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  • 1
    Electronic Resource
    Electronic Resource
    Diversity of bacterial polyhydroxyalkanoic acids (1995)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 128 (1995), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract An overview is provided on the diversity of biosynthetic polyhydroxyalkanoic acids, and all hitherto known constituents of these microbial storage compounds are listed. The occurrence of 91 different hydroxyalkanoic acids reflects the low substrate specificity of polyhydroxyalkanoic acid synthases which are the key enzymes of polyhydroxyalkanoic acid biosynthesis. In addition, the importance of bacterial anabolism and catabolism, which provide the coenzyme A thioesters of the respective hydroxyalkanoic acids as substrates to these PHA synthases, is emphasized.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1995.tb07528.x
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  • 2
    Electronic Resource
    Electronic Resource
    Application of recombinant gene technology for production of polyhydroxyalkanoic acids: Biosynthesis of poly(4-hydroxybutyric acid) homopolyester (1994)
    Springer
    Journal of polymers and the environment 2 (1994), S. 67-74 
    Details
    ISSN: 1572-8900
    Keywords: Polyhydroxyalkanoic acids (PHA) ; 4-hydroxybutyric acid ; homopolyester ; poly(4-hydroxybutyric acid) ; biodegradable polyesters
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Energy, Environment Protection, Nuclear Power Engineering , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract Screening of a large number of bacteria revealed several strains, which utilize 1,4-butanediol and/or 4-hydroxybutyric acid (4HB) as a carbon source for growth and for synthesis of polyhydroxyalkanoic acids (PHA) containing 4HB as one constituent among others (mostly 3-hydroxybutyric acid). However, none of the wild-type strains investigated in this study was able to produce a homopolyester consisting solely of 4HB. Only several poly(3-hydroxybutyric acid)-leaky mutants ofAlcaligenes eutrophus strain JMP222 synthesized poly(4HB) homopolyester, which amounted to approximately 10% (w/w) of the cellular dry matter. If the PHA synthase structural gene ofA. eutrophus strain H16 was expressed in these mutants, the amount of poly(4HB) was increased to approximately 30% (w/w). The occurrence of poly(4HB) was demonstrated by gas chromatographic as well as1H and13C nuclear magnetic resonance spectroscopic analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02074775
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  • 3
    Electronic Resource
    Electronic Resource
    Accumulation of poly(3-hydroxybutyric acid-co-3-hydroxyvaleric acid-co-4-hydroxyvaleric acid) by mutants and recombinant strains ofAlcaligenes eutrophus (1995)
    Springer
    Journal of polymers and the environment 3 (1995), S. 169-175 
    Details
    ISSN: 1572-8900
    Keywords: Alcaligenes eutrophus ; biodegradable polyesters ; 4-hydroxyvaleric acid (4HV) ; PHA synthase ; polyhydroxyalkanoic acid (PHA) ; poly-β-hydroxybutyric acid (PHB)
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology , Energy, Environment Protection, Nuclear Power Engineering , Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Notes: Abstract Alcaligenes eutrophus accumulated a terpolyester of 3-hydroxybutyric acid (3HB), 3-hydroxyvaleric acid (3HV), and 4-hydroxyvaleric acid (4HV) during cultivation with 4HV as carbon and energy source under nitrogen starvation. The polyester accumulated by wild-type strains under these conditions contained 4HV at a molar fraction of approximately 5 mol% only. A catabolic pathway of 4HV was postulated, which included the activation of 4HV to 4HV-CoA and a conversion of 4HV-CoA to 3HV-CoA. Tn5::mob-induced mutants were isolated fromA. eutrophus HF39, which were affected in 4HV and/or valeric acid catabolism. Among 83 mutants were 27 4HV-negative or 4HV-leaky mutants; two mutants were identified which accumulated a terpolyester with a molar fraction of 10.1 to 22.7 mol% 4HV. In addition, a further increase in the molar fraction of 4HV in poly(3HB-co-3HV-co-4HV) and a two- to fourfold increase in the PHA synthase activity were monitored in these mutants or others and also in HF39, if the cells were complemented with the hybrid plasmid pHP1014::PP1, which contained the PHA biosynthesis genes ofA. eutrophus H16. Application of mutagenesis plus recombinant DNA techniques resulted in the accumulation of a terpolyester with up to 30 mol% 4HV and with approximately equimolar fractions of 3HB, 3HV, and 4HV.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF02068468
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  • 4
    Electronic Resource
    Electronic Resource
    Metabolic engineering of Arabidopsis and Brassica for poly(3-hydroxybutyrate- co -3-hydroxyvalerate) copolymer production (1999)
    [s.l.] : Nature America Inc.
