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Characterization of cross‐linked immobilized arylesterase from Gluconobacter oxydans 621H with activity toward cephalosporin C and 7‐aminocephalosporanic acid
Author(s) -
Inmaculada NavarroGonzález,
Francisco GarcíaCarmona
Publication year - 2015
Publication title -
biotechnology progress
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.572
H-Index - 129
eISSN - 1520-6033
pISSN - 8756-7938
DOI - 10.1002/btpr.2178
Subject(s) - cephalosporin c , glutaraldehyde , arylesterase , chemistry , enzyme kinetics , enzyme , immobilized enzyme , kinetics , chromatography , nuclear chemistry , michaelis–menten kinetics , biochemistry , enzyme assay , cephalosporin , active site , pon1 , physics , quantum mechanics , genotype , gene , antibiotics
Cross‐linked enzyme aggregates (CLEAs) were prepared from several precipitant agents using glutaraldehyde as a cross‐linking agent with and without BSA, finally choosing a 40% saturation of ammonium sulfate and 25 mM of glutaraldehyde. The CLEAs obtained under optimum conditions were biochemically characterized. The immobilized enzyme showed higher thermal activity and a broader range of pH and organic solvent tolerance than the free enzyme. Arylesterase from Gluconobacter oxydans showed activity toward cephalosporin C and 7‐aminocephalosporanic acid. The CLEAs had a Kcat /K M of 0.9 M −1 /S −1 for 7‐ACA (7‐aminocephalosporanic acid) and 0.1 M −1 /S −1 for CPC (cephalosporin c), whereas free enzyme did not show a typical Michaelis–Menten kinetics. © 2015 American Institute of Chemical Engineers Biotechnol. Prog. , 32:36–42, 2016