Effect of Pullulanase Debranching of Sago (Metroxylon sagu) Starch at Subgelatinization Temperature on the Yield of Resistant Starch

The effects of pullulanase debranching of sago (Metroxylon sagu) starch in the granular state and subsequent physical treatments on the formation and yield of type III resistant starch (RS 3) have been investigated. Sago starch was enzymatically debranched with pullulanase at 60°C and at pH 5.0 using different enzyme concentrations (24, 30, 40, 50 PUN/g dry starch) which was added to 20% (w/v) starch slurry and incubated for 0 to 48 h. Optimum enzyme concentration of 40 PUN/g dry starch and three debranching times (8, 16 and 24 h) have been selected for subsequent preparation of RS. Granule morphology and molecular weight distribution (MWD) of the debranched and resistant starch were examined. Debranched starch samples showed blurred birefringence patterns, a decrease in amylopectin fraction, an increase in low molecular weight fraction and a broadening of MWD. Debranched starch samples with a maximum RS yield of 7% were obtained at 8 h debranching time. Temperature cycling and incubation at certain temperature and storage time enhanced the formation of RS. Under the conditions used in this study, the optimum conditions to obtain the highest RS yield (11.6%) were 8 h of debranching time, followed by incubation at 80°C for seven days. The MWD analysis showed that RS consisted of material with relatively low degree of polymerization. This study showed that pullulanase treatment of starch in the granular state resulted in limited debranching of amylopectin but the subsequent physical treatments (incubation time/temperature) can be manipulated to promote crystallization and enhance formation of RS 3.     

Synthesis and in vitro digestion of resistant starch type III from enzymatically hydrolysed cassava starch

Resistant starch type III (RS III) was synthesised from cassava starch by autoclaving followed by debranching with pullulanase, at varied concentrations (0.4–12 U g^?1) and times (2–8 h), and recrystallisation (?18 to 90 °C for 1–16 h). The highest RS III yield (22 g/100 g) was obtained at an enzyme concentration of 4 U g^?1 after 8 h incubation, followed by recrystallisation at 25 °C for 16 h. Varying the recrystallisation conditions indicated that higher RS III yields (30–35 g/100 g) could be obtained at 90 °C within 2 h. Thinning cassava starch using α‐amylase prior to debranching using pullulanase did not further increase the RS III content. In vitro digestion data showed that whereas 44% RS III was digested after 6 h, the corresponding value for cassava starch was 89%.     

响应面试验优化晶种诱导-双酶复合法制备RS_3型籼米抗性淀粉工艺

以微波预糊化籼米淀粉为原料,自制RS_3型马铃薯抗性淀粉为晶种,研究RS_3型籼米抗性淀粉的晶种诱导-双酶复合法制备工艺。利用扫描电子显微镜对淀粉颗粒形貌进行表征并研究淀粉的抗酶解性。在单因素试验的基础上,固定其他酶解条件,以RS_3型籼米抗性淀粉产率为响应值,确定晶种添加量、异淀粉酶添加量、普鲁兰酶添加量和普鲁兰酶酶解时间作为影响产率的主要因素,进行Box-Behnken响应面优化试验。得到RS3型籼米抗性淀粉的最佳制备工艺条件为:晶种添加量5%、异淀粉酶添加量8 U/g、普鲁兰酶添加量8 U/g、普鲁兰酶酶解时间3.50 h。在此最佳制备工艺条件下,RS_3型籼米抗性淀粉产率为27.42%,RS3失去原有的淀粉颗粒形貌,表面变得粗糙,结晶结构致密,具有较强抗酶解能力。     

Effect of debranching and storage condition on crystallinity and functional properties of cassava and potato starches

Debranching starch by pullulanase is considered to improve the RS content of starch which is widely used to produce the starch‐based foods with high‐health benefit impacts. In this study, the cassava and potato starches were debranched by pullulanase, followed by an autoclave treatment and storage at −18°C, 4°C, or 25°C to investigate their crystallinity and functional properties. After debranching, the potato starch contained significantly higher CL (35.4 glucose units) than did the cassava starch (32.4 glucose units). The debranched cassava and potato starches after retrogradation at the storage temperatures had a typical B‐type crystalline structure although the native cassava and potato starches exhibited the different crystalline forms (A‐ and B‐type, respectively). The RS contents of the debranched cassava and potato starches significantly improved with higher RS content of the debranched potato starch than that of the debranched cassava starch at the same storage condition. The storage temperature significantly affected the RS formation of the debranched starches with the highest RS content at storage temperature of −18°C (35 and 48% for the debranched cassava and potato starches, respectively). The debranched starches had significantly lower viscosities and paste clarities but higher solubilities than did the native starches. As a result, the debranched cassava and potato starches can be considered for use not only in functional foods with enhanced health benefits but also in pharmaceutical and cosmetic industries.     

