重组人血清白蛋白在Pichia pastoris中分泌表达影响因素的研究  

Study on Factors Influencing Secretion of Recombinant Human Serum Albumin from Pichia pastoris

作　　者：杨晟[1] 黄鹤[1] 章如安[1] 钱美琴 余有浩 钱雪明[1] 袁中一[1] 

机构地区：[1]中国科学院上海生物化学研究所,上海200031 [2]上海第一生化药业公司,上海200080 [3]上海大学生物工程系,上海200180

出　　处：《生物工程学报》2000年第6期675-678,共4页Chinese Journal of Biotechnology

摘　　要：为获得高产稳产重组人血清白蛋白的Pichiapastoris细胞株用于高密度发酵 ,构建了多种分泌表达载体 ,在Pichiapastoris中作表达 ,研究表达载体构建等因素对表达量的影响。发现采用酿酒酵母α性成熟因子前导肽比人血清白蛋白天然前导肽作为分泌信号肽的表达量要高 10 %左右。而载体整合方式、整合拷贝数、5’非翻译区的改造和甲醇利用表型等对表达量无规律性影响。筛选到的高表达株经高密度发酵 ,细胞湿重达 46 0 g/L ,发酵液上清的人血清白蛋白含量为 1 0 g/L。To enhance the expression of recombinant human serum albumin (HSA) in Pichia pastoris, several expression vectors have been constructed and transformed to Pichia pastoris .It was found that the use of α factor from Saccharomyces cerevisiae as a leader sequence was better than that of the native human serum albumin (HSAprepro).Compared with latter,the former (MF α 1prepro) increased the yield of recombinant albumin about 10%.The data also showed that the integrate site,copy number of expression vectors,5' untranslated region and methanol utilization type had no obvious influence on yield of HSA.The high productivity strain obtained by screening GS115(pPIC9\| hsa ) transformants reached cell mass of 460 g/L (Wet) and produced 1 0 g/L HSA in supernatant of culture broth via high density fermentation.

关 键 词：重组人血清白蛋白 高密度发酵 分泌表达 

分 类 号：Q786[生物学—分子生物学]