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Unveiling the crystal structure of thermostable dienelactone hydrolase exhibiting activity on terephthalate esters.
Almeida, Dnane Vieira; Ciancaglini, Iara; Sandano, Ana Luiza Hernandes; Roman, Ellen K B; Andrade, Viviane Brito; Nunes, Ana Bárbara; Tramontina, Robson; da Silva, Viviam Moura; Gabel, Frank; Corrêa, Thamy L R; Damasio, André; Muniz, João Renato Carvalho; Squina, Fabio Marcio; Garcia, Wanius.
Afiliação
  • Almeida DV; Centro de Ciências Naturais e Humanas, Universidade Federal do ABC (UFABC),Santo André, SP, Brazil.
  • Ciancaglini I; Laboratory of Molecular Sciences, University of Sorocaba (UNISO), Sorocaba, SP, Brazil; Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.
  • Sandano ALH; Laboratory of Molecular Sciences, University of Sorocaba (UNISO), Sorocaba, SP, Brazil.
  • Roman EKB; Laboratory of Molecular Sciences, University of Sorocaba (UNISO), Sorocaba, SP, Brazil; Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.
  • Andrade VB; Centro de Ciências Naturais e Humanas, Universidade Federal do ABC (UFABC),Santo André, SP, Brazil.
  • Nunes AB; Laboratory of Molecular Sciences, University of Sorocaba (UNISO), Sorocaba, SP, Brazil; Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.
  • Tramontina R; Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.
  • da Silva VM; Institut de Biologie Structurale (IBS), CEA, CNRS, UGA, 71 Avenue des Martyrs, Grenoble 38000, France.
  • Gabel F; Institut de Biologie Structurale (IBS), CEA, CNRS, UGA, 71 Avenue des Martyrs, Grenoble 38000, France.
  • Corrêa TLR; Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.
  • Damasio A; Laboratory of Enzymology and Molecular Biology of Microorganisms (LEBIMO), Department of Biochemistry and Tissue Biology, Institute of Biology, Universidade Estadual de Campinas (UNICAMP), Campinas, SP, Brazil.
  • Muniz JRC; São Carlos Institute of Physics (IFSC), University of São Paulo (USP), São Carlos, SP, Brazil.
  • Squina FM; Laboratory of Molecular Sciences, University of Sorocaba (UNISO), Sorocaba, SP, Brazil. Electronic address: fabio.squina@prof.uniso.br.
  • Garcia W; Centro de Ciências Naturais e Humanas, Universidade Federal do ABC (UFABC),Santo André, SP, Brazil. Electronic address: wanius.garcia@ufabc.edu.br.
Enzyme Microb Technol ; 180: 110498, 2024 Oct.
Article em En | MEDLINE | ID: mdl-39182429
ABSTRACT
Dienelactone hydrolase (DLH) is one of numerous hydrolytic enzymes with an α/ß-hydrolase fold, which catalyze the hydrolysis of dienelactone to maleylacetate. The DLHs share remarkably similar tertiary structures and a conserved arrangement of catalytic residues. This study presents the crystal structure and comprehensive functional characterization of a novel thermostable DLH from the bacterium Hydrogenobacter thermophilus (HtDLH). The crystal structure of the HtDLH, solved at a resolution of about 1.67 Å, exhibits a canonical α/ß-hydrolase fold formed by eight ß-sheet strands in the core, with one buried α-helix and six others exposed to the solvent. The structure also confirmed the conserved catalytic triad of DHLs formed by Cys121, Asp170, and His202 residues. The HtDLH forms stable homodimers in solution. Functional studies showed that HtDLH has the expected esterase activity over esters with short carbon chains, such as p-nitrophenyl acetate, reaching optimal activity at pH 7.5 and 70 °C. Furthermore, HtDLH maintains more than 50 % of its activity even after incubation at 90 °C for 16 h. Interestingly, HtDLH exhibits catalytic activity towards polyethylene terephthalate (PET) monomers, including bis-1,2-hydroxyethyl terephthalate (BHET) and 1-(2-hydroxyethyl) 4-methyl terephthalate, as well as other aliphatic and aromatic esters. These findings associated with the lack of activity on amorphous PET indicate that HtDLH has characteristic of a BHET-degrading enzyme. This work expands our understanding of enzyme families involved in PET degradation, providing novel insights for plastic biorecycling through protein engineering, which could lead to eco-friendly solutions to reduce the accumulation of plastic in landfills and natural environments.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Estabilidade Enzimática / Hidrolases de Éster Carboxílico Idioma: En Revista: Enzyme Microb Technol Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Estabilidade Enzimática / Hidrolases de Éster Carboxílico Idioma: En Revista: Enzyme Microb Technol Ano de publicação: 2024 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos