Evaluation of BP-M-CPF4 polyhydroxyalkanoate (PHA) synthase on the production of poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) from plant oil using Cupriavidus necator transformants
| Autor: | Min Fey Chek, Manoj Lakshmanan, Choon Pin Foong, Hua Tiang Tan, Kumar Sudesh, Toshio Hakoshima |
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| Rok vydání: | 2020 |
| Předmět: |
Polymers
Cupriavidus necator Mutant 02 engineering and technology Palm Oil Reductase Biochemistry Polyhydroxyalkanoates 03 medical and health sciences Structural Biology Plant Oils Caproates Molecular Biology 030304 developmental biology chemistry.chemical_classification 0303 health sciences 3-Hydroxybutyric Acid Molecular mass biology ATP synthase Chemistry General Medicine 021001 nanoscience & nanotechnology biology.organism_classification Enzyme assay Enzyme Activation Molecular Weight Enzyme Fermentation biology.protein Transformation Bacterial 0210 nano-technology Oxidation-Reduction Acyltransferases Plasmids |
| Zdroj: | International Journal of Biological Macromolecules. 159:250-257 |
| ISSN: | 0141-8130 |
| Popis: | Among the various types of polyhydroxyalkanoate (PHA), poly[(R)-3-hydroxybutyrate-co-(R)-3-hydroxyhexanoate] [P(3HB-co-3HHx)] has a high potential to serve as commercial bioplastic due to its striking resemblance to petroleum-based plastics. In this study, five different genotypes of Cupriavidusnecator transformants harbouring the phaCBP-M-CPF4 gene (including PHB¯4/pBBR1-CBP-M-CPF4) were developed to evaluate the efficiency of 3HHx monomer incorporation. The fraction of 3-hydroxyhexanoate (3HHx) monomer that was incorporated into the PHA synthesized by these C. necator transformants using palm oil as the sole carbon source, was examined. Overall, co-expression of enoyl-CoA hydratase gene (phaJ1) from Pseudomonas aeruginosa, along with PHA synthase (PhaC), increased the 3HHx composition in the PHA copolymer. The differences in the enzyme activities of β-ketothiolase (PhaACn) and NADPH-dependent acetoacetyl-CoA reductase (PhaBCn) of the C. necator mutant hosts used in this study, were observed to alter the 3HHx composition and molecular weight of the PHA copolymer produced. The 3HHx fractions in the P(3HB-co-3HHx) produced by these C. necator transformants ranged between 1 and 18 mol%, while the weight-average molecular weight ranged from 0.7 × 106 to 1.8 × 106 Da. PhaCBP-M-CPF4 displayed a typical initial lag-phase and a relatively low synthase activity in the in vitro enzyme assay, which is thought to be the reason for the higher molecular weights of PHA obtained in this study. |
| Databáze: | OpenAIRE |
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