Cloning and expression of the α-L-arabinofuranosidase gene (ABF2) of Aspergillus niger in Saccharomyces cerevisiae
Date
1996
Authors
Crous J.M.
Pretorius I.S.
Van Zyl W.H.
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Abstract
First-strand cDNA was prepared from mRNA of Aspergillus niger MRC11624 induced on oat spelts xylan. Using the cDNA as a template, the α-L-arabinofuranosidase gene (abfB) was amplified with the polymerase chain reaction technique. The abfB DNA fragment was inserted between the yeast phosphoglycerate kinase I gene promoter (PGK1(P)) and terminator (PGK1(T)) sequences on a multicopy episomal plasmid. The resulting construct PGK1(P)abfB-PGK1(T) was designated ABF2. The ABF2 gene was expressed successfully in Saccharomyces cerevisiae and functional α-L-arabinofuranosidase was secreted from the yeast cells. The ABF2 nucleotide sequence was determined and verified to encode a 449-amino-acid protein (Abf2) that is 9.4% identical to the α-L-arabinofuranosidase B of A. niger N400. Maximum α-L-arabinofuranosidase activities of 0.020 U/ml and 1.40 U/ml were obtained with autoselective recombinant S. cerevisiae strains when grown for 48 h in synthetic and complex medium respectively.
Description
Keywords
complementary dna, messenger rna, article, aspergillus niger, controlled study, dna sequence, enzyme activity, fungus mutant, gene expression, molecular cloning, nonhuman, saccharomyces cerevisiae, Amino Acid Sequence, Aspergillus niger, Base Sequence, Biotechnology, Cloning, Molecular, DNA, Complementary, DNA, Fungal, Gene Expression, Genes, Fungal, Glycoside Hydrolases, Molecular Sequence Data, Saccharomyces cerevisiae, Sequence Homology, Amino Acid, Aspergillus niger, Fungi, Saccharomyces cerevisiae
Citation
Applied Microbiology and Biotechnology
46
3
46
3