Secretion of Trichoderma reesei beta-glucosidase by Saccharomyces cerevisiae.
An intronless form of the bgl1 gene encoding an extracellular beta-glucosidase from Trichoderma reesei was expressed in the yeast Saccharomyces cerevisiae under the control of the yeast GAL1 promoter. Transformation of a yeast strain with this vector resulted in transformants that produce and secrete active beta-glucosidase into the growth medium. Additionally, active recombinant beta-glucosidase protein was shown to be localized predominantly in the periplasmic space by using a p-nitrophenyl beta-D-glycoside hydrolysis assay against fractionated yeast cells. The apparent size of the recombinant enzyme was 10-15 kDa larger than that of the native form. Treatment of the recombinant beta-glucosidase with endoglycosidase-H indicated the apparent increase in size was due to N-linked glycosylation.
Cummings
C
C
Genencor International, 180 Kimball Way, South San Francisco, CA 94080, USA.
Fowler
T
T
D001483:Base Sequence; D003001:Cloning, Molecular; D003470:Culture Media; D005822:Genetic Vectors; D006031:Glycosylation; D006596:Hexosaminidases / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D011401:Promoter Regions, Genetic; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000378:metabolism; D012441:Saccharomyces cerevisiae / Q000235:genetics*; D014170:Transformation, Genetic; D014242:Trichoderma / Q000201:enzymology / Q000235:genetics*; D001617:beta-Glucosidase / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism
D003470:Culture Media; D011994:Recombinant Proteins; D006596:Hexosaminidases; D001617:beta-Glucosidase