Identification of botanical origin of starches by SDS-PAGE analysis of starch granule-associated proteins

Starch granule-associated proteins (SGAPs) were extracted from various starches and analyzed using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with silver staining. The SDS-PAGE results showed that granule-bound starch synthase (GBSS) of approximately 60 kDa was found in most starches, except waxy-type starches. Starches exhibited the presence of proteins specific to the botanical origin, including those of 22 kDa for maize; 160 and 98 kDa for potato; 140, 115, 90, and 80 kDa for wheat. These proteins could be detected from noodles prepared with the corresponding starches. The detection of the specific proteins by SDS-PAGE may be used to identify the origin of starch incorporated in various foods and industrial products.     

Metabolic characteristics in ruminants of the proteins in newly developed hull-less barley varieties with altered starch traits

Recently, new varieties of hull-less barley have been developed with altered carbohydrate traits. To our knowledge, there is no study on metabolic characteristics in ruminants of the proteins in the newly developed hull-less barley varieties. The objectives of this study were to: compare metabolic characteristics of the proteins of zero-amylose waxy (CDC Fibar), low-amylose waxy (CDC Rattan), high-amylose (HB08302), and normal starch (CDC McGwire) hull-less barley. In situ animal trials were carried out to generate the original rumen fermentation data for modeling nutrient supply to dairy cattle by using two dairy nutrition models – Dutch DVE/OEB system and NRC 2001. The major comparisons were made in terms of i) truly absorbed protein in the small intestine (DVE or MP, and ii) degraded protein balance (DPB). The study revealed that zero-amylose waxy hull-less barley was superior (P < 0.05) in both DVE (123 vs. 117, 114, 103 g kg⁻¹ DM) and MP (112 vs. 93, 96, and 87 g kg⁻¹ DM) when hull-less barley was evaluated as a single feed for dairy cattle. All of four hull-less barley varieties had negative DPB (DPB^OEB; −37.4, −17.1, −30.2, and −28.2 g kg⁻¹ DM for normal starch, zero-amylose waxy, waxy, and high-amylose cultivar, respectively), indicating the potential N shortage. In conclusion, the alteration of starch structure in granule provided a relatively balanced energy and protein for microbial synthesis in the rumen. The DVE and DPB predicted by using the DVE/OEB system can be explained (r² > 0.76) by the equivalent parameters, predicted by using the NRC 2001 model. The alteration of starch structure in granule affects metabolic characteristics of the proteins of hull-less barley in ruminants.     

Selection of cultivars for minimization of waste and of water consumption in cassava starch production

When considering the sustainability of a business, deciding on the industrial use of starchy raw materials requires more than just the information on their agricultural productivity and starch yield. The main goal of this work was to investigate ten different cultivars to select for industrial applications seeking to minimize residue generation and water consumption in the production of cassava starch. The cassava cultivars that are richer in starch (22.61-22.89 g 100 g⁻¹) generated the smallest amounts of residues (420.63-423.52 kg ton⁻¹ of cassava roots) and required the smallest amounts of water for processing. There is an inverse relationship between the dry matter content in cassava roots and the amount of solid residues generated. One of the cultivars stood out for showing the following features: high starch yield, little tendency for generation of residues, low requirement of water for processing, easiness in the peeling process, and high content of total solids; therefore such features can suggest its use for starch extraction with wastes minimization.     

Water stress during grain development affects starch synthesis,composition and physicochemical properties in triticale

