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真菌在储存的低变性脱脂大豆粕上的生长和真菌酶对蛋白质和异黄酮糖苷的水解。

Growing of fungi on the stored low denatured defatted soybean meals and the hydrolysis of proteins and isoflavone glycosides by fungal enzymes.

机构信息

State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, PR China; School of Food Science and Technology, Jiangnan University, Wuxi, PR China.

State Key Laboratory of Food Science and Technology, Jiangnan University, Wuxi, PR China; School of Food Science and Technology, Jiangnan University, Wuxi, PR China.

出版信息

Food Res Int. 2023 Jan;163:112261. doi: 10.1016/j.foodres.2022.112261. Epub 2022 Nov 30.

Abstract

Recently, more and more attention has been paid to the effects of fungal contamination and fungal enzymes secreted in raw grain on product quality. As the starting material of protein and active components, the quality of low denatured defatted soybean meals (LDSM) directly determines the qualities of subsequent products. In previous studies, we have revealed that infection with Aspergillus ochraceus protease causes significant hydrolysis of proteins. In this study, growing of fungi on the stored low denatured defatted soybean meals (LDSM) was analyzed by high-throughput sequencing and real-time PCR, which revealed that the abundance of Aspergillus increased significantly after storage. Twenty fungal proteases and 9 fungal glucosidases were found in stored LDSM and zymography showed that the proteases were of serine-type with some cysteine and aspartic activities. Proteolysis of the soybean storage proteins mainly occurred after the hydration of LDSM and the average molecular weight of soy proteins decreased from 57.9 kDa to 30.7 kDa after 60 min's of hydrolysis. Two-dimensional electrophoresis (2-DE) analysis found the polypeptide fragments from soybean 7S and 11S proteins with molecular weight around 10-25 kDa in the hydrated LDSM. Glycosylated isoflavones were hydrolyzed in both dry and hydrated stored LDSM which resulted in significant (p < 0.05) increase in the contents of isoflavone aglycones. This study suggested that fungi contamination be a new factor affecting the properties of LDSM derived soy protein products.

摘要

最近,人们越来越关注生粮中真菌污染和真菌分泌的酶对产品质量的影响。作为蛋白质和活性成分的起始原料,低变性脱脂豆粕(LDSM)的质量直接决定了后续产品的质量。在以前的研究中,我们已经揭示了赭曲霉蛋白酶的感染会导致蛋白质的显著水解。在这项研究中,通过高通量测序和实时 PCR 分析了储存的低变性脱脂豆粕(LDSM)上真菌的生长情况,结果表明,真菌的丰度在储存后显著增加。在储存的 LDSM 中发现了 20 种真菌蛋白酶和 9 种真菌糖苷酶,并且酶谱显示蛋白酶为丝氨酸型,具有一些半胱氨酸和天冬氨酸活性。大豆贮藏蛋白的水解主要发生在 LDSM 水合之后,并且大豆蛋白的平均分子量从 57.9 kDa 降低到水解 60 分钟后的 30.7 kDa。二维电泳(2-DE)分析发现,在水合的 LDSM 中存在分子量约为 10-25 kDa 的大豆 7S 和 11S 蛋白的多肽片段。在干燥和水合储存的 LDSM 中,糖基化异黄酮都被水解,导致异黄酮苷元的含量显著增加(p<0.05)。本研究表明,真菌污染可能是影响 LDSM 衍生大豆蛋白产品性质的一个新因素。

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