Resistant Starch Made from Banana Starch by Autoclaving and Debranching

Resistant starches (RS) were prepared from banana starch by debranching with pullulanase for different times and after autoclaving treatment. The different treatments produced seven RS products, which were tested with respect to available starch (AS), RS and in vitro hydrolysis rate. The control sample (without debranching) had the highest AS (80.5%) and the lowest RS content (9.1%). The samples debranched for 5 h and longer did not show significant differences (α = 0.05) in AS (approximately 72%) and RS (approximately 18%). The RS values obtained in the samples prepared were twice as high as that of the control sample. However, the sample debranched for the longest time had the highest hydrolysis rate, demonstrating that this product has a high digestion rate. Banana starch is a good source for RS preparation by autoclaving due to its high RS content and can be an alternative source in developing countries for obtaining a nutraceutic ingredient for functional food preparation.     

酶脱支处理对玉米缓慢消化淀粉形成的影响

以玉米原淀粉为原料,研究普鲁兰酶脱支处理糊化后制备缓慢消化淀粉(slowly digestible starch,SDS)过程中各影响因素(温度、p H值、酶用量、贮藏及干燥条件)对SDS形成的影响。结果表明,在57.5℃、p H 4.9、酶用量60 U/g的条件下脱支8 h,然后煮沸灭酶30 min,再经4℃冷藏、60℃干燥后,可得SDS含量为31.09%的产品。原淀粉、酶脱支处理样品及脱支并去除快速消化淀粉样品的X射线衍射图谱表明,脱支处理后,玉米淀粉结晶结构由A型向B型转变。因此,通过酶脱支处理提高SDS含量的可能原因是形成了新的结晶结构,SDS含量与结晶的数量和质量有关。采用酶法制备SDS具有较好的工业化应用前景。     

Crystallinity,Thermal and Morphological Characteristics of Resistant Starch Type III Produced by Hydrothermal Treatment of Debranched Cassava Starch

Cassava starch was debranched using pullulanase and the linear glucans recrystallized by incubation at 60°C or by temperature cycling at 120/60°C, and further subjected to heat‐moisture treatment (HMT). Resistant starch (RS III) contents increased from 21.4 g/100 g in the debranched starch (DS) to 67.3 g/100 g in the debranched starch incubated at 60°C (DRS) and 47.8 g/100 g in the debranched starch subjected to temperature cycling (DCS), and further to 84.8 g/100 g and 88.4% g/100 g in HMT‐DRS and HMT‐DCS, respectively. Total crystallinity varied between 31.4‐59.8% and the crystalline type was C in DS and DRS and A in DCS, HMT‐DRS and HMT‐DCS. The melting properties were characterized by broad endotherms, but the exact melting region and enthalpy were dependent on recrystallization method. The main endothermic peaks of DS and DRS occurred at 103.9 and 109.8°C, respectively, whereas DCS exhibited split endotherms at 113.6 and 138.1°C. Heat‐moisture treatment broadened the endotherms and increased their enthalpies. Scanning electron micrographs revealed surface topography differences related to size and aggregation of individual crystalline bodies.     

