Cloning and molecular analysis of the poly(3-hydroxybutyric acid) biosynthetic genes of Thiocystis violacea

  title={Cloning and molecular analysis of the poly(3-hydroxybutyric acid) biosynthetic genes of Thiocystis violacea},
  author={Matthias Liebergesell and Alexander Steinb{\"u}chel},
  journal={Applied Microbiology and Biotechnology},
From a genomic library of Thiocystis violaceae strain 2311 in λL47, two adjacent EcoRI restriction fragments of 5361 base pairs (bp) and of 1978 bp were cloned. The 5361-bp EcoRI restriction fragment hybridized with a DNA fragment harbouring the Alcaligenes eutrophus poly(3-hydroxyalkanoate) (PHA) synthase operon (phbCAB) and restored the ability to synthesize and accumulate PHA in PHA-negative mutants derived from A. eutrophus. The nucleotide sequence analysis of both fragments revealed five… 

Altered composition of Ralstonia eutropha poly(hydroxyalkanoate) through expression of PHA synthase from Allochromatium vinosum ATCC 35206

The results of this study make available a phylogenetically diverse type III phaC and phaE genes, and confirm through kanamycin selection pressure the existence of multiple PHA biosynthesis systems in R. eutropha.

Anaylsis of polyhydroxyalkanoic acid-biosynthesis genes of anoxygenic phototrophic bacteria reveals synthesis of a polyester exhibiting an unusal composition

From genomic libraries of purple sulphur bacteria, fragments were cloned that encoded for proteins involved in the synthesis of poly(3-hydroxyalkanoic acids), PHA, and the most striking result was that a fragment harbouring the PHA-synthase gene of T. pfennigii conferred the ability to synthesize a niosynthetic polyester with this composition.

Characterization of the Polyhydroxyalkanoate Synthase Gene Locus of Rhodobacter Sphaeroides

Enzymatic analysis, nucleotide sequence analysis, hybridization studies and analysis of Tn5-induced PHA-negative mutants provided multiple evidence that phaCRs represents a monocistronic operon and that phACRs is isolated in the genome from other genes known to be relevant for PHA metabolism.

Autoregulator Protein PhaR for Biosynthesis of Polyhydroxybutyrate [P(3HB)] Possibly Has Two Separate Domains That Bind to the Target DNA and P(3HB): Functional Mapping of Amino Acid Residues Responsible for DNA Binding

Direct evidence is obtained that PhaR binds to the target DNA and poly[(R)-3-hydroxybutyrate] [P(3HB)], one of the PHAs, at the same time, by using a quartz crystal microbalance.

Evaluation of promoters for gene expression in polyhydroxyalkanoate-producing Cupriavidus necator H16

Five kinds of promoters were evaluated as tools for regulated gene expression in the PHA-producing bacterium Cupriavidus necator, and plac, Ptac, PphaC, and PphaP mediated constitutive gene expression, among which Ptac was the strongest promoter.

Polyhydroxyalkanoate (PHA) Synthases: The Key Enzymes of PHA Synthesis

Polyhydroxyalkanoic acids represent a rather complex class of polyesters that are synthesized by most genera of bacteria and members of the family Halobacteriaceae of the Archaea and are considered for several applications in the packaging industry, medicine, pharmacy, agriculture and food industry.

Application of enzymatically synthesized short-chain-length hydroxy fatty acid coenzyme A thioesters for assay of polyhydroxyalkanoic acid synthases

Various hydroxyacyl coenzyme A (CoA) thioesters were synthesized from the corresponding hydroxyalkanoic acid and from acetyl-CoA employing the propionate CoA transferase of Clostridium propionicum to provide a convenient assay of poly(3-hydroxybutyrate) synthase.

roteomic analysis of the purple sulfur bacterium andidatus “ Thiodictyon syntrophicum ” strain Cad 16 T solated from Lake Cadagno icola

Details regarding the dark CO2 assimilation of the PSB are disclosed, including three enzymes that could be part of the autotrophic dicarboxylate/4-hydroxybutyrate cycle, normally observed in archaea.



Cloning and nucleotide sequences of genes relevant for biosynthesis of poly(3-hydroxybutyric acid) in Chromatium vinosum strain D.

From a genomic library of Chromatium vinosum strain D in lambda L47, a 16.5-kbp EcoRI-restriction fragment was identified by hybridization with a DNA fragment harboring the operon for Alcaligenes eutrophus poly(3-hydroxyalkanoate) (PHA) synthesis, which restored the ability to synthesize and accumulate PHA in PHA-negative mutants of A. eUTrophus.

Identification and characterization of two Alcaligenes eutrophus gene loci relevant to the poly(beta-hydroxybutyric acid)-leaky phenotype which exhibit homology to ptsH and ptsI of Escherichia coli

DNA fragments which restored the wild-type phenotype to poly(beta-hydroxybutyric acid) (PHB)-leaky mutants derived from strains H16 and JMP222 were cloned and evidence for the involvement of these proteins in regulation of the metabolism of intracellular PHB was obtained and a hypothetical model is proposed.

Molecular analysis of the Alcaligenes eutrophus poly(3-hydroxybutyrate) biosynthetic operon: identification of the N terminus of poly(3-hydroxybutyrate) synthase and identification of the promoter

The translational initiation codon was identified by analysis of the amino acid sequence of a PHB synthase-beta-galactosidase fusion protein, exhibiting striking homology to the Escherichia coli sigma 70 promoter consensus sequence.

Fine structural analysis of the Zoogloea ramigera phbA‐phbB locus encoding β‐ketothiolase and acetoacetyl‐CoA reductase: nucleotide sequence of phbB

N‐terminal protein sequence data obtained by Edman degradation of the purified Mr= 25000 polypeptide were used to identify the correct start of the NADPH‐specific acetoacetyl‐CoA reductase gene in Z. ramigera.

Physiology and molecular genetics of poly(β‐hydroxyalkanoic acid) synthesis in Alcaligenes eutrophus

The Alcaligenes eutrophus genes for β‐ketothiolase, NADPH‐dependent acetoacetyl‐CoA reductase and poly(β‐hydroxybutyric acid) synthase (PHB synthase) which comprise the three‐step PHB‐biosynthetic

Naturally occurring genetic transfer of hydrogen-oxidizing ability between strains of Alcaligenes eutrophus

The results allow the conclusion that A. eutrophus H16 harbors a self-transmissible plasmid designated pHG1, which carries information for hydrogen-oxidizing ability, and that spontaneous Aut- mutants obtained only with strain TF93 were obtained.