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利用表达大肠杆菌谷氨酸脱羧酶的重组谷氨酸棒杆菌进行γ-氨基丁酸的稳健生产。

Robust production of gamma-amino butyric acid using recombinant Corynebacterium glutamicum expressing glutamate decarboxylase from Escherichia coli.

机构信息

Department of Chemical Science and Engineering, Graduate School of Engineering, Graduate School of Science and Technology, Kobe University, 1-1 Rokkodaicho, Nada-ku, Kobe 657-8501, Japan.

出版信息

Enzyme Microb Technol. 2012 Aug 10;51(3):171-6. doi: 10.1016/j.enzmictec.2012.05.010. Epub 2012 Jun 4.

Abstract

Gamma-amino butyric acid (GABA) is a component of pharmaceuticals, functional foods, and the biodegradable plastic polyamide 4. Here, we report a simple and robust system to produce GABA from glucose using the recombinant Corynebacterium glutamicum strain GAD, which expresses GadB, a glutamate decarboxylase encoded by the gadB gene of Escherichia coli W3110. As confirmed by HPLC analysis, GABA fermentation by C. glutamicum GAD cultured at 30°C in GABA Production 1 (GP1) medium containing 50 g/L glucose without the addition of glutamate yielded 8.07 ± 1.53 g/L extracellular GABA after 96 h. Addition of 0.1mM pyridoxal 5'-phosphate (PLP) was found to enhance the production of GABA, whereas Tween 40 was unnecessary for GABA fermentation. Using the optimized GABA Production 2 (GP2) medium, which contained 50 g/L glucose and 0.1mM PLP, fermentation was performed in a flask at 30°C with 10% (v/v) seed culture of C. glutamicum GAD. GABA was produced in the culture supernatant with a yield of 12.37 ± 0.88 g/L after 72 h with a space-time yield of 0.172 g/L/h, which is the highest yield obtained to date for GABA from fermentation with glucose as a main carbon source.

摘要

γ-氨基丁酸(GABA)是药品、功能性食品和可生物降解塑料聚酰胺 4 的组成部分。在这里,我们报告了一种使用重组谷氨酸棒杆菌 GAD 从葡萄糖生产 GABA 的简单而强大的系统,该系统表达了谷氨酸脱羧酶 GadB,这是大肠杆菌 W3110 的 gadB 基因编码的。通过 HPLC 分析证实,在含有 50g/L 葡萄糖的 GABA 生产 1(GP1)培养基中,在 30°C 下培养谷氨酸棒杆菌 GAD ,不添加谷氨酸,经过 96 小时后,细胞外 GABA 的产量为 8.07±1.53g/L。添加 0.1mM 吡哆醛 5'-磷酸(PLP)被发现可以提高 GABA 的产量,而 Tween 40 对于 GABA 发酵不是必需的。使用优化的 GABA 生产 2(GP2)培养基,其中含有 50g/L 葡萄糖和 0.1mM PLP,在 30°C 下用 10%(v/v)谷氨酸棒杆菌 GAD 的种子培养液进行发酵。在培养上清液中生产 GABA,72 小时后产量为 12.37±0.88g/L,时空产率为 0.172g/L/h,这是迄今为止使用葡萄糖作为主要碳源发酵生产 GABA 的最高产量。

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