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受SCO7613基因区域影响的天蓝色链霉菌中聚羟基脂肪酸酯的积累

Polyhydroxyalkanoate accumulation in Streptomyces coelicolor affected by SCO7613 gene region.

作者信息

Demir Öksüz Zeynep, Doruk Tuğrul, Yağci Nevin, Tunca Gedik Sedef

机构信息

Molecular Biology and Genetic Department, Faculty of Science, Gebze Technical University, Kocaeli Turkey.

Molecular Biology and Genetic Department, Faculty of Arts and Science, Ondokuz Mayıs University, Samsun Turkey.

出版信息

Turk J Biol. 2021 Jun 23;45(3):275-286. doi: 10.3906/biy-2011-16. eCollection 2021.

Abstract

Polyhydroxyalkanoate (PHA) is stored as an important carbon and energy source in bacterial cells. For biomedical applications, gram-positive bacteria can be better sources of PHAs, since they lack outer membrane lipopolysaccharide. Although gram-positive A3(2) has been indicated as a high potential PHA producer, C gene that encodes the key enzyme PHA synthase in the metabolic pathway is not determined in its genome. BLAST search results of the GenBank database argued that SCO7613 could specify a putative polyhydroxyalkanoate synthase (PhaC) responsible for PHA biosynthesis. Deduced amino acid sequence of SCO7613 showed the presence of conserved lipase box like sequence, GASAG, in which serine residue was present as the active nucleophile. Present study describes deletion of putative C gene via PCR dependent method. We showed that SCO7613 is not an essential gene in and its deletion affected PHA accumulation negatively although it is not ceased. Transcomplementation abolished the mutant phenotype, demonstrating that the decrease in PHA resulted from the deletion of SCO7613.

摘要

聚羟基脂肪酸酯(PHA)作为重要的碳源和能源存储在细菌细胞中。对于生物医学应用而言,革兰氏阳性菌可能是更好的PHA来源,因为它们缺乏外膜脂多糖。尽管革兰氏阳性菌A3(2)已被表明是一种具有高潜力的PHA生产者,但其基因组中尚未确定编码代谢途径中关键酶PHA合酶的C基因。GenBank数据库的BLAST搜索结果表明,SCO7613可能指定一种负责PHA生物合成的推定聚羟基脂肪酸酯合酶(PhaC)。SCO7613推导的氨基酸序列显示存在保守的脂肪酶盒样序列GASAG,其中丝氨酸残基作为活性亲核试剂存在。本研究描述了通过PCR依赖性方法缺失推定的C基因。我们表明,SCO7613在[具体细菌名称未给出]中不是必需基因,其缺失对PHA积累产生了负面影响,尽管PHA积累并未停止。转互补消除了突变体表型,表明PHA的减少是由SCO7613的缺失导致的。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/d83b/8313934/56ff366da756/turkjbio-45-275-fig001.jpg

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