9种大豆制品中转基因成分定性PCR检测

本研究从豆粕、豆粉、豆腐等9种大豆产品中提取到高质量的DNA,在对内标准基因Lectin进行扩增的基础上,设计相应的特异性引物,建立了稳定的定性PCR反应体系,对35S启动子基因、外源目的基因Cp4-epsps基因进行了扩增,发现豆粕、豆奶粉、豆腐、豆皮、豆豉中含有转基因成分,而其它产品中未发现转基因成分存在.     

Recent progress on biotechnology of soybean proteins and soybean protein food products

Recently it has been recognized that the action of the enzymes contained in soybeans has negative effects on the qualities of soybean protein foods. Based upon these findings, new manufacturing techniques for high quality soybean protein foods have been developed through the control of such enzyme action. For instance, new soybean seeds lacking lipoxygenases have been developed which reduce the production of grassy‐beany flavors. The gel hardness of tofu can also be controlled through the action of the lipoxygenases contained in soybeans. In addition objectionable after‐tastes develop through the hydrolysis of soybean isoflavones by the ß‐glucosidases which are found in soybeans.

Genetic engineering of soybean proteins made a great progress in improving their functional and nutritional qualities through the modifications of genes encoding for soybean storage proteins. The application of a bioreactor for soy sauce is also described as an example of the progress on the biotechnology.     

豆腐渣代替豆粕酿造酱油的研究

该文研究了用豆腐渣代替豆粕生产酱油的工艺，通过试验确定了各生产工序的操作要点等。并对成曲蛋白酶活力、成品的感官指标、理化指标，全氮利用率等进行了对比。     

常见大豆制品中蛋白质的体外消化特性

本研究以常见大豆制品（大豆分离蛋白（soy protein isolate,SPI）、微波大豆、水煮大豆、豆浆、氯化钙豆腐和氯化镁豆腐）为研究对象,采用标准的静态体外消化模型——INFOGEST 2.0,测定不同时间点胃肠消化产物的完整蛋白、分子质量分布、游离氨基浓度和粒径大小,以探究大豆蛋白的消化特性。结果表明,不同加工方式均能促进大豆蛋白的消化。经过胃消化阶段后,几种大豆制品中部分蛋白被消化。而肠消化后,大豆蛋白均被彻底消化为短肽。相较于SPI,大豆经微波和水煮处理后,大豆蛋白的游离氨基浓度明显增加,但微波加热后大豆蛋白消化得更彻底;豆浆中大豆蛋白的消化程度低于氯化钙豆腐和氯化镁豆腐,两种豆腐中大豆蛋白的消化程度没有明显差异。总之,不同加工方式处理大豆均能不同程度地促进大豆蛋白的消化。本研究有助于更好地了解不同加工方式得到的大豆制品中大豆蛋白的消化特性,为人们摄取植物蛋白提供参考。     

酱油渣蛋白质的分离、鉴定和氨基酸组成特征研究

本文以酱油上清液为对照,以过滤-冷冻真空干燥法制备了酱油渣。首先,对比研究了酱油渣的常规理化指标。其次,以SDS-PAGE、MALDI-TOF/TOF MS和HPLC法对酱油渣和酱油上清中蛋白质分子量分布、来源与种类及氨基酸组成特征进行了研究。结果显示酱油渣（湿渣）主要由15.98%的碳水化合物、10.19%的蛋白质、10.10%的食盐和59.50%的水分组成。SDS-PAGE和MALDI-TOF/TOF MS分析结果显示酱油渣蛋白质主要由分子量约为19.3 kDa（a, 59.40%）、31.8 kDa（b, 12.91%）和12.3 kDa（c, 27.69%）的蛋白质亚基组成,经质谱鉴定分子量为19.3 kDa的多肽（a）为Glycinin G4亚基碱性端B3。酱油渣蛋白质氨基酸组成分析表明,与酱油上清中蛋白质相比,酱油渣蛋白质中疏水性氨基酸含量和平均疏水值明显高于酱油上清中蛋白质。上述差异是酱油渣蛋白质氨基酸组成的主要特征,也是酱油渣蛋白质水溶性差和形成的主要原因。     

Effects of Transglutaminase on SDS-PAGE Patterns of Wheat, Soy, and Barley Proteins and their Blends

Transglutaminase (TG) catalyzes the formation of nondisulfide covalent crosslinks between pep‐tide‐bound glutaminyl residues and ε‐amino groups of lysine residues in proteins. TG can be used for polymerizing proteins from 1 or more sources through formation of intermolecular crosslinks. This study investigated, by sodium dodecyl sulfate‐polyacrylamide gel electrophoresis, polymers created by the action of TG on proteins of wheat, soy, barley, wheat‐soy, and wheat‐barley blends. Electrophoretic results showed that with increasing incubation time, the crosslinking reaction is substantially increased, with progressive decrease or disappearance of some protein monomers. Densitometric results showed that soy proteins were the best substrates of TG while barley and wheat proteins were similar in reactivity.     

