Ribosome binding site libraries and pathway modules for shikimic acid synthesis with Corynebacterium glutamicum----中国科学院微生物研究所

科研成果

论文题目:	Ribosome binding site libraries and pathway modules for shikimic acid synthesis with Corynebacterium glutamicum
作者:	Zhang Bo, Zhou Nan, Liu Yi-Ming, Liu Chang, Lou Chun-Bo, Jiang Cheng-Ying, and Liu Shuang-Jiang.
联系作者:
刊物名称:	Microb Cell Fact
期:	14
卷:	1
页:	71
年份:	2015
影响因子:	4.642
论文下载:	http://www.microbialcellfactories.com/content/14/1/71/abstract
摘要:	BACKGROUND: The shikimic acid (SA) pathway is a fundamental route to synthesize aromatic building blocks for cell growth and metabolic processes, as well as for fermentative production of various aromatic compounds. Genes encoding enzymes of SA pathway are not continuous on genome and they are differently regulated. RESULTS: In this study, efforts were made to construct continuous genetic modules of SA pathway that are regulated by a same Ptac promoter. Firstly, aro genes [aroG (NCgl2098), aroB (NCgl1559), aroD (NCgl0408) and aroE (NCgl1567)] from Corynebacterium glutamicum and ribosome binding site (RBS) libraries that were tailored for the above genes were obtained, and the strength of each RBS in the 4 libraries was quantified. Secondly, 9 genetic modules were built up from the RBS libraries, a previously characterized ribozyme insulator (RiboJ) and transcriptional promoter (Ptac) and terminator, and aroG, aroB, aroD and aroE. The functionality and efficiency of the constructed genetic modules were evaluated in C. glutamicum by determination of SA synthesis. Results showed that C. glutamicum RES167DeltaaroK carrying a genetic module produced 4.3 g/L of SA, which was 54 folds higher compared to that of strain RES167DeltaaroK (80 mg/L, without the genetic module) during fermentation in 250-mL flasks. The same strain produced 7.4, and 11.3 g/L of SA during 5-L batch and fed-batch fermentations, respectively, which corresponding to SA molar yields of 0.39 and 0.24 per mole sucrose consumption. CONCLUSION: These results demonstrated that the constructed SA pathway modules are effective in increasing SA synthesis in C. glutamicum, and they might be useful for fermentative production of aromatic compounds derived from SA pathway.