title: Hydrolysis of synthetic polyesters by Clostridium botulinum esterases.
abstract: Two novel esterases from the anaerobe Clostridium botulinum ATCC 3502 (Cbotu_EstA and Cbotu_EstB) were expressed in Escherichia coli BL21-Gold(DE3) and were found to hydrolyze the polyester poly(butylene adipate-co-butylene terephthalate) (PBAT). The active site residues (triad Ser, Asp, His) are present in both enzymes at the same location only with some amino acid variations near the active site at the surrounding of aspartate. Yet, Cbotu_EstA showed higher kcat values on para-nitrophenyl butyrate and para-nitrophenyl acetate and was considerably more active (sixfold) on PBAT. The entrance to the active site of the modeled Cbotu_EstB appears more narrowed compared to the crystal structure of Cbotu_EstA and the N-terminus is shorter which could explain its lower activity on PBAT. The Cbotu_EstA crystal structure consists of two regions that may act as movable cap domains and a zinc metal binding site.
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Konrad Lorenz Strasse 20, Tulln, 3430, Austria.
authors.forename: Veronika
authors.initials: V
authors.lastname: Perz
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Graz, Austria.
authors.forename: Armin
authors.initials: A
authors.lastname: Baumschlager
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Graz, Austria.
authors.forename: Klaus
authors.initials: K
authors.lastname: Bleymaier
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Graz, Austria.
authors.forename: Sabine
authors.initials: S
authors.lastname: Zitzenbacher
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Graz, Austria.|Institute of Molecular Biosciences, University of Graz, Graz, Austria.
authors.forename: Altijana
authors.initials: A
authors.lastname: Hromic
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Graz, Austria.
authors.forename: Georg
authors.initials: G
authors.lastname: Steinkellner
authors.affiliation: Institute of Molecular Biosciences, University of Graz, Graz, Austria.
authors.forename: Andris
authors.initials: A
authors.lastname: Pairitsch
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Graz, Austria.
authors.forename: Andrzej
authors.initials: A
authors.lastname: Łyskowski
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Graz, Austria.|Institute of Molecular Biosciences, University of Graz, Graz, Austria.
authors.forename: Karl
authors.initials: K
authors.lastname: Gruber
authors.affiliation: BASF SE, Ludwigshafen, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Sinkel
authors.affiliation: BASF SE, Ludwigshafen, Germany.
authors.forename: Ulf
authors.initials: U
authors.lastname: Küper
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Konrad Lorenz Strasse 20, Tulln, 3430, Austria. doris.ribitsch@acib.at.|Institute of Environmental Biotechnology, University of Natural Resources and Life Sciences, Vienna, Konrad Lorenz Strasse 20, Tulln, 3430, Austria. doris.ribitsch@acib.at.
authors.forename: Doris
authors.initials: D
authors.lastname: Ribitsch
authors.affiliation: acib-Austrian Centre of Industrial Biotechnology, Konrad Lorenz Strasse 20, Tulln, 3430, Austria.|Institute of Environmental Biotechnology, University of Natural Resources and Life Sciences, Vienna, Konrad Lorenz Strasse 20, Tulln, 3430, Austria.
authors.forename: Georg M
authors.initials: GM
authors.lastname: Guebitz
mesh_terms: D002087:Butyrates / Q000378:metabolism; D020134:Catalytic Domain; D003014:Clostridium botulinum / Q000737:chemistry / Q000201:enzymology* / Q000378:metabolism; D018360:Crystallography, X-Ray; D004950:Esterases / Q000737:chemistry / Q000378:metabolism*; D006868:Hydrolysis; D008958:Models, Molecular; D009596:Nitrophenols / Q000378:metabolism; D011091:Polyesters / Q000378:metabolism*; D011487:Protein Conformation; D013379:Substrate Specificity; D015032:Zinc / Q000378:metabolism
chemical_list: D002087:Butyrates; D009596:Nitrophenols; D011091:Polyesters; C488797:poly(butylene adipate-co-butylene terephthalate); C033592:4-nitrophenyl butyrate; C008642:4-nitrophenyl acetate; D004950:Esterases; D015032:Zinc
keywords: (poly(butylene adipate-co-butylene terephthalate) (PBAT); anaerobic digestion; esterase; polyesterase; polymer hydrolysis; zinc dependent enzyme