Resistant Starch Preparations Research Articles

Green banana resistant starch: A promising potential as functional ingredient against certain maladies.

This review covers the significance of green banana resistant starch (RS), a substantial polysaccharide. The food industry has taken an interest in green banana flour due to its 30% availability of resistant starch and its approximately 70% starch content on a dry basis, making its use suitable for food formulations where starch serves as the base. A variety of processing techniques, such as heat-moisture, autoclaving, microwaving, high hydrostatic pressure, extrusion, ultrasound, acid hydrolysis, and enzymatic debranching treatments, have made significant advancements in the preparation of resistant starch. These advancements aim to change the structure, techno-functionality, and subsequently the physiological functions of the resistant starch. Green bananas make up the highest RS as compared to other foods and cereals. Many food processing industries and cuisines now have a positive awareness due to the functional characteristics of green bananas, such as their pasting, thermal, gelatinization, foaming, and textural characteristics. It is also found useful for controlling the rates of cancer, obesity, and diabetic disorders. Moreover, the use of GBRS as prebiotics and probiotics might be significantly proved good for gut health. This study aimed at the awareness of the composition, extraction and application of the green banana resistant starch in the future food products.

Analysis of the interaction between double-helix starch molecule and α-amylase

Diabetes is a major chronic disease that jeopardises human health. Resistant starch (RS) is important in controlling diabetes. However, the specific relationship between the starch crystalline structure and RS content needs to be further elucidated. In this study, the details of the interactions between different starch molecules (single-amylose and double-helix) and α-amylase were analysed using molecular docking and dynamic simulations. Single-amylose molecules could penetrate deep into the active groove of α-amylase and make full contact with the catalytic triad, and their G3 and G4 glucose residues were firmly bound to the bottom of the active groove throughout the simulation. However, the double-helix molecule in the crystalline region of starch could not fully penetrate to the bottom of the active groove and make full contact with the catalytic triad of α-amylase. This suggests that, without considering other structural factors, the RS content should be positively correlated with relative crystallinity. In addition, starch molecules had strong interactions with α-amylase (approximately 78 kJ/mol) and could form many hydrogen bonds with the amino acid residues of α-amylase such as Thr163, Gln63, and Ile148, which supports the sliding continuum hydrolysis hypothesis of α-amylase. These results explain the intrinsic relationship between the starch hydrolysis efficiency and crystalline structure at the molecular level and provide an important reference for the preparation of RS and elucidation of its mechanism.

Rational Modification of Human Gut Microbiome and Metabolites By Dietary Resistant Starch in Allogeneic Hematopoietic Stem Cell Transplantation: A Feasibility Study

