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Lon蛋白酶基因多拷贝的表达导致天蓝色链霉菌A3(2)抗生素产量增加、渗透压和紫外线胁迫抗性增强。

Expression of Multiple Copies of the Lon Protease Gene Resulted in Increased Antibiotic Production, Osmotic and UV Stress Resistance in Streptomyces coelicolor A3(2).

作者信息

Yilmaz Halil, Yaradir Emine, Tunca Sedef

机构信息

Faculty of Science, Molecular Biology and Genetics Department, Gebze Technical University, Gebze, 41400, Kocaeli, Türkiye.

出版信息

Curr Microbiol. 2024 Dec 17;82(1):43. doi: 10.1007/s00284-024-04021-z.

Abstract

The genus Streptomyces is a group of gram-positive bacteria that exhibit a distinctive growth pattern characterised by elongated, branched hyphae. Streptomyces coelicolor A3(2), which produces at least five different antibiotics, is a model organism that is widely used in genetic studies. There are very few studies in Streptomyces on the ATP-dependent Lon protease, which has very important functions in every organism and is particularly responsible for protein homeostasis. The aim of this study was to construct and characterize a recombinant S. coelicolor strain expressing the lon gene on a multicopy plasmid. For this purpose, the lon gene was first cloned in Escherichia coli under the control of the glycerol-inducible promoter of pSPG, and its expression in S. coelicolor A3(2) cells was demonstrated by RT-qPCR. In contrast with the initial hypothesis, increased lon expression did not affect cell growth seriously. Instead, it increased the cell's tolerance to osmotic and UV stress and led to a significant increase in antibiotic production. The recombinant strain produced 27 times more actinorhodin and 43 times more undecylprodigiosin than the wild-type strain after 120 h of fermentation. To our knowledge, this is the first study to demonstrate the effects of expression of the lon gene on a high copy number plasmid in Streptomyces.

摘要

链霉菌属是一组革兰氏阳性细菌,呈现出独特的生长模式,其特征为菌丝细长且有分支。天蓝色链霉菌A3(2)能产生至少五种不同的抗生素,是一种在遗传学研究中广泛使用的模式生物。在链霉菌中,关于ATP依赖型Lon蛋白酶的研究非常少,该蛋白酶在每种生物体中都具有非常重要的功能,尤其负责蛋白质稳态。本研究的目的是构建并表征在多拷贝质粒上表达lon基因的重组天蓝色链霉菌菌株。为此,首先将lon基因克隆到大肠杆菌中,置于pSPG的甘油诱导型启动子控制下,并通过RT-qPCR证明其在天蓝色链霉菌A3(2)细胞中的表达。与最初的假设相反,lon表达的增加并没有严重影响细胞生长。相反,它提高了细胞对渗透压和紫外线胁迫的耐受性,并导致抗生素产量显著增加。发酵120小时后,重组菌株产生的放线紫红素比野生型菌株多27倍,十一烷基灵菌红素多43倍。据我们所知,这是第一项证明lon基因在链霉菌中高拷贝数质粒上表达的影响的研究。

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