Results
Title

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

Authors

Seok, Ji-Hwan; Kim, Hye-Soo; Hatada, Yuji; Nam, Soo-Wan; Kim, Yeon-Hee

Abstract

α-Agarase hydrolyzes the α-1,3 linkage of agarose yielding agaro-oligosaccharides. It is less well characterized than β-agarase. AgaA gene (2.3 kb ORF), encoding the α-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 α-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 α-agarase was extracellular because each plasmid possesses a signal sequence for the secretory production of α-agarase. In contrast, the Pichia host-vector expression system was unsuitable for the production of recombinant α-agarase. This is the first report of recombinant production of α-agarase in yeast for industrial use.

Subjects

YEAST fungi genetics; AGARASE; GENETIC vectors; RECOMBINANT microorganisms; LEAVENING agents; SACCHAROMYCES cerevisiae

Publication

Biotechnology Letters, 2012, Vol 34, Issue 6, p1041

ISSN

0141-5492

Publication type

Academic Journal

DOI

10.1007/s10529-012-0864-0