The main byproducts and metabolic flux profiling of γ-PGA-producing strain B. subtilis ZJU-7 under different pH values.

Poly-γ-glutamic acid (γ-PGA) is an extracellular anionic polymer with many potential applications. Although microbial fermentation is the common approach to produce γ-PGA, the broth at the latter stage usually becomes very viscous and severely influences the metabolites producing pattern of target microbe. In this study, acetoin and 2,3-butanediol were confirmed to be the main byproducts of γ-PGA-producing strain B. subtilis ZJU-7 (B. subtilis CGMCC1250), and their effects on the cell growth and γ-PGA biosynthesis were further investigated in shake flasks. The outcome indicated that both acetoin and 2,3-butanediol showed clear impairment on γ-PGA production of B. subtilis ZJU-7. Moreover, the extracellular metabolites profiles of fermentation under three different pH values were acquired and the metabolic flux redistribution of pathways related to γ-PGA biosynthesis was calculated based on the collected data. As a result, the metabolic flux favored to distribute toward glycolytic pathway at pH 6.5, in which the ingestion rate of extracellular glutamic acid was higher and the subsequent γ-PGA biosynthesis was enhanced. The present work provided us a deep insight into the metabolic flux control of γ-PGA biosynthesis, which will stimulate some novel metabolic engineering strategies to improve the productivity of γ-PGA in future.