REVIEW
www.starch-journal.com
Recent Advances in Modification Approaches, Health
Benefits, and Food Applications of Resistant Starch
Zhongwei Zhang and Jinsong Bao*
Resistant starch (RS) is a non-digestible carbohydrate that has numerous
biological benefits. However, the amounts of RS are limited in most native
cereal starches, thus developing RS-fortified starch sources and food products
is necessary. This review summarizes the effectiveness of RS improvement by
chemical, enzymatic, and physical modification approaches in starches from
various origins. The functions of RS in regulating the metabolisms and the
influences of RS on the nutritional and textural properties of various foods are
also elucidated. After modification, the molecular structures are altered and
the starch granules became more resistant to enzymatic digestion. However,
the comparison of the RS contents between raw and cooked starch is absent
in many studies. In addition, RS plays an essential role in attenuating
postprandial glycemia, inhibiting glucose and lipid synthesis and promoting
glucose tolerance, insulin sensitivity, and lipid degradation, which are
controlled by the enriched diversity and abundance of the beneficial
microbiota in the colon, together with more short-chain fatty acids. The
incorporation of RS in food products significantly increases nutritional value
without unacceptable quality loss, which provides a new direction for
manufacturing functional foods.
1. Introduction
Starch is classified into rapidly digestible starch (RDS), slowly di-
gestible starch (SDS), and resistant starch (RS) based on its rate of
glucose release and absorption in the gastrointestinal tract.[1] RS
is classified into five types according to their specific preparation
methods: type I resistant starches (RS1) are physically entrapped
starches that protected from hydrolysis by vegetable materials;
type II resistant starches (RS2) are native starches, mainly con-
sisting of high-amylose starch, with a compact crystalline granu-
lar structure that prevents 𝛼-amylase digestion, which are often
Z. Zhang, J. Bao
Yazhou Bay Science and Technology City
Hainan Institute of Zhejiang University
Yazhou Districut, Sanya, Hainan 572025, China
E-mail: jsbao@zju.edu.cn
Z. Zhang, J. Bao
Institute of Nuclear Agricultural Sciences
College of Agriculture and Biotechnology
Zhejiang University
Zijingang Campus, Hangzhou 310058, China
The ORCID identification number(s) for the author(s) of this article
can be found under https://doi.org/10.1002/star.202100141
DOI: 10.1002/star.202100141
Starch - Stärke 2021, 2100141 2100141 (1 of 13)
found in potato; type III resistant starches
(RS3) are formed by recrystallization of
the starch polymers (amylose and amy-
lopectin) during the process of retrogra-
dation after gelatinization; type IV resis-
tant starches are (RS4) chemically modi-
fied starch; type V resistant starch (RS5)
are amylose–lipid complexes formed dur-
ing the heating treatment.[2]
Obesity is also one of the metabolic syn-
drome with high incidence in children and
adolescents over the last few decades. The
occurrence of obesity increases the risks
of diabetes mellitus,[3] which is a chronic
metabolic disease with the characteristics
of fasting and postprandial hyperglycemia
in the body, and type 2 diabetes mellitus
(T2DM) accounts for most cases of dia-
betes. Dietary fiber is able to efficiently reg-
ulate glycemic impact of foods. Higher di-
etary fiber intakes can reduce postprandial
glucose response, ameliorate insulin resis-
tance and glucose tolerance, suppress ap-
petite and modulate energy metabolism in
individuals,[4] leading to the reduced risk of
developing type 2 diabetes, obesity, cardiovascular disease, and
colorectal cancer.[5] RS is a kind of dietary fiber, which resists di-
gestion within the small intestine, reaching the large intestine
intact, and then fermented by intestinal microbiota, producing
health-beneficial materials, such as short-chain fatty acids (SC-
FAs). The gut microbiota has gained widespread concern because
of its crucial role in improving host health and preventing chronic
diseases.[6–11] The correlation between gut microbiota and glu-
cose metabolism, lipid metabolism, and insulin response were
established.[12] In addition, SCFAs have great biological signif-
icance that is associated with the prevention of gastrointestinal
disease and the regulation of metabolic disorders induced by fat
accumulation.[13] RS is considered as a prebiotic fiber in con-
trolling gut microbial ecology and modulating the production of
SCFAs.[14]
RS has been incorporated in the formulations to make func-
tional food products.[15–17] Higher levels of RS in food could re-
duce the glycemic index and retard starch digestibility. Glycemic
index (GI) is an index of the increase in postprandial blood glu-
cose, which is divided into three levels: low GI (≤55), medium
GI (56–69), and high GI (≥70).[18] High GI diet poses a threat
to body health, whereas low GI diet slows down the rate of
glucose release to the blood after digestion, which improves
glycemic control and greatly benefits the diabetic populations.
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com www.starch-journal.com

Figure 1. Overview of starch modification approaches for RS improvement. The modules connected by red line represent the dual modification; the
others connected by blue lines represent multiple modification.

Figure 2. The mechanism for the interaction between caffeic acid and starch molecules. Reproduced with permission.[36] Copyright 2018, Elsevier.

However, traditional staple foods are classified as high GI, so
more efforts have been put in reducing the GI of various prod-
ucts through the substitution of different kinds and amounts
of RS.[19] Glycemic load is a more accurate term to predict
the impact on blood glucose of different types and amounts
of carbohydrate food, which is obtained by multiplying its
GI by the carbohydrate content in the food and then divided
by 100.[20]
2. Modification Approaches for Resistant Starch
Improvement
In order to reduce the rate of native starch digestion, various
modification technologies have been developed to improve the
RS contents, such as chemical,[21–24] enzymatic[25–27] and phys-
ical modifications[28–31] (Figure 1) as well as complexation with
phenolics[32] (Figure 2). Besides, dual modification has attracted
more interests recently, which can further increase RS contents
of singly modified starches.[33–35]
2.1. Chemical Modification
Chemical modifications of starch contain etherification,
esterification, cross-linking, acetylation, oxidation, and
hydroxypropylation.[37] Organic carboxylic acids are less nu-
tritionally harmful, including malic, tartaric, citric, and glutaric
acid, which are usually applied to increase RS contents by an es-
terification reaction between organic acids and starch, leading to
the formation of the substituent groups along the 𝛼-1,4 glycosidic
chains to hinder the enzyme digestion.[38] The effects of acid
modifications on RS contents depend on many attributes, includ-
ing pH, reagent concentration, reaction duration, and degree
of substitution. Ye et al.[39] found the RS content of native rice
starch increased with the higher CA concentration. Corn starch is

