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首页> 外文期刊>Journal of Biotechnology >Expression of hepatitis B surface antigen S domain in recombinant Saccharomyces cerevisiae using GAL1 promoter
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Expression of hepatitis B surface antigen S domain in recombinant Saccharomyces cerevisiae using GAL1 promoter

机译：使用GAL1启动子在重组酿酒酵母中表达乙型肝炎表面抗原S结构域

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摘要

Hepatitis B virus, a member of the hepadnavirus family, causes the acute and chronic diseases of the human liver. The S domain of hepatitis B virus surface antigen (sHBsAg) was expressed under the control of the galactose-inducible GAL1 promoter in recombinant Saccharomyces cerevisiae. Batch fermentation of S. cerevisiae 2805/pdeltaMFalpha-sHBsAg resulted in 4.92gl(-1) dry cell mass and 2.21mgl(-1) sHBsAg concentration. To improve the expression level of sHBsAg, the pdi1 gene encoding protein disulfide isomerase was coexpressed in recombinant S. cerevisiae. Batch fermentation of S. cerevisiae 2805/pdeltaMFalpha-sHBsAg+pPDI using 21gl(-1) glucose and 33gl(-1) galactose resulted in 9.75gl(-1) dry cell mass and 13.3mgl(-1) sHBsAg concentration, which were 2 and 6 times higher than those for S. cerevisiae 2805/pdeltaMFalpha-sHBsAg, respectively. Appearance of three sHBsAgs with different molecular sizes of 24kDa, 34kDa and 40kDa in immunoblot assay and their endoglycosidase treatment indicated that sHBsAg might be expressed in three types of the authentic, MFalpha signal sequence-containing and N-glycosylated MFalpha signal sequence-containing forms. Fed-batch fermentation of recombinant S. cerevisiae 2805 coexpressing the sHBsAg and pdi1 genes was carried out by feeding 600gl(-1) glucose continuously and controlling galactose concentration at around 20gl(-1). As a result, 20.2gl(-1) dry cell mass and 74.4mgl(-1) maximum sHBsAg concentration were obtained, which were 4.1 and 33.7 times higher than those for the batch fermentation of S. cerevisiae 2805/pdeltaMFalpha-sHBsAg, respectively.
机译：乙肝病毒是肝炎病毒家族的成员，可引起人类肝脏的急性和慢性疾病。乙型肝炎病毒表面抗原（sHBsAg）的S结构域在重组酿酒酵母中在半乳糖诱导型GAL1启动子的控制下表达。啤酒酵母2805 / pdeltaMFalpha-sHBsAg的分批发酵产生4.92gl（-1）干细胞质量和2.21mgl（-1）sHBsAg浓度。为了提高sHBsAg的表达水平，在重组酿酒酵母中共表达编码蛋白质二硫键异构酶的pdi1基因。使用21gl（-1）葡萄糖和33gl（-1）半乳糖分批发酵酿酒酵母2805 / pdeltaMFalpha-sHBsAg + pPDI，导致干细胞质量为9.75gl（-1），sHBsAg浓度为13.3mgl（-1）。分别比酿酒酵母2805 / pMF-sHBsAg高2倍和6倍。三种分别具有24kDa，34kDa和40kDa分子大小的sHBsAg的出现及其免疫糖苷内切酶处理表明，sHBsAg可能以三种真实的形式表达，分别是含有MFalpha信号序列和含有N-糖基化的MFalpha信号序列。共表达sHBsAg和pdi1基因的重组酿酒酵母2805的分批补料发酵是通过连续补充600gl（-1）葡萄糖并控制半乳糖浓度在20gl（-1）来进行的。结果，获得了20.2gl（-1）的干细胞质量和74.4mgl（-1）的最大sHBsAg浓度，分别比酿酒酵母2805 / pdeltaMFalpha-sHBsAg的分批发酵分别高4.1和33.7倍。 。

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