Effects of prolamin on the textural and pasting properties of rice flour and starch

Prolamin is a major class of rice proteins but its influence on the physicochemical properties of rice is not clear. Rapid Visco Analyser (RVA) and TA-XT2 TPA textural analyses were performed on rice starch with the addition of prolamin extracted from three rice cultivars (Hitomebore, M103 and Amaroo), and on rice flour with the prolamin removed by propan-2-ol extraction. Addition of prolamin to rice starch was found to cause a significant (P<0.05) increase in RVA breakdown viscosity but significant (P<0.05) decreases in hardness, adhesiveness and gumminess of the starch gel. Similarly, when prolamin was removed from rice flour, exactly the opposite effect was observed. Addition of prolamin to rice starch also caused it to absorb water faster during cooking but the gelatinised starch absorbed less water compared with control samples without prolamin.     

Characterisation and functional properties of Australian rice protein isolates

Albumin, globulin, glutelin and prolamin fractions were isolated from an Australian rice variety (cv. Langi) and characterised by yield, protein content and molecular weight profile using both capillary electrophoresis (SDS-CE) and sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). The influence of pre-extraction enzymatic hydrolysis of starch and heating to 70 °C was also investigated, as was the extraction of the glutelin fraction without prior removal of the albumin and globulin fractions. Pre-extraction treatment affected mainly the albumin fraction, increasing dry matter yield but reducing protein content. SDS-CE was able to separate the protein fractions over a wider molecular weight range than SDS-PAGE, and the peaks from SDS-CE showed slightly higher molecular weight compared to equivalent bands from SDS-PAGE. The glutelin fraction extracted without prior removal of albumin and globulin fractions had different characteristics compared to those obtained by conventional extraction methods. Pre-extraction hydrolysis of starch did not significantly affect the emulsifying, foaming and gelling properties of extracted protein. Although rice glutelin had poor solubility, emulsifying and foaming properties in aqueous systems, it had good gelling properties which could be important for food applications.     

Characterization of rice storage proteins by SE-HPLC and micro z-arm mixer

While the effect of protein content and composition on the functional properties of wheat flour is well studied, our knowledge on the same properties of rice flour is limited. This work was conducted to study the relationship between the dough mixing properties of flour from different rice cultivars and protein content and composition. An efficient sonication-based two-step extraction procedure was applied to isolate rice flour proteins. The size-exclusion HPLC (SE-HPLC) method, originally developed for separating wheat proteins, was applied with some minor modifications in order to study the size distribution of rice flour proteins. Four fractions were distinguished on the SE-HPLC profile and were further characterized by SDS-PAGE. Fractions I–III consisted of glutelins, while fraction IV contained albumin, globulin and prolamin proteins. When rice dough was characterized on the basis of mixing parameters in a micro z-arm mixer, significant differences were observed depending on the protein composition of the flour. Statistical analysis results indicated that the functional properties of the flour from different rice cultivars were associated with the amount of polymeric proteins and their size distribution.     

Antioxidative properties of partially purified barley hordein, rice bran protein fractions and their hydrolysates

Antioxidative properties of proteins from barley and rice bran and their hydrolysates were examined. Three major hordein fractions of barley, B, C and D hordeins, were partially purified by gel filtration. Albumin, globulin, prolamin and glutelin fractions of rice bran were fractioned by the Osborne method. Hydrolysates of these protein fractions were prepared by digesting with pepsin followed by trypsin. Antioxidant properties in terms of antioxidative activity against linoleic acid peroxidation and reducing activity without the lipid adjuvant were investigated. The globulin fraction from rice bran protein revealed the strongest antioxidative activity throughout the incubation time of 7 days (p ≤ 0.05). The albumin fraction of rice bran protein showed the highest reducing activity (6964 μmol of Fe²⁺) followed by globulin, prolamin, glutelin and hordein fractions with activities of 2904, 2017, 1809 and 1333 μmol of Fe²⁺, respectively (p ≤ 0.05). Partially purified C hordein exhibited the highest reducing activity compared with B and D hordeins. Protein hydrolysates obtained after digestion with pepsin and trypsin exhibited much greater antioxidative, as well as reducing, activities than those from before digestion.     

