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

J 2001

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

MARVANOVÁ, Soňa, Yuji NAGATA, Michaela WIMMEROVÁ, Jana SÝKOROVÁ, Kamila HYNKOVÁ et. al.

Basic information

Original name

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

Authors

MARVANOVÁ, Soňa, Yuji NAGATA, Michaela WIMMEROVÁ, Jana SÝKOROVÁ, Kamila HYNKOVÁ and Jiří DAMBORSKÝ

Edition

Journal of Microbiological Methods, 2001, 0167-7012

Other information

Language

English

Type of outcome

Článek v odborném periodiku

Field of Study

10600 1.6 Biological sciences

Country of publisher

Netherlands

Confidentiality degree

není předmětem státního či obchodního tajemství

References:

URL

Impact factor

Impact factor: 1.810

RIV identification code

RIV/00216224:14310/01:00002964

Organization unit

Faculty of Science

UT WoS

000167080200007

Změněno: 13/2/2002 05:58, prof. Mgr. Jiří Damborský, Dr.

Abstract

V originále

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.

Links

ME 276, research and development project

Name: Racionální re-design mikrobiálních enzymů podílejících se na degradaci toxických organických polutantů

Investor: Ministry of Education, Youth and Sports of the CR, Rational re-design of microbial enzymes involved in degradation of toxic organic pollutants.

MSM 143100005, plan (intention)

Name: Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Investor: Ministry of Education, Youth and Sports of the CR, Biomolecular Structure-function Relationships and their role in the Metabolism

Citovat

MARVANOVÁ, Soňa, Yuji NAGATA, Michaela WIMMEROVÁ, Jana SÝKOROVÁ, Kamila HYNKOVÁ and Jiří DAMBORSKÝ. 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, vol. 44, No 1, p. 149-157. ISSN 0167-7012.

@article{352155,
   author = {Marvanová, Soňa and Nagata, Yuji and Wimmerová, Michaela and Sýkorová, Jana and Hynková, Kamila and Damborský, Jiří},
   article_number = {1},
   language = {eng},
   issn = {0167-7012},
   journal = {Journal of Microbiological Methods},
   title = {Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: characterization of the haloalkane dehalogenase mutants},
   url = {http://www.ncbr.chemi.muni.cz/~jiri/ABSTRACTS/jmm01.html},
   volume = {44},
   year = {2001}
}

TY  - JOUR
ID  - 352155
AU  - Marvanová, Soňa - Nagata, Yuji - Wimmerová, Michaela - Sýkorová, Jana - Hynková, Kamila - Damborský, Jiří
PY  - 2001
TI  - Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: characterization of the haloalkane dehalogenase mutants
JF  - Journal of Microbiological Methods
VL  - 44
IS  - 1
SP  - 149
EP  - 149
SN  - 01677012
UR  - http://www.ncbr.chemi.muni.cz/~jiri/ABSTRACTS/jmm01.html
N2  - 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.
ER  -

MARVANOVÁ, Soňa, Yuji NAGATA, Michaela WIMMEROVÁ, Jana SÝKOROVÁ, Kamila HYNKOVÁ and Jiří DAMBORSKÝ. Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: characterization of the haloalkane dehalogenase mutants. \textit{Journal of Microbiological Methods}. 2001, vol.~44, No~1, p.~149-157. ISSN~0167-7012.

Detailed Information on Publication Record