Expression of bovine follicle-stimulating hormone subunits in a Hansenula polymorpha expression system increases the secretion and bioactivity in vivo.
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.
Qian
Weidong
W
Center for Agricultural Biotechnology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Liu
Yueyong
Y
Zhang
Chaozheng
C
Niu
Zhendong
Z
Song
Haolei
H
Qiu
Bingsheng
B
D000818:Animals; D037281:Calnexin / Q000235:genetics / Q000378:metabolism; D002417:Cattle; D002846:Chromatography, Affinity; D003062:Codon / Q000378:metabolism; D037201:Follicle Stimulating Hormone, beta Subunit / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D015292:Glycoprotein Hormones, alpha Subunit / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D006031:Glycosylation; D006639:Histidine / Q000737:chemistry / Q000235:genetics; D009842:Oligopeptides / Q000737:chemistry / Q000235:genetics; D010843:Pichia / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029701:Saccharomyces cerevisiae Proteins / Q000235:genetics / Q000378:metabolism
C080582:CNE1 protein, S cerevisiae; D003062:Codon; D037201:Follicle Stimulating Hormone, beta Subunit; D015292:Glycoprotein Hormones, alpha Subunit; C471213:His-His-His-His-His-His; D009842:Oligopeptides; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D037281:Calnexin; D006639:Histidine