Transglutaminase polymerisation of buckwheat (Fagopyrum esculentum Moench) proteins

Recently, screening of transglutaminase (TGase) treatment on several gluten-free cereals revealed significant improvements on the baking performances of buckwheat flour by promoting protein networks. In this study, the impact of TGase on the protein fractions of buckwheat flour was investigated in order to better understand the activity and specificity of the enzyme. Albumin, globulin, prolamin and glutelin fractions were extracted from the flour and incubated with TGase. Capillary electrophoresis, two dimensional (2D) gel electrophoresis and size exclusion chromatography (SEC) were performed on each fraction. Capillary electrophoresis and 2D gel electrophoresis revealed that buckwheat main storage proteins, i.e. 2S albumin, 13S and 8S globulin, were cross-linked after TGase treatment. SEC showed the presence of high molecular weight (HMW) protein polymers in the TGase-treated albumin and globulin fraction. Analysis of the amino acid composition of the fractions revealed high amounts of glutamine and lysine residues in all fractions. In conclusion, the increase in the average molecular weight of buckwheat proteins and the formation of HMW protein polymers after TGase treatment are responsible for the improved functionality of buckwheat flour in terms of breadmaking potential. The enzyme was revealed to be not fraction specific as all fractions were TGase-reactive.     

Transglutaminase polymerisation of buckwheat (Fagopyrum esculentum Moench) proteins

Recently, screening of transglutaminase (TGase) treatment on several gluten-free cereals revealed significant improvements on the baking performances of buckwheat flour by promoting protein networks. In this study, the impact of TGase on the protein fractions of buckwheat flour was investigated in order to better understand the activity and specificity of the enzyme. Albumin, globulin, prolamin and glutelin fractions were extracted from the flour and incubated with TGase. Capillary electrophoresis, two dimensional (2D) gel electrophoresis and size exclusion chromatography (SEC) were performed on each fraction. Capillary electrophoresis and 2D gel electrophoresis revealed that buckwheat main storage proteins, i.e. 2S albumin, 13S and 8S globulin, were cross-linked after TGase treatment. SEC showed the presence of high molecular weight (HMW) protein polymers in the TGase-treated albumin and globulin fraction. Analysis of the amino acid composition of the fractions revealed high amounts of glutamine and lysine residues in all fractions. In conclusion, the increase in the average molecular weight of buckwheat proteins and the formation of HMW protein polymers after TGase treatment are responsible for the improved functionality of buckwheat flour in terms of breadmaking potential. The enzyme was revealed to be not fraction specific as all fractions were TGase-reactive.     

稻米蛋白质组分及总蛋白质含量与淀粉RVA谱特征值的关系

 利用低谷蛋白含量（low glutelin content, LGC）粳稻品种W1721与大面积应用的杂交粳稻恢复系轮回422构建了重组自交系群体。根据种子总蛋白质的SDS PAGE带型,推断各单株LGC位点的基因型。在排除脂肪的影响下,比较了基因型LGCLGC（谷蛋白含量较醇溶蛋白低）与基因型lgclgc（谷蛋白含量较醇溶蛋白高）在RVA谱特征值上的差异,以及同一基因型内总蛋白质含量与淀粉RVA谱特征值的关系。结果表明基因型LGCLGC的崩解值和回复值的平均值分别显著和极显著大于基因型lgclgc;LGC对崩解值和回复值的相对加性效应分别为3.91%和4.45%,贡献率分别为4.81%和12.81%。LGCLGC型的稻米,总蛋白质含量与崩解值和消减值分别存在显著的负线性回归和正线性回归关系,贡献率分别为9.86%和11.48%;lgclgc型的稻米,总蛋白质含量与消减值和回复值都存在极显著的负线性回归关系,贡献率分别为1341%和27.88%。结合前人研究认为,稻米食味品质受谷蛋白相对于醇溶蛋白的含量以及总蛋白质含量的影响;而总蛋白质含量对食味品质的影响因谷蛋白相对于醇溶蛋白的含量的不同而异。因此,育种工作中应当把谷蛋白相对于醇溶蛋白的含量以及总蛋白质含量同时作为选择指标。     

