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Identification and comparison of cutinases for synthetic polyester degradation

  • Biotechnologically Relevant Enzymes and Proteins
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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.

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Acknowledgments

This work was supported in part by NYU:POLY Seed Fund (RG and JKM), AFOSR DURIP (FA-9550-08-1-0266) (JKM), NSF IUCRC (RG), and NSF GK-12 Fellows grant 0741714 (PJB). We thank Jeremy Minshull and Jonathan Ness from DNA 2.0 for their assistance in generating the cutinase DNA sequences as well as the production and purification of the enzymes.

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Authors and Affiliations

  1. Department of Chemical and Biological Sciences, Polytechnic Institute of New York University, Brooklyn, NY, 11201, USA

    Peter James Baker, Richard Gross & Jin Kim Montclare

  2. Courant Institute of Mathematical Sciences, New York University, New York, NY, 10003, USA

    Christopher Poultney

  3. Institute of Bioengineering, Zhejiang University of Technology, Hangzhou, Zhejiang, 310014, China

    Zhiqiang Liu

  4. Department of Biochemistry, SUNY Downstate Medical Center, Brooklyn, NY, 11203, USA

    Jin Kim Montclare

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  1. Peter James Baker
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  5. Jin Kim Montclare
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Correspondence to Jin Kim Montclare.

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Baker, P.J., Poultney, C., Liu, Z. et al. Identification and comparison of cutinases for synthetic polyester degradation. Appl Microbiol Biotechnol 93, 229–240 (2012). https://doi.org/10.1007/s00253-011-3402-4

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  • DOI: https://doi.org/10.1007/s00253-011-3402-4

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