高产L赖氨酸菌种的选育及其发酵条件的研究-生物工程专业论文.docxVIP

摘要 4 电 本论文根据代谢控制发酵和代谢流分析理论，研究了 L-赖氨酸 高产菌选育及发酵条件。主要研究内容和结果如下: 以黄色短杆菌(Brevibαcterium flavum )FY翩 222 为出发菌株，经硫 酸工乙酶 (DES)和紫外线诱变处理，摇管初筛、摇瓶复筛、遗传标记 验证、单菌落分离和连续传代，筛选出一株 L赖氨酸产生菌株 .. BBCA-31 (丁hr+sar ) 。该菌株在未经优化的摇瓶发酵条件下可产赖 氨酸 96.4g1Lo ( 确定了亲株 BBCA翩 31 和YYCA-21 的原生质体形成和再生及进 行原生质体融合的最佳条件，采用原生质体融合技术最终筛选出一株 句 带有目的遗传标记的 L 赖氨酸高产菌 ZL-231 (Thr+saT + Met干AHVT+AEC T ) 。该商株在未优化条件下可产L-赖氨酸 123.6g1L。 经验证，菌株 ZL-231 的遗传性能稳定。 研究了菌株 ZL-231 的摇瓶分批发酵条件。应用正交设计试验法 优化种子培养基，并对种子培养条件进行了研究;在发酵培养基单因 素试验的基础上，应用晌店面分析试验法，优化出摇瓶发酵培养基的 最佳组成。在优化条件下，赖氨酸高产菌 ZL-231 的赖氨酸产量为 _1 47.3g/L和糖酸转化率为 53.6% ，比初始条件下的122.5g1L 和 48.5% 分别提高了 20.2%和 10.52%。 关键词: L-赖氨酸黄色短杆商诱变育种原生质体融合发酵 Abstract According to the theory of metabolic control fermentation，the dissertation focuses on the breeding of L-lysine high-yielding strain and its fermentation condition. The main research contents and results are as follows: Brevibacerium flavum FY-222 as a starting strain was stepwise mutated with protoplast UV and Diethyl sulfate. One L-lysine high嗣producing strain BBCA-31 (Thr冲SGr) with genetic marker was screened through shake tube primary screening，shake flask compound screening，genetic marker test and single clone isolation and continuous 曲 passage cultivation. 丁his strain could produce L-lysine 96.4g/L a位er fermentation under the fermentation condition without being optimized. Genetic character of strain BBCA-31 was very stable. The protoplast formation rate and regeneration and fusion condition of strain BBCA-31and YYCA-21 was studied in this dissertation. One L-lysine high唰producing strain ZL-231 was screened through protoplast fusion，shake flask compound screening，genetic marker test and single clone isolation and continuous passage cultivation. 丁his strain could produce L-lysine 123.6g/L after fermentation under 阳 fermentation condition without being optimized. Genetic character of strain ZL-231 was very stable. The batch fermentation conditions of strain ZL-231 in shake flas