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| Content Provider | Springer Nature Link |
|---|---|
| Author | Martínez, Virginia Santos, Patricia Gómez García Hidalgo, Javier Hormigo, Daniel Prieto, M. Auxiliadora Arroyo, Miguel Mata, Isabel |
| Copyright Year | 2015 |
| Abstract | 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 27.6 kDa 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. |
| Starting Page | 9605 |
| Ending Page | 9615 |
| Page Count | 11 |
| File Format | |
| ISSN | 01757598 |
| Journal | Applied Microbiology and Biotechnology |
| Volume Number | 99 |
| Issue Number | 22 |
| e-ISSN | 14320614 |
| Language | English |
| Publisher | Springer Berlin Heidelberg |
| Publisher Date | 2015-07-10 |
| Publisher Place | Berlin, Heidelberg |
| Access Restriction | Subscribed |
| Subject Keyword | Streptomyces exfoliatus Polyhydroxyalkanoate depolymerase 3-Hydroxyalkanoic acids Microbiology Microbial Genetics and Genomics Biotechnology |
| Content Type | Text |
| Resource Type | Article |
| Subject | Medicine Applied Microbiology and Biotechnology Biotechnology |
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