Improved E. coli erythromycin a production through the application of metabolic and bioprocess engineering
Improved E. coli erythromycin a production through the application of metabolic and bioprocess engineering
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DOI：
10.1002/btpr.702
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发表时间：
2012-01-01
影响因子：
2.9
通讯作者：
Pfeifer, Blaine A.
中科院分区：
文献类型：
--
作者：
Zhang, Haoran;Skalina, Karin;Pfeifer, Blaine A.
在这份报告中，小规模的培养和生物反应器实验被用来比较和提高抗生素红霉素A的异源生产在一系列工程原型大肠杆菌菌株。原始菌株，称为BAP 1（pBPJW 130、pBPJW 144、pHZT 1、pHZT 2、pHZT 4、pGro 7），被设计为允许从外源添加的丙酸盐进行完全的红霉素A生物合成。然后将该菌株与假设增加最终产品滴度的两种替代品进行比较。菌株TB 3（pBPJW 130、pBPJW 144、pHZT 1、pHZT 2、pHZT 4、pGro 7）是BAP 1的衍生物，其被设计为由于ygfH缺失而增加生物合成途径碳流;而菌株TB 3（pBPJW 130、pBPJW 144、pHZT 1、pHZT 2、pHZT 4 -2、pGro 7）提供了关键脱氧糖糖基转移酶基因的额外拷贝。比较三种菌株的生产，TB 3（pBPJW 130、pBPJW 144、pHZT 1、pHZT 2、pHZT 4、pGro 7）显示出红霉素B（EB）、3-霉糖基红霉素B（ME B）和红霉素A滴度的显著改善。在分批生物反应器生产实验的背景下进一步检测了该菌株，时间过程滴度水平为4 mg/L，代表最终红霉素A滴度增加约7倍。(C)2011年美国化学工程师学会生物技术。程序，2012
In this report, small-scale culture and bioreactor experiments were used to compare and improve the heterologous production of the antibiotic erythromycin A across a series of engineered prototype Escherichia coli strains. The original strain, termed BAP1(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4, pGro7), was designed to allow full erythromycin A biosynthesis from the exogenous addition of propionate. This strain was then compared against two alternatives hypothesized to increase final product titer. Strain TB3(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4, pGro7) is a derivative of BAP1 designed to increase biosynthetic pathway carbon flow as a result of a ygfH deletion; whereas, strain TB3(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4-2, pGro7) provided an extra copy of a key deoxysugar glycosyltransferase gene. Production was compared across the three strains with TB3(pBPJW130, pBPJW144, pHZT1, pHZT2, pHZT4, pGro7) showing significant improvement in erythronolide B (EB), 3-mycarosylerythronolide B (MEB), and erythromycin A titers. This strain was further tested in the context of batch bioreactor production experiments with time-course titers leveling at 4 mg/L, representing an approximately sevenfold increase in final erythromycin A titer. (C) 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012