Starch - Stärke 2021, 2100141 2100141 (2 of 13) © 2021 Wiley-VCH GmbH

www.advancedsciencenews.com
chemically modified in hot acetic anhydride containing dissolved
tartaric acid and RS contents increased with the reaction period
because of the increased new atypical bonds.[40] Thus, optimiza-
tion of the concentration and hydrolysis time of acid modification
is necessary to control the RS contents of starches. The RS con-
tent increased from 28.6% to 87.9% and 18.2% to 74.8% with the
decreasing pH value in malic acid-treated uncooked and cooked
corn starches, respectively.[41] The acid treated starch could resist
the enzymatic digestion and had more RS due to the formation
of ester bonds between starch molecules and malic acid resulted
from higher degree of substitution at lower pH values.[41] How-
ever, heating could destroy ester bonds, leading to lower RS con-
tents in cooked starches. The similar results were also observed
in citric acid (CA) treated potato starch.[42] A comparative study
showed that RS contents of both CA treated corn and sorghum
starches are higher than those of lactic acid (LA) modified
starches, suggesting CA had more distinct effects on lowering
the starch digestibility.[22] The cross-linking modification using
sodium trimetaphosphate and sodium tripolyphosphate in-
creases the RS content of high-amylose maize starch from 66.5%
to 84.7%, as incorporated chemical structure can hinder the ac-
cess to amylase.[23] Similar results are also observed in epichloro-
hydrin cross-linked modified pearl millet starches.[43] Both acety-
lation and oxidation modified kithul palm starches had highest
RS contents due to the introduction of acetyl groups, carbonyl,
and carboxyl groups, respectively, which can hinder the enzyme
hydrolysis.[21]
2.2. Enzymatic Modification
Enzymatic modification is a clean and accurate way to mod-
ify starch digestibility, including 𝛼-amylase, pullulanase and
branching enzymes.[25] The enhanced ordered arrangement of
semi-crystalline structure and the higher proportion of linear
short chains in enzyme modified starch are beneficial to resist
the digestive enzymatic hydrolysis, resulting in the higher RS
contents.[44,45] Pullulanase (PUL) is commonly used to debranch
the starch branched polymers and produce more short chains
by cleaving 𝛼-1, 6 glycosidic linkages in starch.[46] It is reported
that PUL treatment increased RS contents of waxy corn starch
from 13% to 19%, as a result of short linear dextrin generated
by debranching that can promote the formation of crystalline
structures, which are highly resistant to enzymatic digestion. In-
terestingly, RS content peaked when the debranching degree was
41.86% and remained stable, irrespective of higher debranching
degree.[47] Amylosucrase (AS) is a kind of glucosyltransferase,
which is involved in the synthesis of 𝛼-1, 4-glucans and cat-
alyzes the branch chain elongation at the non-reducing ends
of amylopectin.[48] The RS contents of AS modified waxy corn
starches increased from 1.2% to 19.7% with reaction time,[49]
which is attributed to the newly formed B-type crystallite
and elongated amylopectin chains.[48] Cyclodextrin glycosyl-
transferase (CGTase) is a kind of 𝛼-glucanotransferase, which
significantly increased RS contents of waxy maize starch from
8.8% to 35.7% through increasing the proportion of amylopectin
short chains (DP < 13) and producing more cyclodextrins (CDs)
with resistance to digestive enzymes.[50]
Starch - Stärke 2021, 2100141 2100141 (3 of 13)
www.starch-journal.com
2.3. Physical Modification
Physical modification is widely considered as a suitable method
for increasing the content of RS in food industry, since no harm-
ful compounds are involved during processing. Heat moisture
treatment (HMT), a safe and environmental-friendly modifica-
tion method, is widely used in altering the starch multi-level
structures and digestibility. It is a process that starch granules
are treated in a system with limited moisture content and a
temperature between the gelatinization temperature and the
glass transition temperature for a certain period. Many studies
found that RS content increased dramatically in HMT treated
adlay seed starch,[51] rice starch,[28,52,53] and maize starch.[54]
There are various factors contribute to the decreased digestion
in HMT modified starch: the formation of starch–protein com-
plexes, amylose content,[28,55,56] moisture content,[51,53] heating
temperature.[53] Annealing treatment (ANN) is one of the ther-
mal treatments and is expected to introduce interactions and
re-associations of amylose and amylopectin chains, leading to
the alternation in starch structure and improvement of digestive
property. ANN significantly increased the RS contents of foxtail
millet starch from 18.2% to 42.32%,[57] which caused a more
dense structure resistant to enzymatic digestion by reinforcing
the interactions between starch polymers chains and rearranging
the ordered structure in the semi-crystalline region. Autoclaving
treatment (AT) followed by cooling promotes hydration of the
amorphous region in starch granules in a pressure field and
increase its RS contents, which is favorable because of its sim-
plicity and safety.[58] Due to the interaction and rearrangement
of starch chain, higher RS content of cowpea starch was found
in single autoclaving-cooling cycle, while more cycles could
not further improve the RS level.[59] In addition, the suitable
conditions for elevating RS content in rice starch under AT
treatment have been achieved at 30–50% of moisture content,
pH 5–7, 40–80 min of AT treatment and 6–18 h of cooling.[60]
Microwave treatment (MT) refers to impose dielectric heating
and electromagnetic polarization on the hydroxyl groups of
starch granules, which is more efficient to change the structure
and functionality of starch.[61] Microwave heating time was
reported to play a significant role in modulating the digestive
properties of starch. Zhong et al.[62] observed that 1 min of MT
noticeably increased RS content of high amylose maize starch
because of the rearrangement of double helical segments.
Ultrasonic treatment (UT) is a non-thermal method with many
advantages, such as non-use of chemicals, less processing time,
and environment-friendly processing.[63] Ding et al.[64] found that
the level of RS in UT treated RS3 decreased at first, but then
showed an upward trend with increased ultrasonic power. The
explanation is that starch crystalline structures are disrupted un-
der low ultrasonic power, making starch granules more suscepti-
ble to digestive enzymes, whereas high ultrasonic power induces
retrogradation of RS3 and promotes the rearrangement of double
helical structures that are resistant to enzyme digestion. In con-
trast, their recent study displayed the contradict results[65] and the
corresponding reasons are absent.
Effects of different physical modifications on the improvement
of RS contents were compared. Proso millet starch was modi-
fied by HMT, AT, and MT, the RS contents were found to be
significantly increased from 11.5% to 12.3%, 15.5%, and 18.7%,
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com
respectively.[66] ANN had significantly increased the RS content
of RS3 from 46.2% to 61.5%,[67] which enhanced the amylose
chains interactions and the compactness of molecular structure.
Although UT also showed the relatively lower extent of increase
in RS content, ANN had a more pronounced influence on in-
creasing RS content. The more obvious role of ANN was also
found in foxtail millet starch.[57]
2.4. Complexation with Phenolics
Phenolic compounds, a group of natural antioxidants exist in
plant extracts, fruits, vegetables and grains, can interact with
starch to form starch-phenolic complexes with slow digestibility,
which were proposed to decrease starch digestibility by suppress-
ing the enzymatic activities or reinforcing the starch ordered
structures to reduce the accessibility of starch to enzymes.[68] In
terms of the classification of starch-polyphenol complex, it is un-
certain whether it belongs to the group of RS4 or RS5,[69] or a new
type. Recent studies have explored the effects of different pheno-
lic compounds on starch digestibility.[36,70–72] For example, gallic
acid (GA) was reported to increase the RS contents from 15.52%
to 45.93%, followed by the decreased predicted glycemic index
of GA.[70] Young apple is a rich source of polyphenols, where
the content of polyphenols is 10 times as that in fresh apples.
Young apple polyphenols (YAP) can inhibit the activities of 𝛼-
amylase and 𝛼-glucosidase, including chlorogenic acid (CHA),
phlorizin, procyanidin, catechin, epicatechin, caffeic acid, and
quercetin.[73] The addition of YAP, CHA, and phlorizin signifi-
cantly retarded the digestion of the gelatinized wheat starch, the
RS content increased from 25.23% to 47.74% in wheat starch-
YAP complex, 62.26% in wheat starch-CHA complex and 39.55%
in wheat starch-phlorizin complex.[74] Different concentrations of
caffeic acid have various effects on the RS contents starches with
different amylose content,[72,75] e.g., the RS content of waxy maize
starch-complex increased more significantly compared to the na-
tive one. The mechanisms are phenolic compounds complexed
with starch through hydrophobic interaction or hydrogen bonds,
which alter the ordered multi-scale structures and decrease the
accessibility of starch to enzymes, thereby improving RS con-
tents. Since the amount of phenolic compound is also an impor-
tant factor, optimization of phenolics concentration for the for-
mation of desired starch-phenolic complex is necessary.
2.4.1. Dual Modification
Because of the distinct effects of single modification approach
on the improvement of RS content, it is hypothesized the com-
bined treatments could have synergistic effects on lowering the
starch digestibility. The RS content of RS3 developed from dual
enzymatic hydrolysis (𝛼-amylase and pullulanase) and recrystal-
lization treated Canna edulis starch greatly increased from 5.87%
to 48.23%. RS3 with type-B crystallites have more stable double
helices with longer chains than those with A and C patterns,
making them generally more resistant to digestive enzymes.[76]
Transglucosidase (TG) can expand the branched structure by
cleaving and transferring 𝛼-1, 4-glucosidic bonds to form new
𝛼-1, 6-glucosidic bonds, and the process need assistance from
Starch - Stärke 2021, 2100141 2100141 (4 of 13)
www.starch-journal.com
𝛽-amylase (BA) in order to increase the efficiency of catalysis.[77]
The combined treatment can generate new branched chains
with more branch points and shorter chain length, which was
