Enhanced production of recombinant proteins in Corynebacterium glutamicum by constructing a bicistronic gene expression system
Enhanced production of recombinant proteins in Corynebacterium glutamicum by constructing a bicistronic gene expression system
复制标题
通过构建双顺反子基因表达系统提高谷氨酸棒杆菌重组蛋白的产量
DOI：
10.1186/s12934-020-01370-9
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发表时间：
2020-02
影响因子：
6.4
通讯作者：
Bai Zhonghu
中科院分区：
文献类型：
--
作者：
Sun Manman;Gao Xiong;Zhao Zihao;Li An;Wang Yali;Yang Yankun;Liu Xiuxia;Bai Zhonghu
Background
Corynebacterium glutamicum is a traditional food-grade industrial microorganism, in which an efficient endotoxin-free recombinant protein expression factory is under developing in recent years. However, the intrinsic disadvantage of low recombinant protein expression level is still difficult to be solved. Here, according to the bacteria-specific polycistronic feature that multiple proteins can be translated in one mRNA, efforts have been made to insert a leading peptide gene upstream of target genes as an expression enhancer, and it is found that this can remarkably improve the expression level of proteins under the control of inducible tac promoter in C. glutamicum.
Results
In this research, the Escherichia coli (E. coli) tac promoter combined with 24 different fore-cistron sequences were constructed in a bicistronic manner in C. glutamicum. Three strong bicistronic expression vectors were isolated and exhibited high efficiency under different culture conditions. The compatibility of these bicistronic vectors was further validated using six model proteins- aldehyde dehydrogenase (ALDH), alcohol dehydrogenase (ADH), RamA (regulator of acetate metabolism), Bovine interferon-α (BoIFN-α), glycoprotein D protein (gD) of infectious bovine rhinotracheitis virus (IBRV) and procollagen type Ι N-terminal peptide (PΙNP). All examined proteins were highly expressed compared with the original vector with tac promoter. Large-scale production of PΙNP was also performed in fed-batch cultivation, and the highest PΙNP production level was 1.2 g/L.
Conclusion
In this study, the strength of the inducible tac promoter for C. glutamicum was improved by screening and inserting fore-cistron sequences in front of the target genes. Those vectors with bicistronic expression patterns have strong compatibility for expressing various heterogeneous proteins in high yield. This new strategy could be used to further improve the performance of inducible promoters, achieving double competence of inducible control and high yield.
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影响因子：
5.3
作者：
通讯作者：
--
细胞壁蛋白 CspB 和 PBP1a 的双重突变增加了谷氨酸棒杆菌抗体 Fab 片段的分泌。
影响因子：
6.4
作者：
Matsuda Y;Itaya H;Kitahara Y;Theresia NM;Kutukova EA;Yomantas YA;Date M;Kikuchi Y;Wachi M
通讯作者：
Wachi M
影响因子：
6.4
作者：
Zhang B;Zhou N;Liu YM;Liu C;Lou CB;Jiang CY;Liu SJ
通讯作者：
Liu SJ
影响因子：
5
作者：
Michaela Osadská;Hana Boňková;J. Krahulec;S. Stuchlík;J. Turňa
通讯作者：
Michaela Osadská;Hana Boňková;J. Krahulec;S. Stuchlík;J. Turňa
DOI：
10.1089/jir.2013.0139
发表时间：
2015-03
期刊：
Journal of interferon & cytokine research : the official journal of the International Society for Interferon and Cytokine Research
影响因子：
--
作者：
J. Shao;Chong Cao;Jun Bao;Hongtao Liu;Tongquan Peng;Mingchun Gao;Junwei Wang
通讯作者：
J. Shao;Chong Cao;Jun Bao;Hongtao Liu;Tongquan Peng;Mingchun Gao;Junwei Wang