• Journal of Korean Society of Environmental Engineers
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• v.35 no.2
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• pp.109-114
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• 2013

Polyhydroxyalkanoates (PHAs) are synthesized by numerous bacteria as carbon and energy storage compounds and are raw materials for biocompatible plastics. In this paper, the effect of $CO_2$ concentrations on the growth of C. necator and the accumulation of Poly(3-hydroxybutyrate) (P(3HB)) are investigated by increasing the $CO_2$ concentration in the substrate gas mixture. During 6 d cultivation in a nitrogen-present mineral medium, the $CO_2$ concentration did not affect the growth of the cells, while the Poly(3- hydroxybutyrate) (P(3HB)) content decreased with increasing $CO_2$ concentrations from 1% to 20%. During 4 d cultivation in the nitrogen-limited medium, the P(3HB) accumulation was the greatest at 3% $CO_2$; however, the total amount of accumulated P(3HB) was the greatest at 1% $CO_2$, which decreased with increasing $CO_2$ concentrations. The results indicate that the gas mixture with 1% $CO_2$ is the most effective in both growing the cells and accumulating P(3HB) under our experimental conditions.

• 한국생물공학회:학술대회논문집
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• 2000.11a
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• pp.175-178
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• 2000

Biodegradability of the $poly({\beta}-hydroxybutyrate-co-3 -hydroxyvalerate)$ [PHB/V] containing 0, 10 and 15mol% hydroxyvalerate [HV] was studied. Bioderadability of PHB/V was evaluated at $30^{\circ}C$ for 58 days and $55^{\circ}C$ for 33 days by monitoring the time-dependent changes in weight loss(erosion) of aerobic conditions in a temperature-controlled microcosms containing the earthworm cast($30^{\circ}C$) and compost ($55^{\circ}C$). It was found that PHB/V biodegradability occurred with increasing HV monomer concentration from 0 mol% to 15 mol%.

• Journal of Microbiology and Biotechnology
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• v.10 no.5
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• pp.628-631
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• 2000

The timing of poly(3-Hydroxybutyrate) (PHB) biosynthesis was controlled by varying the agitation speed of a stirred tank fermentor during the pH-stat fed-batch culture of recombinant Escherichia coli strain GCSC 6576 harboring pSYL107. Using a concentrated whey solution containing ca. 200 g/l lactose as the nutrient feed, the PHB content was only 57% after 35h due to volumetric limitation of the fermentor. However, by limiting the oxygen by maintaining the agitation speed at 300 rpm, the final PHB content increased to 70% after 70h with a cell concentration of 15 g/l. When the agitation speed was increased up to 500 rpm, a cell concentration of 31 g/l with 80% PHB was obtained after 52h. A further increase in the maximum agitation speed increased the cell concentration, PHB concentration, and PHB productivity, however, the PHB content decreased to 56-58%.

• Biotechnology and Bioprocess Engineering:BBE
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• v.10 no.6
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• pp.603-606
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• 2005

Ralstonia eutropha NCIMB 11599 and ATCC 17699 were grown, and their productions of poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] compared. In flask cultures of R. eutropha NCIMB 11599, cell concentration, P(3HB-co-4HB) concentration and polymer content decreased considerably with increases in the ${\gamma}-butyrolactone$ concentration, and the 4HB fraction was also very low (maximum 1.74 mol%). In fed-batch cultures of R. eutropha NCIMB 11599, glucose and ${\gamma}-butyrolactone$ were fed as the carbon sources, under a phosphate limitation strategy. When glucose was fed as the sole carbon source, with its concentration controlled using an on-line glucose analyzer, 86% of the P(3HB) homopolymer was obtained from 201g/L of cells. In a two-stage fed-batch culture, where the cell concentration was increased to 104g/L, with glucose fed in the first step and constant feeding of ${\gamma}-butyrolactone$, at 6g/h, in the second, final cell concentration at 67h was 106g/L, with a polymer content of 82%, while the 4HB fraction was only 0.7mol%. When the same feeding strategy was applied to the fedbatch culture of R. eutropha ATCC 17699, where the cell concentration was increased to 42 g/L, by feeding fructose in the first step and ${\gamma}-butyrolactone$ (1.5g/h) in the second, the final cell concentration, polymer content and 4HB fraction at 74h were 51g/L, 35% and 32 mol%, respectively. In summary, R. eutropha ATCC 17699 was better than R. eutropha NCIMB 11599 in terms of P(3HB-co-4HB) production with various 4HB fractions.

• Korean Journal of Microbiology
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• v.34 no.4
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• pp.219-224
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• 1998

The cyclic metabolism of poly(3-hydroxyhutyrate-co-3-hydroxyvalerate) synthesized from glucose by Alcaligenes sp. SH-69 in the presence or absence of new carbon substrate was investigated. In batch culture, the content and weight average molecular weight of the copolymer already stored in the cell decreased rapidly when there was no other carbon source available. After the depletion of carbon source, the amount of high molecular weight copolymer decreased more rapidly than that of low molecular weight copolymer, and as a result, average molecular weight distribution shifted to the lower value. The addition of a mixture of glucose and levulinic acid when the initial carbon substrate, glucose, was nearly depleted supported the continual increase in cell mass and the accumulation of poly(3HB-co-3HV) with high molar fraction of 3HV. However, solvent fractionation of the polymer with acetone revealed the degradation of pre-existing polyhydroxyalkanoale (PHA) in parallel with the synthesis of PHA from new carbon substrate. Even though PHAs obtained from each substrate alone were the copolymer of 3HB and 3HV, it was found that the polymer accumulated in the cells grown by sequential feeding was mainly physical mixture of two poly(3HB-co-3HV) copolymers containing different molar fractions of 3HV.