Characterization and In Vitro digestibility of rice protein prepared by enzyme-assisted microfluidization: Comparison to alkaline extraction

Microfluidization followed by density-based separation was employed to extract protein from broken rice by disrupting protein-starch agglomerates. Follow-up enzyme treatments (amylase and glucoamylase) were performed to further improve the purity of the protein-rich fraction. High protein recovery (81.87%) and purity (87.89%) were obtained. The protein composition, solubility, structural properties, and in vitro digestibility of rice proteins prepared by enzyme-assisted microfluidization (EM-RP) and alkaline extraction (AE-RP) were compared. EM-RP was mainly composed of glutelin, which had low solubility and native structure. By contrast, large quantities of prolamin and globulin appeared in the AE-RP except glutelin, leading to the richness of glutamic acid/glutamine, leucine, aromatic and charged amino acids in the AE-RP. Compared to AE-RP, EM-RP showed higher digestibility due to the richness of glutelin (an easy-to-digest protein), as evidenced by higher nitrogen release during pepsin-trypsin digestion. The presence of prolamin (an indigestible protein) in AE-RP decreased protein digestibility although alkaline extraction improved its hydrolysis. These results suggest that enzyme-assisted microfluidization could be an effective technique to non-destructively and selectively extract rice glutelin.     

QTL mapping for crude protein and protein fraction contents in rice (Oryza sativa L.)

Improvement of rice storage proteins is important in rice breeding for high nutritional quality. Seventy-one recombinant inbred lines (RIL) derived from a cross between japonica variety Asominori and indica variety IR24 were used to study the inheritance of crude protein and protein fraction contents in rice. A total of 16 QTL were identified and mapped on eight chromosomes. Several QTL affecting contents of different protein fractions were mapped in the same chromosomal region. In particular, two QTL with a significant contribution were identified to simultaneously affect prolamin and glutelin contents. One QTL denoted as qCP-12 affecting crude protein content (CP) was located in the same region as QTL qGLT-12 affecting glutelin content, in agreement with the positive correlation between glutelin level and protein content. QTL with larger genetic effects were further confirmed using two sets of chromosome segment substitution lines (CSSL), where Asominori and IR24 were used as the recurrent parents. By QTL comparative analysis, two QTL for CP, three for globulin content and one for prolamin content were located in the vicinity of CP QTL previously identified in polished rice. Application of these results in rice breeding is discussed.     

我国江浙地区粳稻品种间的植酸与蛋白质组分差异及其相关性

选用我国江浙地区近期育成的晚粳稻品种（系）29个,对它们在浙江杭州相同栽培条件下籽粒中的植酸、蛋白质含量与其组分差异及其相关性进行了分析。供试29个品种(系)植酸含量的平均值为0.868%,变化幅度为0.699%~1034%,其中秀水系列品种的植酸含量一般较低,而武育粳系列和淮字系列品种（系）的植酸含量则相对较高;供试29个品种材料总蛋白质含量的平均值为8.722%,其含量大致呈正态分布。在4种蛋白质组分中,谷蛋白、球蛋白和清蛋白在品种间的变异系数较大,而醇溶蛋白则相对较小,且在分布频率上也表现出品种类型间的差异特征;除谷蛋白与总蛋白质之间存在着极显著的正相关外,供试29个品种(系)的植酸含量与蛋白质含量及4种蛋白质组分含量间并无密切的关系。     