    Nature biotechnology 17 (1999), S. 1011-1016 
    Details
    ISSN: 1546-1696
    Source: Nature Archives 1869 - 2009
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: [Auszug] Poly(hydroxyalkanoates) are natural polymers with thermoplastic properties. One polymer of this class with commercial applicability, poly(3-hydroxybutyrate- co-3-hydroxyvalerate) (PHBV) can be produced by bacterial fermentation, but the process is not economically competitive with polymer ...
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1038/13711
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  • 5
    Electronic Resource
    Electronic Resource
    Identification of 4-hydroxyvaleric acid as a constituent of biosynthetic polyhydroxyalkanoic acids from bacteria (1992)
    Springer
    Applied microbiology and biotechnology 36 (1992), S. 507-514 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Summary Twenty-four different strains of aerobic Gram-negative bacteria, mainly belonging to the genera Alcaligenes, Paracoccus, Pseudomonas and Methylobacterium, were examined with respect to their ability to utilize 4-hydroxyvaleric acid (4HV), 4-valerolactone (4VL) and 3-hydroxypropionic acid (3HP) as carbon sources for growth and for accumulation of polyhydroxyalkanoic acid (PHA). A gas chromatographic (GC) method for the detection of 3-hydroxyalkanoic acid methyl esters has been extended for the detection of derivatives obtained from the methanolysis of 4-hydroxybutyric acid (4HB) and 4HV. Most of the Alcaligenes species and P. oxalaticus Ox1 accumulated a terpolyester consisting of 3-hydroxybutyric acid (3HB), 3-hydroxyvaleric acid (3HV) and 4HV as constituents from 4HV or 4VL as sole carbon sources in batch, fed-batch or two-stage fed-batch cultures. Poly(3HB-co-3HV-co-4HV) accumulated from 4HV by A. eutrophus strain NCIB 11599 amounted to approximately 50% of the cell dry matter and was composed of 42.0 mol % 3HB, 52.2 mol % 3HV and 5.6 mol % 4HV, respectively. Pseudomonads, which belong to the rRNA homology group I, were not able to incorporate 4HV. With 3HP as carbon source, the GC analysis provided evidence for the presence of 3HP in the PHA of many bacteria. Nuclear magnetic resonance spectroscopic analysis confirmed that, for example, A. eutrophus strain TF93 accumulated poly(3HB-co-3HP) with 98 mol % 3HB and 2 mol % 3HP if the cells were cultivated in the presence of 0.5% (w/v) 3HP.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00170193
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  • 6
    Electronic Resource
    Electronic Resource
    Expression of polyhydroxyalkanoic-acid-biosynthesis genes in methylotrophic bacteria relying on the ribulose monophosphate pathway (1993)
    Springer
    Applied microbiology and biotechnology 40 (1993), S. 284-291 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Four representatives of methylotrophic bacteria relying on the ribulose monophosphate (RMP) pathway were investigated for their capability to synthesize polyhydroxyalkanoic acids (PHA). In Methylophilus methylotrophus B115, Methylobacillus glycogenes strains B121 and B53 and Acetobacter methanolicus B58 no \-ketothiolase, acetoacetyl-coenzyme A (CoA) reductase or PHA synthase could be detected, and hybridization experiments using heterologous DNA probes derived from PHA-biosynthesis genes of Methylobacterium extorquens or Alcaligenes eutrophus gave no evidence for the presence of the corresponding genes in these PHA-negative methylotrophic bacteria. Fragments harbouring a cluster of PHA-biosynthesis genes of A. eutrophus or Chromatium vinosum or isolated PHA synthase structural genes of M. extorquens, Rhodospirillum rubrum or Rhodobacter sphaeroides were mobilized into the RMP pathway bacteria mentioned above. Only transconjugants, which harboured the PHA-biosynthesis genes of A. eutrophus or C. vinosum, expressed active \-ketothiolase, acetoacetyl-CoA reductase and PHA synthase and accumulated poly(3-hydroxybutyric acid) (PHB). Highest amounts of PHB (up to 15% of the cellular dry weight) were accumulated in transconjugants of Methylophilus methylotrophus B115 or of Methylobacillus glycogenes strains B121 and B53 harbouring the PHA-biosynthesis genes of C. vinosum.