Over-expression of the Saccharomyces cerevisiae exo-beta-1,3-glucanase gene together with the Bacillus subtilis endo-beta-1,3-1,4-glucanase gene and the Butyrivibrio fibrisolvens endo-beta-1,4-glucanase gene in yeast

van Rensburg, P.; van Zyl, W.H.; Pretorius, I.S.

Journal of Biotechnology 55(1): 43-53

1997


ISSN/ISBN: 0168-1656
PMID: 9226961
DOI: 10.1016/s0168-1656(97)00059-x
Accession: 009144275

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Abstract
The EXG1 gene encoding the main Saccharomyces cerevisiae exo-beta-1,3-glucanase was cloned and over-expressed in yeast. The Bacillus subtilis endo-1,3-1,4-beta-glucanase gene (beg1) and the Butyrivibrio fibrisolvens endo-beta-1,4-glucanase gene (end1) were fused to the secretion signal sequence of the yeast mating pheromone alpha-factor (MF-alpha-1-S) and inserted between the yeast alcohol dehydrogenase II gene promoter (ADH2p) and terminator (ADH2-T). Constructs ADH2-PMF-alpha-1-S-beg1-ADH2-T and ADH2p-MF-alpha-1-S-end1-ADH2-T, designated BEG1 and END1, respectively, were expressed separately and jointly with EXG1 in S. cerevisiae. The construction of fur1 ura3 S. cerevisiae strains allowed for the autoselection of these multicopy URA3-based plasmids in rich medium. Enzyme assays confirmed that co-expression of EXG1, BEG1 and END1 enhanced glucan degradation by S. cerevisiae.