Allogeneic hematopoietic stem cell transplantation (allo-HCT) is limited by acute graft versus host disease (GVHD). GVHD is the principal cause of non-relapse mortality (NRM) and a major cause of morbidity after allo-HCT. Recent published experimental data demonstrated that altering the microbiome-metabolite axis regulates acute GVHD severity. Specifically, short chain fatty acid (SCFA) butyrate was significantly decreased in the intestinal epithelial cells (IECs) of mice experiencing GVHD, while restoring butyrate levels by increasing intestinal butyrate-producing bacteria, reduced experimental acute GI GVHD severity and mortality. Prebiotics usually refer to indigestible carbohydrates that are metabolized by the intestinal microbiota to produce microbial metabolites, such as SCFAs, that serve as nutrients for IECs. Administration of defined quantities of resistant potato starch (RPS), as a prebiotic, to normal healthy human volunteers promoted increase in butyrogenic bacteria and increased intestinal levels of SCFA butyrate to a greater extent than other commercially available resistant starch preparations tested. These data form the rationale for this study and led to prospectively study the clinical feasibility and impact of the administration of RPS as a prebiotic dietary intervention on the intestinal microbiota and its dependent metabolites in allo-HCT recipients. Adults undergoing human leukocyte antigen-matched, related-donor myeloablative allo-HCT were recruited and received RPS orally daily from day -7 to day 100 after allo-HCT. Stool samples were collected in the OMNIgene-Gut® (DNA Genotek) collection kit. Fecal samples were subjected to 16S rRNA gene sequencing to determine microbiota composition and quantification of fecal SCFAs was performed by liquid chromatography. Blood specimens were collected using standardized protocols. The effect of RPS on plasma SCFAs and > 200 other metabolites was longitudinally assessed using targeted and global metabolomic analyses. Mass spectrometry was utilized for quantification of plasma metabolites. The primary objective was to assess the feasibilty and test the effect of RPS on the structure of the patients' intestinal microbiome. Metrics for the feasibility of this dietary intervention were prespecified by setting a target for ≥ 60 % of patients to adhere to ≥ 70% of scheduled doses. We hypothesized that RPS would be feasible and increase stool butyrate levels as a byproduct of microbial metabolism. Key secondary objectives were to longitudinally evaluate plasma metabolites in recipients of RPS compared to historical controls as well as assess tolerability of RPS in allo-HCT recipients. Ten subjects were enrolled. The primary endpoint was met. Feasibility exceeded the preset goal of ≥ 70% adherence to scheduled dosages in ≥ 60 % of patients as 8 of the 10 patients (80%) received ≥ 70 % of scheduled doses. Intestinal butyrate levels were significantly higher while participants were on RPS as compared to when they were not on RPS (p < 0.0001) (fig. 1A). We observed longitudinal changes in plasma metabolites post allo-HCT compared to baseline independent of whether allo-HCT recipients received RPS (p < 0.0001) (fig. 1B). In RPS recipients, the dominant plasma metabolites were, however, much more stable across timepoints when compared to historic controls suggesting a greater equilibrium in their production and consumption (fig. 1C). The median age of participants was 57 years (range 52-62 years). All subjects received standard GVHD prophylaxis with tacrolimus and methotrexate as well as standard antibiotic prophylaxis with levaquin, and standard neutropenic fever treatment with IV cefepime (90%) or IV vancomycin, No adverse effects/toxicities attributed to RPS were observed and longitudinal specimens were collected successfully (table 1). This study showed that a dietary intervention using RPS in allo-HCT recipients is feasible, and was able to rationally alter the salutary intestinal microbial metabolite SCFA butyrate, despite the utilization of several HCT related medications, including antibiotics. These data demonstrate translation of fundamental discoveries in mouse allo-HCT studies, to testing RPS in healthy humans, to showing that RPS is a feasible microbiome-modifying intervention in allo-HCT recipients, and that a prebiotic strategy can now be applied and tested to mitigate acute GVHD in allo-HCT.

Understanding the Inhibitory Mechanism of Exogenous Protein on Enzymatic Preparation of Resistant Starch

AbstractEnzymatic synthesis of resistant starch strongly depends on the food composition and their interaction with exogenous/endogenous proteins during preparation process. Therefore, the present study systematically investigates the inhibitory effect of egg albumin on pancreatic‐α‐amylase for synthesis of resistant starch from maize flour. The maximum inhibitory concentration of egg albumin is 5 mg mL−1 with % inhibition of 95.15% for pancreatic‐α‐amylase. Lineweaver burk plot reveals that Vmax is constant with and without addition of egg albumin. Whereas Km is increased in presence of albumin to achieve the same Vmax. Increase in the substrate concentration (Km) from 5 to 25 mg mL−1 leads to a decrease in enzyme inhibition. Moreover, the resistant starch content of enzymatically prepared resistant starch with and without albumin as compared to gelatinized maize starch (control) is increased. Scanning electron micrograph reveals interaction of egg albumin with maize starch as thread like aggregation. FTIR and XRD analysis reveal the increase in crystallinity of enzymatically treated maize starch against control. Moreover, egg albumin is negatively associated with enzymatic hydrolysis by preventing its access to maize starch granule and act as a competitive inhibitor. As a result of this interaction decrease in resistant starch yield from maize starch is investigated by enzymatic treatment.