found to double the RS content in modified oat starches.[34] Rice
starches treated sequentially with triple enzymes (BA, TG, and
PUL) exhibited higher RS contents than PUL-modified starch
before and after cooking.[78] Multiple enzymatic treatments led
to the production of more short linear chains (DP 9–11) and
higher relative crystallinity of starch compared with individual
PUL treatment, which further reinforced the enzyme-resistant
structure. Autoclaving was combined with enzyme modifications
to obtain novel modified starches. Besides, amylopectin from
maize dually modified by PUL and AS displayed higher level
of RS than the sample individually treated with amylosucrase,
indicating the debranching treatment could further decrease
the rate of starch digestion.[79] Cyclodextrinase (CDase) belongs
to the 𝛼-amylase family, CDase and CGTase have a synergetic
effect in dual treatment system that CDs constantly generated
by CGTase, followed by hydrolyzed by CDase and then produced
malto-oligosaccharides (MOS).[80] It was reported that CDase
and CGTase increased the RS contents of waxy maize starch by
simultaneous and sequential treatments from 8.8% to 27.5% and
36.9%, respectively. While the simultaneous treatment increased
less RS than sequential treatment and the single CGTase, which
is attributed to more MOS and the higher polydispersity index
of starch hydrolysate.[50]
In terms of dual physical modification, the RS content
in microwave-ultrasound treated A- and B-starch granules in-
creased by 3.46% and 17.36%.[81] RS content in debranched
starch increased from 46.2% to 65.4% after UT and ANN.[67] The
varying degrees of increase in RS content are also associated with
the different orders of physical modifications. For example, UT
prior to ANN had a predominant effect on RS level, because of
the induced high pressure gradients and high local velocities that
damage the starch granules and cleave the long chains, followed
by ANN assemble these short chains to form highly ordered crys-
talline structure, resulting in increased RS contents.[57] Although
there is no significant difference in RS contents of UT or electric
field (ES) individually modified potato starch, ES prior to UT sig-
nificantly increased the RS content of potato starch, which might
be due to relocation between starch and digestive enzyme or the
rearrangement of ordered structure in starch granules.[33]
BA exerted a significant influence on increasing the RS con-
tent of extruded rice starch, rising from 19.89% to 48.74%. The
extrusion is a physical process that starch is subjected to high
temperature and mechanical shearing with limited water. This
pretreatment of rice starch promoted the accessibility of digestive
enzymes to starch, in turn, facilitated the increment of branched
regions in the amylopectin cluster.[82] The mode that physical
modification approaches applied in disrupting starch structures,
followed by enzymatic modification, is likely to mitigate the limi-
tation that native starch granules are slowly hydrolyzed by diges-
tive enzymes and enhance modification efficiency.[83] Dry heat-
ing could promote the flexibility and deformability of starches,
which helps ANN to strengthen the alternatively arranged lamel-
lar structure starch. However, the opposite trends displayed in the
changes of RS contents in normal maize starch and potato starch
dually modified by dry heat and ANN, which is ascribed to the dif-
ference in crystalline structure and chain-length distributions.[84]
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com
Figure 3. The regulatory networks of RS on glucose/lipid metabolism and gut microbiota based on T2DM mice model.[9,12,91] HDL-c, high-density
lipoprotein cholesterol; LDL-c, low-density lipoprotein cholesterol; TG, triglyceride; TC, total cholesterol.
Given that the pretreatment can alter the densely packed struc-
ture of native starch and create a channel for the penetration of
enzymes or chemical substances, more attention should be paid
to the effects of various pretreatment methods on increasing the
efficiency of modification.
Kithul palm starch modified by dual chemical modification
had higher RS content than single modification, as it contained
the functional groups introduced by oxidation and acetylation.[21]
A combination of CA-HMT treatment was found to increase
more RS content in both corn and sorghum starches from
87.96% to 90.19% and from 85.26% to 87.66%, respectively,[22]
which was also observed in wheat starch[85] and rice starches with
different amylose content,[38] CA esterification facilitated the for-
mation of starch citrate by substituting hydroxyl on the starch
chains under the following HMT treatment, hence further en-
hancing the resistance to digestive enzymes.[86] Likewise, various
acids (acetic, lactic, and citric acids) combined with HMT also im-
proved RS yield than individually treated by HMT,[87] indicating
the relatively more pronounced effects of acid modification on
the improvement of starch digestion.
3. Health Benefits
RS, as a dietary fiber, has specific benefits on the prevention of
diabetes, obesity, inflammation, gut health.[5,88–90] The mecha-
nisms for the anti-disease effects of RS are presented in Figure 3.
RS can alter the diversity and enrich the abundance of beneficial
bacteria in the intestine along with the production of short-chain
fatty acids. The changes in gut microbiota could influence the
expression of some glucose and lipid metabolism related genes,
Starch - Stärke 2021, 2100141 2100141 (5 of 13)
www.starch-journal.com
resulting in the reduced glucose and insulin response as well as
lipogenesis.
3.1. Regulation of the Gut Microbiota Metabolism
Gut microbiota composition and diversity have been demon-
strated to be relevant to the occurrence of many human chronic
diseases including obesity, inflammatory bowel diseases, dia-
betes types 1 and 2, and cardiovascular disease.[6] Different
types of RS, derived from various starch sources, can pro-
mote the growth of different types of gut microorganisms,
which are mainly the SCFA-producers. RS from native pea
starch (RS2) contributed mostly to decrease the ratio of Fir-
micutes/Bacteroidetes but increased Bifidobacterium (acetate pro-
ducer), whereas RS from acid-hydrolyzed and pullulanase-
debranched pea starches (RS3) had more pronounced effects on
the expansion of Faecalibacterium (butyrate producer).[92] Some
studies found that Firmicutes/Bacteroidetes ratio is associated with
obesity and diabetes,[11,76,93] while the contradicted views also
exist.[94,95] Qin, et al.[96] also observed the prebiotic properties of
RS3 on modulating the gut microbiota metabolism. They also
found that Bifidobacterium, Collinsella, Romboutsia, Dialister, and
Megamonas were significantly enriched in the RS5 groups in vitro
fecal fermentation, which are responsible for producing butyric
acid and are beneficial for gut health.
Most of the studies focused on in vitro prebiotic effects of
RS on gut microbiota, while little evidence has been found in
vivo. Both the intervention of RS2 from native lentil starch and
RS3 from autoclaved and retrograded lentil starch efficiently
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com
suppressed the effects of the alternation in gut microbial ecology
induced by high fat diet (HFD), with an obvious decreased ratio
of Firmicutes to Bacteroidetes and an increase in the relative abun-
dance of Proteobacteria, especially for RS3 with more significant
effects. A higher ratio of Firmicutes/Bacteroidetes was observed in
the obese populations, indicating the positive effects of RS on the
gastrointestinal tract health.[97] In addition, the obesity associated
microbiota Clostridium, Enterococcus, Streptococcus, and Leuconos-
toc were all significantly decreased in the RS administered group.
The Bacilli species (including Bacillales and Lactobacillales) be-
came the dominant bacteria community in the gut induced by
RS intervention, which are related to the suppression of chronic
inflammation, glucose intolerance, and lipid metabolism in
the obese mice.[93] Canna edulis RS3 (Ce-RS3) intervention
significantly alleviates hyperlipidemia in mice fed with HFD,
which was associated with increased gut microbial diversity,
enriched specific beneficial bacteria, such as SCFAs-producing
bacteria (Alloprevotella, Butyricicoccus, and Ruminiclostridium),
host-beneficial bacteria (Akkermansia and Faecalibaculum) and
reduced inflammatory bacteria (Tyzzerella, Tyzzerella_3, and
Anaerotruncus).[76] This novel RS3 also has anti-diabetic effects
on T2DM rats, which increased the abundance of Prevotella, Ru-
minococcaceae_UCG_005, Ruminococcaceae_NK4A214_group,
Phascolarctobacterium, Lachnospiraceae_UCG_010, Chris-
tensenellaceae_R_7_group, Roseburia, Helicobacter, unclassi-
fied_f_Ruminococcaceae, Ruminococcus and Coprococcus, while
decreased the abundance of Streptococcus and Bacillus.[11]
3.2. Regulation of Short-Chain Fatty Acids
The benefits of RS consumption are derived from the fermenta-
tion by gut microbiota, consequently producing the short-chain
fatty acids. SCFAs contain acetate, butyrate, propionate, which
are proposed to confer gut health benefits and potentially pre-
vent the development of colon cancer and diabetes.[98] Butyrate is
targeted as a beneficial SCFA due to its maintenance of colonic
homeostasis and anti-inflammatory properties.[99] Low acetate-
to-propionate ratio is beneficial to hinder the biosynthesis of
cholesterol and fatty acids in the liver.[100] RS has a noticeable in-
fluence on changing the ratio of SCFAs.[101,102] It is reported that
RS2 could decrease the abundance of pathogen taxa associated
with obesity, inflammation, and aging, but increase the abun-
dance of butyrate producers, resulting in higher butyric acid.[90]
The enrichment of many butyrate-producing bacterial genera, si-
multaneously cooperating with others, leading to the changes in
the metabolic pathways of gut microbiota and the increase in bu-
tyric acid production.[103] Based on mice model, the addition of
RS2 from green banana in HFD significantly increased the to-
tal amount of SCFAs, together with higher levels of acetic acid,
butyric acid, and propanoic acid in mice with obesity.[91] It was
also found that HFD supplemented with RS2 from wheat starch
increased butyric acid produced by more Ruminococcaceae and
Lachnospiraceae, but decreased isobutyric and isovaleric acids in
the cecum and colon of aged rats.[8]
A comparative study showed that both RS isolated from native
(RS2), acid-hydrolyzed and pullulanase-debranched pea starch
