Production of heterologous proteins in yeast with the aid of the Hsp150delta carrier.
Proper folding, and consequently exit from the endoplasmic reticulum (ER) and secretion of heterologous exocytic proteins in yeast can be rescued by fusing the proteins to certain yeast-derived polypeptides. Biologically active mammalian glycoproteins can be produced in Saccharomyces cerevisiae and Pichia pastoris by joining them to a fragment of a natural secretory glycoprotein of S. cerevisiae, Hsp150delta. The performance of the Hsp150delta carrier in both yeasts appears to exceed that of the MFalpha leader, which is widely used in industrial protein production. Here we describe the use of the Hsp150delta carrier in P. pastoris in both shake flask and fermentor cultivations. As a reporter protein we use the periplasmic disulfide-bonded Escherichia coli enzyme beta-lactamase.
Makarow
Marja
M
Department of Applied Chemistry and Microbiology, University of Helsinki, Finland.
Hänninen
Anna-Liisa
AL
Suntio
Taina
T
Bastos
Ricardo Nunes
RN
D001483:Base Sequence; D002352:Carrier Proteins / Q000096:biosynthesis / Q000235:genetics; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005285:Fermentation; D017930:Genes, Reporter; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics*; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics*; D009172:Mycology / Q000379:methods; D010843:Pichia / Q000235:genetics / Q000378:metabolism; D010957:Plasmids / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics*; D012441:Saccharomyces cerevisiae / Q000235:genetics* / Q000378:metabolism*; D029701:Saccharomyces cerevisiae Proteins / Q000096:biosynthesis* / Q000235:genetics*; D014170:Transformation, Genetic; D001618:beta-Lactamases / Q000096:biosynthesis / Q000235:genetics
D002352:Carrier Proteins; D004274:DNA, Recombinant; D006023:Glycoproteins; C074229:HSP150 protein, S cerevisiae; D006360:Heat-Shock Proteins; D011993:Recombinant Fusion Proteins; D029701:Saccharomyces cerevisiae Proteins; D001618:beta-Lactamases