Pea resistant starch preparation with cold-active type I pullulanase from Bacillus megaterium and its potential application in rice noodles

The characterization of PulY103B (pullulanase from Bacillus megaterium) was studied by our group, which showed that the enzyme is a cold-active type I pullulanase with a moderate active pH condition. The potential of PulY103B for resistant starch production in mild conditions (35 °C; pH 6.5) was investigated in this study. The result demonstrated that PulY103B was more efficient in pea resistant starch production with higher growth rate than the other pullulanases reported. The pea resistant starch (PS-PA) prepared with PulY103B had lower swelling power and solubility, and better thermostability. Also, the starch crystallinity pattern was different from the pea resistant starch reported earlier (B + V pattern: 5.9°, 17.0°, 19.2°, 21.9°, and 23.9°). Moreover, the PS-PA was used for preparing rice noodles (the best formula: PS-PA reached 8% rice granule weight). The results showed that the cooking properties of rice noodles were improved significantly (P < 0.05) including higher water absorption, lower cooking loss and breaking rate. The textural properties and its digestibility indicated that the rice noodle with PS-PA was suitable for children and elderly people who need weight loss.

Optimization of Corn Resistant Starch Preparation by Dual Enzymatic Modification Using Response Surface Methodology and Its Physicochemical Characterization.

Corn starch was dually modified using thermostable α-amylase and pullulanase to prepare resistant starch (RS). The concentration of starch liquid, the amount of added thermostable α-amylase, the duration of enzymatic hydrolysis and the amount of added pullulanase were optimized using RSM to increase RS content of the treated sample. The optimum pretreatment conditions were 15% starch liquid, 3 U/g thermostable α-amylase, 35 min of enzymatic hydrolysis and 8 U/g pullulanase. The maximum RS content of 10.75% was obtained, and this value was significantly higher than that of native corn starch. The degree of polymerization (DP) of the enzyme-modified starch decreased compared with that of native starch. The scanning electron microscopy (SEM) and differential scanning calorimetry (DSC) were performed to assess structural changes in native and pretreated starch. The effect of dual enzyme pretreatment on the structure and properties of corn starch was significant. Unlike the untreated one, the pretreated corn starch showed clear pores and cracks. Significant differences in RS contents and structural characterization between starch pretreated and untreated with dual enzymes demonstrated that the dual enzyme modification of corn was effective in enhancing RS contents.

Study on the Preparation and Digestibility of Malic Acid Sweet Potato Starch Ester under Microwave Assistance

AbstractIn this paper, malic acid sweet potato starch ester (MSE) is prepared by the microwave assisted l‐malic acid method, and its digestibility is studied. Through a response surface methodology based on a Box‐Behnken design (BBD), the optimum conditions for the preparation of MSE are obtained as follows: mass ratio (mL‐malic acid: mstarch) of 0.81, pH value of 1.21, microwave time of 6.8 min, and microwave power of 435 W, respectively. Under these conditions, the degree of substitution of MSE is 0.0712. In addition, the freeze‐thaw stability, transparency, and solubility of the MSE are better than the native starch. Microwave assisted l‐malic acid treatment significantly increases the resistant starch (RS) content in starch sample. FTIR reveals a new characteristic peak near 1735 cm–1 in MSE samples, and the relative crystallinity decreases from 33.03% to 21.18%. Differential scanning calorimetry results show that the endothermic peak of MSE decreases. These results indicate that microwave‐assisted l‐malic acid can significantly change the physicochemical properties of sweet potato starch, and it is an effective method for the preparation of RS with heat‐stable resistance.

Preparation of resistant starch by Some Different Physical Methods and Utilization in Muffins Production

Preparation of resistant starch by Some Different Physical Methods and Utilization in Muffins Production

Effect of high hydrostatic pressure treatment on the formation and in vitro digestion of Tartary buckwheat starch/flavonoid complexes

Tartary buckwheat is rich in flavonoids and starch, and the interaction between these two components affects the structural and digestive properties of the food product. In this study, we analyze the effects of thermal gelatinization (GT), ultrasonic treatment (UT), and high hydrostatic pressure treatment (HHP) on the compounding degree of starch and flavonoids in Tartary buckwheat and on the properties of the starch/flavonoid complex system (HBS-BF). Based on scanning electron microscope (SEM) and X-ray diffraction (XRD) analyses, the surface of HBS-BF becomes rough after GT or UT, and many small cavities appear. Comparatively, HHP treatment is less damaging to HBS-BF. Moreover, HHP maintains the original A-type crystal morphology of buckwheat starch in HBS-BF, whereas GT and UT change to V-type. Repeated HHP further improves the crystallinity and digestion resistance of HBS-BF. According to the recorded Fourier transform infrared (FT-IR) spectra, HBS-BF by different methods does not exhibit new covalent bonds.Practical application: The results reported herein promote the application of Tartary buckwheat starch and flavonoids in the food industry by providing a theoretical basis for the development of starch anti-digestion mechanisms and the preparation of resistant starch.

