Construction of an expression system for the secretory production of recombinant 관-agarase in yeast.

국문 초록 (Abstract)

<P>관-Agarase hydrolyzes the 관-1,3 linkage of agarose yielding agaro-oligosaccharides. It is less well characterized than 관-agarase. AgaA gene (2.3 kb ORF), encoding the 관-agarase from Thalassomonas JAMB A33, was subcloned into both a constitutive and an inducible expression vector. Both the constructed plasmids, pVT-AgaA (ADH1 promoter) and pYInu-AgaA (GAL10 promoter), were transformed into Saccharomyces cerevisiae SEY2102 and FY833 and pPIC9-AgaA harboring the AOX1 promoter was transformed into Pichia pastoris GS115. The recombinant 관-agarases were over-expressed with activities from 0.3 to 1.6 unit/ml, the highest being in the SEY2102/pYInu-AgaA transformant. Most of the recombinant 관-agarase was extracellular because each plasmid possesses a signal sequence for the secretory production of 관-agarase. In contrast, the Pichia host-vector expression system was unsuitable for the production of recombinant 관-agarase. This is the first report of recombinant production of 관-agarase in yeast for industrial use.</P>