大豆过敏原11S球蛋白G2中A2链结合表位的预测

采用生物信息学软件NCBI GenBank数据库、PDB数据库、SWISS-MODEL、Deep View 4.1等预测11S球蛋白G2中A2链的二级结构和三级结构模型,使用DiscoTope 2.0 Server软件预测B细胞构象表位,发现构象表位多位于无规则卷曲处。定位B细胞线性结合表位,结果表明11S球蛋白G2中A2链的可能线性表位有：37KPDNRIE43、79PSYTNGP85、114SQQRGRSQRP123、52WNPNNK57、196QQGGSQSQKGKQQE209、241QGENEEED248、267RKPQQEEDDDDEEEQ281、287TDKGC291。此表位预测为后续制肽、免疫等操作找出更准确的肽序,以便建立适合我国大豆过敏人群的食品大豆过敏原的检测技术。     

大米球蛋白铜离子结合肽的鉴定及其螯合活性分析

为分析大米球蛋白与金属元素的结合特性,通过分级提取方法得到4 种大米蛋白,并测定其金属元素含量。选取大米球蛋白经胃蛋白酶酶解,酶解液通过超滤方式进行纯化。将获得的水解液进行质谱检测,并通过软件分析得到23 条多肽序列,分子质量在725～1 409 Da之间。结合已报道的相关研究,分析得到多肽序列QGWSSSSE和YYGGEGSSSEQGY具有潜在金属螯合活性。通过人工合成这两条多肽序列,并选择乙二胺四乙酸（ethylenediaminetetraacetic acid,EDTA）、柠檬酸和金属硫蛋白作为参照物,比较它们与Cu2＋的螯合活性。结果发现YYGGEGSSSEQGY螯合Cu2＋效果比QGWSSSSE好,与Cu2＋螯合的IC20为5.40 mmol/L,并且在低浓度范围与柠檬酸相当,比EDTA和金属硫蛋白差。进一步分析,这两条多肽与其他金属离子之间螯合活性发现,其与Fe2＋结合更强。通过氨基酸分析,连续丝氨酸SS、SSS和SSSS可能与铜的螯合活性相关。     

11S球蛋白酶解产物对猪肉肠品质的影响

以中豆36为原料,提取11S粗蛋白,并采用碱性蛋白酶对其水解,将水解后的产物添加到猪肉肠中。以质构特性、凝胶强度、肉肠得率为评价指标,比较了添加不同水解度和不同添加量的11S酶解蛋白对猪肉肠品质的影响。采用二因素三水平正交试验,确定了肉肠专用蛋白的最佳工艺参数:11S水解度为10%,添加量为1.5%。通过凝胶电泳分析得出:酶解产物主要是分子质量<20ku的多肽。     

The tryptophan content of extractable seed proteins from cultivated legumes,sunflower and Acacia

The total salt-extractable protein from seeds of important cultivated legumes and sunflower kernels has a tryptophan content ranging from 0.91% (by wt) for chickpea (Cicer arietinum) to 1.99% (by wt) for green gram (Vigna radiata). In species with substantial albumin fractions, the tryptophan content of the albumin fraction is higher than that of the globulin fraction by factors ranging between 2.3 (pigeon pea, Cajanus cajan) and 20.3-fold (pea, Pisum sativum, cv. Telephone). Although the protein content of Acacia seeds is relatively low (5–14% of seed weight), the tryptophan content of Acacia seed proteins is usually greater than that of the cultivated legumes (1.50–2.66%, by wt).     

Lack of Globulin Synthesis during Seed Development Alters Accumulation of Seed Storage Proteins in Rice

The major seed storage proteins (SSPs) in rice seeds have been classified into three types, glutelins, prolamins, and globulin, and the proportion of each SSP varies. It has been shown in rice mutants that when either glutelins or prolamins are defective, the expression of another type of SSP is promoted to counterbalance the deficit. However, we observed reduced abundances of glutelins and prolamins in dry seeds of a globulin-deficient rice mutant (Glb-RNAi), which was generated with RNA interference (RNAi)-induced suppression of globulin expression. The expression of the prolamin and glutelin subfamily genes was reduced in the immature seeds of Glb-RNAi lines compared with those in wild type. A proteomic analysis of Glb-RNAi seeds showed that the reductions in glutelin and prolamin were conserved at the protein level. The decreased pattern in glutelin was also significant in the presence of a reductant, suggesting that the polymerization of the glutelin proteins via intramolecular disulfide bonds could be interrupted in Glb-RNAi seeds. We also observed aberrant and loosely packed structures in the storage organelles of Glb-RNAi seeds, which may be attributable to the reductions in SSPs. In this study, we evaluated the role of rice globulin in seed development, showing that a deficiency in globulin could comprehensively reduce the expression of other SSPs.     

