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Structural and functional studies of a Fusarium oxysporum cutinase with polyethylene terephthalate modification potential.

Dimarogona M, Nikolaivits E, Kanelli M, Christakopoulos P, Sandgren M, Topakas E
Biochim Biophys Acta 1850 2308-17 (2015)
Cited: 23 times
EuropePMC logo PMID: 26291558

Abstract

Background

Cutinases are serine hydrolases that degrade cutin, a polyester of fatty acids that is the main component of plant cuticle. These biocatalysts have recently attracted increased biotechnological interest due to their potential to modify and degrade polyethylene terephthalate (PET), as well as other synthetic polymers.

Methods

A cutinase from the mesophilic fungus Fusarium oxysporum, named FoCut5a, was expressed either in the cytoplasm or periplasm of Escherichia coli BL21. Its X-ray structure was determined to 1.9Å resolution using molecular replacement. The activity of the recombinant enzyme was tested on a variety of synthetic esters and polyester analogues.

Results

The highest production of recombinant FoCut5a was achieved using periplasmic expression at 16°C. Its crystal structure is highly similar to previously determined Fusarium solani cutinase structure. However, a more detailed comparison of the surface properties and amino acid interactions revealed differences with potential impact on the biochemical properties of the two enzymes. FoCut5a showed maximum activity at 40°C and pH 8.0, while it was active on three p-nitrophenyl synthetic esters of aliphatic acids (C(2), C(4), C(12)), with the highest catalytic efficiency for the hydrolysis of the butyl ester. The recombinant cutinase was also found capable of hydrolyzing PET model substrates and synthetic polymers.

Conclusion

The present work is the first reported expression and crystal structure determination of a functional cutinase from the mesophilic fungus F. oxysporum with potential application in surface modification of PET synthetic polymers.

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  1. Enzymatic Remediation of Polyethylene Terephthalate (PET)-Based Polymers for Effective Management of Plastic Wastes: An Overview. Maurya A, Bhattacharya A, Khare SK. Front Bioeng Biotechnol 8 602325 (2020)
  2. Fungal Enzymes Involved in Plastics Biodegradation. Temporiti MEE, Nicola L, Nielsen E, Tosi S. Microorganisms 10 1180 (2022)
  3. Progressing Plastics Circularity: A Review of Mechano-Biocatalytic Approaches for Waste Plastic (Re)valorization. Nikolaivits E, Pantelic B, Azeem M, Taxeidis G, Babu R, Topakas E, Brennan Fournet M, Nikodinovic-Runic J. Front Bioeng Biotechnol 9 696040 (2021)
  4. Surface engineering of polyester-degrading enzymes to improve efficiency and tune specificity. Biundo A, Ribitsch D, Guebitz GM. Appl Microbiol Biotechnol 102 3551-3559 (2018)
  5. Bright Side of Fusarium oxysporum: Secondary Metabolites Bioactivities and Industrial Relevance in Biotechnology and Nanotechnology. Ibrahim SRM, Sirwi A, Eid BG, Mohamed SGA, Mohamed GA. J Fungi (Basel) 7 943 (2021)
  6. Recent Advances in Biological Recycling of Polyethylene Terephthalate (PET) Plastic Wastes. Soong YV, Sobkowicz MJ, Xie D. Bioengineering (Basel) 9 98 (2022)
  7. An Overview into Polyethylene Terephthalate (PET) Hydrolases and Efforts in Tailoring Enzymes for Improved Plastic Degradation. Khairul Anuar NFS, Huyop F, Ur-Rehman G, Abdullah F, Normi YM, Sabullah MK, Abdul Wahab R. Int J Mol Sci 23 12644 (2022)
  8. A Review of the Fungi That Degrade Plastic. Ekanayaka AH, Tibpromma S, Dai D, Xu R, Suwannarach N, Stephenson SL, Dao C, Karunarathna SC. J Fungi (Basel) 8 772 (2022)
  9. New approaches for the characterization of plastic-associated microbial communities and the discovery of plastic-degrading microorganisms and enzymes. Viljakainen VR, Hug LA. Comput Struct Biotechnol J 19 6191-6200 (2021)
  10. Fungal Enzymes as Catalytic Tools for Polyethylene Terephthalate (PET) Degradation. Ahmaditabatabaei S, Kyazze G, Iqbal HMN, Keshavarz T. J Fungi (Basel) 7 931 (2021)
  11. Potential Use of Microbial Enzymes for the Conversion of Plastic Waste Into Value-Added Products: A Viable Solution. Tamoor M, Samak NA, Jia Y, Mushtaq MU, Sher H, Bibi M, Xing J. Front Microbiol 12 777727 (2021)

Articles citing this publication (7)

  1. Active Site Flexibility as a Hallmark for Efficient PET Degradation by I. sakaiensis PETase. Fecker T, Galaz-Davison P, Engelberger F, Narui Y, Sotomayor M, Parra LP, Ramírez-Sarmiento CA. Biophys J 114 1302-1312 (2018)
  2. High-level expression and characterization of a novel cutinase from Malbranchea cinnamomea suitable for butyl butyrate production. Duan X, Liu Y, You X, Jiang Z, Yang S, Yang S. Biotechnol Biofuels 10 223 (2017)
  3. Ionic Liquids as Extractants for Nanoplastics. Elfgen R, Gehrke S, Hollóczki O. ChemSusChem 13 5449-5459 (2020)
  4. Versatile Fungal Polyphenol Oxidase with Chlorophenol Bioremediation Potential: Characterization and Protein Engineering. Nikolaivits E, Dimarogona M, Karagiannaki I, Chalima A, Fishman A, Topakas E. Appl Environ Microbiol 84 e01628-18 (2018)
  5. Residue-Specific Incorporation of the Non-Canonical Amino Acid Norleucine Improves Lipase Activity on Synthetic Polyesters. Haernvall K, Fladischer P, Schoeffmann H, Zitzenbacher S, Pavkov-Keller T, Gruber K, Schick M, Yamamoto M, Kuenkel A, Ribitsch D, Guebitz GM, Wiltschi B. Front Bioeng Biotechnol 10 769830 (2022)
  6. Exploring the infiltrative and degradative ability of Fusarium oxysporum on polyethylene terephthalate (PET) using correlative microscopy and deep learning. Cognigni F, Temporiti MEE, Nicola L, Gueninchault N, Tosi S, Rossi M. Sci Rep 13 22987 (2023)
  7. Metagenome-Assembled Genomes of Four Southern Ocean Archaea Harbor Multiple Genes Linked to Polyethylene Terephthalate and Polyhydroxybutyrate Plastic Degradation. Akpudo YM, Bezuidt OK, Makhalanyane TP. Microbiol Resour Announc 12 e0109822 (2023)


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