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二氨基庚二酸途径中的组成型和高基因表达加速了白色链霉菌中 ε-聚赖氨酸的生产。

Constitutive and high gene expression in the diaminopimelate pathway accelerates ε-poly-L-lysine production in Streptomyces albulus.

机构信息

Department of Bioscience, Fukui Prefectural University, Fukui, Japan.

Fukui Bio Incubation Center (FBIC), Fukui Prefectural University, Eiheiji-cho, Fukui, Japan.

出版信息

J Antibiot (Tokyo). 2023 Sep;76(9):522-531. doi: 10.1038/s41429-023-00636-9. Epub 2023 Jun 12.

Abstract

Streptomyces albulus NBRC14147 produces a homopoly(amino acid), ε-poly-L-lysine (ε-PL). Due to its antibiotic activity, thermostability, biodegradability, and non-toxicity to humans, ε-PL is used as a food preservative. In this study, homology searches of diaminopimelate (DAP) pathway genes (dapB and dapE), in an S. albulus genome database, were shown to encode predicted enzymes using dapB or dapE in Escherichia coli strain complementation assays. We observed that dapB and dapE transcriptional levels were weak during ε-PL production stages. Therefore, we strengthened this expression using an ermE constitutive promoter. Engineered strains generated faster growth and ε-PL production rates when compared with the control strain. Moreover, maximum ε-PL yields in S. albulus, where dapB was constitutively expressed, were approximately 14% higher when compared with the control strain. These findings showed that enhanced lysine biosynthetic gene expression generated faster and higher ε-PL production levels.

摘要

白色链霉菌 NBRC14147 产生同聚(氨基酸),ε-聚-L-赖氨酸(ε-PL)。由于其具有抗生素活性、热稳定性、生物降解性和对人体无毒,ε-PL 被用作食品防腐剂。在这项研究中,通过同源搜索白色链霉菌基因组数据库中的二氨基庚二酸(DAP)途径基因(dapB 和 dapE),用 dapB 或 dapE 在大肠杆菌菌株互补测定中编码预测的酶。我们观察到,在 ε-PL 生产阶段,dapB 和 dapE 的转录水平较弱。因此,我们使用 ermE 组成型启动子增强了这种表达。与对照菌株相比,工程菌株的生长和 ε-PL 生产速率更快。此外,当 dapB 组成型表达时,白色链霉菌中 ε-PL 的最大产量比对照菌株约高 14%。这些发现表明,增强赖氨酸生物合成基因的表达可以产生更快和更高的 ε-PL 生产水平。

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