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00170382
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  • 7
    Electronic Resource
    Electronic Resource
    Identification of 4-hydroxyhexanoic acid as a new constituent of biosynthetic polyhydroxyalkanoic acids from bacteria (1994)
    Springer
    Applied microbiology and biotechnology 40 (1994), S. 710-716 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Various aerobic Gram-negative bacteria were analysed for utilizing 4-hydroxyhexanoic acid (4HHx) as a carbon source for growth and for synthesis of polyhydroxyalkanoic acids (PHA). Although many wild types grew on 4HHx, only recombinant strains of the PHA-negative mutants Pseudomonas putida GPp104 and Alcaligenes eutrophus PHB−4, which harboured plasmid pHP1014::E156 with the PHA-biosynthesis genes of Thiocapsa pfennigii, incorporated 4HHx up to a molar fraction of 47 or 1.4%, respectively, into PHA if the cells were cultivated in the presence of 4HHx as sole carbon source and under nitrogen starvation. A terpolyester consisting of 3-hydroxybutyric acid (3HB), 3-hydroxyhexanoic acid (3HHx) and 4HHx was synthesized, as revealed by gas chromatographic analysis of the accumulated polyester and as confirmed by nuclear magnetic resonance spectroscopic analysis of the isolated polyester. 4HHx was also detected in PHA accumulated by Rhodococcus ruber if 4HHx was used as a carbon source. However, it occurred at a molar fraction of maximally 1.3 mol% only beside 3HB, 3-hydroxyvaleric acid and 3HHx.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00173333
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  • 8
    Electronic Resource
    Electronic Resource
    Application of enzymatically synthesized short-chain-length hydroxy fatty acid coenzyme A thioesters for assay of polyhydroxyalkanoic acid synthases (1994)
    Springer
    Applied microbiology and biotechnology 40 (1994), S. 699-709 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract Various hydroxyacyl coenzyme A (CoA) thioesters were synthesized from the corresponding hydroxyalkanoic acid (such as e.g. [3-14C]d-(−)-hydroxybutyric acid, [1-14C]d-lactic acid, [1-14C]l-lactic acid, etc.) and from acetyl-CoA employing the propionate CoA transferase of Clostridium propionicum. Preparative isolation of the thioesters on hydrophobic matrices and analysis by HPLC are reported. These thioesters were subjected to a radiometric or a spectrometric assay of polyhydroxyalkanoic acid (PHA) synthase activity. The latter was based on the release of CoA from, for example, d-(−)-3-hydroxybutyryl-CoA, which was detected spectroscopically at 412 nm by reduction of 5,5′-dithiobis(2-nitrobenzoic acid) and provided a convenient assay of poly(3-hydroxybutyrate) synthase. When [1-14C]lactyl-CoA was used as substrate in a PHA synthase assay employing crude extracts obtained from various wild-type strains, [1-14C]lactyl-CoA was used as a substrate at a rate that was only less than 10−4 of the rate than with [3-14C]d-(−)-3-hydroxybutyryl-CoA or was negligible. One exception was a recombinant strain of Escherichia coli, which overexpressed the PHA synthase complex of Chromatium vinosum and which used [1-14C]d-lactyl-CoA as substrate at a relatively high rate.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00173332
