title: Hydrolysis of cyclic poly(ethylene terephthalate) trimers by a carboxylesterase from Thermobifida fusca KW3.
abstract: We have identified a carboxylesterase produced in liquid cultures of the thermophilic actinomycete Thermobifida fusca KW3 that were supplemented with poly(ethylene terephthalate) fibers. The enzyme hydrolyzed highly hydrophobic, synthetic cyclic poly(ethylene terephthalate) trimers with an optimal activity at 60 degrees C and a pH of 6. V (max) and K (m) values for the hydrolysis were 9.3 micromol(-1) min(-1) mg(-1) and 0.5 mM, respectively. The esterase showed high specificity towards short and middle chain-length fatty acyl esters of p-nitrophenol. The enzyme retained 37% of its activity after 96 h of incubation at 50 degrees C and a pH of 8. Enzyme inhibition studies and analysis of substitution mutants of the carboxylesterase revealed the typical catalytic mechanism of a serine hydrolase with a catalytic triad composed of serine, glutamic acid, and histidine.
authors.affiliation: Department of Microbiology and Bioprocess Technology, Institute of Biochemistry, University of Leipzig, Leipzig, Germany.
authors.forename: Susan
authors.initials: S
authors.lastname: Billig
authors.forename: Thorsten
authors.initials: T
authors.lastname: Oeser
authors.forename: Claudia
authors.initials: C
authors.lastname: Birkemeyer
authors.forename: Wolfgang
authors.initials: W
authors.lastname: Zimmermann
mesh_terms: D000192:Actinomycetales / Q000201:enzymology*; D043182:Carboxylesterase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004795:Enzyme Stability; D006863:Hydrogen-Ion Concentration; D006868:Hydrolysis; D007700:Kinetics; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011092:Polyethylene Glycols / Q000378:metabolism*; D011093:Polyethylene Terephthalates; D017434:Protein Structure, Tertiary; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity; D013696:Temperature
chemical_list: D004269:DNA, Bacterial; D011093:Polyethylene Terephthalates; D011092:Polyethylene Glycols; D043182:Carboxylesterase