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  • 1
    Electronic Resource
    Electronic Resource
    Expression of the Alcaligenes eutrophus poly(β-hydroxybutyric acid)-synthetic pathway in Pseudomonas sp. (1989)
    Springer
    Archives of microbiology 153 (1989), S. 101-104 
    Details
    ISSN: 1432-072X
    Keywords: Poly(β-hydroxybutyric acid) ; PHB ; Biopolymer ; Alcaligenes eutrophus ; Pseudomonas oleovorans ; Fluorescent pseudomonads ; PHB-synthase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The 12.5-kb EcoRI restriction fragment PP1 of Alcaligenes eutrophus strain H16, which encodes for β-ketothiolase, NADP-dependent acetoacetyl-CoA reductase and poly(β-hydroxybutyric acid)-synthase was mobilized to six different species of the genus Pseudomonas belonging to the rRNA homology group I. Pseudomonas aeruginosa, P. fluorescens, P. putida, P. oleovorans, P. stutzeri and P. syringae, which are unable to synthesize and accumulate poly(β-hydroxybutyric acid), PHB, were employed as recipients. Whereas the A. eutrophus PHB-synthetic enzymes were only marginally expressed in P. stutzeri, they were readily expressed in the other species. For example, the specific activity of PHB-synthase was 1.8 U/g protein in transconjugants of P. stutzeri but was between 21 and 77 U/mg protein in transconjugants of the other species. All recombinant strains harboring plasmid pVK101::PP1 except those of P. stutzeri accumulated PHB; the PHB content of the cells grown on gluconate under nitrogen limitation varied between 8 and 24.3% of the cellular dry mass.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00277549
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  • 2
    Electronic Resource
    Electronic Resource
    Formation of poly(3-hydroxyalkanoates) by phototrophic and chemolithotrophic bacteria (1991)
    Springer
    Archives of microbiology 155 (1991), S. 415-421 
    Details
    ISSN: 1432-072X
    Keywords: Poly(3-hydroxybutyric acid) ; PHB ; Biopolymer ; Copolyester ; Phototrophic bacteria ; Chromatiaceae ; Rhodospirillaceae ; Sulfur purple bacteria ; Non-sulfur purple bacteria ; Chemolithotrophic bacteria
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The formation of poly(3-hydroxyalkanoic acid), PHA, by various strains of chemolithotrophic and phototrophic bacteria has been examined. Chemolithotrophic bacteria were grown aerobically under nitrogen-limiting conditions on various aliphatic organic acids. Phototrophic bacteria were grown anaerobically in the light on a nitrogen-rich medium and were subsequently transferred to a nitrogen-free medium containing acetate, propionate, valerate, heptanoate or octanoate as carbon source. All 41 strains investigated in this study were able to synthesize and accumulate PHA. All 11 strains of chemolithotrophic bacteria and all 15 strains belonging to the non-sulfur purple bacteria synthesized a polymer, which contained 3-hydroxy-valerate (3HV) beside 3-hydroxybutyrate (3HB), if the cells were cultivated in the presence of propionate, valerate or heptanoate. Many non-sulfur purple bacteria synthesized copolyesters of 3HB and 3HV even with acetate as carbon source. In contrast, most sulfur purple bacteria did not incorporate 3HV at all. Among 15 strains tested, only Chromatium vinosum strain 1611, C. purpuratum strain BN5500 and Lamprocystis roseopersicina strain 3112 were able to synthesize polyesters containing 3HV with propionate, valerate or heptanoate as carbon source.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00244955
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  • 3
    Electronic Resource
    Electronic Resource
    Incorporation of 2-methyl-3-hydroxybutyric acid into polyhydroxyalkanoic acids by axenic cultures in defined media (1996)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 138 (1996), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Alcaligenes eutrophus and Burkholderia cepacia synthesized and accumulated a terpolyester consisting of 3-hydroxybutyric acid, 3-hydroxyvaleric acid, and 2-methyl-3-hydroxybutyric acid (2Me3HB) if the cells were cultivated in a mineral salts medium containing tiglic acid as the sole carbon source or in combination with gluconic acid. The presence of 1–2 mol% of 2Me3HB in the polyester was confirmed by comparison with chemically synthesized methyl ester of 2Me3HB and by nuclear magnetic resonance spectrometry as well as by gas chromatography/mass spectrometry. This is the first report of the incorporation of 2Me3HB by axenic cultures cultivated under defined conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1996.tb08149.x
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  • 4
    Electronic Resource
    Electronic Resource
