Cloning, expression and characterization of catechol 1,2-dioxygenase from Burkholderia cepacia

Tran Vu Ngoc Thi; Duong Duc Hoang Sinh; Le Thi Ha Thanh; Nguyen Duc Huy; Nguyen Hoang Tue; Masaki Shintani; Kazuhide Kimbara; Nguyen Hoang Loc

doi:10.2323/jgam.2019.06.002

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Cloning, expression and characterization of catechol 1,2-dioxygenase from Burkholderia cepacia

Tran Vu Ngoc Thi, Duong Duc Hoang Sinh, Le Thi Ha Thanh, Nguyen Duc Huy, Nguyen Hoang Tue, Masaki Shintani, Kazuhide Kimbara, Nguyen Hoang Loc

著者情報

Tran Vu Ngoc Thi
Department of Applied Chemistry and Biochemical Engineering, Shizuoka University
Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University
Duong Duc Hoang Sinh
Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University
Le Thi Ha Thanh
Department of Applied Chemistry and Biochemical Engineering, Shizuoka University
Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University
Nguyen Duc Huy
Department of Microbial Biotechnology, Institute of Biotechnology, Hue University
Nguyen Hoang Tue
Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University
Masaki Shintani
Department of Applied Chemistry and Biochemical Engineering, Shizuoka University
Kazuhide Kimbara
Department of Applied Chemistry and Biochemical Engineering, Shizuoka University
Nguyen Hoang Loc
Institute of Bioactive Compounds and Department of Biotechnology, University of Sciences, Hue University

キーワード: Burkholderia cepacia, cat gene, catechol 1,2-dioxygenase, dibenzofuran

ジャーナルフリー早期公開

論文ID: 2019.06.002

DOI https://doi.org/10.2323/jgam.2019.06.002

この記事には本公開記事があります。

The final version of this article is now available: Vol. 66 (2020), No. 3 pp. 188-194

詳細

抄録

The present study reports on the cloning, expression and characterization of catechol 1,2-dioxygenase (CAT) of bacterial strains isolated from dioxin-contaminated soils in Vietnam. Two isolated bacterial strains DF2 and DF4 were identified as Burkholderia cepacia based on their 16S rRNA sequences. Their genes coding CAT was amplified with a specific pair of primers. Recombinant CAT (rCAT) was expressed in E. coli M15 cells and its activity was confirmed by the detection of cis,cis-muconic acid, a product from catechol, by high-performance liquid chromatography (HPLC) analysis. The rCAT of DF4 had an optimal pH and temperature of 7 and 30°C, respectively. Metal ions, such as Zn²⁺ and Mn²⁺, and surfactants, such as SDS, Tween 20 and Triton X100, strongly inhibited enzyme activity, while K⁺ slightly increased the activity.

責任著者(Corresponding author)

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