Extractability and chromatographic separation of rice endosperm proteins

The molecular weight (MW) distribution of proteins extracted with different solvents from defatted rice endosperm was examined by size exclusion-high performance liquid chromatography (SE-HPLC) with 2.0% sodium dodecyl sulfate (SDS) (w/v) as mobile phase. The resulting protein peaks were further characterized by SDS-PAGE. Under the experimental conditions, 2.0% SDS extracted 64% of the proteins. Adding 6.0 M urea resulted in a 15% increase in extractability (up to 79%). With using 20–100 mM NaOH, 70–81% of the proteins were extractable. Maximum extractability was reached with 2.0% SDS, 6.0 M urea and 0.5–1.5% dithiothreitol (DTT). Apparent MW profiles of rice endosperm proteins allowed classification into six fractions of decreasing apparent MW. Fraction VI contained the low MW albumin, globulin, and prolamin protein material. Fractions IV and V originated from α and β glutelin subunits, respectively. The polypeptides of fraction III consisted of an α and a β subunit linked by an intermolecular disulfide bond. The polypeptides of fractions I and II were dimers, trimers or more highly polymerized forms of the (α–β) glutelin subunit dimer in fraction III. While the work confirmed that rice glutelin is composed of polymers of α and β subunits, remarkably, higher MW glutelin aggregates (fractions I–III) only partly dissociated on reduction. Low MW protein material (fraction VI) was entrapped in the aggregated protein network and was released on reduction. The rapid and reproducible SE-HPLC method developed for rice protein separation allows a more quantitative approach than SDS-PAGE.     

Temperature during grain ripening affects the ratio of type-II/type-I protein body and starch pasting properties of rice (Oryza sativa L.)

The starch and protein properties of rice grain are important factors for sake brewing and these properties are reported to be influenced by temperatures during grain ripening. Amylose content, nitrogen content, protein composition, pasting properties measured by a Rapid Visco Analyser (RVA), and their relationship to temperatures during ripening were investigated in a rice cultivar, Yamadanishiki, which was grown under various conditions in the same experimental field. The average temperature after heading was significantly correlated with amylose content and RVA properties, but not with nitrogen content. Under high temperatures during ripening, a decrease in prolamin, which accumulated in type I protein body (PB-I), and an increase in glutelin, which deposits in type II PB (PB-II), were recognized. The ratio of PB-II/PB-I and RVA pasting temperatures were distinctly increased as the temperature became higher. High temperatures during grain ripening would lead to difficulties in digesting steamed rice grains by Aspergillus oryzae, together with ease in digesting rice protein. The average temperature of 11–20 days after heading showed a higher correlation coefficient than that of 1–10 or 21–30 days, implying that temperatures during the middle stage of grain development would be important in determining the rice component that relates to brewing properties.     

Prolamin,a rice protein,augments anti-leukaemia immune response

Rice (Oryza sativa), an important cereal as a staple food worldwide, can be used in gluten-free diets. This study aimed to isolate and characterize the active rice proteins and assess the anti-leukaemia response via in vitro and ex vivo experiments. Temperature-stable protein-containing extracts augmented this effect. Proteomic analysis identified various protein spots known with functions involving metabolism-related, transport, storage, antioxidation, development, and disease-resistance proteins. Among these, storage proteins were the most abundant. To avoid masking the other relatively scanty rice storage proteins, albumin, globulin, glutelin and prolamin were separated and quantified. The human peripheral blood mononuclear cells-conditioned medium (PBMC-CM) prepared from prolamin treatment showed an increment in production of tumor necrosis factor-α. Human leukaemia U937 cells cultured in the presence of prolamin-prepared PBMC-CM were inhibited in growth capacity and were triggered differentiation toward monocytes. Neutralization of prolamin by polyclonal antibody attenuated its activity. Rice prolamin, undetectable by wheat gliadin-specific antibody, has greater anti-leukaemia activity than wheat proteins glutenin and gliadin. In conclusion, rice prolamin is effective in activating human anti-leukaemia immunity and may not induce unwanted inflammatory diseases.     

