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.

Základní údaje

Originální název

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

Autoři

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

Vydání

Journal of Microbiological Methods, 2001, 0167-7012

Další údaje

Jazyk

angličtina

Typ výsledku

Článek v odborném periodiku

Obor

10600 1.6 Biological sciences

Stát vydavatele

Nizozemské království

Utajení

není předmětem státního či obchodního tajemství

Odkazy

URL

Impakt faktor

Impact factor: 1.810

Kód RIV

RIV/00216224:14310/01:00002964

Organizační jednotka

Přírodovědecká fakulta

UT WoS

000167080200007

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

Anotace

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.

Návaznosti

ME 276, projekt VaV

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

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Racionální re-design mikrobiálních enzymů podílejících se na degradaci toxických organických polutantů

MSM 143100005, záměr

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

Investor: Ministerstvo školství, mládeže a tělovýchovy ČR, Strukturně-funkční vztahy biomolekul a jejich role v metabolismu

Citovat

MARVANOVÁ, Soňa, Yuji NAGATA, Michaela WIMMEROVÁ, Jana SÝKOROVÁ, Kamila HYNKOVÁ a 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, roč. 44, č. 1, s. 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 a 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, roč.~44, č.~1, s.~149-157. ISSN~0167-7012.

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