黄杆菌肝素酶Ⅱ重组菌培养条件的优化及高密度发酵  

Culture conditions optimization and high cell density fermentation of recombinant bacteria producing heparinaseⅡ from Flavobacterium heparinum

作　　者：周斌[1] 程咏梅[1] 邓超[2] 刘卫超[1] 陈潮梁 陈敬华[1] 许正宏[1] 

机构地区：[1]江南大学药学院,江苏无锡214122 [2]江南大学无锡医学院,江苏无锡214122

出　　处：《生物工程学报》2014年第4期674-678,共5页Chinese Journal of Biotechnology

基　　金：教育部新世纪优秀人才支持计划(No.NCET-10-0435);教育部博士点基金(No.20110093110008)资助~~

摘　　要：黄杆菌肝素酶Ⅱ(HepⅡ)是一类可特异性切割肝素、硫酸乙酰肝素类分子内连接键的酶。文中对黄杆菌肝素酶Ⅱ重组菌的诱导时机、诱导剂添加量、诱导温度、诱导时间等诱导产酶条件进行优化。经过优化最佳摇瓶发酵产酶条件为:37℃培养重组菌至对数生长前期,添加诱导剂IPTG至终浓度为0.3 g/L,20℃下诱导10 h,酶活达到最高,为570 U/L。在此基础上通过发酵罐高密度培养手段将菌体浓度OD600进一步提高到98,酶活大幅度提高到9 436 U/L,该研究结果为HepⅡ的工业化生产与应用奠定了良好的基础。Heparinase Ⅱ （Hep Ⅱ） from Flavobacterium heparinum is an enzyme that could specifically cleave certain sequence of heparin and heparan sulfate. In this work, fermentation conditions of recombinant heparinase I｜ （His-Hep Ⅱ ） producing bacteria were optimized, including initial induction time, inducer （IPTG） concentration, induction temperature and induction time. The optimum conditions were as follows： cultivating recombinant bacteria to exponential prophase under 37 ℃, then adding IPTG to a final concentration of 0.3 g/L, finally cultivating recombinant bacteria under 20℃for l 0 h. The total crude enzyme activity reached 570 U/L. Based on these results, high cell density fermentation of recombinant bacteria was studied. The final 09600 could reach 98 and the total crude enzyme activity of His-Hep Ⅱ increased to 9 436 U/L.

关 键 词：肝素酶Ⅱ 培养条件优化 高密度发酵 

分 类 号：Q814[生物学—生物工程]