Effect of prolonged exposure to organic solvents on the active site environment of subtisilis Carlsberg (Articolo in rivista)

Type
Label
  • Effect of prolonged exposure to organic solvents on the active site environment of subtisilis Carlsberg (Articolo in rivista) (literal)
Anno
  • 2010-01-01T00:00:00+01:00 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#doi
  • 10.1016/j.molcatb.2010.01.021 (literal)
Alternative label
  • Vibha Bansala; Yamixa Delgado; Ezio Fasoli; Amaris Ferrer; Kai Griebenow; Francesco Secundo; Gabriel L. Barletta. (2010)
    Effect of prolonged exposure to organic solvents on the active site environment of subtisilis Carlsberg
    in Journal of molecular catalysis. B, Enzymatic (Print)
    (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#autori
  • Vibha Bansala; Yamixa Delgado; Ezio Fasoli; Amaris Ferrer; Kai Griebenow; Francesco Secundo; Gabriel L. Barletta. (literal)
Pagina inizio
  • 38 (literal)
Pagina fine
  • 44 (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#numeroVolume
  • 64 (literal)
Rivista
Note
  • ISI Web of Science (WOS) (literal)
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#affiliazioni
  • University of Puerto Rico at Humacao, Department of Chemistry, Humacao 00791, Puerto Rico University of Puerto Rico at Rio Piedras, Department of Chemistry, Puerto Rico (literal)
Titolo
  • Effect of prolonged exposure to organic solvents on the active site environment of subtisilis Carlsberg (literal)
Abstract
  • The potential of enzyme catalysis as a tool for organic synthesis is nowadays indisputable, as is the fact that organic solvents affect an enzyme's activity, selectivity and stability. Moreover, it was recently realized that an enzyme's initial activity is substantially decreased after prolonged exposure to organic media, an effect that further hampers their potential as catalysts for organic synthesis. Regrettably, the mechanistic reasons for these effects are still debatable. In the present study we have made an attempt to explain the reasons behind the partial loss of enzyme activity on prolonged exposure to organic solvents. Fluorescence spectroscopic studies of the serine protease subtilisin Carlsberg chemically modified with polyethylene glycol (PEG-SC) and inhibited with a dansyl fluorophore, and dissolved in two organic solvents (acetonitrile and 1,4-dioxane) indicate that when the enzyme is initially introduced into these solvents, the active site environment is similar to that in water; however prolonged exposure to the organic medium causes this environment to resemble that of the solvent in which the enzyme is dissolved. Furthermore, kinetic studies show a reduction on both Vmax and KM as a result of prolonged exposure to the solvents. One interpretation of these results is that during this prolonged exposure to organic solvents the active-site fluorescent label inhibitor adopts a different binding conformation. Extrapolating this to an enzymatic reaction we argue that substrates bind in a less catalytically favorable conformation after the enzyme has been exposed to organic media for several hours. (literal)
Prodotto di
Autore CNR

Incoming links:


Prodotto
Autore CNR di
Http://www.cnr.it/ontology/cnr/pubblicazioni.owl#rivistaDi
data.CNR.it