ArticleApplied and environmental microbiology2002
Enhanced accumulation and changed monomer composition in polyhydroxyalkanoate (PHA) copolyester by in vitro evolution of Aeromonas caviae PHA synthase.
Article in Applied and environmental microbiology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 24 papers.
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Who cites it
24 citing papers in PubMed, 125 citations in OpenAlex.
- Recent Advances of Controllable Production of PHA Biosynthesis in Chain Length, Monomers and Special Functional Groups.Current microbiology · 2026Review
- Monitoring proliferation and material production of Cupriavidus necator H16 using cell count and volume measurement.Scientific reports · 2025Article
- Metabolic Engineering Strategies for Enhanced Polyhydroxyalkanoate (PHA) Production inPolymers · 2025Review
- Structures of Polyhydroxyalkanoate Synthase PhaC from Aeromonas caviae, Producing Biodegradable Plastics.Angewandte Chemie (International ed. in English) · 2025Article
- Toward the production of block copolymers in microbial cells: achievements and perspectives.Applied microbiology and biotechnology · 2024Review
- Exploring Class I polyhydroxyalkanoate synthases with broad substrate specificity for polymerization of structurally diverse monomer units.Frontiers in bioengineering and biotechnology · 2023Article
- A review on poly(3-hydroxybutyrate-Frontiers in bioengineering and biotechnology · 2022Review
- The Synthesis, Characterization and Applications of Polyhydroxyalkanoates (PHAs) and PHA-Based Nanoparticles.Polymers · 2021Review
- Tools for the discovery of biopolymer producing cysteine relays.Biophysical reviews · 2021Review
- Considerable increase in Poly(3-hydroxybutyrate) production viaBioImpacts : BI · 2021Article
- Engineering ofFrontiers in bioengineering and biotechnology · 2021Article
- Engineered biosynthesis of biodegradable polymers.Journal of industrial microbiology & biotechnology · 2016Review
- MtgA Deletion-Triggered Cell Enlargement of Escherichia coli for Enhanced Intracellular Polyester Accumulation.PloS one · 2015Article
- Directed evolution and structural analysis of NADPH-dependent Acetoacetyl Coenzyme A (Acetoacetyl-CoA) reductase from Ralstonia eutropha reveals two mutations responsible for enhanced kinetics.Applied and environmental microbiology · 2013Article
- Characterization of site-specific mutations in a short-chain-length/medium-chain-length polyhydroxyalkanoate synthase: in vivo and in vitro studies of enzymatic activity and substrate specificity.Applied and environmental microbiology · 2013Article
- Mutations derived from the thermophilic polyhydroxyalkanoate synthase PhaC enhance the thermostability and activity of PhaC from Cupriavidus necator H16.Journal of bacteriology · 2012Article
- Characterization of the highly active polyhydroxyalkanoate synthase of Chromobacterium sp. strain USM2.Applied and environmental microbiology · 2011Article
- Bacterial polymers: biosynthesis, modifications and applications.Nature reviews. Microbiology · 2010Review
- Engineered cyanophycin synthetase (CphA) from Nostoc ellipsosporum confers enhanced CphA activity and cyanophycin accumulation to Escherichia coli.Applied and environmental microbiology · 2006Article
- Engineering of chimeric class II polyhydroxyalkanoate synthases.Applied and environmental microbiology · 2004Article
Corrections and comments
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Authors and funding
3 authors at 2 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
By in vitro evolution experiment, we have first succeeded in acquiring higher active mutants of a synthase that is a key enzyme essential for bacterial synthesis of biodegradable polyester, polyhydroxyalkanoate (PHA). Aeromonas caviae FA440 synthase, termed PhaC(Ac), was chosen as a good target for evolution, since it can synthesize a PHA random copolyester of 3-hydroxybutyrate and 3-hydroxyhexanoate [P(3HB-co-3HHx)] that is a tough and flexible material compared to polyhydroxybutyrate (PHB) homopolyester. The in vitro enzyme evolution system consists of PCR-mediated random mutagenesis targeted to a limited region of the phaC(Ac) gene and screening mutant enzymes with higher activities based on two types of polyester accumulation system by using Escherichia coli for the synthesis of PHB (by JM109 strain) (S. Taguchi, A. Maehara, K. Takase, M. Nakahara, H. Nakamura, and Y. Doi, FEMS Microbiol. Lett. 198:65-71, 2001) and of P(3HB-co-3HHx) [by LS5218 [fadR601 atoC(Con)] strain]. The expression vector for the phaC(Ac) gene, together with monomer-supplying enzyme genes, was designed to synthesize PHB homopolyester from glucose and P(3HB-co-3HHx) copolyester from dodecanoate. Two evolved mutant enzymes, termed E2-50 and T3-11, screened through the evolution system exhibited 56 and 21% increases in activity toward 3HB-coenzyme A, respectively, and consequently led to enhanced accumulation (up to 6.5-fold content) of P(3HB-co-3HHx) in the recombinant LS5218 strains. Two single mutations in the mutants, N149S for E2-50 and D171G for T3-11, occurred at positions that are not highly conserved among the PHA synthase family. It should be noted that increases in the 3HHx fraction (up to 16 to 18 mol%) were observed for both mutants compared to the wild type (10 mol%).
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