Optimization of the Kudzu Resistant Starch Preparation Based on Debranching and Recrystallization and its Structural Properties

Kudzu-resistant starch was prepared by pullulanase debranching recrystallization. Based on single factor experiments, the response surface optimization method was used to analyze the optimal conditions for the preparation of kudzu-resistant starch. The results showed that the highest yield of 90.46% was obtained at the enzymatic reaction time of 4 h, temperature of 56°C, and amount of enzyme being 57 U/g. After debranching, the content of the resistant starch was higher than native starch. The crystal structure of the debranched starch changed from A-type to a mixture of B+V-type. The results of scanning electron microscope showed that the surface of the native starch granule was smooth and varied in size, showing a spherical or irregular polyhedral shape, while the debranched starch granule was irregular in shape and size and the surface was rough and gully. The granule morphology of the modified starch granule was also changed.

Rearranged supramolecular structure of resistant starch with polymorphic microcrystals prepared in high-solid enzymatic system

Resistant starch (RS) is widely used as a new type of dietary fiber, but it is still a limitation of controllable formation of microstructure and polymeric architecture for preparing single-component RS (pure-phase). In this study, RS was prepared using maize starch with two ratios of amylose to amylopectin (RAA1:1 and RAA1:2) and four pullulanase concentrations (5, 15, 25, 35 U/g) in a high-solid dynamic reaction environment of bio-extrusion. Based on the long- and short-range orders reflected by the glycosidic bonds (13C NMR), chain-length distributions (SEC/FACE), single-/double-helices (FTIR), lamellas (SAXS) and crystalline pattern (XRD), the rearranged supramolecular structure of starch and their relationships with physicochemical properties (DSC and RVA) was illuminated. There were two morphologies of RS were created: 1) disordered fusion-like structure displayed by RAA1:1-P15 and RAA1:2-P35, which had the high contents of RS (54.95 and 54.19%) and short amylopectin (DP ≤ 24) (70.52% and 60.20%). The short (A chains) and extremely short (DP < 6) amylopectin could contribute to form submicro-crystallite with B1 chains in the extruded matrix at a low moisture level; 2) thinner, scalier and layered-like structure displayed by RAA1:2-P15 that was the transition state from thick layered strips of large granules. Its SDS content was high (33.61%) and owned the larger rearranged dLorentz (16.97 nm). Overall, the entangled regime of amylose and amylopectin in the modifications of starch with different structural units or micro-domains in this study will provide a new idea and reference to explore the preparation of novel RS.

Effect of Two Combined Functional Additives on Yoghurt Properties.

The aim of the research was the analysis of yoghurts enriched with blue honeysuckle berries dry polyphenolic extract and new preparation of resistant starch. The additives were introduced individually at concentration 0.1% (w/v) and in mixture at final concentration of 0.1 and 0.2% of both components. Yogurt microflora, pH, and its physicochemical and antioxidant properties were examined over 14 days of storage under refrigerated conditions. Studies showed that both substances can be successfully used in yoghurt production. Yoghurt microflora es. S. thermophilus and Lb. delbrueckii subsp. bulgaricus counts appeared to be higher in samples supplemented with these additives comparing to control yoghurt by 3–8%. More stimulating effect on their growth, especially on S. thermophilus, revealed resistant starch. Addition of this polysaccharide improved also the rheological properties of yogurts, which showed higher viscosity than samples produced without it. Addition of honeysuckle berries preparation significantly influenced the yogurts’ color, giving them deep purple color, and their antioxidant potential. During storage, contents of anthocyanin and iridoid compounds were decreasing, but antioxidant activity in the products remained stable.

