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

Volumn 44, Issue 2, 2001, Pages 149-157

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

^a MASARYK UNIVERSITY   (Czech Republic)

^b TOHOKU UNIVERSITY   (Japan)

Author keywords

Colorimetric assay; Experimental design; Microplate; Multivariate; Principal component analysis

Indexed keywords

HALOALKANE DEHALOGENASE; MICROBIAL ENZYME; UNCLASSIFIED DRUG;

ARTICLE; COLORIMETRY; CONTROLLED STUDY; ENZYME SPECIFICITY; ENZYME STRUCTURE; GAS CHROMATOGRAPHY; METHODOLOGY; NONHUMAN; PHYSICAL CHEMISTRY; POINT MUTATION; PRINCIPAL COMPONENT ANALYSIS; PRIORITY JOURNAL; SCREENING; SPHINGOMONAS PAUCIMOBILIS; VALIDATION PROCESS;

COLORIMETRY; HYDROGEN-ION CONCENTRATION; HYDROLASES; MULTIVARIATE ANALYSIS; MUTATION; PROTEIN ENGINEERING; SPHINGOMONAS; SUBSTRATE SPECIFICITY;

SPHINGOMONAS PAUCIMOBILIS;

EID: 0035284071     PISSN: 01677012     EISSN: None     Source Type: Journal    
DOI: 10.1016/S0167-7012(00)00250-5     Document Type: Article

References (16)

1. Analysis of the reaction mechanism and substrate specificity of haloalkane dehalogenases by sequential and structural comparisons
2. Some biochemical properties and classification of a range of bacterial haloalkane dehalogenases
3. Alternation of the substrate range of haloalkane dehalogenase by site-directed mutagenesis
4. A colorimetric assay for detecting haloalkane dehalogenase activity
5. Identification of the catalytic triad in the haloalkane dehalogenase from Sphingomonas paucimobilis UT26
6. Construction and characterization of histidine-tagged haloalkane dehalogenase (LinB) of a new substrate class from a γ-hexachlorocyclohexane-degrading bacterium, Sphingomonas paucimobilis UT26
7. Protein Engineering
8. Site-directed mutagenesis and oxygen isotope incorporation studies of the nucleophilic aspartate of haloalkane dehalogenase
9. Activation of an Asp-124→Asn mutant of haloalkane dehalogenase by hydrolytic deamidation of asparagine
10. Histidine 289 is essential for hydrolysis of the alkyl-enzyme intermediate of haloalkane dehalogenase
11. Kinetic characterization and X-ray structure of a mutant of haloalkane dehalogenase with higher catalytic activity and modified substrate range
12. Haloalkane dehalogenases: Steady-state kinetics and halide inhibition
13. Dehalogenases applied to industrial-scale biocatalysis
14. Principal Component Analysis