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PHB synthase from Streptomyces aureofaciens NRRL 2209
1Ecologie Microbienne, Bat.405, University Claude Bernard Lyon 1, 69622 Villeurbanne Cedex, France. rchyd@yahoo.com
FEMS Microbiology Letters
|December 29, 2004
Summary
Researchers identified a novel polyhydroxyalkanoate (PHA) synthase gene, phaC(Sa), from Streptomyces aureofaciens. This gene aids in PHB biosynthesis in engineered E. coli, showing significant enzyme activity.
Area of Science:
- Microbiology
- Molecular Biology
- Biotechnology
Background:
- Polyhydroxyalkanoes (PHAs) are biodegradable polyesters with diverse applications.
- Identifying novel PHA synthase genes is crucial for improving PHA production efficiency.
Purpose of the Study:
- To sequence and analyze a DNA fragment from Streptomyces aureofaciens involved in PHA biosynthesis.
- To characterize the putative PHA synthase gene (phaC(Sa)) and its expression in recombinant Escherichia coli.
Main Methods:
- Genomic DNA sequencing of a 4.9 kb Sau3A I fragment from Streptomyces aureofaciens.
- Analysis of the phaC(Sa) gene for its coding potential and GC content.
- Expression studies in recombinant E. coli to detect PHA synthase activity.
Main Results:
- A 2 kb putative PHA synthase gene, phaC(Sa), was identified, encoding a 63.5 kDa protein.
- The gene exhibited high GC content (79.1%) and expressed under its native promoter.
- Significant PHA synthase activity was confirmed in recombinant E. coli.
- This represents the first reported PHA synthase from Streptomyces with a serine active nucleophile.
Conclusions:
- The identified phaC(Sa) gene is a functional PHA synthase from Streptomyces.
- Its proximity to a regulatory gene suggests a potential regulatory mechanism for PHA production.
- This discovery offers new possibilities for engineering microbial hosts for enhanced PHA synthesis.