Qian Weidong, Liu Yueyong, Zhang Chaozheng, Niu Zhendong, Song Haolei, Qiu Bingsheng
Center for Agricultural Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Protein Expr Purif. 2009 Dec;68(2):183-9. doi: 10.1016/j.pep.2009.07.008. Epub 2009 Jul 25.
Bovine follicle-stimulating hormone (bFSH), a pituitary gonadotropin, is a heterodimer hormone that consists of a common alpha-subunit non-covalently associated with the hormone-specific beta-subunit. Unfortunately, expression levels of recombinant bFSH or its subunits are invariably low. We report here the secretory expression of biologically active bFSHalpha and bFSHbeta subunit in the methylotrophic yeast Hansenula polymorpha. A slightly higher level of expression of recombinant bFSH subunits was achieved by using the Saccharomyces cerevisiae-derived calnexin (ScCne1) as a chaperone in engineered H. polymorpha strains. The preliminary data also suggested that bFSH subunits expressed in H. polymorpha appeared to be less-glycosylated. This isoform had been shown to be 80% increase in in vivo bioactivity compared with the hyperglycosylated Pichia pastoris-derived recombinant bFSHalpha/beta. More sophisticated applications of bFSH would profit from the assembled less-glycosylated heterodimer.
牛卵泡刺激素（bFSH）是一种垂体促性腺激素，是一种异源二聚体激素，由一个共同的α亚基与激素特异性β亚基非共价结合组成。不幸的是，重组bFSH或其亚基的表达水平总是很低。我们在此报告在甲基营养型酵母多形汉逊酵母中生物活性bFSHα和bFSHβ亚基的分泌表达。通过在工程化的多形汉逊酵母菌株中使用酿酒酵母来源的钙连接蛋白（ScCne1）作为伴侣蛋白，实现了重组bFSH亚基的稍高水平表达。初步数据还表明，在多形汉逊酵母中表达的bFSH亚基糖基化程度似乎较低。与高糖基化的毕赤酵母来源的重组bFSHα/β相比，这种异构体在体内生物活性方面已显示出增加了80%。bFSH更复杂的应用将受益于组装的低糖基化异源二聚体。
