Expression of the STA2 glucoamylase gene of Saccharomyces cerevisiae in brewers' yeast

Expression of the STA2 glucoamylase gene of Saccharomyces cerevisiae in brewers' yeast

Two fragments of DNA containing the Saccharomyces cerevisiae STA2 glucoamylase gene, with differing lengths of 5' non-coding DNA, were separately subcloned into a yeast centromeric plasmid. Of these two subclones, only the shorter one (containing 127 base-pairs of 5' non-coding DNA) was ab...

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Bibliographic Details
Published in:Journal of the Institute of Brewing Vol. 103; no. 1; pp. 35 - 39
Main Authors: Lyness, C.A, Meaden, P.G
Format: Journal Article
Language:English
Published: Oxford, UK Blackwell Publishing Ltd 01-01-1997
Institute of Brewing
Subjects:
Biological and medical sciences
Biotechnology
brewers yeast
clones
DNA
enzyme activity
Fundamental and applied biological sciences. Psychology
gene expression
Genetic engineering
Genetic technics
genetic transformation
glucan 1,4-alpha-glucosidase
Glucoamylase
Methods. Procedures. Technologies
Modification of gene expression level
Saccharomyces cerevisiae
STA2 gene
structural genes
subclones
yeast
Recombinant microorganism
Glucan 1,4-α-glucosidase
Yeast
Genetic transformation
Enzyme
Gene expression
Fungi
Enzymatic activity
Glycosidases
Hydrolases
Ascomycetes
Brewer yeast
Recombinant protein
Saccharomyces cerevisiae
Vector
Thallophyta
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Description
Summary:Two fragments of DNA containing the Saccharomyces cerevisiae STA2 glucoamylase gene, with differing lengths of 5' non-coding DNA, were separately subcloned into a yeast centromeric plasmid. Of these two subclones, only the shorter one (containing 127 base-pairs of 5' non-coding DNA) was able to confer glucoamylase production on a standard laboratory strain of S. cerevisiae. The longer subclone (containing 465 bp of 5' non-coding DNA) did, however, confer glucoamylase production on a strain of S. cerevisiae lacking a functional STA10 gene (which encodes a repressor of STA2 gene expression). All-yeast plasmids lacking bacterial DNA were constructed from the two STA2 subclones for the transformation of a lager brewing yeast. Only the shorter STA2 subclone conferred glucoamylase activity on this yeast. The level of enzyme activity was comparable to that produced by the same yeast strain containing STA2 expressed from the PGK1 (that is, PGK1) promoter.
Bibliography:ark:/67375/WNG-JXX86WD8-L
istex:A228E51220E6172F50D21EA21D1CA939FCAC348E
ArticleID:JIB935
ISSN:0046-9750
2050-0416
DOI:10.1002/j.2050-0416.1997.tb00935.x