Triticale, a man-made cereal crop developed from a cross between wheat and rye, has excellent agronomic traits to produce starch for bioindustrial applications. The effects of different levels of water stress on expression of starch synthesis genes and starch composition and physicochemical properties were investigated in this study. Three triticale varieties from 5 days post-anthesis were treated with three levels of water stress: low water stress (LWS) at 55–60% of soil moisture, moderate water stress (MWS) at 30–35% soil moisture and severe water stress (SWS) at 10–15% soil moisture. Water stress led to a significant reduction in seed weight at SWS (35–45%). A decrease in starch content was noticeable from MWS onwards and the values were decreased by 42–55% at SWS across all varieties. Such decrease was associated with the reduced expression of starch synthesis genes at 19 days of water stress (DWS). MWS favoured an increase of amylose proportion in triticale starch and it was accompanied by a significant up-regulation of GBSSI expression throughout the grain development. Triticale starch synthesized under water stress showed a reduced population of small granules and an increase of A-type to B-type ratio. SWS caused pitting on starch granules but did not alter the biconcave disc shape of mature granules. An inverse relationship between water stress and a range of starch gelatinization temperature was observed and the MWS environment specifically decreased the peak temperature (Tp) and increased the enthalpy. Our results signify that starch morphology, composition and physicochemical properties in triticale grains could be altered if triticale is grown under drought conditions.     

Dynamic water vapour sorption in gluten and starch films

Water sorption of gluten and wheat starch films as a function of water activity was studied using gravimetric step-change sorption experiments. Films of different thicknesses were used with the aim to vary the characteristic diffusion time and to get insights in the contribution of the polymer-chain rearrangement in the sorption behaviour. It is shown that both starch and gluten are in the glassy state for a water activity a_w below 0.9. From comparison of the dynamical sorption curves with a Fickian diffusion model, it is shown that water diffusion in gluten films seems Fickian for a_w < 0.7, and non-Fickian for a_w > 0.7, while for starch films, non-Fickian sorption behaviour is observed for a_w > 0.1. The results show that polymer-chain rearrangement and the stress built up in the matrix play an important role in the sorption dynamics of these films. Even when the material is in the glassy state matrix relaxation phenomena play a role in the sorption behaviour of starch and gluten.     

Effects of inhibition of starch branching enzymes on starch ordered structure and component accumulation in developing kernels of rice

Three rice varieties, Te-qing (TQ), Wu-xiang 9915 (WX) and Guang-ling-xiang-nuo (GLXN) with different amylose contents, and their derived lines with inhibition of starch branching enzyme I and IIb (SBEI/IIb) (GLXN-, WX- and TQ-SBEI/IIb^–) were used to investigate and compare the starch ordered structure and component accumulation in developing kernels. The starches in developing kernels of TQ, WX and GLXN all showed a typical A-type crystallinity and had similar short-range ordered structure, but their relative crystallinities gradually decreased in TQ and WX and had no significant change in GLXN with kernel development. For wild-type rices, starch components gradually increased in developing kernels. With inhibition of SBEI/IIb, the accumulation of amylose and amylopectin long branch-chains in kernels was slightly affected, but amylopectin short branch-chains seriously decreased, resulting in a significantly lower dry weight and starch content in kernel. The B-type crystallinity in developing starch gradually accumulated with kernel development of TQ-, Wx- and GLXN-SBEI/IIb^– lines. The inhibition of SBEI/IIb decreased the relative crystallinity but increased the short-range ordered structure of starches. The effects of inhibition of SBEI/IIb on starch ordered structure and component accumulation mainly appeared after 10 days after flowering, and were the most conspicuous for indica rice TQ. Though the inhibition of SBEI/IIb influenced the ordered structure and component accumulation of starch, the starch content and components in developing kernels all showed significantly positive correlations with kernel dry weight.     

青稞幼苗期叶片中 SBEⅡa基因的克隆及其表达分析

为进一步研究淀粉分支酶(Starch branching enzyme,SBE)在青稞叶片淀粉代谢中的作用,以青稞品种喜马拉雅2号和康青1号为材料,通过同源克隆技术分离 SBEⅡa基因,并对分离得到的 SBEⅡa基因进行生物信息学分析,同时利用实时荧光定量PCR(qRT-PCR)技术和淀粉分支酶活性测定试剂盒分析了在高温、低温和盐胁迫条件下 SBEⅡa基因的表达情况和SBEⅡa蛋白酶活性的变化情况。结果表明,从喜马拉雅2号和康青1号克隆得到的 SBEⅡa基因ORF区为2466 bp,编码821个氨基酸。生物信息学分析表明,该基因编码的蛋白质分子量为92.09 kDa,预测等电点(pI)为5.4,是酸性蛋白;该蛋白酶的不稳定系数为38.03,属于稳定蛋白;参试材料与大麦的亲缘关系最近,与细江蓠的亲缘关系最远。qRT-PCR和蛋白酶活性测定结果表明, SBEⅡa基因在不同环境下表达水平和SBEⅡa蛋白酶活性变异具有高度的正相关性,在不同环境和不同材料之间没有明显差异。     

