Cheng Hui, Chen Yuanyuan, Zhu Yaxin, Cao Rong, Xu Guoqiang, Zhang Xiaomei, Shi Jinsong, Xu Zhenghong
Key Laboratory of Industrial Biotechnology of Ministry of Education, Jiangnan University, Wuxi 214122, Jiangsu, China.
National Engineering Laboratory for Cereal Fermentation Technology, Jiangnan University, Wuxi 214122, Jiangsu, China.
Sheng Wu Gong Cheng Xue Bao. 2020 Feb 25;36(2):295-308. doi: 10.13345/j.cjb.190071.
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.
γ-聚谷氨酸(γ-PGA)广泛应用于食品加工、化妆品生产、医药工业等领域。目前,发酵过程中使用的生产菌株通常依赖谷氨酸,这导致成本增加。在本研究中,报道了一种从糖类从头生产γ-PGA的方法。为此,从天然γ-PGA生产菌株枯草芽孢杆菌(ATCC 6051-U)中克隆了γ-聚谷氨酸合酶基因簇pgsBCA,并在谷氨酸棒杆菌ATCC 13032中组成型和诱导型表达。只有pgsBCA的诱导型表达才能在不添加任何谷氨酸的情况下,从葡萄糖产生滴度为1.43 g/L的γ-PGA。通过优化诱导条件,接种后2小时诱导时间和0.8 mmol/L的IPTG浓度,产量进一步提高到1.98 g/L。此外,为了获得更高滴度的γ-PGA,pgsBCA在谷氨酸棒杆菌F343中诱导表达,该菌株具有极高的谷氨酸生产能力。以葡萄糖为底物,在摇瓶中最终γ-PGA产量达到10.23 g/L,在5-L发酵罐中达到20.08 g/L。重组菌株F343产生的γ-PGA的重均分子量(Mw)比枯草芽孢杆菌产生的高34.77%。本研究提供了一种直接从糖类生产γ-PGA的新方法,并有望在未来拓展γ-PGA的新应用。