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Identification of an alkaline‐tolerant cyclodextrin‐metabolizing bacterium and characterization of its cyclodextrinase gene
Author(s) -
Kaulpiboon Jarunee,
Rimphanitchayakit Vichien,
Pongsawasdi Piamsook
Publication year - 2004
Publication title -
journal of basic microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.58
H-Index - 54
eISSN - 1521-4028
pISSN - 0233-111X
DOI - 10.1002/jobm.200410412
Subject(s) - bacteria , cyclodextrin , identification (biology) , gene , chemistry , biochemistry , microbiology and biotechnology , characterization (materials science) , biology , genetics , botany , materials science , nanotechnology
Bacillus circulans A11, an alkaline‐tolerant cyclodextrin‐metabolizing bacterium isolated from South‐East Asian soil, was reidentified as Paenibacillus sp. A11 based on 16S rRNA gene sequence comparison, G + C content and cellular fatty acid composition. Levels of similarity of the 16S rRNA gene between strain A11 and the Paenibacillus species were 90–99%, while similarity with Bacillus circulans was only 86%. The major cellular fatty acid was anteiso‐C 15:0 which accounted for 59.3% of the total cellular fatty acids and the G+C content was 50.3 mol%. The CDase gene coding for this enzyme was cloned into E. coli . The open reading frame of the CDase gene was 1,959 bp encoding a CDase of 653 amino acid residues. At maximum growth, the specific activity of the recombinant CDase from E. coli was higher than that of Paenibacillus sp. A11. By SDS‐PAGE, the translation product of the recombinant gene showed the same mobility as the purified CDase from the original strain. CDase from both Paenibacillus sp. A11 and E. coli produced glucose and maltose as dominant end‐products of β‐CD hydrolysis. The ratio of maltose to glucose was 1:2. (© 2004 WILEY‐VCH Verlag GmbH & Co. KGaA, Weinheim)