Quantitative screening of hydrolase libraries using pH indicators: Identifying active and enantioselective hydrolases

Chemistry - A European Journal

Volumn 4, Issue 11, 1998, Pages 2324-2331

  Janes, Lana E ^a   Löwendahl, A Christina ^a   Kazlauskas, Romas J ^a  

^a MCGILL UNIVERSITY   (Canada)

Author keywords

Enantiomeric resolution; Enzyme catalysis; Lipases; Screening; Synthetic methods

Indexed keywords

HYDROLASE; INDICATOR; TRIACYLGLYCEROL LIPASE;

ARTICLE; CATALYSIS; ENANTIOMER; ENZYME ACTIVITY; ENZYME SPECIFICITY; PH; SCREENING; STEREOSPECIFICITY; SYNTHESIS;

EID: 0031796722     PISSN: 09476539     EISSN: None     Source Type: Journal    
DOI: 10.1002/(SICI)1521-3765(19981102)4:11<2324::AID-CHEM2324>3.0.CO;2-I     Document Type: Article

References (32)

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22. Ref. [5], p. 80. The extinction coefficients change slightly upon addition of cosolvent and should be determined experimentally.
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24. The pathlength in a 96-well plate depends on the volume of the solution in the well, since the light passes from the top of the plate through the solution. Thus, the maximum indicator concentration varies with the solution volumes. With other acid-base indicators, poor water solubility can also limit the maximal concentration. The 4-nitrophenol concentration in our solutions (0.45 mM or 0.006%) was well below its solubility limit, 0.08%: see ref. [19], p. 92.
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28. a of solketal. Other esters (for example, 4-nitrophenyl esters) may release more than one proton per ester group hydrolyzed and should be accounted for in Equation (1).
29. Other researchers have also estimated enantioselectivity by separately measuring the rates of hydrolysis of the pure enantiomers. For example: G. Zandonella, L. Haalck, F. Spener, K. Faber, F. Paltauf, A. Hermetter, Chirality 1996, 8, 481-489; M. T. Reetz, A. Zonta, K. Schimossek, K. Liebeton, K.-E. Jaeger, Angew. Chem. 1997, 109, 2961-2963; Angew. Chem. Int. Ed. Engl. 1997, 36, 2830-2832.
30. Other researchers have also estimated enantioselectivity by separately measuring the rates of hydrolysis of the pure enantiomers. For example: G. Zandonella, L. Haalck, F. Spener, K. Faber, F. Paltauf, A. Hermetter, Chirality 1996, 8, 481-489; M. T. Reetz, A. Zonta, K. Schimossek, K. Liebeton, K.-E. Jaeger, Angew. Chem. 1997, 109, 2961-2963; Angew. Chem. Int. Ed. Engl. 1997, 36, 2830-2832.
31. Other researchers have also estimated enantioselectivity by separately measuring the rates of hydrolysis of the pure enantiomers. For example: G. Zandonella, L. Haalck, F. Spener, K. Faber, F. Paltauf, A. Hermetter, Chirality 1996, 8, 481-489; M. T. Reetz, A. Zonta, K. Schimossek, K. Liebeton, K.-E. Jaeger, Angew. Chem. 1997, 109, 2961-2963; Angew. Chem. Int. Ed. Engl. 1997, 36, 2830-2832.
32. Ref. [19], p. 94.