(RS3) significantly increased the amounts of acetate, butyrate,
propionate, and total SCFA after 24 h of in vitro fecal fermen-
Starch - Stärke 2021, 2100141 2100141 (6 of 13)
www.starch-journal.com
tation. Large number of dense aggregates in the highly ordered
structures of RS have a higher resistance to the amylolysis by gut
microbiota. More importantly, due to its structural heterogeneity,
RS3 shows better performance in producing SCFAs, so it is more
suitable for SCFAs production.[92] In addition, RS5 was reported
to produce the largest amount of acetic acid, propionic acid, and
butyric acid due to the higher expression of the genes related to
starch degradation and metabolism, followed by RS2 and RS3,
whereas RS3 produced higher amount of lactic acid than RS2 and
RS5.[96] A recent study also showed that RSs with small structural
differences had divergent and highly specific effects on the gut
microbiome to produce SCFAs. Furthermore, the fermentation
of four chemically modified starches (RS4) enhanced the over-
all production of SCFAs, but induced different changes in pH,
gas, and the degree of increment in each SCFA production.[104]
Thus, decreasing the degree of ordered structures in RS by vari-
ous modification approaches is an effective strategy to modulate
the SCFAs production.
3.3. Regulation of Glucose Metabolism
Postprandial glycaemia is proposed to be associated with chronic
diseases, such as type 2 diabetes, cardiovascular disease, and
several types of cancer.[105] Insulin resistance is one of the
obesity-related disorders, as identified by abnormal higher con-
centration of insulin which cannot maintain a normal glycemia
and sufficient glucose utilization in insulin target tissues.[106]
Enhanced activity and secretion of insulin is beneficial to alle-
viate diabetic symptoms and pancreatic repair. Many authors
highlight the health effects of RS on improving the glycemic con-
trol and insulin sensitivity.[8,12,107–110] VERSAFIBE 1490 resistant
starch (RS4), a distarch phosphate derived from potato, contains
phosphodiester cross-links in the distarch phosphate molecules
that reduce swelling and enzyme accessibility, which was found
to have hyperglycemia effects. A group of healthy adults con-
sumed high fiber cookies containing RS4 had the postprandial
intravenous blood and capillary glucose as well as the maximum
glucose concentration during a period of two hours, resulting in
the significantly decreased postprandial serum insulin concen-
tration compared with the panel consumed control cookies.[111]
Although the perspectives about the role of gender and baseline
insulin sensitivity in the effects of RS on insulin sensitivity were
not universal, these two factors should also be considered in the
analysis of glucose metabolism.[107] The following studies fur-
ther confirmed the role of distarch phosphate in attenuating the
postprandial glycemia in muffin[110] and breakfast cereal bars.[19]
Some animal studies also demonstrated the health benefits
of RS in some chronic diseases, including obesity, type 2 dia-
betes, and inflammation. The anti-diabetes effects of different
RSs (RS2, RS3) have been previously reported.[11,91,112,113] RS en-
capsulated with Ganoderma lucidum spores (EGLS) exerted hy-
poglycemic effects separately on T2DM rats. During the 28 days
experimental period, the blood glucose level in the EGLS treated
T2DM rats was constantly lower than the control and decreased
by 21.9% from the third week. In addition, the expressions of glu-
coneogenesis regulation-related genes (G6PC1) were downregu-
lated, while glycogen synthesis-related genes (GS2 and GYG1)
and insulin-induced genes (Insig1 and Insig2) were upregulated.
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com
Glucose metabolism consists of glycogen synthesis and gluco-
neogenesis. It is suggested that the hypoglycemic effects of EGLS
might be associated with the limited insulin secretion and the
enhancement of the glucose homeostasis by inhibiting glycogen
synthesis and hepatic glucose production. Thus, EGLS could be
considered as a valuable dietary ingredient to alleviate diabetes.[9]
The anti-obesity effects of RS2 from native lentil starch and
RS3 from autoclaved and retrograded lentil starch on mice with
HFD-induced obesity were compared. Following 6 consecutive
weeks, the obese mice in the two RS administered groups dis-
played decreased blood glucose after 6 weeks of treatment. Com-
pared with the RS2 intervention, the RS3 treatment had more
remarkable effects in reducing blood glucose along with control-
ling body weight because of its resistance structure with higher
crystallinity and double helix content.[93] Future studies can uti-
lize prebiotic properties of RS from modified starch to modulate
specific intestinal flora to improve health.
The benefits of oat RS intervention for rats with HDF induced
type 2 diabetes were also investigated. After 9 weeks of adminis-
tration, oat RS significantly decreased the level of fasting blood
glucose, glucose concentration, insulin resistance and glycated
haemoglobin, while promoted the proliferation and function of
pancreatic 𝛽-cells, indicating RS had anti-diabetic effects.[12] It
also had anti-inflammatory effects with the inhibition of cytokine
release in blood, together with the suppressed mRNA expres-
sion of TNF-𝛼, IL-6, and IL-1𝛽.[12] Proinflammatory cytokines are
mainly secreted from adipose tissues associated with the regu-
lation of glucose metabolism.[114] The changes in metabolic re-
sponses were found to be controlled by gut microbiota. Oat RS
increased genus Clostridium and Butyricicoccus and decreased
genus Bacteroides, Lactobacillus, Oscillospira and Ruminococcus.
Among them, genus Clostridium was negatively but genus Bac-
teroides, Lactobacillus, Oscillospira and Ruminococcus and Butyric-
occus were positively correlated with the parameters of diabetes
and inflammation.[12]
3.4. Regulation of Lipid Metabolism
Serum lipid compositions include triglyceride (TG), total choles-
terol (TC), high-density lipoprotein cholesterol (HDL-c), and low-
density lipoprotein cholesterol (LDL-c). Obesity is associated with
the dysregulation of lipid metabolism increases serum triglyc-
erides, hence reducing HDL-c level.[115] The analysis of lipid
metabolism of obese mice induced by HFD showed that RS
supplementation decreased the levels of TG, TC, LDL-c, and in-
creased HDL-c to the levels that were comparable to the control
group after 6 weeks, indicating RS intervention could attenuate
hyperlipidemia in the obese mice.[93] Rosado et al.[91] found that
the addition of 15% green banana RS in HFD reduced the accu-
mulation of hepatic steatosis, TG and cholesterol, also increased
the protein level expression of AMPK phosphorylation, ABCG5
and ABCG8 but decreased that of HMGCoA-R and FAS. AMPK,
HMGCoA-R, and FAS are involved in lipid synthesis, ABCG5 and
ABCG8 are sterol transporters in liver responsible for inhibiting
deposition of dietary sterols, indicating that green banana RS re-
duced the risk of liver lipogenesis through downregulating the
protein levels involved in lipid synthesis but strengthening the
expression of sterol transport proteins related to lipid excretion.
Starch - Stärke 2021, 2100141 2100141 (7 of 13)
www.starch-journal.com
Previous studies based on HFD-induced hyperlipidemic mice
model, showed that the RS from Canna edulis,[76] buckwheat,[116]
and sorghum[117] displayed anti-hyperlipidemic activities.
Likewise, EGLS was reported to have similar effects on modu-
lating the lipid metabolisms in T2DM rats by suppressing the ex-
pression of lipogenesis associated genes, such as ACC, FAS, Acly,
and Fads1. In addition, Acox1 was significantly upregulated, to-
gether with the upregulation of Insig1 and Insig2 in EGLS-treated
rats. Acox1 is the rate-limiting enzyme of lipid degradation, Insig1
and Insig2 play an important role in the regulation of intracellular
cholesterol, thus the anti-hyperlipidemic effect of EGLS was re-
lated to the promotion of lipid oxidation and the maintenance of
cholesterol homeostasis, which might be related to the improved
composition of fecal microbial community.[9]
4. Food Application
RS is considered as a potential functional ingredient in making
fiber-rich food products.[118] Compared with other dietary fibers,
RS has little influence on the appearance, texture, and sensory
properties of the end-products.[119] This review is an update of the
previous reviews about the food applications of RS,[120,121] includ-
ing bread, noodle, biscuit, and pasta. The changes in nutritional,
textural, and sensory properties induced by the incorporation of
different RSs in various food products are summarized in Table 1.
4.1. Biscuit
Biscuits are convenient food with a long shelf life, mainly com-
prise flour, fats and sugar, which belong to medium to high GI
categories.[132] Despite the importance of sugar in textural forma-
tion of biscuits, excessive intake of sugar can trigger the risk of
type 2 diabetes. Different types of RS can have various influences
on the quality of end-products. A comparative study about the ef-
fect of Hylon VII (RS2), Novelose330 (RS3), Fibersym (RS4) as
well as cross-linked corn or wheat starches (RS4) supplementa-
tion on the cookie quality showed that more than half supplemen-
tation with cross-linked wheat starch resulted in the improved
cookie quality and had no adverse effects on the textural proper-
ties of cookies, despite other RSs provoked the deterioration in
cookie quality. All the RSs increased the total dietary fiber and
decreased the GI of cookies, and those supplemented with cross-
linked wheat starch had the largest values of total dietary fiber
(16–32%) and the lowest GI (77.2), suggesting that this novel
ingredient can enhance the nutritional properties without com-
promising with quality loss.[17] Compared with gluten-containing
foods, gluten free (GF) foods have lower dietary fiber content,
higher GI, and total fat content.[133] The improved nutritional
properties of GF biscuits enriched with RS derived from ANN
modified white sorghum starch were reported, the content of to-
tal dietary fiber increased along with the starch hydrolysis index
decreased with higher concentration of RS. Unfortunately, the
RS substitution increased the hardness of biscuits and decreased
the scores for all the sensory attributes. The sensory test was
conducted by using a nine-point hedonic scale, where 9 = like
extremely and 1 = dislike extremely, and the scores were more
than five for the overall acceptability, suggesting the RS-enriched
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com www.starch-journal.com