酶解法制备荞麦抗性淀粉的工艺优化

为确定荞麦粉制备抗性淀粉的工艺条件,采用普鲁兰酶酶解脱支法,并通过单因素和正交试验研究了影响抗性淀粉得率的因素。结果表明：影响抗性淀粉得率的因素主次顺序依次为荞麦粉浓度、普鲁兰酶用量、酶解时间和酶解温度。酶解法制备荞麦抗性淀粉的适宜工艺条件为荞麦粉浓度5 g/（100 mL）、普鲁兰酶用量7.2 PUN/g、酶解温度45℃、酶解时间8 h,在此条件下测得的抗性淀粉含量为15.82%。与原粉相比,普鲁兰酶酶解脱支与湿热法相结合制备荞麦抗性淀粉使其抗性淀粉含量显著提高。     

Effect of Cooling,and Freezing on the Digestibility of Debranched Rice Starch and Physical Properties of the Resulting Material

Ten percent non‐waxy and waxy starch suspensions were debranched with pullulanase followed by heating and cooling (1 °C) to crystallize and/or gel. Products with a range of textures can be made depending on the type (waxy and non‐waxy) of starch used. The water holding capacity was 35% and 84% for waxy and non‐waxy cooled debranched starch, respectively, at 4 h of cooling and did not change. The hardness of the debranched waxy and non‐waxy starch continued to increase beyond 24 h up to 45 g and 245 g of force, respectively. The particle size of precipitates of non‐waxy and waxy debranched starch was 45 μm and 4 μm after 4 h of cooling and did not change. Cooling of debranched non‐waxy starch at 1 °C for 12 h without agitation decreased digestibility by 59%; with stirring digestibility decreased by 42% after 24 h of cooling. Freezing of debranched cooled waxy and non‐waxy starch does not effect the decreases in digestibility. Particle size of debranched, cooled/freeze‐thawed, dried, and milled starch affects digestibility.     

Structural characterizations and digestibility of debranched high-amylose maize starch complexed with lauric acid

Structural characterizations and digestibility of debranched high-amylose maize starch complexed with lauric acid (LA) were studied. The cooked starch was debranched by using pullulanase and then complexed. Light microscopy showed that the lipids complexed starches had irregularly-shaped particles with strong birefringence. Gel-permeation chromatograms revealed that amylopectin degraded to smaller molecules during increasing debranching time, and the debranch reaction was completed at 12 h. Debranching pretreatment and prolonged debranching time (from 2 h to 24 h) could improve the formation of starch lipids complex. X-ray diffraction pattern of the amylose–lipid complexes changed from V-type to a mixture of B- and V-type polymorphs and relative crystallinity increased as the debranching time increased from 0 to 24 h. In DSC thermograms, complexes from debranched starch displayed three separated endotherms: the melting of the free lauric acid, starch–lipid complexes and retrograded amylose, respectively. The melting temperature and enthalpy changes of starch–lipid complex were gradually enhanced with the increasing of debranching time. However, no significant enthalpy changes were observed from retrograded amylose during the starch–lipid complex formation. Rapidly digestible starch (RDS) content decreased and resistant starch (RS) content increased with the increasing of debranching time, while the highest slowly digestible starch (SDS) content was founded at less debranching time of 2 h. The crystalline structures with dense aggregation of helices from amylose-LA complex and retrograded amylose could be RS, while SDS mostly consisted of imperfect packing of helices between amylopectin residue and amylose or LA.     

紫山药抗消化淀粉的制备及水解特性*

以酶解-压热法制备紫山药抗消化淀粉,考察了淀粉乳浓度、普鲁兰酶用量、酶解时间、压热时间对制备淀粉中抗消化淀粉含量的影响,通过正交试验和方差分析明确影响因素的重要性并优化工艺条件;比较分析了糊化淀粉、压热淀粉以及酶解-压热法制备淀粉的水解动力学。结果表明:酶解-压热法制备紫山药抗消化淀粉的含量随各因素水平的增加呈先增加后减小的趋势,优化的条件为:淀粉乳质量分数20%、普鲁兰酶用量8 U/g、酶解12 h、以120℃压热处理40 min 2次时,制备抗消化淀粉样品纯度为96.67%,其中抗消化淀粉含量为47.85%;水解特性研究表明:与糊化、压热法相比,酶解-压热法制备抗消化淀粉的水解率、水解指数与血糖指数均显著降低,具有更好的抗消化性。     

