γ-Polyglutamic acid production in Corynebacterium glutamicum using sugar by one-step fermentation

Abstract:

γ-polyglutamic acid (γ-PGA) is widely used in food processing, cosmetic production, medicinal industry, etc. Currently, the production strains used in fermentation process are commonly glutamic acid-dependent, which results in extra cost. In this study, a de novo way of producing γ-PGA from sugars was reported. To this end, the γ-polyglutamate synthase gene cluster pgsBCA was cloned from the natural γ-PGA-producing strain Bacillus subtilis (ATCC 6051-U), and was constitutively and inducibly expressed in Corynebacterium glutamicum ATCC 13032. Only inducible expression of pgsBCA can lead to the generation of γ-PGA with a titer of 1.43 g/L from glucose, without any supplementation of glutamic acid. The production was further elevated to 1.98 g/L upon optimization of the induction conditions with the induction time at 2 h post-inoculation and the IPTG concentration of 0.8 mmol/L. Moreover, to achieve a higher titer of γ-PGA, pgsBCA was inducibly expressed in C. glutamicum F343, which shows a paramount glutamate production capacity. The final γ-PGA production reached 10.23 g/L in shake flasks and 20.08 g/L in a 5-L fermentor using glucose as the substrate. The weight-average molecular weight (Mw) of γ-PGA from recombinant strain F343 showed 34.77% higher than that produced by B. subtilis. This study provides a novel way of producing γ-PGA from sugars directly and potentiates new applications of γ-PGA in the future.

Key words:γ-polyglutamate, Corynebacterium glutamicum, one-step fermentation, sugar, pgsBCA