Identification and functional characterization of a potential l-Homoserine exporter in Corynebacterium glutamicum

Xiaodi Liu , Xiangyu Zhu , Wenxin Jiang , Huanmin Du
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Abstract

Exporter protein systems play a crucial role in the efficient production of valuable chemicals. However, the lack of active exporters significantly limits the application of industrial bio-based production, making the identification and utilization of novel exporters highly important. In this study, we discovered a novel l-Homoserine exporter, Cg0701, in Corynebacterium glutamicum through homology analysis. First, tolerance assays revealed that the cg0701 overexpression strain (CgH-2) exhibited a 10.45% increase in cell growth compared to the control when cultivated with 30 g/L-Homoserine. Additionally, export assays demonstrated that the l-Homoserine export capacity of CgH-2 increased by approximately 30%. Furthermore, genomic overexpression of cg0701 in an l-Homoserine-producing chassis also enhanced both tolerance and export activity. As a result, the recombinant strain CgH-11 produced 10.79 g/L-Homoserine in shake flask cultures and 48.72 g/L in a 5 L fermenter, representing improvements of 19.89% and 24.44%, respectively. In summary, our results indicate that Cg0701 is a novel l-Homoserine exporter in C. glutamicum, enriching our understanding of amino acid export systems and providing a valuable target for the construction of l-Homoserine microbial cell factories.

Abstract Image

谷氨酸棒状杆菌l-同型丝氨酸潜在输出菌的鉴定和功能表征
出口蛋白系统在有效生产有价值的化学品方面起着至关重要的作用。然而,缺乏活跃的出口商极大地限制了工业生物基生产的应用,使得新的出口商的识别和利用非常重要。本研究通过同源性分析,在谷氨酸棒状杆菌中发现了一个新的l-同型丝氨酸输出基因Cg0701。首先,耐受性实验表明,当添加30 g/ l的同型丝氨酸培养时,cg0701过表达菌株(CgH-2)的细胞生长比对照增加了10.45%。此外,出口分析表明,CgH-2的l-同型丝氨酸出口能力提高了约30%。此外,cg0701基因在l-同型丝氨酸产生基质中的基因组过表达也增强了耐受性和出口活性。结果表明,重组菌株CgH-11在摇瓶培养中产生10.79 g/L的同型丝氨酸,在5 L发酵罐中产生48.72 g/L,分别提高了19.89%和24.44%。综上所述,我们的研究结果表明Cg0701是谷氨酰胺中一个新的l-纯丝氨酸输出系统,丰富了我们对氨基酸输出系统的认识,并为l-纯丝氨酸微生物细胞工厂的建设提供了一个有价值的靶点。
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CiteScore
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