多拷贝 Lon 蛋白酶基因的表达提高了链霉菌 A3(2) 的抗生素产量、抗渗透压和抗紫外线能力。

IF 2.3 3区 生物学 Q3 MICROBIOLOGY Current Microbiology Pub Date : 2024-12-17 DOI:10.1007/s00284-024-04021-z
Halil Yilmaz, Emine Yaradir, Sedef Tunca
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引用次数: 0

摘要

链霉菌属是一组革兰氏阳性细菌,表现出独特的生长模式,其特征是细长的分枝菌丝。colicololstreptomyces A3(2)是一种模式生物,被广泛用于遗传研究,它可以产生至少五种不同的抗生素。链霉菌中对atp依赖性Lon蛋白酶的研究很少,而这种蛋白酶在每个生物体中都具有非常重要的功能,特别是对蛋白质稳态负责。本研究的目的是构建并鉴定在多拷贝质粒上表达lon基因的重组大肠杆菌菌株。为此,在甘油诱导的pSPG启动子的控制下,首先在大肠杆菌中克隆了lon基因,并通过RT-qPCR证实了其在S. coelicolor A3(2)细胞中的表达。与最初的假设相反,lon表达增加并不严重影响细胞生长。相反,它增加了细胞对渗透和紫外线胁迫的耐受性,并导致抗生素产量显著增加。经过120 h的发酵,重组菌株的放线菌素产量是野生型菌株的27倍,十一酰芥子素产量是野生型菌株的43倍。据我们所知,这是第一次在链霉菌的高拷贝数质粒上证明lon基因表达的影响。
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Expression of Multiple Copies of the Lon Protease Gene Resulted in Increased Antibiotic Production, Osmotic and UV Stress Resistance in Streptomyces coelicolor A3(2).

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.

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来源期刊
Current Microbiology
Current Microbiology 生物-微生物学
CiteScore
4.80
自引率
3.80%
发文量
380
审稿时长
2.5 months
期刊介绍: Current Microbiology is a well-established journal that publishes articles in all aspects of microbial cells and the interactions between the microorganisms, their hosts and the environment. Current Microbiology publishes original research articles, short communications, reviews and letters to the editor, spanning the following areas: physiology, biochemistry, genetics, genomics, biotechnology, ecology, evolution, morphology, taxonomy, diagnostic methods, medical and clinical microbiology and immunology as applied to microorganisms.
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