Identification and characterization of a dextranase gene of Streptococcus criceti.

Identification and characterization of a dextranase gene of Streptococcus criceti. Research Abstract Details 

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Identification and characterization of a dextranase gene of Streptococcus criceti. Abstract Text:

Haruki Tamura,Arisa Yamada,Hirohisa Kato,Haruki Tamura,Arisa Yamada,Hirohisa Kato,

The dextranase gene, dex, was identified in Streptococcus criceti strain E49 by degenerate PCR and sequenced completely by the gene-walking method. A sequence of 3,960 nucleotides was determined. The dex gene encodes a 1,200-amino acid protein, which has a calculated molecular mass of 128,129.91 and pI of 4.15 and is predicted to be a cell-surface protein. The deduced amino acid sequence of dex showed homology to S. downei dextranase (63.9% identity). Phylogenetic analysis revealed the similarity of the deduced amino acid sequence of dextranases in S. criceti, S. sobrinus, and S. downei. A recombinant form of the protein with six histidine residues tagged in the C-terminus was partially purified and showed dextranase activity on blue-dextran sodium dodecyl sulfate-polyacrylamide gel electrophoresis (BD-SDSPAGE) followed by renaturation. We also detected dextranase activity in S. criceti cell extracts and culture supernatant by renatured BD-SDS-PAGE, whereas no dextranase activity of the cells was observed on blue-dextran brain heart infusion (BD-BHI) agar plates. Furthermore, PCR-based mutations of dextranase indicated that a deletion mutant of the C-terminal region could hydrolyze blue dextrans and that the D453E mutation, W793L mutation, and double mutations (W793L and deletion of the C-terminal region) resulted in a loss of dextranase activity. These findings suggest that Asp-453 and Trp-793 residues of S. criceti dextranase are critical to the enzyme's activity.

Identification and characterization of a dextranase gene of Streptococcus criceti. Publishing Authors By Initials

H Tamura,A Yamada,H Kato,H Tamura,A Yamada,H Kato,

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Identification and characterization of a dextranase gene of Streptococcus criceti. Journal Published:

PUBLICATION TYPE: Research Support, Non-U.S. Gov

Journal: Microbiology and immunology

VOLUME: 51

Page Numbers: 721-32

Journal Abbreviation: Microbiol. Immunol.

ISSN: 0385-5600

DAY: 20

MONTH: 08

YEAR: 2007

Identification and characterization of a dextranase gene of Streptococcus criceti. Information

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LANGUAGE: eng

NlmUniqueID: 7703966

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Grant and Affiliation Information for Identification and characterization of a dextranase gene of Streptococcus criceti.

AFFILIATION: Department of Dental Pharmacology, School of Dentistry, Iwate Medical University, Morioka, Iwate 020-8505, Japan. htamura@iwate-med.ac.jp

Country: Japan

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MEDLINETA: Microbiol Immunol

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