Many heterologous transformation studies have been carried out using the Cupriavidus necator PHB strain to investigate the expression characteristics of various polyhydroxyalkanoate (PHA) synthase enzymes. In this study, we generated a recombinant C. necator PHB strain by transforming a plasmid (pMRC03) harbouring the synthetic phaC2 gene of Pseudomonas putida CA-3. Under conditions favourable for expression of the phaC2 gene, canola oil was used as carbon source for the synthesis of PHAs. The expressed synthase polymerised monomers of 3-hydroxybutyrate (3-HB), 3-hydroxyvalerate (3-HV) and 3-hydroxyhexanoate (3-HHx) in the recombinant C. necator PHB (pMRC03) strain. We then co-expressed the phaC2 gene with the native phaC1 gene in wild type Cupriavidus necator H16 (C. necator H16 (pMRC03)). This co-expression produced a PHA blend of 3-HB, 3-HV, 3-HHx and 3-hydroxyoctanoate (3-HO) monomers in the presence of canola oil. Gas chromatography analysis revealed the presence of 94mol% 3-HB, 1mol% 3-HV, 4mol% 3-HHx and 1mol% 3-HO in a tetra-polymer. Thus, we confirmed that a synthetic phaC2 gene encoding the synthase enzyme is functionally active with substrates ranging from short to medium chain length PHAs.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.nbt.2017.08.001 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
S211 as a microbial cell factory for direct bioconversion of waste cooking oil into medium-chain-length polyhydroxyalkanoates.
3 Biotech
September 2024
APVA-LR16ES20, National School of Engineers of Sfax (ENIS), University of Sfax, Sfax, Tunisia.
The present study examines the use of waste cooking oil (WCO) as a substrate for medium-chain-length polyhydroxyalkanoates (mcl-PHA) production by S211. The genome analysis revealed that the S211 strain has a mcl-PHA cluster () encoding two class II PHA synthases (PhaC1 and PhaC2) separated by a PHA depolymerase (PhaZ), a transcriptional activator (PhaD) and two phasin-like proteins (PhaFI). Genomic annotation also identified a gene encoding family I.
View Article and Find Full Text PDFCultivation driven transcriptomic changes in the wild-type and mutant strains of .
Comput Struct Biotechnol J
December 2024
Department of Biomedical Engineering, Faculty of Electrical Engineering and Communication, Brno University of Technology, Brno, Czech Republic.
Purple photosynthetic bacteria (PPB) are versatile microorganisms capable of producing various value-added chemicals, e.g., biopolymers and biofuels.
View Article and Find Full Text PDFEngineering HT66 for the Biosynthesis of Copolymers Containing 3-Hydroxybutyrate and Medium-Chain-Length 3-Hydroxyalkanoates.
J Agric Food Chem
April 2024
State Key Laboratory of Microbial Metabolism, School of Life Sciences and Biotechnology, Shanghai Jiao Tong University, Shanghai 200240, China.
Polyhydroxyalkanoates (PHAs) are promising alternatives to petroleum-based plastics, owing to their biodegradability and superior material properties. Here, the controllable biosynthesis of scl--mcl PHA containing 3-hydroxybutyrate (3HB) and mcl 3-hydroxyalkanoates was achieved in HT66. First, key genes involved in fatty acid β-oxidation, the fatty acid biosynthesis pathway, and the -- operon were deleted to develop a chassis strain.
View Article and Find Full Text PDFMaximization of 3-hydroxyhexanoate fraction in poly(3-hydroxybutyrate-co-3-hydroxyhexanoate) using lauric acid with engineered Cupriavidus necator H16.
Int J Biol Macromol
January 2024
Department of Biological Engineering, College of Engineering, Konkuk University, Seoul, Republic of Korea. Electronic address:
Article Synopsis
- Researchers are working to improve the mechanical properties of polyhydroxyalkanoates (PHAs) by increasing the mole fraction of 3-hydroxyhexanoate (3HHx) in P(3HB-co-3HHx).
- Instead of genetic modification, they found that using coconut oil as a substrate produced the highest 3HHx mole fraction due to its high lauric acid content.
- By mixing lauric acid with other oils, they were able to control the 3HHx mole fraction between 9% and 31.1%, resulting in improved P(3HB-co-3HHx) with better molecular properties than those derived from gene-deleted bacteria.
Enhancing Production of Medium-Chain-Length Polyhydroxyalkanoates from sp. SG4502 by Enhancer Insertion.
Polymers (Basel)
May 2023
International Cooperation Research Center of China for New Germplasm Breeding of Edible Mushrooms, Jilin Agricultural University, Changchun 130118, China.
sp. SG4502 screened from biodiesel fuel by-products can synthesize medium-chain-length polyhydroxyalkanoates (mcl-PHAs) using glycerol as a substrate. It contains a typical PHA class II synthase gene cluster.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!