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米曲霉 3.042 株与米曲霉 100-8 株的比较蛋白质组分析:探索不同酱油风味的产生。

Comparative proteome analysis of Aspergillus oryzae 3.042 and A. oryzae 100-8 strains: Towards the production of different soy sauce flavors.

机构信息

Key Laboratory of Food Nutrition and Safety (Tianjin University of Science & Technology), Ministry of Education, TEDA,Tianjin, China.

出版信息

J Proteomics. 2012 Jul 16;75(13):3914-24. doi: 10.1016/j.jprot.2012.04.056. Epub 2012 May 24.

Abstract

Aspergillus oryzae plays a central role in soybean fermentation, particularly in its contribution to the flavor of soy sauce. We present a comparative assessment of the intracellular differences between wild-type strain 3.042 and mutant strain A100-8, at the proteome level. 522 different protein spots were identified by MALDI-TOF MS, with 134 spots being confirmed by MALDI-TOF MS/MS. Of these, 451 were differentially expressed proteins (DEPs). There was at least a two-fold increase for 288 spots, and at least a two-fold decrease for 163 spots, in strain A100-8 when compared to 3.042. Further analysis showed that 63 of the more abundant proteins were involved in glycolysis and the citrate cycle; 43 more abundant proteins and 10 less abundant proteins were related to amino acid biosynthesis and metabolism; two of the more abundant proteins were involved in vitamin biosynthesis; and five of the more abundant proteins and four of the less abundant proteins were related to secondary metabolites. Moreover, quantitative real time PCR showed that the mRNA expression levels of six typical genes we selected were consistent with changes in protein expression. We postulate that there may be a relationship between DEPs and the flavor formation mechanism in A. oryzae.

摘要

米曲霉在大豆发酵中起着核心作用,特别是在酱油风味的形成方面。我们在蛋白质组水平上对野生型菌株 3.042 和突变型菌株 A100-8 之间的细胞内差异进行了比较评估。通过 MALDI-TOF MS 鉴定了 522 个不同的蛋白质斑点,通过 MALDI-TOF MS/MS 鉴定了 134 个斑点。其中,451 个为差异表达蛋白(DEPs)。与 3.042 相比,A100-8 中的 288 个斑点的表达水平至少增加了两倍,163 个斑点的表达水平至少减少了两倍。进一步分析表明,63 个丰度较高的蛋白参与糖酵解和柠檬酸循环;43 个丰度较高的蛋白和 10 个丰度较低的蛋白与氨基酸的生物合成和代谢有关;两个丰度较高的蛋白与维生素的生物合成有关;5 个丰度较高的蛋白和 4 个丰度较低的蛋白与次生代谢物有关。此外,定量实时 PCR 显示,我们选择的六个典型基因的 mRNA 表达水平与蛋白表达变化一致。我们推测 DEPs 与米曲霉风味形成机制之间可能存在关系。

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