Barley genotype expressing “stay-green”-like characteristics maintains starch quality of the grain during water stress condition

The identification of “stay-green” in sorghum and its positive correlation with yield increases has encouraged attempts to incorporate “stay-green”-like traits into the genomes of other commercially important cereal crops. However, knowledge on the effects of “stay-green” expression on grain quality under extreme physiological stress is limited. This study examines impacts of “stay-green”-like expression on starch biosynthesis in barley (Hordeum vulgare L.) grain under mild, severe, and acute water stress conditions induced at anthesis. The proportions of long amylopectin branches and amylose branches in the grain of Flagship (a cultivar without “stay-green”-like characteristics) were higher at low water stress, suggesting that water stress affects starch biosynthesis in grain, probably due to early termination of grain fill. The changes in long branches can affect starch properties, such as the rates of enzymatic degradation, and hence its nutritional value. By contrast, grain from the “stay-green”-like cultivar (ND24260) did not show variation in starch molecular structure under the different water stress levels. The results indicate that the cultivar with “stay-green”-like traits has a greater potential to maintain starch biosynthesis and quality in grain during drought conditions, making the “stay-green”-like traits potentially useful in ensuring food security.     

西藏冬青稞胚乳淀粉粒发育特征研究

为了解青稞胚乳淀粉及淀粉粒形成与生长特征,以西藏育成青稞品种冬青8号为材料,对籽粒发育过程中胚乳灌浆速度、淀粉积累量及淀粉粒的产生、二维面积、粒径变化进行了分析。结果表明,胚乳发育过程中,干物质和淀粉积累量呈S型曲线,两者呈正相关。花后5d,青稞胚乳中可零星见到粒径和二维面积较小的A型小淀粉粒;花后10~15d,A型小淀粉粒数量显著增大,但粒径和二维面积增加较慢;花后20d,B型小淀粉粒可能已经形成,与A型小淀粉粒组成一个复合群体;花后25d,形成了粒径10μm的A型淀粉粒、粒径5~10μm的B型淀粉粒和粒径5μm的C型淀粉粒;花后30~55d,不同粒径的淀粉粒数量和二维面积逐渐增加,其中,C型淀粉粒数量不断增加直至籽粒成熟。青稞成熟籽粒胚乳中,淀粉粒的粒径为1.45~44.76μm,二维面积为3.56~1 750.8μm2;A型淀粉粒形状为近圆形,表面光滑,个别表面附着有C型淀粉粒,B型淀粉粒形状有近圆形、椭圆形多种类型,C型淀粉粒形状比B型淀粉粒的变异更大。     

青藏高原青稞的淀粉特性

为给优质专用青裸品种的选育提供淀粉特性方面的参考依据,对122份来自青藏高原的青稞品种（系）的总淀粉含量和直链淀粉含量以及淀粉粘度性状进行了测定,并对这些品质性状进行了相关性分析和评价。结果表明,供试材料的淀粉特性差异较大,总淀粉含量变幅为51.26％～66．70％,平均59．89％;峰值粘度变幅为194～1135BU,平均710BU;直链淀粉含量和峰值粘度间存在极显著的相关性。在供试材料中,NB63—1、藏青80、康青3号等5个品种的总淀粉含量都超过了65％;北青三号和阿青4号的直链淀粉含量小于15%;而康青6号、拉萨勾芒等4个品种的直链淀粉含量在29％以上;NB63—1、康青3号、北青6号等4个品种的糊化温度在59℃以下,峰值粘度又都在1000BU以上。这些品种可用作未来优质专用青稞选育的种质材料。     

