重组酪氨酸酚裂解酶发酵工艺的研究  

Study on fermentation process for recombinant tyrosine phenol lyase

作　　者：汤晓玲 杨健 王志超 索慧 郑仁朝 TANG Xiaoling;YANG Jian;WANG Zhichao;SUO Hui;ZHENG Renchao(Institute of Biological Engineering, Zhejiang University of Technology, Hangzhou 310014)

机构地区：[1]浙江工业大学生物工程学院,浙江杭州310014

出　　处：《工业微生物》2018年第6期1-6,共6页Industrial Microbiology

基　　金：浙江省自然科学基金项目;青年基金(LQ17B060004)

摘　　要：左旋多巴是治疗帕金森氏病的首选药物,生物酶法合成左旋多巴具有工艺简单、条件温和、立体选择性高和环境友好等优点。本论文以实验室前期构建的表达具核梭杆菌(Fusobacterium nucleatum) TPL (Fn-TPL)的重组大肠杆菌为基础,采用单因素实验通过对5 L发酵罐发酵工艺优化以及补料策略的研究,确定了分批发酵的工艺参数:pH 6. 5,诱导温度30℃,诱导剂乳糖20 g/L。在5 L发酵罐中,进一步研究了10 mL/h、20 mL/h、30 mL/h三个速率的恒速流加对菌体生物量和TPL酶活的影响。结果表明,补料速率为20 mL/h时,生物量最高为30. 43 g dcw/L,体积酶活最高为9 420 U/L,较摇瓶发酵培养活力提高了3. 3倍。Levodopa(3,4-dihydroxyphenyl-l-alanine,L-DOPA)is the first choice of drug for the treatment of Parkinson’s disease.Compared with plant extraction and chemical synthesis,biosynthesis of L-DOPA has the advantages of simple process,mild reaction condition,high stereoselectivity and environmental friendliness.In this study,the fermentation process was optimized for the recombinant E.coli strain harboring TPL from Fusobacterium nucleatum(Fn-TPL)in a 5 L fermenter.The process parameters for batch fermentation were determined as follows:pH 6.5,induction temperature 30℃and inducer lactose 20 g/L.Fed-bath process with constant feeding was further applied and the results indicated that with 20 mL/h of constant feeding strategy,both the biomass and volumetric activity of Fn-TPL reached highest,which were 30.43 g dcw/L and 9 420 U/L,respectively.

关 键 词：酪氨酸酚裂解酶 左旋多巴 发酵工艺优化 分批补料培养 

分 类 号：TQ925[轻工技术与工程—发酵工程] TQ460.1