    Cloning and characterization of the Alcaligenes eutrophus 2-oxoglutarate dehydrogenase complex (1996)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 136 (1996), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Nucleotide sequence analysis of a 3.3-kb genomic EcoRI fragment and of relevant subfragments of a genomic 13.2-kb SmaI fragment of Alcaligenes eutrophus, which were identified by using a dihydrolipoamide dehydrogenase-specific DNA probe, revealed the structural genes of the 2-oxoglutarate dehydrogenase complex in a 7.5-kb genomic region. The genes odhA (2850 bp), odhB (1248 bp), and odhL (1422 bp), encoding 2-oxoglutarate dehydrogenase (El), dihydrolipoamide succinyltransferase (E2), and dihydrolipoamide dehydrogenase (E3), respectively, occur co-linearly in one gene cluster downstream of a putative −35 / −10 promoter in the order odhA, odhB, and odhL. In comparison to other bacteria, the occurrence of genes for two E3 components for the pyruvate as well as for the 2-oxoglutarate dehydrogenase complexes is unique. Heterologous expression of the A. eutrophus odh genes in E. coli XL1-Blue and in the kgdA mutant Pseudomonas putida JS347 was demonstrated by the occurrence of protein bands in electropherograms, by spectrometric detection of enzyme activities, and by phenotypic complementation, respectively.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1996.tb08054.x
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  • 5
    Electronic Resource
    Electronic Resource
    Utilization of methylacetoin by the strict anaerobe Pelobacter carbinolicus and consequences for the catabolism of acetoin (1988)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 55 (1988), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract Pelobacter carbinolicus strain GraBd1 fermented methylacetoin, which is a good carbon source for growth (μ= 0.16 h−1) of this strict anaerobic bacterium, to acetone, acetate and ethanol (main products), acetoin, 2,3-butanediol and methylbutanediol (minor products). During growth on 2,3-butanediol, acetoin and methyl-acetoin the formation of a protein exhibiting acetoin: DCPIP oxidoreductase activity is induced. This enzyme amounts to a substantial portion of the soluble proteins. In vitro, it cleaves acetoin into acetate and acetaldehyde but reacts also with diacetyl or methylacetoin. We discussed four different models for the degradation of acetoin in the cells and came to the conclusion that in vivo the oxidative-thiolytic acetoin cleavage model is most probably realized in P. carbinolicus.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1988.tb02796.x
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  • 6
    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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  • 7
    Electronic Resource
    Electronic Resource
    The Alcaligenes eutrophus ldh structural gene encodes a novel type of lactate dehydrogenase (1993)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 112 (1993), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract The lactate dehydrogenase gene, ldh, of Alcaligenes eutrophus H16 was identified on a 14-kbp EcoRI restriction fragment of a genomic library in the cosmid pHC79 by hybridization with a 50-mer synthetic oligonucleotide which was derived from the N-terminal amino acid sequence of the purified enzyme. Recombinant strains of Escherichia coli JM83, which harboured a 2.0-kbp Pst I subfragment in pUC9-1, expressed LDH at a high level, if ldh was downstream from and colinear to the E. coli lac promoter. The nucleotide sequence of a region of 4245 bp revealed several open reading frames which might represent coding regions. One represented the ldh gene. The amino acid sequence deduced from ldh exhibited 29% and 36% identity to the L-malate dehydrogenase of Methanothermus fervidus and to the putative translation product of an E. coli sequence of unknown function, respectively. The ldh was separated by short intergenic regions from two other open reading frames: ORF5 was located downstream of and colinear to ldh, and its putative translational product revealed 38 to 56% amino acid identity to penicillin-binding proteins. ORF3 was located upstream of and colinear to ldh, and its putative gene translational product represented a hydrophobic protein. A sequence, which resembled the A. eutrophus alcohol dehydrogenase promoter, was detected upstream of ORF3, which most probably represents the first transcribed gene of an operon consisting of ORF3, ldh and ORF5.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.1993.tb06453.x
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  • 8
    Electronic Resource
    Electronic Resource
