Identification and comparison of cutinases for synthetic polyester degradation

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.

[1]  F. Hasan,et al.  Biological degradation of plastics: a comprehensive review. , 2008, Biotechnology advances.

[2]  J. Sussman,et al.  An electrostatic mechanism for substrate guidance down the aromatic gorge of acetylcholinesterase. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[3]  R. Baptista,et al.  Purification and identification of cutinases from Colletotrichum kahawae and Colletotrichum gloeosporioides , 2007, Applied Microbiology and Biotechnology.

[4]  M. Aires-Barros,et al.  Cutinase: from molecular level to bioprocess development. , 1999, Biotechnology and bioengineering.

[5]  C. Soares,et al.  Effect of immobilization support, water activity, and enzyme ionization state on cutinase activity and enantioselectivity in organic media , 2004, Biotechnology and bioengineering.

[6]  C. Pace,et al.  Conformational stability and activity of ribonuclease T1 with zero, one, and two intact disulfide bonds. , 1988, The Journal of biological chemistry.

[7]  K. Toshima,et al.  Perspectives for synthesis and production of polyurethanes and related polymers by enzymes directed toward green and sustainable chemistry , 2006, Applied Microbiology and Biotechnology.

[8]  B. Mooney,et al.  The second green revolution? Production of plant-based biodegradable plastics. , 2009, The Biochemical journal.

[9]  M. Aires-Barros,et al.  Immobilization of a recombinant cutinase by entrapment and by covalent binding , 1996, Applied Biochemistry and Biotechnology.

[10]  H. Maeda,et al.  Purification and characterization of a biodegradable plastic-degrading enzyme from Aspergillus oryzae , 2005, Applied Microbiology and Biotechnology.

[11]  D. Rice,et al.  Catalysis by Glomerella cingulata cutinase requires conformational cycling between the active and inactive states of its catalytic triad. , 2009, Journal of molecular biology.

[12]  Hyo-Seel Seo,et al.  Enhanced degradation and toxicity reduction of dihexyl phthalate by Fusarium oxysporum f. sp. pisi cutinase , 2007, Journal of applied microbiology.

[13]  U. Bornscheuer,et al.  Improved biocatalysts by directed evolution and rational protein design. , 2001, Current opinion in chemical biology.

[14]  M. R. Egmond,et al.  Fusarium solani pisi cutinase. , 2000, Biochimie.

[15]  J. Cabral,et al.  Sol-gel encapsulation: an efficient and versatile immobilization technique for cutinase in non-aqueous media. , 2006, Journal of biotechnology.

[16]  Torben Vedel Borchert,et al.  Industrial enzyme applications. , 2002, Current opinion in biotechnology.

[17]  Norma J Greenfield,et al.  Analysis of Circular Dichroism Data , 2004, Numerical Computer Methods, Part D.

[18]  G. L. Butterfoss,et al.  Structural and functional studies of Aspergillus oryzae cutinase: enhanced thermostability and hydrolytic activity of synthetic ester and polyester degradation. , 2009, Journal of the American Chemical Society.

[19]  S. Petersen,et al.  How do lipases and esterases work: the electrostatic contribution. , 2001, Journal of biotechnology.

[20]  P. Kolattukudy,et al.  Depolymerization of a hydroxy fatty acid biopolymer, cutin, by an extracellular enzyme from Fusarium solani f. pisi: isolation and some properties of the enzyme. , 1973, Archives of biochemistry and biophysics.

[21]  P. Fernandes,et al.  Biosynthesis of ethyl caproate and other short ethyl esters catalyzed by cutinase in organic solvent , 2009 .

[22]  C. Fan,et al.  Diversity of cutinases from plant pathogenic fungi: differential and sequential expression of cutinolytic esterases by Alternaria brassicicola , 1998 .

[23]  Maria Teresa Neves-Petersen,et al.  Protein electrostatics: a review of the equations and methods used to model electrostatic equations in biomolecules--applications in biotechnology. , 2003, Biotechnology annual review.