Structural characteristics and prebiotic activities of resistant starch from Solanum tuberosum: Kufri Bahar, a popular Indian tuber variety

The present study reports the effect of enzymatic debranching and heating-cooling treatment on morphology, crystallinity and thermal properties of resistant starch preparations from Solanum tuberosum, variety Kufri Bahar. Further, the proliferative and protective effect of resistant starch on viability of probiotic cultures in fermented milk during storage at refrigeration was also investigated. The results indicated that enzymatic treatment resulted in more crystalline form of resistant starch granules with an increased thermal stability. A strain specific prebiotic activity of potato resistant starch was observed with a higher proliferative activity towards Lactobacillus paracasei CD4 among the tested probiotic strains. In synbiotic fermented milk preparation, the supplementation of enzymatic treated resistant starch increased the viability of L. paracasei CD4 under refrigeration up to 14 days as compared to control (p < 0.05).

Study on the structure and digestibility of high amylose Tartary buckwheat (Fagopyrum tataricum Gaertn.) starch-flavonoid prepared by different methods.

Tartary buckwheat (Fagopyrum tataricum Gaertn.) is the only food rich in flavonoid bioactive substances in grains. Studies have shown that flavonoids interaction with amylose has an important impact on the physical and chemical properties and structure of starch. In this study, Tartary buckwheat was used as a raw material. It was then threshed with pullulanase, and a high amylose Tartary buckwheat starch flavonoid complex (HBS/BF) was prepared by physical mixing (PM), water bath treatment (WT), acid-base precipitation (AP), microwave treatment (MT), and ultrasonic treatment (UT); the physical and chemical properties were then evaluated. The results show that HBS/BF-UT and HBS/BF-MT have a higher iodine binding rate than HBS/BF-PM; X-ray diffraction results show that HBS/BF-AP has a V-type crystal form, but the relative crystallinity was reduced. Fourier infrared spectroscopy showed that there is no new covalent bond between Tartary buckwheat starch and flavonoids. In vitro digestion showed that adding flavonoid significantly increased the digestibility of Tartary buckwheat starch. PRACTICAL APPLICATION: These results will provide a theoretical basis for further starch anti-digestion mechanisms and the preparation of resistant starch. These steps will provide insights into the application of Tartary buckwheat starch and flavonoids in the food industry.

Using a novel hyperthermophilic amylopullulanase to simplify resistant starch preparation from rice starches

• A hyperthermophilic amylopullulanase is recombinantly expressed in E. coli. • The enzyme is able to efficiently improve resistant starch yield from rice starches. • Pregelatinization of starches and enzymatic hydrolysis were combined into one step. A glycoside hydrolase family 57 protein, designated APU_Aquae, from hyperthermophilic bacterium Aquifex aeolicus was expressed in Escherichia coli . APU_Aquae was biochemically defined as a novel bifunctional amylopullulanase, which exhibited efficient activity towards rice starches. It is extremely thermostable with half-life of 7 h at 100 °C. Therefore, APU_Aquae was used to simplify the process of enzymatic debranching treatment in resistant starch type III (RS3) preparation from rice starches. Starch pregelatinization and enzymatic hydrolysis were combined into one step. It took only 7 h, which was much shorter than that of other enzymatic treatments. RS3 content of treated rice starches was 40.5 ± 0.7%. APU_Aquae treatment resulted in more medium branch chains B1 with degree of polymerization (DP) values 13–24, which might be one main reason to promote the RS3 formation. This study provides the new design for the industrial RS3 production.

Effect of debranching and temperature-cycled crystallization on the physicochemical properties of kudzu (Pueraria lobata) resistant starch

Resistant starch from Kudzu (Pueraria lobata) was prepared by debranching and subsequent recrystallization under isothermal and cycled temperature conditions. The granule morphology of the resistant starch was irregular in shape and size with rough surface. All the resistant starch samples exhibited B + V-type crystalline structure. The peripheral regions of native starch were better organized than that of the resistant starch samples. The solubility and swelling degree of native starch and resistant starch samples increased with the increase of temperature. The water holding capacity of native starch was much higher in comparison with resistant starch when the temperature was above 80 °C. The resistant starch showed better transmittance and freeze-thaw stability than that of native starch. These results suggest that kudzu starch is a potential resource for resistant starch preparation.