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  • 9
    Electronic Resource
    Electronic Resource
    Cloning and characterization of the Methylobacterium extorquens polyhydroxyalkanoic-acid-synthase structural gene (1993)
    Springer
    Applied microbiology and biotechnology 39 (1993), S. 309-317 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract A cosmid gene bank of partially EcoRI-digested genomic DNA from Methylobacterium extorquens IBT no. 6 was screened for DNA fragments restoring polyhydroxyalkanoic-acid (PHA) accumulation in the PHA-negative mutant Alkaligenes eutrophus H16 PHB−4. The M. extorquens PHA-synthase structural gene phaC Mex was mapped on a 23-kbp EcoRI fragment by complementation studies, by hybridization experiments with heterologous DNA probes from A. eutrophus H16 encoding for phaA, phaB and phaC and by nucleic acid sequence analysis. Evidence for the presence of genes for a β-ketothiolase or an acetoacetyl-coenzyme A reductase on this fragment was not obtained. The nucleotide sequence of a 3.7-kbp region was obtained. It contained the entire 1.815-kbp phaC Mex plus approximately each 900-bp upstream and downstream of phaC Mex. PhaC Mex encoded a protein of 605 amino acods with a relative molecular mass (Mr) of 66742, which exhibited 38.1% amino acid identity with the A. eutrophus PHA synthase. Determination of the N-terminal amino acid sequence of an Mr 65 000 protein, which was enriched concomitantly with the purification of PHA granules in sucrose gradients, revealed a sequence that was identical with the amino acid sequence deduced from the most probable translation start codon except for a valine, which was obviously removed post-translationally. Enzyme analysis, which was done with the native gene and a phaC Mex ‘-’lacZ fusion gene, gave no evidence for expression of phaC Mex in Escherichia coli.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00192084
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  • 10
    Electronic Resource
    Electronic Resource
    Identification of 4-hydroxyhexanoic acid as a new constituent of biosynthetic polyhydroxyalkanoic acids from bacteria (1994)
    Springer
    Applied microbiology and biotechnology 40 (1994), S. 710-716 
    Details
    ISSN: 1432-0614
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Process Engineering, Biotechnology, Nutrition Technology
    Notes: Abstract. Various aerobic Gram-negative bacteria were analysed for utilizing 4-hydroxyhexanoic acid (4HHx) as a carbon source for growth and for synthesis of polyhydroxyalkanoic acids (PHA). Although many wild types grew on 4HHx, only recombinant strains of the PHA-negative mutants Pseudomonas putida GPp104 and Alcaligenes eutrophus PHB−4, which harboured plasmid pHP1014 : : E156 with the PHA-biosynthesis genes of Thiocapsa pfennigii, incorporated 4HHx up to a molar fraction of 47 or 1.4%, respectively, into PHA if the cells were cultivated in the presence of 4HHx as sole carbon source and under nitrogen starvation. A terpolyester consisting of 3-hydroxybutyric acid (3HB), 3-hydroxyhexanoic acid (3HHx) and 4HHx was synthesized, as revealed by gas chromatographic analysis of the accumulated polyester and as confirmed by nuclear magnetic resonance spectroscopic analysis of the isolated polyester. 4HHx was also detected in PHA accumulated by Rhodococcus ruber if 4HHx was used as a carbon source. However, it occurred at a molar fraction of maximally 1.3 mol% only beside 3HB, 3-hydroxyvaleric acid and 3HHx.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/s002530050054
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