title: Expression, Glycosylation, and Secretion of an Aspergillus Glucoamylase by Saccharomyces cerevisiae.
abstract: A strain of Saccharomyces cerevisiae capable of simultaneous hydrolysis and fermentation of highly polymerized starch oligosaccharides was constructed. The Aspergillus awamori glucoamylase enzyme, form GAI, was expressed in Saccharomyces cerevisiae by means of the promoter and termination regions from a yeast enolase gene. Yeast transformed with plasmids containing an intron-free recombinant glucoamylase gene efficiently secreted glucoamylase into the medium, permitting growth of the transformants on starch as the sole carbon source. The natural leader sequence of the precursor of glucoamylase (preglucoamylase) was processed correctly by yeast, and the secreted enzyme was glycosylated through both N- and O-linkages at levels comparable to the native Aspergillus enzyme. The data provide evidence for the utility of yeast as an organism for the production, glycosylation, and secretion of heterologous proteins.
authors.forename: M A
authors.initials: MA
authors.lastname: Innis
authors.forename: M J
authors.initials: MJ
authors.lastname: Holland
authors.forename: P C
authors.initials: PC
authors.lastname: McCabe
authors.forename: G E
authors.initials: GE
authors.lastname: Cole
authors.forename: V P
authors.initials: VP
authors.lastname: Wittman
authors.forename: R
authors.initials: R
authors.lastname: Tal
authors.forename: K W
authors.initials: KW
authors.lastname: Watt
authors.forename: D H
authors.initials: DH
authors.lastname: Gelfand
authors.forename: J P
authors.initials: JP
authors.lastname: Holland
authors.forename: J H
authors.initials: JH
authors.lastname: Meade