Table 1. Application of RS as a functional ingredient in various food products.

Food products Substitution Nutritional properties Textural properties Sensory properties References
percentage/Ingredients

Cookie 0%, 25%, 50%, and 75% RS4 Increased total dietary fiber content, Decreased hardness – [17]
from cross-linked wheat and decreased in vitro GI
corn starches
0%, 25%, 50%, and 75% RS2 Increased total dietary fiber content, Decreased hardness –
(Hylon VII) decreased in vitro GI
0%, 25%, 50%, and 75% RS3 Increased total dietary fiber content, Increased hardness –
(Novelose330) decreased in vitro GI
0%, 25%, 50%, and 75% RS4 Increased total dietary fiber content, Decreased hardness –
(Fibersym) decreased in vitro GI
Gluten-free 0%, 15%, 30%, and 45% RS Increased total dietary fiber content and Increased hardness Lower scores for [122]
biscuit from annealed white RS content, decreased starch appearance, texture,
sorghum starch hydrolysis index flavor, taste and
overall acceptability
Gluten-free 0%, 4.4%, 8.8% RS2 – Decreased hardness and – [123]
biscuit (Hi-Maize260) fracturability
Noodle 1%, 5%, and 10% RS3 from Increase SDS and RS levels Decreased tensile strength, – [124]
HMT modified waxy, normal extensibility, hardness
and high-amylose maize and chewiness, different
starches changes in adhesiveness

Food products Substitution Health benefits Textural properties Sensory properties References
percentage/Ingredients

Noodle 20%, 40%, and 60% RS3 from
wheat starch
20%, 40%, and 60% RS3 from
corn starch
Noodle 10%, 20%, 30%, and 40% RS4
(barley starch citrate)
White salted 10%, 20%, 30%, 40%, and 50%
noodle RS4 from cross-linked
phosphorylated wheat starch
White salted 4.8%, 9%, 13%, and 15.3%
noodle unripe banana flour
Bread 15%, 20%, and 25%
phosphorylated cross-linked
RS4
– Decreased hardness Higher scores for [125]
appearance and
taste, lower scores
for thickness
–
– – Lower scores for [126]
transparency,
firmness,
slipperiness, and
tooth packing
– Decreased tensile strength, – [16]
extensibility,
cohesiveness and
springiness
Increased RS content Decreased hardness, Increased scores for [127]
cohesiveness, shearing color and roughness,
force, and tensile decreased scores for
strength firmness and
elasticity
Increased total dietary fiber content, Increased firmness – [15]
values of bile acid-binding capacity
and mineral bioavailability,
decreased GI

Food products Substitution Health benefits Textural properties Sensory properties References
percentage/Ingredients

Bread 20% RS4 from wheat starch Increased ratio of lactic – – [7]
bacteria/enterobacteria, daily intake
and apparent absorption of calcium,
total bone mineral content, bone
mineral density and bone weight
(Continued)

Starch - Stärke 2021, 2100141 2100141 (8 of 13) © 2021 Wiley-VCH GmbH

www.advancedsciencenews.com www.starch-journal.com

Table 1. (Continued).

Food products Substitution Health benefits Textural properties Sensory properties References
percentage/Ingredients

Gluten-free 7% RS4 from phosphorylated – Increased hardness, Increased bitterness, [128]

bread corn starch chewiness, cohesiveness firmness, chewiness
and gumminess and dryness
Pasta 0%, 5%, and 10% RS4 from Increased total dietary fiber content, RS Decreased firmness – [129]
phosphorylated cross-linked content, antioxidant activity and P
wheat starch content, decreased GI, K, and Mg
content
Spaghetti 15%, 20%, and 25% RS4 Increased total dietary fiber content, Increased stickiness and Decreased hardness, [130]
bile acid binding capacity and decreased firmness adhesiveness and
mineral bioavailability, decreased GI bulkiness

Food products Substitution Health benefits Textural properties Sensory properties References
percentage/Ingredients

Gluten-free 0%, 5%, 10%, 15% RS from Increased total dietary fiber content and Increased firmness Increased scores for [131]
pasta annealed white sorghum RS content, decreased starch decreased stickiness texture, decreased
starch hydrolysis index scores for
appearance and
overall acceptance

biscuits were still well-acceptable.[122] The opposite results were
shown in the changes in the physical properties of GF biscuits
prepared by the replacement of a blend of buckwheat, sorghum,
and lentil flours with RS2 (Hi-Maize260). The biscuits had de-
creased hardness and fracturability due to the lower protein con-
tent caused by RS replacement, and the soft and crispy texture
were preferred by consumers.[123] Additionally, sucrose replace-
ment is an efficient way to reduce the sugar level in biscuits but
has a detrimental influence on textural properties. RS2 substitu-
tion was demonstrated to successfully remedy the problems of
inferior quality.[123]
4.2. Noodle
Noodles are one of the main traditional wheat-based foods con-
sumed in many Asian countries. In order to increase the health
benefits, traditional wheat flour is replaced by RS to manufacture
novel healthy noodles. HMT modified maize starches (RS3) with
different amylose content were added in wheat flour, which in-
creased the peak viscosity but decreased the setback of the blend,
also weakened the strength of gluten network, leading to the in-
ferior tensile properties with lower tensile strength and extensi-
bility and softer texture of final noodles with lower hardness and
chewiness. Interestingly, the discrepancy in adhesiveness of noo-
dles depends on different addition ratios and amylose content
of maize starches, which is attributed to the different abilities
of amylose and amylopectin leaching.[124] The effects of wheat
flour replaced with 20%, 40%, and 60% of wheat resistant starch
and corn resistant starch (RS3) on noodle quality was investi-
gated. The hardness of noodle showed a noticeable decreasing
trend with the higher levels of RS, which is attributed to the
lower gluten content in wheat flour supplemented with RS. Al-
though wheat flour replaced by 40% RS achieved the highest
acceptability scores, noodles with 60% wheat RS and corn RS
achieved the lowest grade, indicating that excessive addition of
RS could exert adverse effects on the quality of food products.[125]
Future studies should focus on the optimization of RS concen-
trations added in food formulations to improve their sensory
properties.
RS4 was reported to adversely affect the textural and sensory
properties of noodle because of its physical properties. Starch
noodle made from the wheat starch replaced by different con-
centrations of barley starch citrate (RS4) exhibited lower val-
ues of sensory parameters (transparency, firmness, slipperiness,
and tooth packing). Additionally, the researcher suggested noo-
dle incorporated with 20% of barley starch citrate could have de-
sired quality.[126] Higher substitution of hard red winter wheat
flour with RS4 (cross-linked phosphorylated wheat starch) led
to the lower values of extensibility, cohesiveness, and springi-
ness but had no significant effect on hardness in cooked white
salt noodles. Also, the supplementation of 2–8% of vital wheat
proteins can merely compensate for part of textural deficiency,
which did not noticeably affect the cohesiveness and springi-
ness of cooked noodles.[16] The quality of cooked noodles is in-
fluenced by the degree of starch gelatinization and the forma-
tion of a protein network from wheat flours, while the gluten
networks was weakened by of RS4 with lower swelling power,
resulting in the deteriorative quality of noodles. Unripe banana
flour is a rich source of RS content, Liu et al.[127] found that its
addition significantly enhanced the RS contents in white salted
noodles. The incorporation also reduced the hardness, cohesive-
ness, shearing force, and tensile strength of white salted noodles,
which is ascribed by weakened gluten strength of flour and un-
gelatinized starches induced by the incorporation of unripe ba-
nana flour. In addition, it also enhanced color and roughness
of noodle, while decreased the scores for firmness and elastic-
ity, which could be complemented by adding an optimal amount
of wheat gluten flour (2.5–4.7%) so as to produce superior quality
of noodles.[127]

