Enhanced production of recombinant proteins in Corynebacterium glutamicum using a molecular chaperone.

IF 0.8 4区 生物学 Q4 BIOTECHNOLOGY & APPLIED MICROBIOLOGY Journal of General and Applied Microbiology Pub Date : 2023-06-22 DOI:10.2323/jgam.2022.10.002
Yali Wang, Xiuxia Liu, Ye Li, Yankun Yang, Chunli Liu, Robert J Linhardt, Fuming Zhang, Zhonghu Bai
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引用次数: 1

Abstract

Protein synthesis in Corynebacterium glutamicum is critical for applications in biotechnology and medicine. However, the use of C. glutamicum for protein production is limited by its low expression and aggregation. To overcome these limitations, a molecular chaperone plasmid system was developed in this study to improve the efficiency of recombinant protein synthesis in C. glutamicum. The effect of molecular chaperones on target protein synthesis (Single-chain variable fragment, Scfv) under three different promoter strengths was tested. In addition, the plasmid containing the molecular chaperone and target protein was verified for growth stability and plasmid stability. This expression model was further validated using two recombinant proteins, human interferon-beta (Hifn) and hirudin variant III (Rhv3). Finally, the Rhv3 protein was purified, and analysis of Rhv3 activity confirmed that the use of a molecular chaperone led to an improvement in test protein synthesis. Thus, the use of molecular chaperones is believed to will improve recombinant proteins synthesis in C. glutamicum.

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利用分子伴侣增强谷氨酸棒状杆菌重组蛋白的产生。
谷氨酸棒状杆菌的蛋白质合成对生物技术和医学的应用具有重要意义。然而,谷氨酰胺在蛋白质生产中的应用受到其低表达和低聚集的限制。为了克服这些局限性,本研究开发了一种分子伴侣质粒系统,以提高谷氨酰胺重组蛋白的合成效率。研究了三种不同启动子强度下分子伴侣对靶蛋白(单链可变片段,Scfv)合成的影响。此外,还验证了含有分子伴侣和靶蛋白的质粒的生长稳定性和质粒稳定性。利用人干扰素- β (Hifn)和水蛭素变体III (Rhv3)两种重组蛋白进一步验证了该表达模型。最后,纯化了Rhv3蛋白,并对Rhv3活性进行了分析,证实了分子伴侣的使用导致了测试蛋白合成的改善。因此,分子伴侣的使用被认为将改善重组蛋白在谷氨酰胺中的合成。
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来源期刊
Journal of General and Applied Microbiology
Journal of General and Applied Microbiology 生物-生物工程与应用微生物
CiteScore
2.40
自引率
0.00%
发文量
42
审稿时长
6-12 weeks
期刊介绍: JGAM is going to publish scientific reports containing novel and significant microbiological findings, which are mainly devoted to the following categories: Antibiotics and Secondary Metabolites; Biotechnology and Metabolic Engineering; Developmental Microbiology; Environmental Microbiology and Bioremediation; Enzymology; Eukaryotic Microbiology; Evolution and Phylogenetics; Genome Integrity and Plasticity; Microalgae and Photosynthesis; Microbiology for Food; Molecular Genetics; Physiology and Cell Surface; Synthetic and Systems Microbiology.
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