大豆球蛋白的纤维聚集体行为及其稳定性研究

为了明确蛋白质的纤维聚集行为,本研究以大豆球蛋白(soy globulin,11S)为原料,从亚基层面对酸性条件下热诱导的11S纤维聚集过程进行跟踪,监测蛋白及其亚基的水解过程、结构变化及其稳定性。结果表明,11S的纤维化是一个多步骤的过程,包括多肽链的水解、自组装成淀粉样纤维聚集结构及逐渐生长成宏观可见的具有扭曲螺旋结构的纤维聚集体。与11S纤维化过程的单指数增长相比,酸性亚基的纤维化过程存在迟滞期。酸性亚基在纤维化聚集的初期主要贡献于纤维聚集的成核过程,碱性亚基的加入改变其纤维聚集进程。蛋白质的纤维化过程会增加11S在等电点处的溶解度,降低中性和酸性pH下的溶解度。此外,碱性环境(pH值10.0)会导致11S纤维聚集体全部溶解、宏观纤维长度变小、结构发生改变。以上研究结果旨在为合理利用蛋白纤维化聚集体作为新的功能性食品配料提供理论依据。     

化学发光法对燕麦贮藏蛋白质抗氧化活性的评价

燕麦营养价值高,且蛋白含量丰富,球蛋白作为其主要的贮藏蛋白质,其必需氨基酸比例合理。目前,有很多燕麦蛋白加工方面的研究,而对于其生物活性的探讨较少。用三种化学发光体系对燕麦球蛋白和清蛋白的体外抗氧化能力进行研究,发现在清除超氧阴离子和保护DNA损伤方面球蛋白的效果优于清蛋白,其半数抑制率分别为0.85 mg/mL和7.3 mg/mL,而清蛋白有较好的清除羟基自由基的作用,其半数抑制率为2.26 mg/mL。结果说明,两种燕麦贮藏蛋白质有较弱的抗氧化活性。     

11S球蛋白水解产物功能特性及其对猪肉肠质构影响

以中豆36为原料,提取11S球蛋白,用菠萝蛋白酶对其水解,研究不同水解度酶解产物的持水性、吸油性、乳化性、乳化稳定性及疏水性等功能特性;结果表明,水解度为5%时11S球蛋白酶解产物吸油性、持水性、乳化活性最高;并将不同水解度酶解蛋白添加到猪肉肠中,水解度5%酶解蛋白可明显改善猪肉肠质构特性。     

Allergenicity of proteolytic hydrolysates of the soybean 11S globulin

ABSTRACT:  A substantial portion of the human population has immune hypersensitivities to various food materials. Soybean is one of the most common foods involved in such hypersensitivity reactions, especially in younger children. In this study, we investigated the effect of peptic and chymotryptic hydrolysis on the allergenicity of the 11S soybean globulin, which is the primary soybean allergen. The 11S globulin is composed of both acidic and basic polypeptides, and we found that the acidic polypeptide was effectively hydrolyzed, while basic polypeptide was more resistant to hydrolysis. The 11S globulin hydrolysate was size-fractionated by gel filtration, and 9 of the fractions obtained were tested for allergenicity against sera from 6 soybean-allergenic patients. The overall allergenicity of soybean 11S globulin was reduced by peptic and chymotryptic hydrolysis, although a gel filtration fraction with a major peptide of 20 kDa was highly immunoreactive. Hydrolyzed fragments of less than about 20 kDa were not immunoreactive.     

黑豆蛋白的分级提取及黑豆花色苷的成分鉴定

以黑豆为原料,添加不同浓度(0、5、10、15、20、25 mmol/L)的不同二价阳离子(Ca~(2+)、Mg~(2+)和Zn~(2+))分级提取黑豆蛋白,经冷冻干燥后用凯氏定氮法测得蛋白质量浓度,并对所得蛋白级分(7S、11S)的纯度和得率进行分析。结果表明,黑豆提取过程中所加入二价阳离子的类型及浓度显著影响7S和11S球蛋白的得率和纯度。利用综合平衡法,最终选定添加10 mmol/L的Mg2+分级提取黑豆球蛋白,得到7S蛋白纯度和得率分别为(86.29±3.25)%和(2.90±0.14)%,11S蛋白纯度和得率分别为(87.42±3.30)%和(9.11±0.28)%。用乙醇法提取黑豆花色苷,测得黑豆中总花色苷含量为(0.58±0.03)mg/g,总酚含量为(2.22±0.12)mg/g,对黑豆花色苷进行高效液相色谱-串联质谱分析,其组成成分为飞燕草色素-3-O-葡萄糖苷、矢车菊色素-3-O-葡萄糖苷、牵牛花色素-3-O-葡萄糖苷和锦葵花色素-3-O-葡萄糖苷。