Stabilizing leaf and branch compost cutinase (LCC) withglycosylation: Mechanism and effect on PET hydrolysis

Authors
Shirke, A.N.
White, C.
Englaender, J.A.
Sophie Zwarycz, A.
Butterfoss, G.L.
Linhardt, Robert J.
Gross, R.A.
ORCID
https://orcid.org/0000-0003-2219-5833
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Other Contributors
Issue Date
2018
Keywords
Biology , Chemistry and chemical biology , Chemical and biological engineering , Biomedical engineering
Degree
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Full Citation
Stabilizing leaf and branch compost cutinase (LCC) withglycosylation: Mechanism and effect on PET hydrolysis, A. N. Shirke, C. White, J. A. Englaender, A. Sophie Zwarycz, G. L. Butterfoss, R. J. Linhardt, R. A. Gross, Biochemistry, 57, 1190−1200, 2018.
Abstract
Cutinases are polyester hydrolases that show a remarkable capability to hydrolyze polyethylene terephthalate (PET) to its monomeric units. This revelation has stimulated research aimed at developing sustainable and green cutinase-catalyzed PET recycling methods. Leaf and branch compost cutinase (LCC) is particularly suited toward these ends given its relatively high PET hydrolysis activity and thermostability. Any practical enzymatic PET recycling application will require that the protein have kinetic stability at or above the PET glass transition temperature (Tg, i.e., 70 °C). This paper elucidates the thermodynamics and kinetics of LCC conformational and colloidal stability. Aggregation emerged as a major contributor that reduces LCC kinetic stability. In its native state, LCC is prone to aggregation owing to electrostatic interactions. Further, with increasing temperature, perturbation of LCC’s tertiary structure and corresponding exposure of hydrophobic domains leads to rapid aggregation. Glycosylation was employed in an attempt to impede LCC aggregation. Owing to the presence of three putative N-glycosylation sites, expression of native LCC in Pichia pastoris resulted in the production of glycosylated LCC (LCC-G). LCC-G showed improved stability to native state aggregation while increasing the temperature for thermal induced aggregation by 10 °C. Furthermore, stabilization against thermal aggregation resulted in improved catalytic PET hydrolysis both at its optimum temperature and concentration.
Description
Biochemistry, 57, 1190−1200
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Department
The Linhardt Research Labs.
The Shirley Ann Jackson, Ph.D. Center for Biotechnology and Interdisciplinary Studies (CBIS)
Publisher
Relationships
The Linhardt Research Labs Online Collection
Rensselaer Polytechnic Institute, Troy, NY
https://harc.rpi.edu/
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