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Cloning and characterization of a dextranase gene from Lipomyces starkeyi and its expression in Saccharomyces cerevisiae
Author(s) -
Kang HeeKyoung,
Kim Seung Heuk,
Park JiYoung,
Jin XingJi,
Oh DeokKun,
Soo Kang Seong,
Kim Doman
Publication year - 2005
Publication title -
yeast
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.923
H-Index - 102
eISSN - 1097-0061
pISSN - 0749-503X
DOI - 10.1002/yea.1311
Subject(s) - biology , cloning (programming) , saccharomyces cerevisiae , dextranase , gene , molecular cloning , gene expression , microbiology and biotechnology , genetics , biochemistry , enzyme , computer science , programming language
A dextranase‐encoding cDNA from L. starkeyi KSM22 was isolated and characterized. The 2052 bp cDNA fragment ( lsd1 ) harbouring the dextranase gene exhibited one open reading frame (ORF) composed of 1824 bp flanked by a 41 bp 5′‐UTR and a 184 bp 3′‐UTR, including a 27 bp poly(A) tail. The lsd1 gene contains no introns. The open reading frame encodes a 608 amino acid polypeptide (LSD1) with a 67.6 kDa predicted molecular mass. There was a 77% deduced amino acid sequence identity between the LSD1 dextranase and the dextranase from Penicillium minioluteum . The primary structure of LSD1 dextranase exhibits distant similarity with the enzymes of the glycosyl hydrolase family 49 that comprises Penicillium dextranase. The optimum pH of LSD1 was 6.0 and the optimum temperature was 37 °C. LSD1 dextranase activity was substantially abolished by exposure to 1 m M Hg 2+ , Ag 3+ and Mn 2+ . LSD1 exhibited high hydrolysing activity towards dextran (100%), soluble starch (22%) and mutan (8%). Copyright © 2005 John Wiley & Sons, Ltd.