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UV-ARTP 复合诱变育种提高解淀粉芽孢杆菌产大环内酯的能力,并通过蛋白质组学揭示代谢变化。

UV-ARTP compound mutagenesis breeding improves macrolactins production of Bacillus siamensis and reveals metabolism changes by proteomic.

机构信息

Department of Food Science and Engineering, College of Light Industry and Food Engineering, Guangxi University, Nanning, Guangxi 530004, China.

Guangxi Key Laboratory of Marine Environmental Science, Beibu Gulf Marine Research Center, Guangxi Academy of Sciences, Nanning 530007, China.

出版信息

J Biotechnol. 2024 Feb 10;381:36-48. doi: 10.1016/j.jbiotec.2023.12.011. Epub 2024 Jan 6.

Abstract

Macrolactins are a type of compound with complex macrolide structure which mainly be obtained through microbiological fermentation now. They have excellent antifungal, antibacterial and antitumor activity. In order to improve macrolactins production, Bacillus siamensis YB304 was used as the research object, and a mutant Mut-K53 with stable genetic characters was selected by UV-ARTP compound mutagenesis. The yield of macrolactins was 156.46 mg/L, 3.95 times higher than original strain. The metabolic pathway changes and regulatory mechanism of macrolactins were analyzed by quantitative proteomics combined with parallel reaction monitoring. This study revealed that 1794 proteins were extracted from strain YB304 and strain Mut-K53, most of them were related to metabolism. After UV-ARTP compound mutagenesis treatment, the expression of 628 proteins were significantly changed, of which 299 proteins were significantly up-regulated. KEGG pathway analysis showed that differentially expression proteins mainly distributed in biological process, cellular component, and molecular function processing pathways. Such as utilization of carbon sources, glycolysis pathway, and amino acid metabolism pathway. Furthermore, key precursor substances such as acyl-CoA and amino acids of macrolactin biosynthesis are mostly up-regulated, which are one of the main reasons for increased production of macrolactin.This study will provide a new way to increase the yield of macrolactins through mutagenesis breeding and proteomics.

摘要

大环内酯类化合物是一种具有复杂大环内酯结构的化合物,目前主要通过微生物发酵获得。它们具有优异的抗真菌、抗菌和抗肿瘤活性。为了提高大环内酯类化合物的产量,以芽孢杆菌 YB304 为研究对象,采用 UV-ARTP 复合诱变技术选育出遗传性状稳定的突变株 Mut-K53。突变株 Mut-K53 的大环内酯产量为 156.46mg/L,比原始菌株提高了 3.95 倍。通过定量蛋白质组学结合平行反应监测技术分析了大环内酯代谢途径的变化和调控机制。本研究从菌株 YB304 和 Mut-K53 中提取了 1794 种蛋白质,其中大部分与代谢有关。经 UV-ARTP 复合诱变处理后,有 628 种蛋白质的表达发生了显著变化,其中 299 种蛋白质表达显著上调。KEGG 通路分析表明,差异表达蛋白主要分布在生物过程、细胞成分和分子功能处理途径中。如碳源利用、糖酵解途径和氨基酸代谢途径。此外,大环内酯生物合成的关键前体物质如酰基辅酶 A 和氨基酸大多上调,这也是大环内酯产量增加的主要原因之一。本研究将为通过诱变育种和蛋白质组学提高大环内酯类化合物的产量提供新的途径。

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