title: Influence of surface charge, binding site residues and glycosylation on Thielavia terrestris cutinase biochemical characteristics.
abstract: Cutinases are esterases of industrial importance for applications in recycling and surface modification of polyesters. The cutinase from Thielavia terrestris (TtC) is distinct in terms of its ability to retain its stability and activity in acidic pH. Stability and activity in acidic pHs are desirable for esterases as the pH of the reaction tends to go down with the generation of acid. The pH stability and activity are governed by the charged state of the residues involved in catalysis or in substrate binding. In this study, we performed the detailed structural and biochemical characterization of TtC coupled with surface charge analysis to understand its acidic tolerance. The stability of TtC in acidic pH was rationalized by evaluating the contribution of charge interactions to the Gibbs free energy of unfolding at varying pHs. The activity of TtC was found to be limited by substrate binding affinity, which is a function of the surface charge. Additionally, the presence of glycosylation affects the biochemical characteristics of TtC owing to steric interactions with residues involved in substrate binding.
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, CBIS 4105, 110 8th Street, Troy, NY, 12180, USA.|Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.
authors.forename: Abhijit N
authors.initials: AN
authors.lastname: Shirke
authors.affiliation: Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.|Department of Biological Sciences, Rensselaer Polytechnic Institute, J-Rowl, 3C07, 110 8th Street, Troy, NY, 12180, USA.
authors.forename: Danielle
authors.initials: D
authors.lastname: Basore
authors.affiliation: Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.|Department of Biomedical Engineering, Rensselaer Polytechnic Institute, Troy, NY, USA.
authors.forename: Samantha
authors.initials: S
authors.lastname: Holton
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, CBIS 4105, 110 8th Street, Troy, NY, 12180, USA.|Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.
authors.forename: An
authors.initials: A
authors.lastname: Su
authors.affiliation: Center for Genomics and Systems Biology, New York University, New York, NY, USA.
authors.forename: Evan
authors.initials: E
authors.lastname: Baugh
authors.affiliation: Center for Genomics and Systems Biology, New York University Abu Dhabi, Abu Dhabi, United Arab Emirates.
authors.forename: Glenn L
authors.initials: GL
authors.lastname: Butterfoss
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, CBIS 4105, 110 8th Street, Troy, NY, 12180, USA.|Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA.|Department of Biological Sciences, Rensselaer Polytechnic Institute, J-Rowl, 3C07, 110 8th Street, Troy, NY, 12180, USA.
authors.forename: George
authors.initials: G
authors.lastname: Makhatadze
authors.affiliation: Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA. bystrc@rpi.edu.|Department of Biological Sciences, Rensselaer Polytechnic Institute, J-Rowl, 3C07, 110 8th Street, Troy, NY, 12180, USA. bystrc@rpi.edu.|Department of Computer Science, Rensselaer Polytechnic Institute, Troy, NY, USA. bystrc@rpi.edu.
authors.forename: Christopher
authors.initials: C
authors.lastname: Bystroff
authors.affiliation: Department of Chemistry and Chemical Biology, Rensselaer Polytechnic Institute, CBIS 4105, 110 8th Street, Troy, NY, 12180, USA. grossr@rpi.edu.|Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, USA. grossr@rpi.edu.|Department of Biological Sciences, Rensselaer Polytechnic Institute, J-Rowl, 3C07, 110 8th Street, Troy, NY, 12180, USA. grossr@rpi.edu.
authors.forename: Richard A
authors.initials: RA
authors.lastname: Gross
mesh_terms: D000595:Amino Acid Sequence; D001665:Binding Sites; D002265:Carboxylic Ester Hydrolases / Q000737:chemistry*; D002384:Catalysis; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism; D006031:Glycosylation; D006863:Hydrogen-Ion Concentration; D011091:Polyesters / Q000737:chemistry; D017434:Protein Structure, Tertiary; D020037:Sordariales / Q000201:enzymology*; D013499:Surface Properties
chemical_list: D011091:Polyesters; D002265:Carboxylic Ester Hydrolases; C019952:cutinase
keywords: Biochemical characterization; Cutinase; Glycosylation; Polyester hydrolysis; Substrate binding; Surface charge