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Titolo:
DIRECTED EVOLUTION OF AN ESTERASE FOR THE STEREOSELECTIVE RESOLUTION OF A KEY INTERMEDIATE IN THE SYNTHESIS OF EPOTHILONES
Autore:
BORNSCHEUER UT; ALTENBUCHNER J; MEYER HH;
Indirizzi:
UNIV STUTTGART,INST TECH BIOCHEM,ALLMANDRING 31 D-70569 STUTTGART GERMANY UNIV STUTTGART,INST IND GENET D-70569 STUTTGART GERMANY UNIV HANNOVER,INST ORGAN CHEM D-30167 HANNOVER GERMANY
Titolo Testata:
Biotechnology and bioengineering
fascicolo: 5, volume: 58, anno: 1998,
pagine: 554 - 559
SICI:
0006-3592(1998)58:5<554:DEOAEF>2.0.ZU;2-J
Fonte:
ISI
Lingua:
ENG
Soggetto:
RANDOM MUTAGENESIS; ESCHERICHIA-COLI; NUCLEOTIDE-SEQUENCE; 3-HYDROXY ESTERS; ORGANIC-SOLVENTS; LIPASE;
Keywords:
DIRECTED EVOLUTION; ESTERASE; EPOTHILON; PSEUDOMONAS FLUORESCENS; STEREOSELECTIVITY; MUTATOR STRAIN;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Science Citation Index Expanded
Citazioni:
20
Recensione:
Indirizzi per estratti:
Citazione:
U.T. Bornscheuer et al., "DIRECTED EVOLUTION OF AN ESTERASE FOR THE STEREOSELECTIVE RESOLUTION OF A KEY INTERMEDIATE IN THE SYNTHESIS OF EPOTHILONES", Biotechnology and bioengineering, 58(5), 1998, pp. 554-559

Abstract

The directed evolution of an esterase from Pseudomonas fluorescens using the mutator strain Epicurian coli XL1-Red was investigated. Mutants were assayed for their ability to hydrolyze a sterically hindered 3-hydroxy ester, which can serve as a building block in the synthesis ofepothilones. Screening was performed by plating esterase producing colonies derived from mutation cycles onto minimal media agar plates containing indicator substances (neutral red and crystal violet). Esterase-catalyzed hydrolysis of the 3-hydroxy ester (ethyl or glycerol ester) was detected by the formation of a red color due to a pH decrease caused by the released acid. Esterases isolated from positive clones were used in preparative biotransformations of the ethyl ester. One variant containing two mutations (A209D and L181V) stereoselectively hydrolyzed the ethyl ester resulting in 25% ee for the remaining ester. (C) 1998 John Wiley & Sons, Inc.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 27/11/20 alle ore 02:20:15