[24]  B. Hauer,et al.  Heterologous expression, characterization and site-directed mutagenesis of cutinase CUTAB1 from Alternaria brassicicola , 2010, Applied Microbiology and Biotechnology.

[25]  A. Sali,et al.  Comparative protein structure modeling of genes and genomes. , 2000, Annual review of biophysics and biomolecular structure.

[26]  Per Jambeck,et al.  Conservation of electrostatic properties within enzyme families and superfamilies. , 2003, Biochemistry.

[27]  J. Cabral,et al.  Triglyceride hydrolysis and stability of a recombinant cutinase fromFusarium solani in AOT-iso-octane reversed micelles , 1995, Applied biochemistry and biotechnology.

[28]  Pari Skamnioti,et al.  Cutinase and hydrophobin interplay , 2008, Plant signaling & behavior.

[29]  Margarida Casal,et al.  Tailoring cutinase activity towards polyethylene terephthalate and polyamide 6,6 fibers. , 2007, Journal of biotechnology.

[30]  Simon J. Bennett,et al.  From petrochemical complexes to biorefineries? The past and prospective co-evolution of liquid fuels and chemicals production in the UK , 2009 .

[31]  Richard A. Gross,et al.  Cutinase-Catalyzed Hydrolysis of Poly(ethylene terephthalate) , 2009 .

[32]  K. S. Shashidhara,et al.  Conformational and Functional Transitions in Class II α-mannosidase from Aspergillus fischeri , 2010, Journal of Fluorescence.

[33]  P. Kolattukudy,et al.  Hydrolysis of plant cuticle by plant pathogens. Purification, amino acid composition, and molecular weight of two isozymes of cutinase and a nonspecific esterase from Fusarium solani f. pisi. , 1975, Biochemistry.

[34]  E. Alexov,et al.  Combining conformational flexibility and continuum electrostatics for calculating pK(a)s in proteins. , 2002, Biophysical journal.

[35]  Gabriela Alves Macedo,et al.  Cutinases: properties and industrial applications. , 2009, Advances in applied microbiology.

[36]  Gabriela Alves Macedo,et al.  Chapter 4 Cutinases , 2009 .

[37]  C. Cambillau,et al.  Fusarium solani cutinase is a lipolytic enzyme with a catalytic serine accessible to solvent , 1992, Nature.

[38]  S. Petersen,et al.  The thermal stability of the Fusarium solani pisi cutinase as a function of pH , 2001, Journal of biomedicine & biotechnology.

[39]  Valerie Daggett,et al.  The present view of the mechanism of protein folding , 2003, Nature Reviews Molecular Cell Biology.

[40]  M Czjzek,et al.  Atomic resolution (1.0 A) crystal structure of Fusarium solani cutinase: stereochemical analysis. , 1997, Journal of molecular biology.

[41]  C. Pichot,et al.  Activity, conformation and dynamics of cutinase adsorbed on poly(methyl methacrylate) latex particles. , 2003, Journal of biotechnology.

[42]  Kolattukudy Pe,et al.  Hydrolysis of plant cuticle by plant pathogens. Properties of cutinase I, cutinase II, and a nonspecific esterase isolated from Fusarium solani pisi. , 1975, Biochemistry.

[43]  R. Kotek,et al.  Enzymatic hydrolysis of PTT polymers and oligomers. , 2008, Journal of biotechnology.

[44]  R. Vinopal,et al.  Fusarium polycaprolactone depolymerase is cutinase , 1996, Applied and environmental microbiology.

[45]  David L. Wheeler,et al.  GenBank , 2015, Nucleic Acids Res..

[46]  R. Hatti-Kaul,et al.  Synthesis and production of polyhydroxyalkanoates by halophiles: current potential and future prospects , 2010, Applied Microbiology and Biotechnology.

[47]  E. Alexov,et al.  Incorporating protein conformational flexibility into the calculation of pH-dependent protein properties. , 1997, Biophysical journal.

[48]  K. P. Murphy,et al.  The molecular basis of cooperativity in protein folding. Thermodynamic dissection of interdomain interactions in phosphoglycerate kinase. , 1992, Biochemistry.