高产谷胱甘肽新菌种的选育及其工艺条件的研究-生物化工专业论文.docxVIP

高产谷胱甘肽新菌种的选育 及其工艺条件的研究 摘要 以乙硫氨酸和氯化锌为筛选标记，对出发菌株SL21通过紫外线 诱变处理，运用推理育种技术，选育到一株抗乙硫氨酸和氯化锌的突 变株HY78，该突变株经摇瓶发酵谷胱甘肽的产量为76．Omg／L，干细 胞谷胱甘肽的含量为10．Omg／g，分别比出发菌株SL21提高268．9％ 和263．O％。该菌株经10次传代培养，谷胱甘肽产量仅下降了13．0 ％，是一株性状较稳定的优良菌株。 运用单因素实验和正交实验设计对突变株HY78产谷胱甘肽的发 酵条件进行了研究，研究表明该突变株产谷胱甘肽的最佳发酵条件为 培养温度30C、初始pH7．0、摇床转速180 r／min、接种龄18h、接 种量10％，摇瓶装液量30ml／250m1；最佳培养基组成是：糖蜜409， 牛肉膏209，Cys·HCI 1．59，MgS04·7H。0 99，KH2P04 lg，ZnSO。 5mg，水1000 ml，结果表明突变株HY78在该优化条件下摇瓶发酵， 发酵液中谷胱甘肽的含量可达182．6mg／L，较优化前提高了140．3％。 在摇瓶培养的基础上，通过分批培养和补料分批培养，在3．7L 自动生物发酵罐中对突变株HY78生物合成谷胱甘肽进行了扩大培 养，结果表明分批培养较摇瓶培养谷胱甘肽的产量下降了15．9％， 仅有157．6mg／L，而在补料分批培养中，由于流加了培养液，谷胱甘  肽的产量较分批培养提高了59．7％，为251．7mg／L。 关键词：藤黄八叠球菌，谷胱甘肽，高产菌株，发酵条件，扩大培养  —————————————————————————————————-—————_—————————————，————————一一堂竺兰兰圭兰兰主竺兰—— BREEDING OF NOVEL OVERPRODUING GLUTATHIONE STRAIN AND STUDYING ON ITS FERN匝NT芦汀ION CONDITIONS ABSTRACT A Ethionine—．and Zinc Chloride．．resistant mutant HY78 was obtained from its parem strain SL21 by UV treatment and rational screening．The glutathione productivity of the mutant reached 76．0medL by flask culture．which was 268．9％ higher than the parent strain．And the glutathione content in the dried ceils reached 10．Omg／L．which was 263．0％higher than the parent strain．A descend ofonly 13．O％ in the glutathione yield of the mutant WaS observed after ten times of subculture． Therefore，the obtained mutant is a stable strain． Studies were carried out on the fermentation condition through Single Factor design and Orthogonal Test design，which showed that the optimum fermentation condition was culture temperature 30℃，initial pH7．0，rotate speed 1 80r／min， inoculum age 1 8h，inoculum size 1 0％，medium volumes 30ml／250m1．And the optimum medium composition was molasses 409，beef extract 209，Cys。HCl 1．59， MgS04·7H20 99，KH2P04 l g，ZnS04 5mg，water 1 000ml，In this condition the ghitathione content carrie tO 182．6mg／L．which wag 140_3％hi．gher than that of non—optimized medium． In a 3．7L scale batch bioreactoL the rules of glutathione fermentation were