Digestibility of starch fractions in wholegrain rolled oats

A practical in vitro method was tested for measuring differences in rapidly digested starch (RDS measured at 20 min), slowly digested starch (SDS between 20 and 120 min) and inaccessible digestible starch (IDS as carbohydrate digested after homogenising at 120 min to release inaccessible starch and digesting for a further 40 min) in wholegrain cereal samples retaining some grain structure, using in vitro pancreatic digestion after a “gastric” pepsin–HCl pre-treatment. The persistence of digestive capacity, the influence of homogenising on digestive capacity, and the benefit of adding supplementary amyloglucosidase after 120 min digestion were tested as effects on capacity to digest a further 2.5 g portion of finely ground wholegrain added to selected 120 min digests. The results showed that sufficient digestive capacity remained to digest the IDS released by homogenising the wholegrains after 120 min prior digestion. The method measured all fractions with good precision (CV < 6%), and was shown to be capable of measuring a wide degree of variation in starch fractions of differing digestibility in rolled oats from an oat breeding population, so may be useful in detecting the effects of variations in grain structure that influence the glycemic impact of wholegrain products.     

Rheology and texture of starch gels containing iodine

The objective of this study was to investigate the textural and rheological properties of starch gels containing different levels of iodine. The research goal is to understand the architectural organization of granules by utilizing iodine as a marker. Corn (CS), wheat (WS), potato (PS) and tapioca (TS) starch gels containing varying concentrations of iodine were prepared in a Rapid Visco Analyser (RVA). The gels were analysed by using a rheometer for small-deformation properties and also by using a texture analyser for large deformation properties following storage for 1 or 7 days. With increasing concentration of iodine, the firmness of gels decreased and the extent of decrease varied for the different starches. The extent of change in complex modulus (G*) of the gels was highest in the A-type starch, CS, but was different from WS, which is also an A-type starch. The G* for PS gels (B-type) decreased with the addition of 0.1% iodine but there was no significant difference upon increasing the iodine concentration. The G* for the C-type TS gels was lower than 15 Pa. These observations suggest a difference in the microstructure of the starch gels formed, likely resulting from differential leaching of polymers.     

The effect of gluten on the retrogradation of wheat starch

The retrogradation of amylopectin in a wheat starch and a wheat starch/gluten (10:1) blend prepared by extrusion and containing 34% water (wet weight basis) was studied using X-ray diffraction, differential scanning calorimetry and NMR relaxometry during storage at constant water content and temperature (25 °C). For both samples, amylopectin ‘fully’ retrograded after 2–3 days storage, i.e. the different parameters monitored with time to follow the retrogradation had reached their maximum value, and crystallised predominantly into the A polymorph. Under the experimental conditions used, there was no evidence of any significant effects of the presence of gluten on the kinetics, extent or polymorphism of amylopectin retrogradation.     

糯大麦与非糯大麦种子萌发和胚乳消亡的比较研究

为探明糯大麦与非糯大麦种子萌发和胚乳消亡的差异,以典型糯大麦品种白青稞和甘垦5号以及非糯大麦品种扬农啤10号和苏裸麦1号为材料,采用组织化学染色、树脂半薄切片和光学显微镜观察等方法,研究了糯大麦与非糯大麦胚乳的消亡和淀粉体形态变化。结果表明,在种子萌发过程中,糯大麦种子萌发较好,发芽率较高;糯大麦和非糯大麦颖果胚乳的消亡过程基本相同,呈现出一定的时序性,均是从近胚端的腹部开始消耗,逐渐延伸至背部,最后沿腹沟延伸至远胚端;在大麦种子萌发过程中,近糊粉层中淀粉体最先消亡,小淀粉体早于大淀粉体被消耗;与非糯大麦相比,糯大麦胚乳淀粉体的消亡时间较早、消亡速率较快。概言之,糯大麦与非糯大麦种子萌发过程中胚乳的消亡规律基本相同,但糯大麦种子萌发好,胚乳消亡快。     

Hydrolysis of granular corn starch with controlled pore size

Native corn (Zea mays L.) starch granules were hydrolyzed using glucoamylase at 50 °C for 1–8 h. The degree of hydrolysis over time was analyzed by the concentration of glucose released into solution. The pore sizes of hydrolyzed starch granules increased gradually with the degree of hydrolysis, as evidenced by scanning electron micrographs. It was deduced that every pore on the surface of granules was formed by hydrolysis of one enzyme molecule, so the size of pores distributed on the surface of starch granules was almost homogeneous for the same hydrolysis time. The specific surface area (S_BET), porosity, adsorptive capacity and mean pore radius of porous starch granules were determined to analyze the effect of digestion time on granule properties.     