High-pressure treatment effects on proteins in soy milk

Effects of high-pressure treatment on the modifications of soy protein in soy milk were studied using various analytical techniques. Blue shifts of λ_max could be observed in the fluorescence spectra. Spectrofluorimetry revealed that the soy protein exhibited more hydrophobic regions after high-pressure treatment. Electrophoretic analysis showed the change of soy protein clearly and indicated that soy proteins were dissociated by high pressure into subunits, some of which associated to aggregate and became insoluble. High-pressure denaturation occurred at 300 MPa for β-conglycinin (7S) and at 400 MPa for glycinin (11S) in soy milk. High pressure-induced tofu gels could be formed that had gel strength and a cross-linked network microstructure. This provided a new way to process soy milk for making tofu gels.     

基于LF-NMR方法分析电子束辐照技术影响大豆浓缩蛋白粉水分的变化

本研究借助低场核磁共振技术(low field nuclear magnetic resonance,LF-NMR)测定经不同电子束辐照(electron beam irradiation,EBI)剂量处理后的大豆浓缩蛋白粉的水分分布信息,并通过热变性实验和傅里叶变换红外光谱(Fourier transform infrared,FT-IR)初步探究了EBI对大豆浓缩蛋白粉水分变化的影响及其变化机制。结果表明,大豆浓缩蛋白粉中的主要水组分为与大豆蛋白分子紧密结合的结合水(T_(21),占97.0%±0.7%)。随着EBI剂量的增加,结合水和总水分含量均先增大后减小,在5.40 kGy时达到最大值。经热变性实验和FT-IR光谱图分析得知EBI处理不会改变大豆浓缩蛋白粉的变性温度,但是能引起结合水中的O-H基团与氨基酸中的C=O所形成的分子内H键以及分子间H键振动增强。这些结果不仅表明EBI可通过改变大豆蛋白分子内和分子间的H键作用来影响其水分分布及含量,还为进一步探究经EBI处理的蛋白类食品中的水分迁移机制奠定了基础。     

The impact of Maillard cross-linking on soy proteins and tofu texture

In this paper, we explore the use of Maillard cross-linking agents (formaldehyde, glyceraldehyde and glutaraldehyde) in tofu, to probe the relationship between cross-linking of soy proteins and tofu texture. 1–2 mM glutaraldehyde treatment resulted in an acceptable product with altered microstructure. At 2 mM glutaraldehyde, the fracture force was increased if the reagent was added before soymilk boiling, but decreased if it was added after boiling, when more cross-linking was observed due to the denaturation of the proteins. The fracture forces and microstructures are hard to rationalise in terms of cross-linking, suggesting that other factors, such as non-cross-linking protein glycation, may be responsible for the observed changes. Taken together with the results reported in the previous paper, these findings suggest that protein cross-linking agents may change the functional properties of tofu via non-cross-linking modifications of the sidechains of the amino acid residues, changing their isoelectric point and their gelation properties.     

酱油原料脂肪利用率研究实验报告

本实验选用大豆、豆粕、豆粕和菜粕混合料三组蛋白质原料,采用固态无盐发酵工艺酿造酱油,通过检测酱油中的含油量和氨基酸态氮含量,计算原料的脂肪利用率和蛋白质利用率,结果表明：大豆、豆粕、混合料所酿酱油的脂肪利用率分别为6.15%、35.6%、36.9%;蛋白质利用率分别为56.3%、62.29%、60.38%。     

3‐Chloropropane‐1,2‐diol (3‐MCPD) in Soy Sauce: A Review on the Formation,Reduction, and Detection of This Potential Carcinogen

Soy sauce, a dark‐colored seasoning, is added to enhance the sensory properties of foods. Soy sauce can be consumed as a condiment or added during the preparation of food. There are 3 types of soy sauce: fermented, acid‐hydrolyzed vegetable protein (acid‐ HVP), and mixtures of these. 3‐Chloropropane‐1,2‐diol (3‐MCPD) is a heat‐produced contaminants formed during the preparation of soy sauce and was found to be a by‐product of acid‐HVP‐produced soy sauce in 1978. 3‐MCPD has been reported to be carcinogenic, nephrotoxic, and reproductively toxic in laboratory animal testing and has been registered as a chemosterilant for rodent control. 3‐MCPD is classified as a possible carcinogenic compound, and the maximum tolerated limit in food has been established at both national and international levels. The purpose of this review is to provide an overview on the detection of 3‐MCPD in soy sauce, its toxic effects, and the potential methods to reduce its concentration, especially during the production of acid‐HVP soy sauce. The methods of quantification are also critically reviewed with a focus on efficiency, suitability, and challenges encountered in analysis.     