嗜冷普鲁兰酶制备玉米抗性淀粉工艺优化及其表征

以玉米淀粉为原料,采用嗜冷普鲁兰酶脱支处理和压热处理相结合的方式制备玉米抗性淀粉,考察了玉米淀粉乳质量分数、耐高温α-淀粉酶添加量、嗜冷普鲁兰酶添加量、嗜冷普鲁兰酶作用时间对抗性淀粉得率的影响,采用正交试验对压热-酶解法制备玉米抗性淀粉的工艺参数进行了优化。采用扫描电子显微镜、X-射线衍射和差示扫描量热仪对玉米抗性淀粉形貌、晶体结构、热特性进行了观察与分析。结果表明,制备玉米抗性淀粉的最佳工艺条件为：玉米淀粉乳质量分数18%、耐高温α-淀粉酶添加量7 U/g、嗜冷普鲁兰酶添加量10 U/g、嗜冷普鲁兰酶作用时间9 h。在最佳条件下,玉米抗性淀粉得率为16.84%。玉米淀粉经复合酶法处理后,抗性淀粉形成了致密的层状晶体结构,表面形态结构呈现出不同于玉米原淀粉A型晶体结构的V型晶体结构;玉米抗性淀粉的起始温度、峰值温度、终止温度和相变焓值分别为117.07、140.69、153.03 ℃和1 858.12 J/g,均高于玉米原淀粉。     

微波辅助酶法制备碎米抗性淀粉的工艺研究

研究以碎米为原料微波辅助酶法制备抗性淀粉的工艺。通过单因素和正交试验,获取最佳工艺条件：淀粉浆添加量25g/mL、微波时间90s、普鲁兰酶添加量4.0U/g 干淀粉、酶解时间2h、回生时间20h。在此条件下抗性淀粉得率为21.81%。本实验可为碎米抗性淀粉的制备提供一定参考。     

响应面法优化玉米抗性淀粉制备工艺

研究普鲁兰酶法制备玉米抗性淀粉的工艺。在单因素试验基础上,采用响应曲面法研究pH值、反应温度、反应时间和加酶量对抗性淀粉得率的影响,优化玉米抗性淀粉制备工艺,建立各因素与抗性淀粉得率关系的数学回归模型。确定最佳的制备工艺条件为普鲁兰酶加酶量12.8ASPU/g、反应时间32h、反应温度46.2℃、pH5.0。在该制备条件下,抗性淀粉得率为46.2%。     

响应面法优化板栗抗性淀粉制备工艺

为了提高板栗抗性淀粉含量,并获得抗性淀粉制备方法的最适工艺参数,本研究优化了压热—普鲁兰酶法制备板栗抗性淀粉的工艺,在单因素试验基础上,采用响应面法研究淀粉悬浮液质量分数、普鲁兰酶添加量、酶解时间和冷凝时间对抗性淀粉得率的影响,建立各因素与抗性淀粉得率关系的数学回归模型。最终根据实际工艺操作确定最佳的制备工艺条件为淀粉悬浮液质量分数11.00%,酶添加量9 PUN/g、酶解时间10 h、冷凝时间15 h。在该制备条件下,测得抗性淀粉得率为64.90%,基本符合理论预测值(65.70%)。试验证明,响应面法能够提高板栗抗性淀粉的制备率。     

The impact of coupled acid or pullulanase debranching on the formation of resistant starch from maize starch with autoclaving–cooling cycles

Maize starch was treated by autoclaving–cooling cycles, coupled with acid or pullulanase hydrolysis to prepare resistant starch (RS). Debranching of retrograded or gelatinized maize starch with acid or pullulanase was studied to show the corresponding impact on RS formation. When maize starch was treated with three autoclaving–cooling cycles and retrograded maize starch was hydrolyzed at room temperature, with 0.1 mol L⁻¹ citric acid for 12 h, analysis results showed that debranching of citric acid was helpful in RS formation for RS yield increased from 8.5 to 11%. Debranching of gelatinized or retrograded maize starch at 60 °C with pullulanase at addition level of 3 PUN g⁻¹ starch showed a more favorable effect on RS formation. When gelatinized maize starch was hydrolyzed by pullulanase for 12 h and then treated with two autoclaving–cooling cycles, RS yield increased to 23.5%. If retrograded maize starch subjected to one autoclaving–cooling cycle was hydrolyzed by pullulanase for 10 h and then followed by two autoclaving–cooling cycles, RS yield elevated to 32.4%. The debranching effect of pullulanase on retrograded maize starch to help RS formation is obvious and most effective, indicating this treatment is applicable in RS preparation to increase the RS yield.     

