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Differential protein expression of a streptomycin-resistant Streptomyces albulus mutant in high yield production of ε-poly-l-lysine: a proteomics study

机译:ε-聚L-赖氨酸高产量生产中抗链霉素抗性链霉菌蛋白突变体的差异蛋白质表达:蛋白质组学研究

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ε-Poly- L -lysine (ε-PL), produced by Streptomyces albulus , is an excellent antimicrobial agent which has been extensively used in the field of food and medicine. In our previous study, we have improved ε-PL production by S. albulus M-Z18 through iterative introduction of streptomycin resistance. To decipher the overproduction mechanism of high-yielding mutant S. albulus SS-62, we conducted a comparative proteomics analysis between S. albulus SS-62 and its parent strain S. albulus M-Z18. Approximately 11.5% of the predicted S . albulus proteome was detected and 401 known or putative regulatory proteins showed statistically differential expression levels. Expression levels of proteins involved in ε-PL precursor metabolism and energy metabolism, and proteins in the pathways related to transcriptional regulation and translation were up-regulated. It was indicated that mutant SS-62 could not only strengthen the ε-PL precursor metabolism and energy metabolism but also tune the pathways related to transcriptional regulation and translation, suggesting a better intracellular metabolic environment for the synthesis of ε-PL in mutant SS-62. To confirm these bioinformatics analyses, qRT-PCR was employed to investigate the transcriptional levels of pls , frr and hrd D and their transcription levels were found to have increased more than 4-fold. Further, overexpression of pls and frr resulted in an increase in ε-PL titer and the yield of ε-PL per unit cell. This report not only represents the first comprehensive study on comparative proteomics in S . albulus , but it would also guide strain engineering to further improve ε-PL production.
机译:由链霉菌蛋白生成的ε-聚-L-碘碱(ε-PL)是一种优异的抗微生物剂,其已被广泛用于食品和药物领域。在我们以前的研究中,我们通过迭代引入链霉素抗性,通过S.Bolulus M-Z18改善了ε-PL生产。为了破译高产突变体SS-62的过生产机制,我们在S.Bolulus SS-62及其亲本菌株S.Bolulus M-Z18之间进行了对比较蛋白质组学分析。预计的大约11.5%。检测蛋白蛋白质组,401名已知或推定的调节蛋白显示出统计学上差异表达水平。上调ε-PL前体代谢和能量代谢和能量代谢的蛋白质的表达水平,以及与转录调节相关的途径和翻译中的蛋白质。结果表明,突变体SS-62不仅可以增强ε-PL前体代谢和能量代谢,还可以调整与转录调节和翻译相关的途径,表明突变体SS中的ε-PL的细胞内代谢环境更好62。为了确认这些生物信息学分析,使用QRT-PCR来研究PLS,FRR和HRD D的转录水平,并且它们的转录水平被发现增加了4倍以上。此外,PLS和FRR的过度表达导致ε-PL滴度的增加和每单位电池的ε-p的产率。本报告不仅代表了S中对比较蛋白质组学的第一综合研究。白蚁,但还将引导应变工程进一步改善ε-pl生产。

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