Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: Characterization of the haloalkane dehalogenase mutants

Soa Marvanová; Yuji Nagata; Michaela Wimmerová; Jana Sýkorová; Kamila Hynková; Jirí Damborský

doi:10.1016/S0167-7012(00)00250-5

Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: Characterization of the haloalkane dehalogenase mutants

Soa Marvanová, Yuji Nagata, Michaela Wimmerová, Jana Sýkorová, Kamila Hynková, Jirí Damborský

LIF - Molecular and Chemical Life Science

Research output: Contribution to journal › Article › peer-review

21 Citations (Scopus)

Abstract

The pH indicator dye-based colorimetric method and multivariate experimental design were used for the systematic biochemical characterization of the broad-specificity enzymes haloalkane dehalogenases. Halogenated compounds for characterization of the enzymes were selected using Principal Component Analysis. The substrates were characterised by 24 physico-chemical and structural descriptors. Thirty-four substrates were selected for testing out of 194 halogenated compounds. Relative activities determined using the optimised colorimetric microplate assay were validated against the catalytic constants determined by gas chromatography. The applicability of the assay was tested with F151L, F154L and F169L mutants of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26.

Original language	English
Pages (from-to)	149-157
Number of pages	9
Journal	Journal of Microbiological Methods
Volume	44
Issue number	2
DOIs	https://doi.org/10.1016/S0167-7012(00)00250-5
Publication status	Published - 2001 Mar 1

Keywords

Colorimetric assay
Experimental design
Microplate
Multivariate
Principal component analysis

ASJC Scopus subject areas

Microbiology
Molecular Biology
Microbiology (medical)

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Marvanová, S., Nagata, Y., Wimmerová, M., Sýkorová, J., Hynková, K., & Damborský, J. (2001). Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: Characterization of the haloalkane dehalogenase mutants. Journal of Microbiological Methods, 44(2), 149-157. https://doi.org/10.1016/S0167-7012(00)00250-5

Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay : Characterization of the haloalkane dehalogenase mutants. / Marvanová, Soa; Nagata, Yuji; Wimmerová, Michaela; Sýkorová, Jana; Hynková, Kamila; Damborský, Jirí.

In: Journal of Microbiological Methods, Vol. 44, No. 2, 01.03.2001, p. 149-157.

Research output: Contribution to journal › Article › peer-review

Marvanová, S, Nagata, Y, Wimmerová, M, Sýkorová, J, Hynková, K & Damborský, J 2001, 'Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: Characterization of the haloalkane dehalogenase mutants', Journal of Microbiological Methods, vol. 44, no. 2, pp. 149-157. https://doi.org/10.1016/S0167-7012(00)00250-5

Marvanová S, Nagata Y, Wimmerová M, Sýkorová J, Hynková K, Damborský J. Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: Characterization of the haloalkane dehalogenase mutants. Journal of Microbiological Methods. 2001 Mar 1;44(2):149-157. https://doi.org/10.1016/S0167-7012(00)00250-5

Marvanová, Soa ; Nagata, Yuji ; Wimmerová, Michaela ; Sýkorová, Jana ; Hynková, Kamila ; Damborský, Jirí. / Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay : Characterization of the haloalkane dehalogenase mutants. In: Journal of Microbiological Methods. 2001 ; Vol. 44, No. 2. pp. 149-157.

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abstract = "The pH indicator dye-based colorimetric method and multivariate experimental design were used for the systematic biochemical characterization of the broad-specificity enzymes haloalkane dehalogenases. Halogenated compounds for characterization of the enzymes were selected using Principal Component Analysis. The substrates were characterised by 24 physico-chemical and structural descriptors. Thirty-four substrates were selected for testing out of 194 halogenated compounds. Relative activities determined using the optimised colorimetric microplate assay were validated against the catalytic constants determined by gas chromatography. The applicability of the assay was tested with F151L, F154L and F169L mutants of the haloalkane dehalogenase from Sphingomonas paucimobilis UT26.",

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Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: Characterization of the haloalkane dehalogenase mutants

Abstract

Keywords

ASJC Scopus subject areas

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