Cloning and characterization of a dextranase gene fromLipomyces starkeyi and its expression inSaccharomyces cerevisiae
| Autor: | Hee-Kyoung Kang, Seung Heuk Kim, Doman Kim, Jiyoung Park, Deok-Kun Oh, Seong Soo Kang, Xing-Ji Jin |
|---|---|
| Rok vydání: | 2005 |
| Předmět: |
animal structures
Molecular Sequence Data Bioengineering Saccharomyces cerevisiae Biology Applied Microbiology and Biotechnology Biochemistry Dextranase activity Substrate Specificity Complementary DNA Genetics Amino Acid Sequence Cloning Molecular Peptide sequence Penicillium minioluteum Dextranase Base Sequence Molecular mass Temperature Protein primary structure Sequence Analysis DNA Hydrogen-Ion Concentration Molecular biology Recombinant Proteins Open reading frame Saccharomycetales Biotechnology |
| Zdroj: | Yeast. 22:1239-1248 |
| ISSN: | 1097-0061 0749-503X |
| DOI: | 10.1002/yea.1311 |
| Popis: | 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 mM Hg2+, Ag3+ and Mn2+. LSD1 exhibited high hydrolysing activity towards dextran (100%), soluble starch (22%) and mutan (8%). Copyright © 2005 John Wiley & Sons, Ltd. |
| Databáze: | OpenAIRE |
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