A modeling approach in the interpretation of starch pasting properties

In this work, starch pasting curves were reinterpreted with suitable models to show deficiency of widely used starch pasting parameters in the literature. The aim was to attract scientific attentions to the behavior and dynamics of the curve with respect to process parameters instead of values on the curve. Mainly, pasting curve was separated into four parts combining sigmoidal gelatinization model, exponential thinning model and Arrhenius temperature model. For the gelatinization part, starch swelling rate, water binding capacity and starch resistivities to heat and shear might be compared by the proposed model parameters. Amylose and damaged starch contents of starches had a good relation with Arrhenius and exponential model parameters. Activation energy values gave clues about retrogradation rate of starches and changed between 7142 and 17,327 kJ/mol.     

Effects of starch composition and type of non-solvent on the formation of starch nanoparticles and improvement of curcumin stability in aqueous media

In this study, we evaluated the effects of amylopectin/amylose ratio and non-solvent type on starch nanoparticle formation including the average particle size, polydispersity index, size distribution, and nanoparticle morphology using dynamic light scattering (DLS) and scanning electron microscopy (SEM). The most uniform particles were obtained from normal corn starch with ethanol. The average particle size was 98.8 ± 1.8 nm using DLS while combination of size distribution study and SEM images showed that particle size ranged between 60 and 90 nm. A bimodal distribution was observed with two defined groups of nanoparticles when waxy corn starch (Amioca) was nanoparticulated with ethanol. SEM images of freeze dried samples and DLS size distribution curves of fresh samples showed that high amylose starch including Hylon V and Hylon VII gave uniform, spherical and small nanoparticles in the size range of 20–60 and 15–50 nm, respectively. The smallest nanoparticles were fabricated by precipitation with methanol, followed by ethanol and the largest nanoparticles were formed using acetone. Re-dispersion of nanoparticles was good when nanoparticles were fabricated using ethanol and acetone especially for Amioca, whereas redispersion of samples in aqueous PBS solution, precipitated using methanol was difficult especially in the case of Hylon VII. Stability of curcumin in the presence of 1 mg/ml native starch nanoparticles was much higher (83.7 ± 3.1%) than curcumin in phosphate buffered saline at pH 7.0 (5.5 ± 1.5%) over 10 days at ambient temperature. Interaction between iodine-potassium iodide solution and starch nanoparticles showed that the helical structures of amylose and amylopectin molecules remain in the nanoparticles and curcumin may interact with these helical structures giving it the stability which is not observed in water.     

Kinetics of starch digestion and functional properties of twin-screw extruded sorghum

The time-course of starch digestion in twin-screw extruded milled sorghum grain was investigated using an in-vitro procedure based on glucometry. The sorghum grains were hammer-milled, and extruded at three levels each of moisture and screw speed. Irrespective of the extrusion conditions, extruded and non-extruded milled sorghum grain exhibited monophasic digestograms, and the modified first-order kinetic and Peleg models adequately described the digestograms. Extrusion increased the rate of digestion by about ten times compared with non-extrudates. Starch gelatinisation varied in the extrudates, and microscopy revealed a mixture of raw, gelatinised and destructured starch and protein components in the extrudates. Starch digestion parameters significantly (p < 0.05) correlated with extruder response and various functional properties of the extrudates. Extrusion conditions for maximum starch gelatinisation in milled sorghum grain for fastest digestion as an efficient animal feed were interpolated, as well as the conditions for directly-expanded extrudates with potential for human food, where minimum starch digestion is desired.