以葡萄糖为原料直接发酵生产γ聚谷氨酸的研究.pdfVIP

以葡萄糖为原料直接发酵生产γ-聚谷氨酸的研究 * 张慧莉 张绪 石峰 徐志南 岑沛霖 （浙江大学化学工程与生物工程学系，浙江杭州 310027） 摘要：γ-聚谷氨酸（γ-PGA）是一种可以通过微生物合成的均聚谷氨酸高分子化合物，在食品、医药、水处 理和农业中有广泛的应用前景。为了避免在γ-聚谷氨酸的发酵生产中加入L-谷氨酸，本文探讨将谷氨酸的 发酵生产与谷氨酸聚合相耦联，实现直接利用葡萄糖生产 γ- 聚谷氨酸。首先开展将谷氨酸棒杆菌 Corynebacterium glutamicum S9114 与具有γ-聚谷氨酸合成能力的枯草芽孢杆菌Bacillus subtilis ZJU7 进行混 合培养，研究了接种量、培养基、温度和溶氧对混合培养体系发酵产率的影响，经过优化最终γ-聚谷氨酸 -1 产量达到32.8g ·l 。然后以L-谷氨酸生产菌Brevibacterium flavum 和枯草芽孢杆菌Bacillus subtilis ZJU7 为亲本，利用原生质体融合技术构建可自身合成和聚合谷氨酸的融和菌株，考察了两亲本的原生质体形成、 再生和融合的最优条件，对所获融合子进行了筛选和分析。 关键词：聚谷氨酸 混合培养 原生质体融合 Studies on Microbial Production of γ-PGA from glucose ZHANG Huili, ZHANG Xu, SHI Feng, XU Zhinan, CEN Peilin (School of Materials Science Chemical Engineering，Zhejiang University, Hangzhou 310027 ，Zhejiang ，China) Abstract: Poly-γ-glutamic acid (γ-PGA) is a kind of water-soluble and biodegradable anionic polypeptide that is made of D- and L-glutamic acid units linked by amide linkages between α-amino and γ-carboxylic acid groups with various applications in medicine, food and agriculture. In order to avoid the addition of L-glutamic acid in the medium, the coupled bioprocess of the glutamine producing and polymerization process was explored. Firstly, a co-cultivation system of Corynebacterium glutamicum and Bacillus subtilis was established and the co-cultivation conditions, such as strain ratio, temperature and oxygen concentration, were optimized. A γ-PGA productivity of 32.8 g ·l-1 was achieved in the optimized conditions. Secondly, a method of PEG induced protoplast fusion was developed for obtaining stains with abilities of producing glutamine and polymerization. The conditions of protoplast forming and regeneration were optimized and the fusants were screened and analyzed. Key words: poly γ-glutamic acid; co-cultivation; protoplast fusion 引言