高产谷氨酸产生菌的工艺研究.docxVIP

高产谷氨酸产生菌的工艺研究Study on technology of high yielding glutamic acid producing strains13生工杨雅娴前言工业发酵研究和开发的主要目标之一是建立一种能实现高产、低成本的可行过程，生产条件优化是改进现有菌种产量的重要方法。本文对发酵法生产谷氨酸的试验流程进行优化，探讨发酵生产过程中的关键影响因素，发现谷氨酸发酵生产中，溶解氧水平是发酵生产的限制瓶颈，也是得到高产谷氨酸的关键因素之一。因为低溶氧更有利于产乳酸，乳酸产量增加将直接导致谷氨酸产量减少。试验以糖质原料为出发，结合谷氨酸产量跟踪，确定了菌株由“生长型”转化为“生产型”的时间，从而确定了“分瓶”培养的方法。发酵前期注重菌体的生长培养，发酵后期减少液体量以增大溶解氧来进一步提高谷氨酸产量。通过这种培养，得到一套较为完整的谷氨酸摇瓶发酵的方案。同时，对发酵过程中培养基及培养条件进行优化，得到较好的谷氨酸产量。Industrial fermentation research and development one of the main objectives is to establish which can realize high yield, low cost feasible process, optimization of production conditions is important for improved strains of existing production methods. This article to optimize the testing process of the glutamic acid production by fermentation, explores the key influencing factors of fermentation found in glutamate fermentation, the level of dissolved oxygen is the limit of production bottlenecks, is also one of the key factors of high yielding glutamic acid. Due to low dissolved oxygen is more conducive to production, will lead directly to an increase in production of glutamic acid production of lactic acid. Experiments with sugar raw material based on the combination of glutamic acid production tracking, strains were determined by growing into production time, so as to determine the bottle method of training. Fermentation early focus on bacterial growth, late fermentation to further reduce the amount of liquid to increase dissolved oxygen increasing glutamic acid production. Through this training, get a more complete fermentation of glutamic acid solution. Same time, to optimize fermentation culture medium and culture conditions, get a better production of glutamic acid.一、谷氨酸棒杆菌的诱变选育1.1 试验方法 1.1.1菌种纯化谷氨酸棒杆菌 W3 平板（完全培养基）划线分离。32 ℃，培养 24～30 h，挑取单菌落，继续纯化培养，如此反复三次，保证菌落纯后保藏。1.1.2菌种保藏用斜面保藏培养基，4 ℃保藏。 2.3.3 斜面活化 W3 菌株(分离纯化后)接入斜面培养基，32 ℃培养 24 h。 1.1.3种子液摇床培养条件 32 ℃，200 r/min。 2.3.5 种子培养取活化菌种两环接入装有 30 m L/250 m L 种子培养液的三角瓶中，摇床培养 16 h。1.1.4.1紫外诱变处理（1）菌种