Characterization of Soy Protein Concentrate Produced by Membrane Ultrafiltration

The functionality of membrane processed soy concentrate was very similar to soy flour in terms of solubility and water hydration capacity. The high emulsifying activity index of soy flour is believed to be reflective of its higher solubility, while surface hydrophobicity is believed to be responsible for an equally high emulsifying activity index in acid precipitated soy isolate. The proteins of soy flour and membrane soy concentrate seem to have most of their hydrophobic residues buried in the interior, while they are exposed in acid precipitated soy isolate. Heating resulted in a decrease in solubility but improved the hydration capacity and emulsifying activity of both soy flour and membrane soy concentrate. The essential amino acid profile of concentrate was comparable to current commercial isolates manufactured by acid precipitation. The majority of the polypeptides present in soy flour were observed to be present in the concentrate. The membrane soy concentrate was determined to have the least soybean aroma when compared to both soy flour and acid precipitated soy isolate.     

5种不同酱油抗氧化活性的对比分析

该研究对比分析了1种黑豆酱油和4种黄豆酱油中总酚、总黄酮、色深物质、氨基酸态氮、无盐固形物、还原糖、总糖、总酸和游离氨基酸等成分的含量及两类酱油的DPPH自由基清除能力、ABTS自由基清除能力、还原力和金属离子螯合能力。此外,采用相关性分析、聚类分析和主成分分析法分析了5种酱油的抗氧化活性成分与抗氧化活性之间的内在联系。结果表明,除氨基酸态氮、无盐固形物、游离氨基酸外,黑豆酱油的其他6种活性成分含量均显著高于其余4种黄豆酱油（p<0.05）;除金属离子螯合能力外,黑豆酱油的其他3种抗氧化活性均显著高于其余4种黄豆酱油（p<0.05）;相关性分析结果表明酱油中总酚、总黄酮和色深物质是造成5种酱油抗氧化活性存在显著差异的主要原因,聚类分析结果和主成分分析结果均表明黑豆酱油的综合品质高于其余4种黄豆酱油。因此黑豆酱油的综合品质显著高于其余4种黄豆酱油。上述研究结果可为黑豆酱油抗氧化活性的深入研究和消费者选择合适的酱油提供数据支撑和科学依据。     

大豆高压短时蒸煮显著提高酱油二次沉淀蛋白降解率

酱油二次沉淀严重影响其外观质量和商品价值。本研究以大豆低压长时(0.24 MPa/18 min)蒸煮工艺为对照,采用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和基质辅助激光解吸附电离串联飞行时间质谱(MALDI-TOF/TOF MS)等方法研究了大豆高压短时(0.54MPa/14min)蒸煮工艺对酱油二次沉淀蛋白和二次沉淀生成量的影响。实验结果表明大豆球蛋白G4中的B3亚基(20 ku)和G1中的A1a亚基(35 ku)是酱油二次沉淀蛋白的主要成分,采用大豆高压短时蒸煮工艺制备的酱油(样品)比其对照酱油中B3亚基减少65.38%,样品中A1a亚基被完全降解;样品中二次沉淀含量比其对照减少72.69%。同时,采用大豆高压短时蒸煮工艺可提高酱油原料蛋白质利用率9.37%、氨基酸态氮14.14%、总氮10.74%、总糖22.36%、还原糖27.87%和无盐固形物12.38%(p0.05)。因此,采用大豆高压短时蒸煮工艺可显著提高酱油二次沉淀蛋白降解率、减少酱油二次沉淀生成量和提高酱油滋味物质含量,同时改善酱油外观质量和滋味。     

Changes in myofibrillar structure of silver carp (Hypophthalmichthys molitrix) as affected by endogenous proteolysis under acidic condition

Effect of endogenous proteolysis on myofibrillar conformational changes under acidic condition was investigated by spectroscopic techniques. The result of intrinsic tryptophan fluorescence showed that tryptophan residues were exposed to polar environment by acid denaturation of protein and participation of endogenous proteolysis. Extrinsic fluorescence indicated that average surface hydrophobicity (S_hANS) decreased by reduction of pH but increased by endogenous proteolysis. Second‐derivative UV‐spectroscopy implied that the decrease of S_hANS under acidic condition may be due to burying of some aromatic amino acid residues into hydrophobic cores induced by acid‐induced aggregation, such as tyrosine, while S_hANS increased by exposure of some hydrophobic amino acids due to the effect of endogenous proteases. Raman spectra verified the changes in microenvironment of tryptophan and tyrosine residues, and showed that aliphatic amino acids were mainly exposed to polar solvent by both acid denaturation and endogenous proteolysis. In addition, new disulphide bonds were less likely to form by acid‐induced aggregation and proteolytic cleavage, while random coils increased and smaller water domains were found.     

