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基于 2D-DIGE 和 MALDI-TOF/TOF-MS 的不同糖基上培养的游动放线菌比较蛋白质组分析。

Comparative proteome analysis of Actinoplanes utahensis grown on various saccharides based on 2D-DIGE and MALDI-TOF/TOF-MS.

机构信息

Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China; Engineering Research Center of Bioconversion and Biopurification of the Ministry of Education, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, PR China; The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou 310014, PR China.

Key Laboratory of Bioorganic Synthesis of Zhejiang Province, College of Biotechnology and Bioengineering, Zhejiang University of Technology, Hangzhou 310014, PR China; Engineering Research Center of Bioconversion and Biopurification of the Ministry of Education, Zhejiang University of Technology, Hangzhou, Zhejiang 310014, PR China; The National and Local Joint Engineering Research Center for Biomanufacturing of Chiral Chemicals, Zhejiang University of Technology, Hangzhou 310014, PR China.

出版信息

J Proteomics. 2021 May 15;239:104193. doi: 10.1016/j.jprot.2021.104193. Epub 2021 Mar 20.

Abstract

Comparative proteomes of Actinoplanes utahensis ZJB-03852 grown on various saccharides (glucose, maltotriose, maltose, glucose + maltose) were analyzed using 2D-DIGE and MALDI-TOF/TOF-MS. Acarbose was detected in all groups except in the glucose only culture. The abundance of acarbose synthesis proteins AcbV, AcbK, AcbL and AcbN was highest in the medium containing mixed glucose and maltose. The accumulation of Zwf and Xpk1 in acarbose-producing media indicated that the cyclitol moiety of acarbose was derived from pentose phosphate pathway. The elevation of GlnA supported that glutamine was a good nitrogen source of the nitrogen-atom in acarbose synthesis. SIGNIFICANCE: Non-insulin-dependent diabetes mellitus, also known as Type II diabetes, constitutes >90% of the diabetes mellitus worldwide. Acarbose is clinically utilized to treat Type II diabetes, but the fermentation process of acarbose-producing Actinoplanes is usually accompanied with structural analogues of acarbose. In this study, we compared the proteomics of Actinoplanes utahensis ZJB-03852 grown on various saccharides by 2D-DIGE and MALDI-TOF/TOF-MS. Our findings highlighted the importance of key proteins in the formation of acarbose and its analogues when A. utahensis was cultivated in various saccharides. These results revealed fundamental data to elucidate the complexity of formation of acarbose analogues.

摘要

使用 2D-DIGE 和 MALDI-TOF/TOF-MS 分析了 Actinoplanes utahensis ZJB-03852 在不同糖(葡萄糖、麦芽三糖、麦芽糖、葡萄糖+麦芽糖)上生长的比较蛋白质组。除仅葡萄糖培养物外,所有组均检测到阿卡波糖。含有混合葡萄糖和麦芽糖的培养基中阿卡波糖合成蛋白 AcbV、AcbK、AcbL 和 AcbN 的丰度最高。阿卡波糖产生培养基中 Zwf 和 Xpk1 的积累表明,阿卡波糖的环醇部分来自戊糖磷酸途径。GlnA 的升高支持了谷氨酰胺是阿卡波糖合成中氮原子的良好氮源。意义:非胰岛素依赖型糖尿病,也称为 2 型糖尿病,构成了全球 90%以上的糖尿病。阿卡波糖临床上用于治疗 2 型糖尿病,但产阿卡波糖放线菌的发酵过程通常伴随着阿卡波糖的结构类似物。在这项研究中,我们通过 2D-DIGE 和 MALDI-TOF/TOF-MS 比较了 Actinoplanes utahensis ZJB-03852 在不同糖上生长的蛋白质组。我们的发现强调了当 A. utahensis 在各种糖中培养时,形成阿卡波糖及其类似物的关键蛋白的重要性。这些结果揭示了阐明阿卡波糖类似物形成复杂性的基本数据。

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