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.
出处
《生物工程学报》
CAS
CSCD
北大核心
2000年第6期675-678,共4页
Chinese Journal of Biotechnology
关键词
重组人血清白蛋白
高密度发酵
分泌表达
Human serum albumin, Pichia pastoris, α factor secretion signal, high density fermentation