Starch - Stärke 2021, 2100141 2100141 (9 of 13) © 2021 Wiley-VCH GmbH

www.advancedsciencenews.com
4.3. Bread
Bread is one of the staple foods consumed in Western countries,
which is a high GI food because the conditions of bread baking is
suitable for complete starch gelatinization.[134] Given that RS is
beneficial to decrease postprandial glycaemia, its incorporation
in bread is a common strategy to develop products with low GI.
Aribas et al.[15] found the increased substitution of wheat flour
with RS4 (phosphorylated cross-linked) enhanced the nutritional
properties of bread by increasing the amount of total dietary fiber,
bile acid-binding capacity, mineral bioavailability values and de-
creasing GI. Bile acid binding is one of the most important char-
acteristics of dietary fiber. RS binds to bile acids to hinder their re-
absorption, hence removing them from the body, which is consid-
ered as cholesterol-lowering effect.[135] They suggested the bread
samples supplemented with more than 20% RS4 can be classified
as medium or low GI food. The consumption of bread containing
RS4 was reported to improve the calcium absorption and bone
strength of rats, which were strongly correlated with the better
intestinal balance (higher lactobacillus/enterobacteria ratio).[7] In
terms of the physical properties of bread, the supplementation of
RS4 largely compensated for the deficiency in textural and sen-
sory properties caused by bran, as evidence by the breads supple-
mented with higher proportion of RS4 had greater loaf volume,
brighter color, and higher firmness. According to the storage ex-
periment, the shelf-life of bread was also found to be extended
by RS4, which is attributed to the highly cross-linked structure of
RS4 can decrease the mobility of starch molecules and substan-
tially reduce the rate of retrogradation, hence delaying bread stal-
ing. In contrast, Arp et al.[136] reported the quality loss in bread
doughs incorporated with RS2, then they used two modified cel-
luloses (MC) as additives to improve the rheological properties
of bread dough enriched with 30% RS2 (Hi-Maize260).[137] The
elasticity of bread dough decreased with the increased levels of
two MCs, suggesting the addition of MC softened the dough. The
researchers supposed that improved dough viscoelasticity was as-
cribed to the hydrocolloid–protein interaction but the underlying
mechanism is unclear.
For GF bread with low fiber contents, the fortification of GF
bread with RS4 (phosphorylated corn starch) can improve its nu-
tritional properties, but it had a detrimental effect on the textu-
ral and sensory properties of final product, which was relatively
drier, harder, more cohesive and chewable due to more ordered
structure of starch matrix.[128] In the purpose of increasing the
consumer acceptance of healthy bread, the effects of two veg-
etable proteins (hemp protein and soy protein) on the GF bread
dough with RS2 were compared, doughs with exceed 9% hemp
protein displayed the decreased dough consistency that was close
to the rheological properties of wheat flour dough, and the yield
loaves with higher peak higher indicated the improved texture.
Thus, hemp protein can be considered as a functional ingredient
in making GF bread with RS.[138]
4.4. Pasta
Pasta is one of the prevalently consumed wheat flour-based food
in the western countries since the ancient times.[139,140] Owing
to the low levels of nutrients (dietary fiber, mineral, vitamin, and
Starch - Stärke 2021, 2100141 2100141 (10 of 13)
www.starch-journal.com
phenolics) in pasta, recent studies attempted to improve the nu-
tritional properties of pasta with incorporation of RS. The addi-
tion of RS4 (phosphorylated cross-linked wheat starch) in pasta
decreased its firmness, which is possibly because RS4 increased
the content of dietary fiber and disrupted the protein-starch ma-
trix in pasta.[129] Besides, it was found to achieve higher bile acid
binding capacity and mineral bioavailability but lower GI com-
pared with the control in spaghetti.[130] Although the RS4 can
reduce GI, it exerted no adverse effects on sensory properties
due to its mild flavor. The RS4 supplementation with different
levels improved texture with increased stickiness and decreased
firmness, and sensory properties with decreased hardness, adhe-
siveness and bulkiness of spaghetti due to the disrupted protein
matrix.[130]
GF pasta prepared by rice flour substituted with different lev-
els of RS from annealed white sorghum starch had higher total
dietary fiber, RS contents, and lower starch hydrolysis index with
increasing levels of RS. In terms of textural properties, the values
of firmness increased but those of stickiness decreased, springi-
ness was only improved by the highest level of RS, which was still
not comparable to wheat flour-based pasta. In the sensory test,
color, aroma, or taste did not change significant, the scores for
texture attribute increased. Although appearance and the overall
acceptability scores distinctly decreased, they were above the min-
imal acceptable value.[131] In another study, the differences be-
tween RS2 (Hi-Maize 260) and unripe plantain flour with 50–70%
RS in improving the nutritional properties of GF spaghetti were
compared. RS2 increased more total dietary fibers than unripe
plantain flour in uncooked spaghetti, while made similar per-
formances in lowering GI, consequently transferring spaghetti
into medium GI food.[141] It is strongly demonstrated that unripe
plantain flour is an alternative dietary fiber to RS, and the similar
functions were also found in chickpea,[142] cassava and banana
flour.[143]
5. Conclusion
Recent studies explored the effects of various modification ap-
proaches used in starches from different origins for the improve-
ment of RS contents and starch digestibility, mainly including
chemical, enzymatic, physical modification, and complexation
with phenolics. Compared with single modification, dual mod-
ification shows more pronounced effects on increasing the RS
contents. The effects of different combination of modification
treatments on the functionality of various starch sources have
been understood, but most studies neglected the changes in
starch digestion, which should be further uncovered. In addi-
tion, health benefits of RS have been increasingly acknowledged
through analyzing the diversity and composition of gut micro-
biota both in vitro and in vivo fermentation based on human and
mice models and the production of different SCFAs. Besides, RS
can also mitigate the adverse effects of HFD on the glucose and
lipid metabolism, the molecular mechanisms and the relation-
ship with gut microbiota are presented. The increased abundance
of beneficial bacteria positively affected the hepatic metabolism,
leading to the positive effects on diabetes, inflammation, hy-
perglycemia, and hyperlipidemia. However, many studies only
focused on the changes of RS in gut microbiota or metabolic
products, more comprehensive mechanisms of the role of RS
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com
in metabolism pathway should be further elucidated. Moreover,
the intervention of RSs with different addition ratios, physico-
chemical properties and molecular structures exerts various ef-
fects on the texture and sensory properties of food products, thus
it is necessary to optimize the concentration of RS added in the
formulations of food products based on various flour, such as
