Overexpression of a bacterial chymotrypsin: Application for L-amino acid ester hydrolysis (Articolo in rivista)

Type

• Articolo in rivista (Classe)
• Prodotto della ricerca (Classe)

Label

• Overexpression of a bacterial chymotrypsin: Application for L-amino acid ester hydrolysis (Articolo in rivista) (literal)

Anno

• 2011-01-01T00:00:00+01:00 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi

• 10.1016/j.enzmictec.2011.06.009 (literal)

Alternative label

• Volonte, Federica; Pisanelli, Ines; D'Arrigo, Paola; Viani, Fiorenza; Molla, Gianluca; Servi, Stefano; Pollegioni, Loredano (2011)
  Overexpression of a bacterial chymotrypsin: Application for L-amino acid ester hydrolysis
  in Enzyme and microbial technology
  (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori

• Volonte, Federica; Pisanelli, Ines; D'Arrigo, Paola; Viani, Fiorenza; Molla, Gianluca; Servi, Stefano; Pollegioni, Loredano (literal)

Pagina inizio

• 560 (literal)

Pagina fine

• 566 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume

• 49 (literal)

Rivista

• Enzyme and microbial technology (Rivista)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#pagineTotali

• 7 (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroFascicolo

• 6-7 (literal)

Note

• ISI Web of Science (WOS) (literal)
• ISI Web of Science (WoS) (literal)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni

• University of Insubria; Polytechnic University of Milan; Polytechnic University of Milan; Consiglio Nazionale delle Ricerche (CNR) (literal)

Titolo

• Overexpression of a bacterial chymotrypsin: Application for L-amino acid ester hydrolysis (literal)

Abstract

• In this work, a reliable protocol was designed to rapidly express and purify a microbial chymotrypsin(ogen) as a useful alternative to using animal proteases. The cDNA encoding for chymotrypsinogen from the deuteromycete Metarhizium anisopliae (chy1) was overexpressed in an Origami2(DE3) E. coli strain deficient in thioredoxin reductase and glutathione reductase activities, thus possibly allowing disulfide exchange. By using a quick purification protocol, in which the hexahistidine tag was added at the C-terminal end of the protease, the recombinant CHY1 protein could be purified in a single step on an Ni-NTA column as a mixture of 19.5- and 15-kDa mature active forms and did not require further activation/maturation steps. This expression and purification procedure offers an easier and faster means of producing recombinant CHY1 chymotrypsin than that previously described for Pichia pastoris. The kinetic properties could be characterized and CHY1 chymotrypsin was demonstrated to efficiently catalyze N-acetylated L-phenylalanine and L-tyrosine methyl ester hydrolysis. (C) 2011 Elsevier Inc. All rights reserved. (literal)

Prodotto di

• Istituto di chimica del riconoscimento molecolare (ICRM) (Istituto)
• Applicazioni sintetiche della biocatalisi (PM.P03.009.002) (Modulo)

Autore CNR

• FIORENZA VIANI (Unità di personale interno)
• STEFANO SERVI (Unità di personale esterno)

Insieme di parole chiave

• Keywords of "Overexpression of a bacterial chymotrypsin: Application for L-amino acid ester hydrolysis" (Insieme di parole chiave)

Incoming links:

Autore CNR di

• FIORENZA VIANI (Unità di personale interno)
• STEFANO SERVI (Unità di personale esterno)

Prodotto

• Istituto di chimica del riconoscimento molecolare (ICRM) (Istituto)
• Applicazioni sintetiche della biocatalisi (PM.P03.009.002) (Modulo)

Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#rivistaDi

• Enzyme and microbial technology (Rivista)

Insieme di parole chiave di

• Keywords of "Overexpression of a bacterial chymotrypsin: Application for L-amino acid ester hydrolysis" (Insieme di parole chiave)