Effects of protein oxidation on gelatinization characteristics during rice storage

Gliadin and glutelin are major rice storage proteins. In this study, we evaluated changes in these proteins to determine the influence of pasting properties on rice storage. Notably, the physical and chemical properties of these proteins changed steadily. We analyzed protein oxidation indexes and determined correlations between the protein oxidation index and gelatinization property index. The results showed that significant oxidation of gliadin and glutelin occurred during the process of rice storage (P < 0.05). Protein oxidation had a significant impact on pasting properties. For glutelin, changes in the structure and function of the protein during rice storage had a significant impact on the five rice pasting property indexes, including peak viscosity (PV), holding strength (HS), breakdown, final viscosity (FV), and setback (SB; P < 0.05). In particular, the levels of carbonyl compounds and active sulfur and the surface hydrophobicity of glutelin were significantly correlated with rice pasting property indexes (P < 0.05). However, gliadin only significantly affected three indexes, i.e., PV, HS, and FV (P < 0.05). Thus, these findings suggested that the carbonyl compounds, active sulfur, and surface hydrophobicity of glutelin could be used as sensitive indexes for changes in rice quality evaluation.     

醋制对黑豆蛋白含量及多肽ACE抑制活性的影响

为考察醋制对黑豆蛋白含量及多肽ACE抑制活性的影响,为对黑豆醋浸过程中蛋白组分及含量变化进行分析,并对相应ACE抑制肽活性进行测定。黑豆经醋浸不同时间后,采用顺序抽提法提取清蛋白、球蛋白、醇溶蛋白和谷蛋白,并采用凯氏定氮法及SDS-PAGE电泳进行定量和定性分析;各蛋白组分经胃蛋白酶水解,经超滤(截留分子量3 k D)后收集滤液,获得多肽组分,RP-HPLC法进行ACE抑制活性评价。结果表明:经醋浸14 d后,黑豆总蛋白含量变化幅度小于±0.5%;清蛋白含量由55.13%(0 d)降低至9.61%(14 d);球蛋白由7.04%(0 d)增加到20.34%(14 d);醇溶蛋白由0.81%(0 d)增加到1.72(14 d);谷蛋白由21.11%(0 d)增加到59.45%(14 d),含量最高。醋浸处理降低了清蛋白源多肽的ACE抑制活性,但提高了球蛋白、醇溶蛋白及谷蛋白多肽的ACE抑制活性,其中,谷蛋白多肽抑制率由18.18%(0 d)增加至37.58%(14 d),抑制活性升高。醋浸可改变黑豆各蛋白组分的含量和对应多肽的ACE抑制活性,其中,谷蛋白含量及其多肽ACE抑制活性均有增加。研究结果表明可进一步采用分离纯化技术从谷蛋白多肽中获得高活性降压肽。     

郑麦1860品质特性及面条加工适用性分析

为明确郑麦1860的品质及加工适用性,对2020-2021年河南省29个地点收获的郑麦1860的籽粒性状、磨粉品质、面团流变学特性、淀粉糊化特性和面条加工品质指标进行了测定与分析。结果表明,郑麦1860为优质中强筋面条小麦品种,特点是籽粒大、容重高、出粉率高,面条弹性好、色泽白亮、褐变较轻。在不同地点间郑麦1860的多数品质性状变异程度小,品质表现较稳定;相比较而言,其粉质参数、拉伸参数变异程度较大,淀粉糊化特性和面筋特性变异程度次之,籽粒品质性状、磨粉品质和面条加工参数变异程度较小。籽粒硬度、粒径、容重与出粉率呈显著正相关;容重、籽粒蛋白质含量、面粉灰分含量与面条总评分呈显著负相关;面粉色泽L^*值、吸水率、峰值黏度、低谷黏度、终值黏度、回生值与面条总评分呈显著正相关。综上所述,籽粒蛋白质含量、面粉色泽和淀粉糊化特性是影响郑麦1860面条品质的主要因素。郑麦1860制作的面条感官评分高、品质稳定,在豫北、豫东和豫南等区域种植均适合制作优质面条。     