rice flour, chickpea flour, and green banana flour. Apart from the
wheat-flour based products, rice flour can also be used to produce
RS-fortified foods such as rice noodle and rice bread. Due to a
consensus that nutritional properties are commonly improved in
these functional foods, the specific changes in gut microbiota and
the amounts of metabolites are lack in most studies. Overall, the
consumption of various RS enriched healthy foods is expected to
address the high incidence of chronic diseases.
Acknowledgements
This work was financially supported by the Natural Science Foundation
of Zhejiang Province (LZ21C130003) and Fanxing Science Foundation of
Zhejiang University.
Conflict of Interest
The authors declare no conflicts of interest.
Keywords
food applications, gut microbiota, resistant starches, starch modification
Received: June 6, 2021
Revised: July 8, 2021
Published online:
[1] H. N. Englyst, S. M. Kingman, J. H. Cummings, Eur. J. Clin. Nutr.
1992, 46, 33.
[2] F. Jiang, C. Du, W. Jiang, L. Wang, S.-k. Du, Int. J. Biol. Macromol.
2020, 150, 1155.
[3] S. T. Nyberg, G. D. Batty, J. Pentti, M. Virtanen, L. Alfredsson, E.
I. Fransson, M. Goldberg, K. Heikkila, M. Jokela, A. Knutsson, M.
Koskenvuo, T. Lallukka, C. Leineweber, J. V. Lindbohm, I. E. H. Mad-
sen, L. L. M. Hanson, M. Nordin, T. Oksanen, O. Pietilainen, O.
Rahkonen, R. Rugulies, M. J. Shipley, S. Stenholm, S. Suominen, T.
Theorell, J. Vahtera, P. J. M. Westerholm, H. Westerlund, M. Zins, M.
Hamer, A. Singh-Manoux, J. A. Bell, J. E. Ferrie, M. Kivimaki, Lancet
Public Health 2018, 3, e490.
[4] M. Champ, A. M. Langkilde, F. Brouns, B. Kettlitz, Y. L. Collet, Nutr.
Res. Rev. 2003, 16, 71.
[5] S. Lockyer, A. P. Nugent, Nutr. Bull. 2017, 42, 10.
[6] Z. A. Bendiks, K. E. B. Knudsen, M. J. Keenan, M. L. Marco, Nutr.
Res. 2020, 77, 12.
[7] M. J. Correa, L. Giannuzzi, A. R. Weisstaub, A. Zuleta, C. Ferrero, Int.
J. Food Sci. Technol. 2020, 55, 239.
[8] R. L. Hughes, W. H. Horn, P. Finnegan, J. W. Newman, M. L. Marco,
N. L. Keim, M. E. Kable, Nutrients 2021, 13, 645.
[9] Y. Jiang, N. Zhang, Y. Zhou, Z. Zhou, Y. Bai, P. Strappe, C. Blanchard,
Food Sci. Biotechnol. 2021, 30, 755.
[10] E. S. Lee, E. J. Song, Y. D. Nam, T. G. Nam, H. J. Kim, B. H. Lee, M.
J. Seo, D. H. Seo, Int. J. Biol. Macromol. 2020, 145, 235.
Starch - Stärke 2021, 2100141 2100141 (11 of 13)
www.starch-journal.com
[11] C. Zhang, S. Ma, J. Wu, L. Luo, S. Qiao, R. Li, W. Xu, N. Wang,
B. Zhao, X. Wang, Y. Zhang, X. Wang, Pharmacol. Res. 2020, 159,
104985.
[12] Y. Zhu, L. Dong, L. Huang, Z. Shi, J. Dong, Y. Yao, R. Shen, J. Funct.
Foods. 2020, 69, 103939.
[13] S. A. Zaman, S. R. Sarbini, Crit. Rev. Biotechnol. 2016, 36, 578.
[14] E. Fuentes-Zaragoza, E. Sanchez-Zapata, E. Sendra, E. Sayas, C.
Navarro, J. Fernandez-Lopez, J. A. Perez-Alvarez, Starch/Stärke
2011, 63, 406.
[15] M. Aribas, K. Kahraman, H. Koksel, Cereal Chem. 2020, 97, 163.
[16] C. Hsieh, L. Wang, B. Xu, P. A. Seib, Y. Shi, J. Sci. Food Agric. 2020,
100, 5334.
[17] K. Kahraman, E. Aktas-Akyildiz, S. Ozturk, H. Koksel, J. Cereal Sci.
2019, 90, 102851.
[18] S. W. Rizkalla, F. Bellisle, G. Slama, Br. J. Nutr. 2002, 88, 255.
[19] Y. J. Du, Y. H. Wu, D. Xiao, G. Guzman, M. L. Stewart, V. Gourineni,
B. Burton-Freeman, I. Edirisinghe, Food Funct. 2020, 11, 2231.
[20] T. Penlioglou, V. Lambadiari, N. Papanas, Nutrition 2021, 89,
111234.
[21] C. Sudheesh, K. V. Sunooj, J. George, Int. J. Biol. Macromol. 2019,
125, 1084.
[22] F. Shaikh, T. M. Ali, G. Mustafa, A. Hasnain, Int. J. Biol. Macromol.
2019, 135, 314.
[23] S. Wang, B. Zhang, T. Chen, C. Li, X. Fu, Q. Huang, J. Agric. Food
Chem. 2019, 67, 13728.
[24] B. W. Wolf, L. L. Bauer, G. C. Fahey, J. Agric. Food Chem. 1999, 47,
4178.
[25] L. Cai, Y.-C. Shi, Carbohydr. Polym. 2010, 79, 1117.
[26] M. M. Kasprzak, H. N. Laerke, F. H. Larsen, K. E. B. Knudsen, S.
Pedersen, A. S. Jorgensen, Int. J. Mol. Sci. 2012, 13, 929.
[27] H. Jiang, M. Miao, F. Ye, B. Jiang, T. Zhang, Int. J. Biol. Macromol.
2014, 65, 208.
[28] Z. Sui, T. Yao, X. Ye, J. Bao, X. Kong, Y. Wu, Starch/Stärke 2017, 69,
1600164.
[29] L. Jayakody, R. Hoover, Carbohydr. Polym. 2008, 74, 691.
[30] J. Jin, H. Lin, A. E. A. Yagoub, S. Xiong, L. Xu, C. C. Udenigwe, J. Sci.
Food Agric. 2020, 100, 3498.
[31] N. Li, L. Wang, S. Zhao, D. Qiao, C. Jia, M. Niu, Q. Lin, B. Zhang,
Food Hydrocoll. 2020, 103, 105690.
[32] T. Xu, X. Li, S. Ji, Y. Zhong, J. Simal-Gandara, E. Capanoglu, J. Xiao,
B. Lu, Trends Food Sci. Technol. 2021, 111, 12.
[33] M. Cao, Q. Gao, Int. J. Biol. Macromol. 2020, 150, 637.
[34] A. Shah, F. A. Masoodi, A. Gani, B. Ashwar, Int. J. Biol. Macromol.
2018, 106, 140.
[35] A. O. Ashogbon, Food Chem. 2021, 342, 128325.
[36] M. Li, C. Pernell, M. G. Ferruzzi, Food Hydrocoll. 2018, 77, 843.
[37] Y. J. Wang, L. F. Wang, Carbohydr. Polym. 2003, 52, 207.
[38] H. Pham Van, V. Ngo Lam, P. Nguyen Thi Lan, Food Chem. 2016,
191, 67.
[39] J. Ye, S. Luo, A. Huang, J. Chen, C. Liu, D. J. McClements, Food Hy-
drocoll. 2019, 92, 135.
[40] M. V. Tupa, L. Altuna, M. L. Herrera, M. L. Foresti, Starch/Stärke
2020, 72, 1900300.
[41] C. J. Lee, J. H. Na, J.-Y. Park, P. S. Chang, Molecules 2019, 24, 10.
[42] S. Y. Lee, K. Y. Lee, H. G. Lee, Int. J. Biol. Macromol. 2018, 107, 1235.
[43] A. K. Siroha, K. S. Sandhu, Int. J. Food Prop. 2018, 21, 1371.
[44] A. Evans, D. B. Thompson, Cereal Chem. 2004, 81, 31.
[45] R. Hoover, Y. Zhou, Carbohydr. Polym. 2003, 54, 401.
[46] P. Rudeekulthamrong, K. Sawasdee, J. Kaulpiboon, Biotechnol. Bio-
process Eng. 2013, 18, 778.
[47] W. Liu, Y. Hong, Z. Gu, L. Cheng, Z. Li, C. Li, Food Hydrocoll. 2017,
67, 104.
[48] A. Rolland-Sabaté, P. Colonna, G. Potocki-Véronèse, P. Monsan, V.
Planchot, J. Cereal Sci. 2004, 40, 17.
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com
[49] H. Zhang, X. Zhou, J. He, T. Wang, X. Luo, L. Wang, R. Wang, Z.
Chen, Food Chem. 2017, 220, 413.
[50] H. Ji, X. Li, Y. Bai, Y. Shen, Z. Jin, Int. J. Biol. Macromol. 2021, 180,
187.
[51] H. Wang, J. Ding, N. Xiao, X. Liu, Y. Zhang, H. Zhang, Food Chem.
2020, 318, 126489.
[52] H. Wang, Y. Liu, L. Chen, X. Li, J. Wang, F. Xie, Food Chem. 2018, 242,
323.
[53] H. Pham Van, V. T. Binh, P. H. Y. Nhi, P. Nguyen Thi Lan, Int. J. Biol.
Macromol. 2020, 154, 1.
[54] Y. Chen, Q. Yang, X. Xu, L. Qi, Z. Dong, Z. Luo, X. Lu, X. Peng, Car-
bohydr. Polym. 2017, 177, 232.
[55] X. Yang, C. Chi, X. Liu, Y. Zhang, H. Zhang, H. Wang, Int. J. Biol.
Macromol. 2019, 139, 785.
[56] H. Xie, J. Gao, X. Xiong, Q. Gao, Food Hydrocoll. 2018, 83, 213.
[57] A. S. Babu, R. J. Mohan, R. Parimalavalli, Food Chem. 2019, 271, 457.
[58] L. Bravo, P. Siddhuraju, F. Saura-Calixto, J. Agric. Food Chem. 1998,
46, 4667.
[59] N. Ratnaningsih, Suparmo, E. Harmayani, Y. Marsono, Int. J. Biol.
Macromol. 2020, 142, 191.
[60] Y. Zheng, Z. Wei, R. Zhang, Y. Deng, X. Tang, Y. Zhang, G. Liu, L. Liu,
J. Wang, N. Liao, M. Zhang, Food Sci Nutr 2020, 8, 2383.
[61] C. Bilbao-Sáinz, M. Butler, T. Weaver, J. Bent, Carbohydr. Polym.
2007, 69, 224.
[62] Y. Zhong, W. Liang, H. Pu, A. Blennow, X. Liu, D. Guo, Int. J. Biol.
Macromol. 2019, 137, 870.
[63] Y. Monroy, S. Rivero, M. A. García, Ultrason. Sonochem. 2018, 42,
795.
[64] Y. Ding, F. Luo, Q. Lin, Food Chem. 2019, 294, 248.
[65] Y. Ding, Y. Liang, F. Luo, Q. Ouyang, Q. Lin, Carbohydr. Polym. 2020,
228, 115350.
[66] M. Zheng, Y. Xiao, S. Yang, H. Liu, M. Liu, S. Yaqoob, X. Xu, J. Liu,
Food Sci. Nutr. 2020, 8, 735.
[67] R. Chang, H. Lu, X. Bian, Y. Tian, Z. Jin, Food Hydrocoll. 2021, 110,
