谷氨酸棒杆菌的天然启动子及其在L-赖氨酸生产中的应用。
Native promoters of Corynebacterium glutamicum and its application in L-lysine production.
作者信息
Shang Xiuling, Chai Xin, Lu Xuemei, Li Yuan, Zhang Yun, Wang Guoqiang, Zhang Chen, Liu Shuwen, Zhang Yu, Ma Jiyin, Wen Tingyi
机构信息
CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 1 Beichen West Road, Chaoyang District, Beijing, 100101, China.
Beijing Zhongke EPPEN Biotechnology Co., Ltd, Beijing, 100085, China.
出版信息
Biotechnol Lett. 2018 Feb;40(2):383-391. doi: 10.1007/s10529-017-2479-y. Epub 2017 Nov 21.
OBJECTIVE
To identify useful native promoters of Corynebacterium glutamicum for fine-tuning of gene expression in metabolic engineering.
RESULTS
Sixteen native promoters of C. glutamicum were characterized. These promoters covered a strength range of 31-fold with small increments and exhibited relatively stable activity during the whole growth phase using β-galactosidase as the reporter. The mRNA level and enzymatic activity of the lacZ reporter gene exhibited high correlation (R = 0.96) under the control of these promoters. Sequence analysis found that strong promoters had high similarity of the -10 hexamer to the consensus sequence and preference of the AT-rich UP element upstream the -35 region. To test the utility of the promoter library, the characterized native promoters were applied to modulate the sucCD-encoded succinyl-CoA synthetase expression for L-lysine overproduction.
CONCLUSIONS
The native promoters with various strengths realize the efficient and precise regulation of gene expression in metabolic engineering of C. glutamicum.
目的
鉴定谷氨酸棒杆菌中用于代谢工程中基因表达微调的有用天然启动子。
结果
对谷氨酸棒杆菌的16个天然启动子进行了表征。这些启动子的强度范围为31倍，增量较小，以β-半乳糖苷酶为报告基因，在整个生长阶段表现出相对稳定的活性。在这些启动子的控制下，lacZ报告基因的mRNA水平和酶活性表现出高度相关性（R = 0.96）。序列分析发现，强启动子的-10六聚体与共有序列具有高度相似性，并且在-35区域上游富含AT的UP元件具有偏好性。为了测试启动子文库的实用性，将表征的天然启动子应用于调节编码琥珀酰辅酶A合成酶的sucCD表达，以过量生产L-赖氨酸。
结论
具有不同强度的天然启动子实现了谷氨酸棒杆菌代谢工程中基因表达的高效和精确调控。
Zotero 插件