• Microbiology and Biotechnology Letters
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• v.24 no.1
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• pp.92-100
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• 1996

For the production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(P(3HB-co-3HV)) from butyric acid and valeric acid, 10 strains of bacteria capable of producing P(3HB-co-3HV) were isolated from soil. Among them, the strain HJ-067 showed the best ability of producing P(3HB-co-3HV), and was indentified as a Azotobacter sp. For the production of P(3HB-co-3HV), the optimum concentrations of butyric and valeric acid were 3.0g/l, respectively. The most effective nitrogen source was $(NH_4)_{2}SO_4$ at an optimum concentration of 0.75g/l, which was equivalent to 21.36 in C/N ratio. Deficiency of the cationic metal ions ($Zn^{2+},\;Co^{2+},\;Mn^{2+}$) in the proguction medium had stimulating effect on P(3HB-co-3HV) accumulation, especially in the manganese. deficient medium. The optimum temperature for P(3HB-co-3HV) production was 27$^{\circ}C$ and the optimum initial pH was 7.0. Under the optimum conditions, 1.82g/l of P(3HB-co-3HV) and 3.00g/l of dry biomass were produced after 36 hour cultivation, and the P(3HB-co-3HV) yield and HV% were 60.60% (w/w), 15.92%, respectively.

• KSBB Journal
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• v.18 no.5
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• pp.404-407
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• 2003

Two recombinant Escherichia coli strains, GCSC6576 harboring a plasmid pSYL107 containing the Ralstonia eutropha polyhydroxyalkanoate (PHA) biosynthesis genes and a fadR atoC mutant LS5218 harboring a plasmid pJC4 containing the Alcaligenes latus PHA biosynthesis genes were compared for their ability to synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [P(3HB-co-3HV) from whey. The 3HV fraction could be increased by acetic acid induction and oleic acid supplementation in flask cultures of recombinant E. coli GCSC6576. With the pH-stat fed-batch culture of recombinant E. coli LS5218, we obtained a cell concentration, a P(3HB-co-3HV) concentration, a P(3HB-co-3HV) content, and a 3HV fraction of 31.8 g/L, 10.6 g/L, 33.4%, and 6.26 mol%, respectively in 39 h.

• Journal of Applied Biological Chemistry
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• v.42 no.1
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• pp.1-6
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• 1999

Poly-${\beta}$-hydroxybutyrate (PHB) and enzymes related PHB metabolism have been measured in nitrogen-fixing symbiosis of chickpea and cowpea plants. Bacteroids from chickpea and cowpea contained PHB to 0.8% and 43% of their dry weight, respectively, whereas the free-living cells CC 1192 and I 16 produced $285{\pm}55mg$ and $157{\pm}18mg$ of PHB g (dry weight)$^{-1}$. To further understand why chickpea bacteroids contained little PHB, the enzyme activities of PHB metabolism (3-ketothiolase, acetoacetyl-CoA reductase, PHB depolymerase, and 3-hydroxybutyrate dehydrogenase), the TCA cycle (malate dehydrogenase, citrate synthase, and isocitrate dehydrogenase), and related reactions (malic enzyme, pyruvate dehydrogenase, and glutamate:2-oxoglutarate transaminase) were compared in extracts from chickpea and cowpea bacteroids and the respective free-living bacteria. Significant differences were observed between chickpea and cowpea bacteroids and between the bacteroid and free-living forms of CC 1192, with respect to the capacity for some of these reactions. It is indicated that a greater potential for oxidizing malate to oxaloacetate in chickpea bacteroids could be a factor that favors the utilization of acetyl-CoA in TCA cycle rather than for PHB synthesis.

• Microbiology and Biotechnology Letters
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• v.19 no.1
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• pp.94-99
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• 1991

- The further study for the identification of the previously reported pink-pigmented facultative methylotrophic bacterium (PPFM) GL-10 was carried out. The PPFM GL-10 was Gram nagative, rod, and motile by a single polarly inserted flagellum. The colonies were smooth, pink, circular, along with convex with entire margin. The isolate could utilize C1 compounds and a variety of multicarbon substrates as sole carbon and energy source. The isolate was obligately aerobic, and exhibited both catalase and oxidase activities. The deoxyribonucleic acid base composition was 65-67 mol% guanine plus cytosine. The isolate was mostly identical with Methylobacterium extorquens and named Methylobacterium sp. strain GL-10. Methylobacterium GL-10 accumulated a copolyester of 3-hydroxybutyrate and 3-hydroxyvalerate (poly-3HB/3 HV) when grown in nitrogen-free culture media containing sodium propionate as substrate at the second polyester accumulation stage. The composition of copolyester, as determined from $^1h$ NMR spectra, was 23 mol% of 3-hydroxyvalerate (3HV).

• Textile Coloration and Finishing
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• v.7 no.1
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• pp.51-57
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• 1995

Poly-$\beta$-hydroxybutyrate(PHB) was biosynthesized using Alcaligenes sp. FL-027. Alcaligenes sp. FL-027 was cultivated by fed-batch methods, in order to promote cell growth and PHB accumulation with carbon source. The cells were first grown at 3$0^{\circ}C$ on the fermentor. The structure of biosynthesized PHB is investigated by the NMR, IR. The crystalline portions were identified through the use of DSC and X-ray diffractometer. The melting point was about 16$0^{\circ}C$ and the diffraction peaks of (020) and (110) were shown at 13$^{\circ}$ and 17$^{\circ}$, respectively.