106141.
[68] C. Chi, X. Li, Y. Zhang, L. Chen, L. Li, Z. Wang, Food Funct. 2017, 8,
720.
[69] T. J. Gutierrez, J. Tovar, Trends Food Sci. Technol. 2021, 109, 711.
[70] C. Chi, X. Li, Y. Zhang, L. Chen, F. Xie, L. Li, G. Bai, Food Hydrocoll.
2019, 89, 821.
[71] J. H. Dupuis, R. Tsao, R. Y. Yada, Q. Liu, LWT – Food Sci. Technol.
2017, 85, 218.
[72] M. Yu, B. Liu, F. Zhong, Q. Wan, S. Zhu, D. Huang, Y. Li, Food Hy-
drocoll 2021, 114, 106544.
[73] D. Li, L. Sun, Y. Yang, Z. Wang, X. Yang, T. Zhao, T. Gong, L. Zou, Y.
Guo, J. Funct. Foods. 2019, 56, 127.
[74] D. Li, Y. L. Yang, X. Yang, X. Y. Wang, C. Guo, L. J. Sun, Y. R. Guo,
Food Funct. 2021, 12, 1983.
[75] M. Han, W. Bao, Y. Wu, J. Ouyang, Int. J. Biol. Macromol. 2020, 162,
922.
[76] C. Zhang, M. Qiu, T. Wang, L. Luo, W. Xu, J. Wu, F. Zhao, K. Liu, Y.
Zhang, X. Wang, Food Chem. 2021, 351, 129340.
[77] H. Li, Y. Gui, J. Li, Y. Zhu, B. Cui, L. Guo, Int. J. Biol. Macromol. 2020,
144, 500.
[78] H. Li, J. Li, Y. Xiao, B. Cui, Y. Fang, L. Guo, Int. J. Biol. Macromol.
2019, 136, 1228.
[79] H. Zhang, R. Wang, Z. Chen, Q. Zhong, Food Hydrocoll. 2019, 95,
195.
[80] H. Ji, Y. Bai, X. Li, D. Zheng, Y. Shen, Z. Jin, Carbohydr. Polym. 2020,
237, 116137.
[81] K. Zhang, D. Zhao, D. Guo, X. Tong, Y. Zhang, L. Wang, Int. J. Biol.
Macromol. 2021, 183, 481.
[82] B. E. Hickman, S. Janaswamy, Y. Yao, J. Agric. Food Chem. 2009, 57,
7005.
Starch - Stärke 2021, 2100141 2100141 (12 of 13)
www.starch-journal.com
[83] J. Ye, C. Liu, S. Luo, X. Hu, D. J. McClements, Food Res. Int. 2018,
111, 590.
[84] C. Chi, X. Li, P. Lu, S. Miao, Y. Zhang, L. Chen, Int. J. Biol. Macromol.
2019, 137, 554.
[85] M. Li, Y. Xie, H. Chen, B. Zhang, Food Chem. 2019, 272, 523.
[86] E. Olsson, C. Menzel, C. Johansson, R. Andersson, K. Koch, L. Järn-
ström, Carbohydr. Polym. 2013, 98, 1505.
[87] L. d. O. Maior, V. S. de Almeida, B. R. V. Barretti, V. C. Ito, C. Ben-
inca, I. M. Demiate, E. Schnitzler, M. A. D. S. Carvalho Filho, L. G.
Lacerda, J. Therm. Anal. Calorim. 2021, 143, 265.
[88] S. K. Gill, M. Rossi, B. Bajka, K. Whelan, Nat. Rev. Gastroenterol.
Hepatol. 2021, 18, 101.
[89] M. Meenu, B. Xu, Crit. Rev. Food Sci. Nutr. 2019, 59, 3019.
[90] Y. Zhang, L. Chen, M. Hu, J. J. Kim, R. Lin, J. Xu, L. Fan, Y. Qi, L.
Wang, W. Liu, Y. Deng, J. Si, S. Chen, Aging 2020, 12, 9173.
[91] C. P. Rosado, V. H. C. Rosa, B. C. Martins, A. C. Soares, I. B. Santos,
E. B. Monteiro, N. Moura-Nunes, C. A. da Costa, A. D. P. Mulder, J.
B. Daleprane, Int. J. Biol. Macromol. 2020, 145, 1066.
[92] D. Zhou, Z. Ma, X. Hu, J. Agric. Food Chem. 2021, 69, 5392.
[93] J. Xu, Z. Ma, X. Li, L. Liu, X. Hu, Food Funct. 2020, 11, 1982.
[94] M. M. Finucane, T. J. Sharpton, T. J. Laurent, K. S. Pollard, PLoS One
2014, 9, e84689.
[95] C. The Human Microbiome Project, Nature 2012, 486, 207.
[96] R. Qin, J. Wang, C. Chao, J. Yu, L. Copeland, S. Wang, S. Wang, J.
Agric. Food Chem. 2021, 69, 3209.
[97] D. Mariat, O. Firmesse, F. Levenez, V. D. Guimarăes, H. Sokol, J.
Doré, G. Corthier, J. P. Furet, BMC Microbiol. 2009, 9, 123.
[98] A. Dura, D. J. Rose, C. M. Rosell, J. Agric. Food Chem. 2017, 65, 4651.
[99] M. M. Soliman, M. M. Ahmed, A.-e. Salah-eldin, A. A.-A. Abdel-Aal,
J. Vet. Sci. 2011, 12, 319.
[100] C. Chen, Q. Huang, X. Fu, R. H. Liu, Food Funct. 2016, 7, 4637.
[101] T. V. Maier, M. Lucio, L. H. Lee, N. C. VerBerkmoes, C. J. Brislawn,
J. Bernhardt, R. Lamendella, J. E. McDermott, N. Bergeron, S. S.
Heinzmann, J. T. Morton, A. González, G. Ackermann, R. Knight, K.
Riedel, R. M. Krauss, P. Schmitt-Kopplin, J. K. Jansson, mBio 2017,
8, e01343.
[102] Y. Duan, Y. Wang, Q. Liu, H. Dong, H. Li, D. Xiong, J. Zhang, Sci.
Rep. 2019, 9, 6464.
[103] N. T. Tran, Y. Tang, Z. Z. Li, M. Zhang, X. B. Wen, H. Y. Ma, S. K. Li,
Front Microbiol 2020, 11, 14.
[104] J. M. Erickson, J. L. Carlson, M. L. Stewart, J. L. Slavin, Foods 2018,
7, 18.
[105] S. Sieri, V. Krogh, Nutr. Metab. Cardiovasc. Dis. 2017, 27, 18.
[106] M. Arrese, A. Riquelme, A. Soza, Ann. Hepatol. 2010, 9, S112.
[107] L. M. Sanders, M. R. Dicklin, O. M. Palacios, C. E. Maki, M. L. Wilcox,
K. C. Maki, J. Hum. Nutr. Diet. 2021, 34, 94.
[108] T. H. T. Wong, J. C. Y. Louie, Starch – Stärke 2017, 69, 1600205.
[109] J. X. Wang, X. Si, W. T. Shang, Z. K. Zhou, P. Strappe, C. Blanchard,
Starch – Stärke 2017, 69, 209.
[110] M. L. Stewart, J. P. Zimmer, Nutrition 2018, 53, 59.
[111] M. L. Stewart, J. P. Zimmer, Nutrients 2017, 9, 237.
[112] H. Sun, X. H. Ma, S. Q. Zhang, D. Zhao, X. Liu, Int. J. Biol. Macromol.
2018, 110, 276.
[113] C. Zhang, L. Dong, J. Wu, S. Qiao, W. Xu, S. Ma, B. Zhao, X. Wang,
Biomed. Pharmacother. 2020, 128, 110350.
[114] Y. Wang, S.-Y. Lin, Y.-H. Chuang, C.-J. Chen, K.-C. Tung, W. H.-H.
Sheu, Am. J. Physiol. Endocrinol. Metab. 2010, 300, E155.
[115] K. G. Alberti, P. Z. Zimmet, Diabetic Med 1998, 15, 539.
[116] Y. Zhou, S. Zhao, Y. Jiang, Y. Wei, X. Zhou, J. Food Sci. 2019, 84, 2674.
[117] Y. Ge, C. Wei, W. Wang, L. Cao, J. Food Biochem. 2020, 44, 10.
[118] L. Laguna, A. Salvador, T. Sanz, S. M. Fiszman, LWT – Food Sci. Tech-
nol. 2011, 44, 737.
[119] E. Fuentes-Zaragoza, M. J. Riquelme-Navarrete, E. Sanchez-Zapata,
J. A. Perez-Alvarez, Food Res. Int. 2010, 43, 931.
© 2021 Wiley-VCH GmbH
www.advancedsciencenews.com
[120] S. Tian, Y. Sun, Int. J. Biol. Macromol. 2020, 149, 424.
[121] D. Bede, L. Zaixiang, Starch – Stärke 2021, 73, 2000139.
[122] M. Cervini, A. Frustace, G. D. Garrido, G. Rocchetti, G. Giuberti,
Heliyon 2021, 7, e06562.
[123] M. Di Cairano, M. C. Caruso, F. Galgano, F. Favati, N. Ekere, F.
Tchuenbou-Magaia, Eur. Food Res. Technol. 2021, 247, 707.
[124] M. Ma, Z. Li, F. Yang, H. Wu, W. Huang, Z. Sui, H. Corke, Int. J. Food
Sci. Technol. 2021.
[125] K. Pourmohammadi, E. Abedi, M. J. Amiri, M. H. Daneshgar, L. Torri,
J. Food. Meas. Charact. 2019, 13, 3149.
[126] S. Punia, A. K. Siroha, K. S. Sandhu, M. Kaur, Int. J. Biol. Macromol.
2019, 130, 595.
[127] J. Liu, M. Meenu, B. Xu, J. Food Process. Preserv. 2020, 44, e14513.
[128] M. Kiumarsi, M. Shahbazi, S. Yeganehzad, D. Majchrzak, O. Lieleg,
B. Winkeljann, Food Chem. 2019, 277, 664.
[129] E. Yaver, N. Bilgiçli, Food Chem. 2021, 357, 129758.
[130] M. Aribas, K. Kahraman, H. Koksel, J. Funct. Foods. 2020, 65,
103778.
[131] M. Cervini, A. Gruppi, A. Bassani, G. Spigno, G. Giuberti, Foods
2021, 10, 908.
Starch - Stärke 2021, 2100141 2100141 (13 of 13)
www.starch-journal.com
[132] R. G. M. van der Sman, S. Renzetti, Crit. Rev. Food Sci. Nutr. 2019,
59, 2225.
[133] J. S. Woomer, A. A. Adedeji, Crit. Rev. Food Sci. Nutr. 2021, 61, 14.
[134] A. Fardet, F. Leenhardt, D. Lioger, A. Scalbert, C. Remesy, Nutr. Res.
Rev. 2006, 19, 18.
[135] S. Simsek, S. N. El, Carbohydr. Polym. 2012, 90, 1204.
[136] C. G. Arp, M. J. Correa, C. Ferrero, Food Bioprocess Technol 2018, 11,
1149.
[137] C. G. Arp, M. J. Correa, C. Ferrero, Food Hydrocoll. 2021, 112, 106302.
[138] N. Baldino, I. Carnevale, F. Laitano, F. R. Lupi, S. Curcio, D. Gabriele,
Eur. Food Res. Technol. 2020, 246, 397.
[139] S. B. Kumar, P. Prabhasankar, Trends Food Sci. Technol. 2014, 35, 32.
[140] K. Tazrart, C. Lamacchia, F. Zaidi, M. Haros, J. Food Compos. Anal.
2016, 47, 8.
[141] E. Agama-Acevedo, L. A. Bello-Perez, G. Pacheco-Vargas, J. Tovar, S.
G. Sayago-Ayerdi, J. Food Process. Preserv. 2019, 43, 14012.
[142] D. E. Garcia-Valle, L. A. Bello-Perez, J. Tovar, J. Sci. Food Agric. 2021,
101, 2869.
[143] A. Rachman, M. A. Brennan, J. Morton, C. S. Brennan, J. Food Pro-
cess. Preserv. 2019, 43, e14084.
© 2021 Wiley-VCH GmbH