巴斯德毕赤酵母发酵生产重组人溶菌酶  

Fermentation of recombinant human lysozyme in Pichia pastoris

作　　者：陈晶晶[1] 陈劲春[1] 

机构地区：[1]北京化工大学生命科学与技术学院,北京100029

出　　处：《北京化工大学学报（自然科学版）》2006年第6期34-37,共4页Journal of Beijing University of Chemical Technology(Natural Science Edition)

摘　　要：对巴斯德毕赤酵母GS115菌株在工业培养基中发酵和分离条件进行了研究。结果表明:将传统工业培养基中碳源和氮源的质量浓度分别降低至35 g/L和5 g/L时,酵母生长期的细胞密度与传统培养基的相差不大,但培养时间可减少4 h,节省了生产成本;维持诱导期间的pH值为4.0对目标蛋白的表达最有利,目标蛋白的表达量占酵母分泌蛋白总量的70%,比原来增加了近30%;分离纯化过程,采用将强阳离子(SPFF)与弱阴离子(DEAE)交换层析柱偶联的分离方法,达到除杂,并浓缩目标蛋白的作用,目标蛋白产量约为50 mg/L;将目标蛋白酶切后,质量酶活性为8.56×10-3mol/(s.g)。Human lysozyme, sourced from human body fluids, has fewer side effects when injected into the human body and is therefore safer in practice. Here we report a study of fermentation conditions and purification of recombinant human lysozyme in Pichia pastoris GS115. （1） When the carbon and nitrogen sources were simultaneously reduced from 40 g/L to 35 g/L and from 10 g/L to 5 g/L respectively, the cell density was not significantly reduced compared with that in the traditional medium. However, the culture time could be shortened by about 4 hours. （2） During the induction period of the yeast cells, it was found that the target protein yield could be increased by about 30% by maintaining the medium at pH 4.0. （3） For chromatographic purification of the target protein, a coupled column of SP-Sepharose FF strong cation-exchange resin and DEAE Sepharose FF weak anion-exchange resin was employed to obtain the purified target protein with a concentration of about 50mg/L. （4） After digestion with enterokinase, the target protein showed bioactivity of about 8.56×10^-3 mol/（s·g）.

关 键 词：人溶菌酶 毕赤酵母 发酵 分离纯化 

分 类 号：Q786[生物学—分子生物学]