    Effect of poly(3-hydroxybutyrate) (PHB) content on the starvation-survival of bacteria in natural waters (1995)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology ecology 16 (1995), S. 0 
    Details
    ISSN: 1574-6941
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Abstract The effect of poly(3-hydroxybutyrate) (PHB) content on the survival of wild-type strains and PHB negative mutants of Bacillus megaterium and Alcaligenes eutrophus in natural waters was studied. The survival strategy of B. megaterium was dominated by the development of resistant forms, but the number of the wild-type vegetative cells was higher than that of PHB mutant strain. In some environmental conditions the mutant spores needed a heat shock for germination, a fact that suggests, for the first time, that PHB plays a role in this phenomenon. Survival of A. eutrophus wild-type strain in all experiments was higher compared to the PHB mutant, and differences were significant. In raw river water, survival of both species was lower than in sterile river water.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6941.1995.tb00273.x
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  • 9
    Electronic Resource
    Electronic Resource
    Novel precursor substrates for polythioesters (PTE) and limits of PTE biosynthesis in Ralstonia eutropha (2003)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 221 (2003), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: A novel class of biopolymers referred to as polythioesters (PTE) was recently detected when the polyhydroxyalkanoate (PHA) accumulating bacterium Ralstonia eutropha was cultivated in the presence of 3-mercaptopropionic acid (3MP) or 3,3′-thiodipropionic acid (TDP). In this study, 3,3′-dithiodipropionic acid (DTDP) and 3-mercaptovaleric acid (3MV) were identified as two additional precursor carbon sources for in vivo biosynthesis of PTE in R. eutropha. Biosynthesis of copolymers of 3-hydroxybutyrate (3HB) and 3MP, which contributed 19–25% of cell dry matter, was compared referring to the different precursor substrates. Using DTDP as carbon source, which is probably cleaved into two molecules 3MP, yielded an about 2.3-fold higher molar 3MP content of the copolyester than TDP, which is probably cleaved into only one molecule 3MP. Furthermore, cultivation of R. eutropha in the presence of 3MV resulted in biosynthesis of copolymers consisting predominantly of 3HB with low amounts of 3MV and 3-hydroxyvalerate, each contributing less than 5 mol% of the constituents. In contrast, 4-mercaptobutyric acid could be not incorporated into PHAs, although – as documented in this study – five different strategies, various precursor substrates, R. eutropha and also a recombinant strain of Escherichia coli were employed. Therefore, this study not only extended the range of substrates suitable for PTE biosynthesis and also the range of PTE constituents in R. eutropha, it also demonstrates limits for PTE biosynthesis in this bacterium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1016/S0378-1097(03)00185-X
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  • 10
    Electronic Resource
    Electronic Resource
    Conversion of the nitrogen content in liquid manure into biomass and polyglutamic acid by a newly isolated strain of Bacillus licheniformis (2003)
    Oxford, UK : Blackwell Publishing Ltd
    FEMS microbiology letters 218 (2003), S. 0 
    Details
    ISSN: 1574-6968
    Source: Blackwell Publishing Journal Backfiles 1879-2005
    Topics: Biology
    Notes: Extensive spreading of liquid manure onto agricultural fields causes eutrophication of ground and surface water and also pollution of the atmosphere due to the high ammonium nitrogen content. A poly(γ-glutamic acid) (PGA)-producing strain of Bacillus licheniformis was isolated in this study and investigated for its ability to reduce the ammonium nitrogen by converting ammonium into biomass and PGA as depot forms of nitrogen. In batch cultivations swine manure and an optimized mineral salts medium were used for PGA production. For example the cultivation of B. licheniformis strain S2 in liquid manure, which was modified by adding of 18 g citrate and 80 g glycerol l−1 and exhibited a carbon to nitrogen ratio of 15.5:1, led to severe reduction of the ammonium content from 2.83 to 0.1 g l−1 and to the production of 0.16 g PGA and 7.5 g cell dry mass l−1 within 410 h. Approximately 28% (w/w) of the total nitrogen was converted into cellular biomass, whereas 0.1% (w/w) was used for the production of PGA. In addition, approximately 33% (w/v) of the original ammonium was lost by stripping.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1111/j.1574-6968.2003.tb11495.x
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