Improved E. coli erythromycin a production through the application of metabolic and bioprocess engineering

✍ Scribed by Haoran Zhang; Karin Skalina; Ming Jiang; Blaine A. Pfeifer

• Publisher: American Institute of Chemical Engineers
• Year: 2011
• Tongue: English
• Weight: 926 KB
• Volume: 28
• Category: Article
• ISSN: 8756-7938
• DOI: 10.1002/btpr.702

✦ Synopsis

Abstract

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. © 2011 American Institute of Chemical Engineers Biotechnol. Prog., 2012