高直链淀粉突变体Goamy2的淀粉糊化特性及淀粉结构分析

以高直链淀粉及突变体材料Goamy2及其野生型亲本Ilpumbyeo米粉为材料,比较了它们在理化性质和糊化特性上的差别,并通过体积排阻色谱(Size Exclusion Chromatography,SEC)分析其水溶性和不溶性淀粉组成结构差别。从冷水溶性成分来看,Goamy2主要以小分子DP141～DP2(DP为聚合度)为主,而Ilpumbyeo中带分支的链DP4100～DP64也就是相对大分子所占的比例较高,达到33.3%。从热水溶性淀粉成分来看,突变体Goamy2直链淀粉和支链淀粉的比例大致为4∶1,而Ilpumbyeo直链淀粉和支链淀粉比例大致为1∶1。至于两个材料的热水溶性支链淀粉链长分布,长短链比例差别较大,Goamy2长短链比率为0.67,而Ilpumbyeo为0.32。从热水不溶性成分来看,两个材料均以支链淀粉为主,直链淀粉所占的比例很低,均不到5%。热水不溶性支链成分的链长分布中,野生型亲本Ilpumbyeo以短链为主,短链所占的比例高达72.5%,而突变体Goamy2短链和长链几乎各占一半。     

Effects of added water and retrogradation on starch digestibility of cooked rice flours with different amylose content

Flours derived from rice varieties with different amylose content possess distinct physicochemical and molecular properties. The aim of this study was to determine optimal processing conditions for preparing rice flour-based foods with reduced starch digestibility. To do so, we evaluated the in vitro starch digestibility of rice flours with five varieties. Reducing the amount of water (from 10-fold to 4-fold of rice flour) used for cooking rice flour lowered its starch digestibility, and the magnitude of the decrease was positively correlated with amylose content. When retrogradation of cooked rice flour proceeded for 7 days, the digestibility of high-amylose rice flours declined rapidly in the first 3 days, whereas the digestibility of low-amylose rice flours declined continuously. Our analysis also demonstrated that the chain length distribution of starch molecules and the final and setback viscosity pasting properties were the most important parameters affecting the digestibility of rice flours. Based on our results, it appears possible to reduce rice starch digestibility by establishing optimum processing conditions for different varieties. We suggest a 7-fold addition of water and retrogradation for 1 day for high-amylose rice varieties and a 4-fold addition of water with 3 days of retrogradation for low-amylose rice.     

Difference of Phytic Acid Content and its Relation to Four Protein Composition Contents in Grains of Twenty-nine japonica Rice Varieties from Jiangsu and Zhejiang Provinces, China

Twenty-nine japonica rice varieties collected from Jiangsu and Zhejiang Provinces, China were planted in Hangzhou, China, to investigate the phytic acid content in brown rice and its frequency distribution as well as the correlation among the contents of phytic acid, total protein and four protein compositions in brown rice. The phytic acid content in brown rice ranged from 0.699% to 1.034%, with a mean of 0.868% for the 29 tested rice varieties. Xiushui series rice varieties generally exhibited lower phytic acid level than Wuyujing and Huai series rice varieties. A rough normal distribution, with a mean of 8.722%, was observed for the total protein contents in the tested varieties. Of the four protein compositions, the glutelin, globulin and albumin contents had larger coefficient of variation than the prolamin content, although the difference in prolamin content was genotype-dependent. No significant correlation was found between the phytic acid and four protein composition contents, whereas the total protein content was significantly and positively related to the glutelin content in brown rice.     

Effect of germination time on physicochemical properties of brown rice flour and starch from different rice cultivars

The present work was designed to obtain information on the effect of germination time on the selected physicochemical properties of brown rice flour and starch prepared from three different rice cultivars. Changes in total starch, amylose and amylopectin contents of flour, amylopectin/amylose ratio and molecular weight of starch, gelatinization, pasting, rheological, and morphological properties of flour and starch during 5 days of germination were investigated. Significant changes of pasting and rheological properties of brown rice flour were found during germination, but only small changes of these properties could be found in isolated starch. Scanning electron micrographs of flour showed that the continuous matrix structure of flour was highly destroyed after germination and scanning electron micrographs of isolated starch showed that after three days of germination, pits and holes were discovered on the surface of some starch granules. Germination had little effect on the average molecular weight of starch, but the polydispersity value in germinated brown rice (2–5 days germination) was higher than that in non-germinated brown rice. The changes observed in physicochemical properties of brown rice flour and starch after germination provided a crucial basis for understanding flour and starch modification mechanisms with potential applications for an industrial scale.