caa a22 4500 605501114 CHVBK 20210128100559.0 cr unu---uuuuu 210128e20151101xx s 000 0 eng 10.1007/s00253-015-6780-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6780-1 Novel extracellular medium-chain-length polyhydroxyalkanoate depolymerase from Streptomyces exfoliatus K10 DSMZ 41693: a promising biocatalyst for the efficient degradation of natural and functionalized mcl-PHAs [Elektronische Daten] [Virginia Martínez, Patricia de Santos, Javier García-Hidalgo, Daniel Hormigo, M. Prieto, Miguel Arroyo, Isabel de la Mata] Cloning and biochemical characterization of a novel extracellular medium-chain-length polyhydroxyalkanoate (mcl-PHA) depolymerase from Streptomyces exfoliatus K10 DSMZ 41693 are described. The primary structure of the depolymerase (PhaZSex2) includes the lipase consensus sequence (serine-histidine-aspartic acid) which is known for serine hydrolases. Secondary structure analysis shows 7.9% α-helix, 43.9% β-sheet, 19.4% β-turns, and 31.2% random coil, suggesting that this enzyme belongs to the α/β hydrolase fold family, in agreement with other PHA depolymerases and lipases. The enzyme was efficiently produced as an extracellular active form in Rhodococcus and purified by two consecutive hydrophobic chromatographic steps. Matrix-assisted laser desorption-time-of-flight (MALDI-TOF) analysis of the purified enzyme revealed a monomer of 27.6kDa with a midpoint transition temperature of 44.2°C. Remarkably, the activity is significantly enhanced by low concentrations of nonionic and anionic detergents and thermal stability is improved by the presence of 10% glycerol. PhaZSex2 is an endo-exohydrolase that cleaves both large and small PHA molecules, producing (R)-3-hydroxyoctanoic acid monomers as the main reaction product. Markedly, PhaZSex2 is able to degrade functionalized polymers containing thioester groups in the side chain (PHACOS), releasing functional thioester-based monomers and oligomers demonstrating the potentiality of this novel biocatalyst for the industrial production of enantiopure (R)-3-hydroxyalkanoic acids. Springer-Verlag Berlin Heidelberg, 2015 Streptomyces exfoliatus nationallicence Polyhydroxyalkanoate depolymerase nationallicence 3-Hydroxyalkanoic acids nationallicence Martínez Virginia Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut de Santos Patricia Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut García-Hidalgo Javier Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut Hormigo Daniel Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut Prieto M. Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain aut Arroyo Miguel Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut de la Mata Isabel Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9605-9615 0175-7598 99:22<9605 2015 99 253 https://doi.org/10.1007/s00253-015-6780-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6780-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Martínez Virginia Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut NATIONALLICENCE 700 1- de Santos Patricia Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut NATIONALLICENCE 700 1- García-Hidalgo Javier Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut NATIONALLICENCE 700 1- Hormigo Daniel Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut NATIONALLICENCE 700 1- Prieto M. Department of Environmental Biology, Centro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, Madrid, Spain aut NATIONALLICENCE 700 1- Arroyo Miguel Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut NATIONALLICENCE 700 1- de la Mata Isabel Department of Biochemistry and Molecular Biology, Facultad de Biología, Universidad Complutense de Madrid, Madrid, Spain aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/22(2015-11-01), 9605-9615 0175-7598 99:22<9605 2015 99 253