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

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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.

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Acknowledgments

This research was supported by the Basic Science Research Program through the National Research Foundation of Korea (NRF) funded by the Ministry of Education, Science and Technology (2009-0075943 and 2010-0002360). J. H. Seok is the recipient of a graduate fellowship from the Ministry of Education through the Brain Korea 21 Project.

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Correspondence to Yeon-Hee Kim.

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Seok, JH., Kim, HS., Hatada, Y. et al. Construction of an expression system for the secretory production of recombinant α-agarase in yeast. Biotechnol Lett 34, 1041–1049 (2012). https://doi.org/10.1007/s10529-012-0864-0

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