Biochemical characterization of broad-specificity enzymes using multivariate experimental design and a colorimetric microplate assay: characterization of the haloalkane dehalogenase mutants
Jazyk angličtina Země Nizozemsko Médium print
Typ dokumentu hodnotící studie, časopisecké články, práce podpořená grantem
PubMed
11165344
DOI
10.1016/s0167-7012(00)00250-5
PII: S0167701200002505
Knihovny.cz E-zdroje
- MeSH
- hydrolasy genetika metabolismus MeSH
- kolorimetrie metody MeSH
- koncentrace vodíkových iontů MeSH
- multivariační analýza MeSH
- mutace * MeSH
- proteinové inženýrství metody MeSH
- Sphingomonas enzymologie genetika MeSH
- substrátová specifita MeSH
- Publikační typ
- časopisecké články MeSH
- hodnotící studie MeSH
- práce podpořená grantem MeSH
- Názvy látek
- haloalkane dehalogenase MeSH Prohlížeč
- hydrolasy MeSH
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.
Citace poskytuje Crossref.org
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