Deletion of the aroK gene is essential for high shikimic acid accumulation through the shikimate pathway in E. coli
Deletion of the aroK gene is essential for high shikimic acid accumulation through the shikimate pathway in E. coli
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DOI：
10.1016/j.biortech.2012.05.100
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发表时间：
2012-09-01
影响因子：
11.4
通讯作者：
Zhou, Changlin
中科院分区：
文献类型：
--
作者：
Chen, Kai;Dou, Jie;Zhou, Changlin
莽草酸（Shikimic acid，SA）是一种重要的代谢中间体，具有广泛的商业用途。本研究利用反义RNA干扰技术和基因缺失技术对SA产生菌DHPYA-T7的aroK基因进行了改造。在该菌株中，缺失aroL、ptsHIcrr和ydiB基因，并且过表达tktA、glk、aroE和aroB基因。DHPYA-T7衍生菌株的摇瓶培养表明，aroK基因缺失（DHPYAAS-T7）后SA积累增加了2.69倍，反义RNA干扰（DHPYAS-T7）后增加了1.29倍。此外，DHPYAAS-T7中的莽草酸激酶活性是菌株DHPYAS-T7中的0.21倍。在10-L发酵中，菌株DHPYAAS-T7中SA积累增加到1850 mg L-1，这是菌株DHPYAS-T7中SA积累的1.5倍。这些结果表明，DHPYA-T7中的aroK基因失活导致高SA积累，特别是当这种失活由染色体缺失引起时。(C)2012爱思唯尔有限公司保留所有权利。
Shikimic acid (SA) is an important metabolic intermediate with diverse commercial applications. In this work, antisense RNA interference and gene deletion were carried out to inactivate the aroK gene in an SA-producing Escherichia coli strain, DHPYA-T7. In this strain, the aroL, ptsHIcrr and ydiB genes are deleted, and the tktA, glk, aroE and aroB genes are overexpressed. Flask cultivations of the DHPYA-T7 derivative strains showed that the accumulation of SA increased 2.69-fold after aroK gene deletion (DHPYAAS-T7) and 1.29-fold after antisense RNA interference (DHPYAS-T7). Furthermore, the activity of shikimate kinase in DHPYAAS-T7 was 0.21-fold of that in strain DHPYAS-T7. In a 10-L fermentation, SA accumulation increased to 1850 mg L-1 in strain DHPYAAS-T7, which is a 1.5-fold increase over that in strain DHPYAS-T7. These results demonstrate that aroK gene inactivation in DHPYA-T7 leads to high SA accumulation, especially when this inactivation is caused by chromosomal deletion. (C) 2012 Elsevier Ltd. All rights reserved.