添加酱渣对黄豆酱品质影响的研究

为研究添加酱渣对黄豆酱品质的影响,在发酵过程中添加不同比例酱渣,比较黄豆酱中氨基酸态氮、色差、质构、综合感官 评价和风味物质等品质指标变化。 结果表明,随着酱渣添加量在1%～9%范围内增加,黄豆酱中氨基酸态氮含量呈下降趋势,当酱 渣添加量＜3%时,氨基酸态氮含量（1.14 g/100 g）较对照组（1.18 g/100 g）无显著差异（P＞0.05）;当酱渣添加量为1%时,黄豆酱a*值（15.80）和L*值（24.80）最大,且硬度、稠度、黏性指数均与对照组无显著差异（P＞0.05）;添加酱渣后,黄豆酱的风味物质种类及综合 感官评分均高于对照组,当酱渣添加量为7%和3%时,黄豆酱中风味物质种类最多（33种）和综合感官评分最高（75.42分）。 综合考虑, 添加3%酱渣不会降低黄豆酱品质,且在风味物质及综合感官评价方面更具优势,该研究结果可为酱渣在黄豆酱生产中的应用提供 参考。     

黄豆酱油与黑豆酱油抗氧化活性及风味物质的比较

本文研究了相同工艺下,黄豆、黑豆两种蛋白原料制备传统高盐稀态酱油的差异。比较两种酱油基本成分及抗氧化活性的差异,并利用顶空-固相微萃取(HS-SPME)和气相色谱-质谱联用(GC-MS)分析两种酱油挥发性风味物质。结果表明,黑豆酱油总氮、氨基态氮、还原糖等显著高于黄豆酱油(p0.05),黄豆酱油鲜味氨基酸含量更高。黑豆酱油抗氧化活性高于黄豆酱油,相关性分析表明总酚、总黄酮及类黑精是酱油抗氧化活性的重要物质基础。两种酱油挥发性风味物质种类及含量差异显著(p0.05),共鉴定出98种物质,黄豆酱油、黑豆酱油总风味物质种类分别有63种、59种,总峰面积分别为297.6×10~7、213.6×10~7。综合分析表明黄豆酱油醇香突出,黑豆酱油醇、酯及醛酮类主要挥发性成分较均衡,风味协调。     

Suppression of Umami Aftertaste by Polysaccharides in Soy Sauce

Umami is one of 5 basic tastes that make foods savory and palatable. The umami aftertaste is a long‐lasting taste sensation that is important for Japanese broth (dashi) utilized for various Japanese foods. Soy sauce is usually added when making dashi‐based dishes; however, different soy sauces produce distinct effects on the umami aftertaste. In this study, we attempted to identify the substances that cause the suppression of the umami aftertaste in soy sauce by combining sensory analysis, size fractionation, chemical analysis, and enzymatic treatment. The suppressive substance was revealed to be polysaccharides with molecular weights between 44900 and 49700. The results of acid hydrolysis and enzymatic treatment suggested that the polysaccharides were cellulose. These results indicate that a type of water‐soluble cellulose derived from soybean, wheat, or microorganisms has a suppressive effect on the umami aftertaste of soy sauce. Future studies should focus on developing a strategy that regulates the amount of these polysaccharides generated during soy sauce production, to maintain or enhance the umami aftertaste.     

分子间作用力对大豆蛋白凝胶形成的影响

本实验用大豆分离蛋白模拟传统豆腐的凝胶形成过程,研究分子间作用力对大豆蛋白凝胶形成过程及质构特性的影响。结果表明,大豆蛋白凝胶的形成过程受多种分子间作用力的影响,其中静电作用、疏水相互作用和氢键对石膏凝固大豆蛋白凝胶的形成具有重要的影响。     

酱油酿造过程中微生物及生物酶的研究进展

酱油是以大豆或豆粕、小麦粉或麸皮为原料,依靠微生物发酵而生产的一种液态调味品。在发酵过程中,微生物对原料中的蛋白、淀粉等营养物质进行分解,此过程起主导作用的是微生物所分泌产生的生物酶。当前国产酱油采用米曲霉沪酿3.042（Aspergillus oryzae 3.042）进行发酵,利用其产生的碱性和中性蛋白酶把原料中的蛋白分解为 氨基酸和多肽,为酱油提供以鲜味为主的多种滋味,但仅以米曲霉单菌种酿造的酱油存在原料利用率低、风味相对差等问题。随着消费者对酱油品质要求的提高,学术界和生产企业正在通过微生物诱变、多菌种发酵、生物酶制剂应用等多种方式改善发酵过程中生物酶的种类和活性,以进一步提升酿造酱油的品质。该文重点综述了酱油酿造过程中的关键微生物、生物酶及其研究进展和在酱油中的应用,以期对利用微生物、生物酶制剂提升酱油品质提供理论指导。