3种大米淀粉脱支前后结构及流变特性

通过普鲁兰酶处理糯米、粳米和籼米淀粉,研究酶水解对3 种大米淀粉结构和流变特性的影响。结果表明,经普鲁兰酶处理后3 种大米淀粉的结晶度降低,无定型区域增加;链长分布结果表明3 种大米淀粉的精细结构相似,水解反应对较短的侧链更有效,糯米淀粉更易被酶解;脱支淀粉和天然大米淀粉的傅里叶变换红外光谱没有明显差异,—OH的伸缩振动吸收峰相对增强;添加普鲁兰酶后,淀粉糊黏度急剧下降,糯米淀粉黏度下降最快,较容易被水解;流变学特性表明淀粉颗粒分子间缔合、排列松散,运动性增强,溶解度和持水力有所增强。糯米淀粉对普鲁兰酶处理较其他两种大米淀粉更为敏感。结论：脱支处理改善了淀粉凝胶性能,增强了淀粉的流动性。     

Production of Trehalose from Starch by Thermostable Enzymes from Sulfolobus acidocaldarius

The optimum conditions for the production of trehalose from starch were investigated using two thermostable enzymes, maltooligosyl trehalose synthase (MTSase) and maltooligosyl trehalose trehalohydrolase (MTHase), from Sulfolobus acidocaldarius ATCC 33909. The optimum pH was 5.5 and the optimum temperature was 55—57°C using isoamylase from Pseudomonas amyloderamosa as a debranching enzyme. The addition of CGTase to the reaction mixture during the saccharification process caused an increase in trehalose and a decrease in maltose and maltotriose. Isoamylase was better than pullulanase as a debranching enzyme. The yield of trehalose was independent of the type of starch used. Under optimum conditions, the yield of trehalose from corn starch at 30% concentration was more than 82%.     

Production and physicochemical properties of resistant starch from hydrolysed wrinkled pea starch

The content and physicochemical properties of resistant starches (RS) from wrinkled pea starch obtained by different molecular mass reduction processes were evaluated. Native and gelatinised starches were submitted to acid hydrolysis (2 m HCl for 2.5 h) or enzymic hydrolysis (pullulanase, 40 U g^?1 for 10 h), followed by hydrothermal treatment (autoclaving at 121 °C for 30 min), refrigeration (4 °C for 24 h) and lyophilisation. Native starch showed RS and total dietary fibre contents of 39.8% and 14.3%, respectively, while processed ones showed values from 38.5% to 54.6% and from 22.9% to 37.1%, respectively. From these, the highest contents were among acid‐modified starches. Processed starches showed endotherms between 144 and 166 °C, owing to the amylose retrogradation. Native and processed starches showed low viscosity, which is inversely proportional to the RS concentration in samples. The heat treatment promoted an increase in the water absorption index. The pea starch is a good source for obtaining resistant starch by acid hydrolysis.     

Characterization of Dextrins obtained by Enzymatic Treatment of Cationic Potato Starch

The substitution pattern of cationic potato starches was studied using starch hydrolyzing enzymes and a characterization of the hydrolysis products. Native and cationic starch samples were hydrolyzed with pullulanase, isoamylase, and α‐amylase and the molecular‐weight distributions of the resulting dextrins were studied using gel permeation chromatography. Isoamylase hydrolyzed the native potato starch sample readily, whereas hydrolysis with pullulanase was incomplete. Pullulanase hydrolyzed, however, cationic starch with higher DS (degree of substitution) more efficiently than isoamylase. The hydrolysis products obtained with pullulanase were separated according to charge using cation‐exchange chromatography into one unbound and two bound fractions. The unbound fraction possessed an increasing number of short chains from amylopectin with increasing DS of the starch sample. The bound material contained amylose and dextrins with sizes corresponding to the long B‐chains. The high portion of uncharged dextrins after α‐amylolysis suggested that the substitution pattern, on the molecular level, was non‐random. The composition of the unbound and bound material, obtained by ion‐exchange chromatography of α‐amylase treated starches, suggested a more intense fragmentation with increasing DS of the sample. Possibly, the substituents influence substrate conformation and thereby alter the hydrolysis patterns. It is concluded that a thorough understanding of the enzymatic hydrolysis patterns is of ultimate importance in structural studies of modified starch.