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Titolo:
Directed evolution of an esterase: Screening of enzyme libraries based on pH-Indicators and a growth assay
Autore:
Bornscheuer, UT; Altenbuchner, J; Meyer, HH;
Indirizzi:
Univ Stuttgart, Inst Tech Biochem, D-70569 Stuttgart, Germany Univ Stuttgart Stuttgart Germany D-70569 hem, D-70569 Stuttgart, Germany Univ Stuttgart, Inst Ind Genet, D-70569 Stuttgart, Germany Univ StuttgartStuttgart Germany D-70569 net, D-70569 Stuttgart, Germany Univ Hannover, Inst Organ Chem, D-30167 Hannover, Germany Univ Hannover Hannover Germany D-30167 n Chem, D-30167 Hannover, Germany
Titolo Testata:
BIOORGANIC & MEDICINAL CHEMISTRY
fascicolo: 10, volume: 7, anno: 1999,
pagine: 2169 - 2173
SICI:
0968-0896(199910)7:10<2169:DEOAES>2.0.ZU;2-C
Fonte:
ISI
Lingua:
ENG
Soggetto:
PSEUDOMONAS-FLUORESCENS; RECOMBINANT ESTERASE; 3-HYDROXY ESTERS; ORGANIC-SOLVENTS; ENANTIOSELECTIVITY; EPOTHILONES;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Life Sciences
Citazioni:
25
Recensione:
Indirizzi per estratti:
Indirizzo: Bornscheuer, UT Univ Stuttgart, Inst Tech Biochem, Allmandring 31, D-70569Stuttgart, Germany Univ Stuttgart Allmandring 31 Stuttgart Germany D-70569 y
Citazione:
U.T. Bornscheuer et al., "Directed evolution of an esterase: Screening of enzyme libraries based on pH-Indicators and a growth assay", BIO MED CH, 7(10), 1999, pp. 2169-2173

Abstract

In order to resolve a sterically hindered 3-hydroxy ethyl ester, which wasnot accepted as substrate by 20 wild-type hydrolases, a directed evolutionof an esterase from Pseudomonas fluorescens (PFE) was performed. Mutationswere introduced using the mutator strain Epicurian coli XL1-Red. Enzyme libraries derived from seven mutation cycles were assayed on minimal media agar plates supplemented with pH indicators (neutral red and crystal violet),thus allowing the identification of active esterase variants by the formation of a red color caused by a pH decrease due to the released acid. A further selection criteria was introduced by using the corresponding glycerol ester, because release of the carbon source glycerol facilitates growth on minimal media. By this strategy, one double mutant (A209D and L181V) of PFE was identified, which hydrolyzed the 3-hydroxy ethyl ester in a stereoselective manner (25% ee for the remaining ester, E similar to 5). (C) 1999 Elsevier Science Ltd. All rights reserved.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 02:06:06