield Kittipongpatana, 2013
treatment
Oxidation Normal corn Injection of O3, H2O2, O2 in • Enzymatic hydrolysis by Chung et al., 2008, Simsek
Rice alkaline conditions for a-amylase et al., 2012
Pinto bean 30 min • Slightly increase in bean
starch
Acetylation Rice Treatment with vinyl acetate • Around 16% vinyl acetate led Sha et al., 2012, Simsek et al.,
Canna (30
C, 5 h) to higher amount of RS 2012; Juansang et al., 2012
Normal corn
Hydroxypropylation Canna Addition of propylene oxide in • Long treatment (40 h) Juansang et al., 2012, Chung
Normal corn alkaline conditions Followed • Decrease of RS compared to et al., 2008, Han and
Waxy corn by gelatinization (40min) initial amount Bemiller, 2007
Citric Acid treatment Normal 12 h reaction with 0.1 or • Decrease of RS2 amount Sun et al., 2012, Shin et al.,
Waxy 0.01 M citric acid followed • Long procedure time 2007, Xie and Liu, 2004
Hylon VII by oven heating • Only suitable for high
OR amylose cereals
Non-autoclaved method: 16 h
reaction þ 9 h heating at
120
C

hydrochloric acid) leaves behind a higher ratio of crystalline parts which are more difficult for enzymes to access. In table 2 some of
the main chemical modifications are shown.
Enzymatic treatments focus on debranching the a-1-6 amylopectin bonds by pullulanase and isoamylase which results in
rearranging the structure later in the retrogradation process (Reddy et al., 2013). Other enzymes such as a and b-amylase are
used to hydrolyze the starch amorphous regions and leave a tightly packed crystalline structure behind. Consequently, the aim
of using debranching enzymes is to hydrolyze amylopectin branch chains and provide more linear parts (Cai and Shi, 2010). In
this regard, subsequent retrogradation will produce higher levels of RS3 with double helical structures stabilized by hydrogen
bonds. However, in order to achieve maximum enzyme performance, optimal pH, temperature and time of reaction should be
precisely controlled. Table 3 shows the main enzyme processes in enzymatic treatment of starchy crops.

Table 3 Enzymatic treatments for the production of resistant

Enzymatic method Starch type Main enzyme process Remarks Reference

b-amylase corn 20 h of b-amylase reaction • Long preparation procedure Hickman et al., 2009
Wheat followed by 3 autoclave • Two-fold increase in RS
cycles
b-Amylase and corn starch b-Amylase in conjunction with • Optimum amount of enzyme Zhang and Jin, 2011
Pullulanase Pullulanase concentration is of importance
Isoamylase Waxy corn 1g/100 g starch dry basis • Pre-cooking is needed Cai and Shi, 2010
Waxy wheat Isoamylase (50
C, 24 h) Long post-treatment (48 h)
Waxy potato followed by retrogradation • Significantly increased RS3
Pullulanase Gelatinized red Pullulanase, 40U/g,10 h • Thermal treatment is needed Reddy et al., 2013, van
kidney bean followed by heat treatment at • Long post-treatment (24h/25
C) Hung et al., 2012
Cassava 50
C
potato
4 Resistant Starch Preparation Methods

Table 4 Combination treatments for increasing resistant starch

Source Procedure %RS (g/100 g) Remarks Reference

Taro corm autoclaving, enzymatic
debranching,
retrogradation and oven drying
processes for two times
(60
C)
Maize starch twin-screw extrusion, cooking,
mild acid hydrolysis,
hydrothermal treatments
(110
C)
Cassava starch enzymatic debranching, steam
cooking, drying (45
C) and
freezing (28
C)
High amylose corn starch storage in 23
C then drying at
(Hylon V and VII) 50
C after each extrusion
cooking with high moisture
feed for 3 cycles
High-amylose corn starch Hydrothermal Pressure
(15 MPa for 2 h) followed by
retrogradation (120
C/24h,
4
C)
35.1  1.9% (dry basis)
increased from 11% to 20%
19–20
Hylon V: from 43 to 40
Hylon VII: from 53 to 45.1
At 100
C reached to 27%
• A combination of methods in Simsek and El, 2012
two cycles resulted in 16-fold
increase in RS amount
• Acid modified normal-maize Hasjim and Jane, 2009
starch (AMMS) produced
a greater RS
• Refrigeration of cassava Abioye et al., 2017
starch gave rise to higher RS
compared to freezing storage
• RS3 made from Hylon VII Masatcioglu et al., 2017
had high emulsion stability
• Repeated autoclaving-
storage cycles are necessary
to increase the RS3 content
to the desired levels
• The hydrothermal pressure Pu et al., 2013
resulted in a significant
increase in the total RS
content if the samples were
gelatinized at temperatures
below 120
C

Regarding that a combination of mentioned methods could also increase the RS amount, most of them ended with retrogradation
processes resulting in RS3 increments. The conditions and percentage of produced RS3 in the finished product are summarized in
table 4.

Conclusion

Resistant starch is an exceptional prebiotic with numerous additional health benefits. It also brings technological positive effects for
food products as a functional ingredient. However, processing conditions, especially heat treatment, can attenuate its content.
Consequently, there are ways in order to isolate and increase the amount of resistant starch with the aim of increasing its capability
to be resistant during food processes.
Chemical reagents could increase the amount of resistant starch type four in starchy foods, however, long reaction times and
some health concerns about the dosage of chemicals and the residue are limiting factors. Enzymatic debranching is shown to be
more effective due to the high concentration of produced resistant starch type three. It also seems to be an essential step when
used with other methods, such as heat and cool cycles. Though, it needs a controlled environment, effective enzyme activity and
subsequent heat treatments. Although the heating–cooling cycles could increase the resistant starch type three in produced products,
there are several factors that needed to be investigated including the rate of cooling/heating process, number of cycles and various
temperatures. Additionally, the combination effect of fatty acid addition and other methods for producing resistant starch type five
has not been considered yet.

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