title: Identification and comparison of cutinases for synthetic polyester degradation.
abstract: Cutinases have been exploited for a broad range of reactions, from hydrolysis of soluble and insoluble esters to polymer synthesis. To further expand the biotechnological applications of cutinases for synthetic polyester degradation, we perform a comparative activity and stability analysis of five cutinases from Alternaria brassicicola (AbC), Aspergillus fumigatus (AfC), Aspergillus oryzae (AoC), Humicola insolens (HiC), and the well-characterized Fusarium solani (FsC). Of the cutinases, HiC demonstrated enhanced poly(ε-caprolactone) hydrolysis at high temperatures and under all pH values, followed by AoC and AfC. Both AbC and FsC are least stable and function poorly at high temperatures as well as at acidic pH conditions. Surface charge calculations and phylogenetic analysis reveal two important modes of cutinase stabilization: (1) an overall neutral surface charge within the "crowning area" by the active site and (2) additional disulfide bond formation. These studies provide insights useful for reengineering such enzymes with improved function and stability for a wide range of biotransformations.
authors.affiliation: Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, Brooklyn, NY 11201, USA.
authors.forename: Peter James
authors.initials: PJ
authors.lastname: Baker
authors.forename: Christopher
authors.initials: C
authors.lastname: Poultney
authors.forename: Zhiqiang
authors.initials: Z
authors.lastname: Liu
authors.forename: Richard
authors.initials: R
authors.lastname: Gross
authors.forename: Jin Kim
authors.initials: JK
authors.lastname: Montclare
mesh_terms: D000595:Amino Acid Sequence; D001203:Ascomycota / Q000201:enzymology*; D001711:Biotransformation; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D002942:Circular Dichroism; D004220:Disulfides; D004795:Enzyme Stability; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008969:Molecular Sequence Data; D010802:Phylogeny; D011091:Polyesters / Q000378:metabolism*; D011487:Protein Conformation; D017386:Sequence Homology, Amino Acid; D055672:Static Electricity
chemical_list: D004220:Disulfides; D011091:Polyesters; D002265:Carboxylic Ester Hydrolases; C019952:cutinase