A high-throughput colorimetric assay for screening halohydrin dehalogenase saturation mutagenesis libraries

Journal of Biotechnology

Volumn 147, Issue 3-4, 2010, Pages 164-168

  Tang, Lixia ^a   Li, Yang ^a   Wang, Xiong ^a  

^a UNIVERSITY OF ELECTRONIC SCIENCE AND TECHNOLOGY OF CHINA   (China)

Author keywords

Colorimetric assay; Halohydrin dehalogenase; High throughput screening; Microplate format; PH indicator

Indexed keywords

COLORIMETRIC ASSAYS; DEHALOGENASE; HIGH THROUGHPUT SCREENING; MICROPLATES; PH INDICATOR;

ENZYMES; ETHANOL; MUTAGENESIS; PH; PHENOLS;

COLORIMETRY;

1,3 DICHLORO 2 PROPANOL; 4 NITRO 2 BROMO 1 PHENYLETHANOL; ALCOHOL DERIVATIVE; ENZYME VARIANT; HALOHYDRIN DEHALOGENASE; HYDROLASE; PHENOLSULFONPHTHALEIN; PROPANOL; PROTON; UNCLASSIFIED DRUG; BUFFER; DYES, REAGENTS, INDICATORS, MARKERS AND BUFFERS; MUTANT PROTEIN;

ARTICLE; COLORIMETRY; CONTROLLED STUDY; ENZYME ACTIVITY; ENZYME KINETICS; ENZYME MECHANISM; HIGH THROUGHPUT SCREENING; NONHUMAN; PH MEASUREMENT; PRIORITY JOURNAL; RING CLOSING METATHESIS; SITE DIRECTED MUTAGENESIS; WILD TYPE; ENZYME ASSAY; ENZYMOLOGY; GENETICS; KINETICS; METABOLISM; METHODOLOGY; MUTAGENESIS; MUTATION; PH; POLYACRYLAMIDE GEL ELECTROPHORESIS; RHIZOBIUM; SPECTROSCOPY;

BUFFERS; COLORIMETRY; ELECTROPHORESIS, POLYACRYLAMIDE GEL; ENZYME ASSAYS; HIGH-THROUGHPUT SCREENING ASSAYS; HYDROGEN-ION CONCENTRATION; HYDROLASES; INDICATORS AND REAGENTS; KINETICS; MUTAGENESIS; MUTANT PROTEINS; MUTATION; PHENOLSULFONPHTHALEIN; RHIZOBIUM; SPECTRUM ANALYSIS;

EID: 78349291583     PISSN: 01681656     EISSN: 18734863     Source Type: Journal    
DOI: 10.1016/j.jbiotec.2010.04.002     Document Type: Article

References (15)

1. Bergmann J.G., Sanik J. Determination of trace amounts of chlorine in naphtha. Anal. Chem. 1957, 29:241-243.
2. Chapman E., Wong C.H. A pH sensitive colorimetric assay for the high throughput screening of enzyme inhibitors and substrates: a case study using kinases. Bioorg. Med. Chem. 2002, 10:551-555.
3. de Jong R.M., Tiesinga J.J.W., Rozeboom H.J., Kalk K.H., Tang L., Janssen D.B., Dijkstra B.W. Structure and mechanism of a bacterial haloalcohol dehalogenase: a new variation of the short-chain dehygrogenase/reductase fold without an NAD(P)H binding site. EMBO J. 2003, 22:4933-4944.
4. Fox R.J., Davis C.S., Mundorff E.C., et al. Improving catalytic function by ProSAR-driven enzyme evolution. Nat. Biotechnol. 2007, 25:338-344.
5. ® multisite-directed mutagenesis kit. BioTechniques 2002, 33:1158-1165.
6. Holloway P., Knoke K.L., Trevors J.T., Lee H. A colorimetric assay for detecting haloalkane dehalogenase activity. Biotech. Bioeng. 1998, 59:520-523.
7. Lutje Spelberg J.H., Tang L., van Gelder M., Kellogg R.M., Janssen D.B. Exploration of the biocatalytic potential of a halohydrin dehalogenase using chromogenic substrates. Tetrahedron: Asymm. 2002, 13:1083-1089.
8. Lutje Spelberg J.H., Tang L., Kellogg R.M., Janssen D.B. Enzymatic dynamic resolution of epihalohydrins. Tetrahedron: Asymm. 2004, 15:1095-1102.
9. Majeric-Elenkov M., Tang L., Meetsma A., Hauer B., Janssen D.B. Formation of enantiopure 5-substituted oxazolidinones through enzyme-catalysed kinetic resolution of epoxides. Organic Lett. 2008, 10:2417-2420.
10. Neidleman, S.L., Amon, W.F. Jr., Geigert, J., 1981. Preparation of epoxides and glycols from gaseous alkenes. US patent. No 4 284 723 (to Cetus).
11. Parikh M.R., Matsumura I. Site-saturation mutagenesis is more efficient than DNA shuffling for the directed evolution of β-fucosidase from β-galactosidase. J. Mol. Biol. 2005, 352:621-628.
12. Price J.A. A colorimetric assay for measuring phospholipase A2 degradation of phosphatidylcholine at physiological pH. J. Biochem. Biophys. Methods 2007, 70:441-444.
13. Reetz M.T., Carballeira J.D. Iterative Saturation Mutagenesis (ISM) for rapid directed evolution of functional enzymes. Nat. Protoc. 2007, 2:891-903.
14. Tang L., van Merode A.E.J., Lutje Spelberg J.H., Fraaije M.W., Janssen D.B. Steady-state kinetics and tryptophan fluorescence properties of halohydrin dehalogenase from Agrobacterium radiobacter, Roles of W139 and W249 in the active site and halide-induced conformational change. Biochemistry 2003, 42:14057-14065.
15. Van Hylckama Vlieg J.E.T., Tang L., Lutje Spelberg J.H., Smilda T., Poelarends G.J., Bosma T., van Merode A.E.J., Fraaije M.W., Janssen D.B. Halohydrin dehalogenases are structurally and mechanistically closely related to short-chain dehydrogenases/reductases. J. Bacteriol. 2001, 183:5058-5066.