Anti-digestion properties of amylosucrase modified waxy corn starch

Amylosucrase is a valuable enzymatic tool for the preparation of resistant starch (RS). This study investigated the morphology, thermal properties and molecular structure of the RS components resulting from the digestion of amylosucrase modified starches (MSs). Morphology of starch samples suggests that the surface of MSs is partially attacked by the digestion enzymes. Besides, the peak melting temperature of RSs shows a considerable increase compared with that of the corresponding MSs. Branch chain length distributions data reveals that the long branch chains of MSs are prone to being hydrolyzed, and the branch chains with degree polymerization of 22–32 are the largest populations among the RS components. Moreover, the RS content in MSs decreases significantly after thermal treatment. Overall results demonstrates that the existence of crystalline structure with thermostable double helices is responsible for the anti-digestion properties of modified starches.

Gluten free rice cookies with resistant starch ingredients from modified waxy rice starches: Nutritional aspects and textural characteristics

Experimental gluten-free (GF) rice cookies were formulated with 100% rice flour (CTR) or by substituting 50% of rice flour with native waxy rice starch (WRS) or with three different resistant starch (RS) ingredients obtained from debranched, annealed or acid and heat-moisture treated WRS (RSa, RSb and RSc, respectively). Chemical composition, in vitro starch digestibility and physical and textural characteristics were carried out. Among cookies, RSa-cookies had the highest total dietary fibre content, the lowest rapidly digestible starch and the highest RS contents. All the three RS preparations have proved effective in increasing the proportion that tested as RS with respect to native WRS. However, the estimated RS loss for each applied RS ingredients caused by the baking process followed the order of RSa < RSc < RSb. Last, the lowest vitro glycaemic index value was measured for RSa-cookies. Among cookies, differences in colour and hardness were reported. The partial replacement of commercial rice flour with RSa could contribute to formulate GF cookies with higher dietary fibre content and likely slowly digestible starch properties more than equivalent amounts of RSb and RSc.

Investigation of debranching pattern of a thermostable isoamylase and its application for the production of resistant starch

Debranching enzymes contribute to the enzymatic production of resistant starch (RS) by reducing substrate molecular weight and increasing amylose yield. In the present study, the action pattern of a thermostable isoamylase-type debranching enzyme on different types of starch was investigated. The molecular weight distribution, glycosidic bond composition and contents of oligosaccharides released were monitored by various liquid chromatography techniques and nuclear magnetic resonance spectroscopy (NMR). These analyses showed that the isoamylase could specifically and efficiently attack α-1,6-glucosidic linkages at branch points, leaving the amylose favored by other amylolytic enzymes. Its ability to attack side chains composed of 1–3 glucose residues differentiates it from other isoamylases, a property which is also ideal for the RS preparation process. The enzyme was used as an auxiliary enzyme in the hydrolytic stage. The highest RS yield (53.8%) was achieved under the optimized conditions of 70 °C and pH 5.0, using 7 U isoamylase per g starch and 2 NU amylase per g starch. These data also help us better understand the application of isoamylase for preparation of other products from highly branched starch materials.

An innovative approach for significantly increasing enzyme resistant starch type 3 content in high amylose starches by using extrusion cooking

In this study effects of extrusion cooking on enzyme resistant starch (RS) formation in high amylose corn starches (Hylon V and VII) and the functional properties of RS preparations were investigated. Native starches were extruded at 50, 60, 70% feed moisture contents, at constant screw speed (100 rpm) and barrel temperature (140 °C). Among these samples, the highest RS contents were observed at 60% feed moisture. Therefore, feed moisture in the second and third extrusion cycles was set at 60%. There were significant increases in RS contents of both Hylon V and Hylon VII after the second extrusion cycle (p < 0.05). After the third extrusion, the RS levels reached to 40.0 and 45.1% for Hylon V and Hylon VII, respectively. Substantial loss of birefringence in these samples indicated that the increases in RS were mainly due to RS3 formation. The RS samples produced by extrusion did not have high emulsion capacity, but the ones produced from Hylon VII had high emulsion stability. Although, decreases in L* and increases in b* values of extruded samples were significant as compared to respective native starches, the changes were not substantial. Therefore, their incorporation is not expected to cause major changes in the colour of end-products.