• 대한환경공학회지
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• 제35권2호
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• pp.109-114
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• 2013

Polyhydroxyalkanoates (PHAs)는 다양한 미생물이 세포 내에 저장하는 에너지 저장물질로 생체적합성 플라스틱의 원료 물질이다. 본 연구에서는 Cupriavidus necator를 사용하여 독립영양배양조건으로 poly(3-hydroxybutyrate) (P(3HB))를 생산함에 있어서 $CO_2$ 농도가 미생물의 성장과 P(3HB)의 축적에 미치는 영향을 확인하였다. 첫 번째 질소원을 포함한 배지에서의 6일 간의 배양 단계에서 C. necator의 균체수는 $CO_2$ 농도와 관계없이 증가했지만, 균체 내에 축적된 P(3HB)의 양은 $CO_2$ 농도가 1-20%로 증가함에 따라 감소하였다. 두 번째 질소원이 제거된 배지에서의 4일 간의 배양 단계에서 P(3HB)의 축적량은 3% $CO_2$에서 가장 많았다. 하지만 전체 10일 간의 배양기간 동안 합성된 총 P(3HB)의 양은 1, 3, 10, 20% 순으로 나타났다. 따라서 본 연구에서는 $CO_2$를 사용하여 P(3HB)를 축적할 때 1-20% $CO_2$ 중에서 1% $CO_2$가 가장 효율적임을 확인하였다.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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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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• 제10권5호
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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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• 제10권6호
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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.

• 미생물학회지
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• 제34권4호
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• pp.219-224
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• 1998

Alcaligenes sp. SH-69에서 탄소원의 고갈 또는 새로운 탄소원의 첨가시 포도당을 단일탄소원으로 하여 생합성된 poly(3-hydroxybutyrate-co-3-hydroxyvalerate) [poly(3HB-co-3HV)] 공중합체의 물질대사 변화 양상을 조사하였다. 회분배양 과정 중 탄소원이 고갈된 후에는 세포 내에 축척되었던 poly(3HB-co-3HV) 공중합체의 분해가 일어나 고분자 함량의 감소와 함께 무게평균 분자량도 감소하였다. 분해 과정 중에는 저분자량의 PHA에 비하여 상대적으로 고분자량의 PHA양이 크게 감소하여 평균 분자량 분포가 보다 낮은 방향으로 이동되는 양상을 보였다. 이에 반하여 1차 탄소원(기질)으로 사용된 포도당의 고갈 직후 2차 기질로서 포도당과 함께 3HV의 전구물질인 levulinic acid를 혼합기질로 첨가해 주었을 경우, 세포 건체량의 지속적인 증가와 아울러 3HV 함량이 높은 poly(3HB-co-3HV) 공중합체의 합성이 이루어졌다. 그러나 아세톤을 이용한 고분자의 분획 실험 결과, 2차 기질로부터의 poly(3HB-co-3HV) 공중합체의 생합성과 1차 기질로부터 생합성된 공중합체의 분해가 동시에 일어나며, 또한 각 기질로부터 생합성된 고분자가 단지 3HV의 함량이 다른 poly(3HB-co-3HV) 공중합체임에도 불구하고 혼합물 형태로 존재하고 있음을 확인하였다.

• 한국미생물·생명공학회지
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• 제24권1호
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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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• 제18권5호
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• pp.404-407
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• 2003

R. eutropha의 PHA 생합성 유전자를 포함하는 플라스미드 pSYL107을 가진 재조합 대장균 GCSC6576과 A. latus PHA 생합성 유전자를 포함하는 플라스미드 pJC4를 가진 fadR atoC 돌연변이주 재조합 대장균 LS5218의 유청으로부터 P(3HB-co-3HV) 합성을 비교하였다. 재조합 대장균 GCSC6576(pSYL107)의 플라스크 배양에서 acetic acid induction과 oleic acid의 첨가는 3HV 함량을 증가시켰다. 재조합 대장균 LS5218의 pH-stat 유가식 배양결과, 39시간에 균체농도 31.8 g/L, P(3HB-co-3HV) 농도 10.6 g/L, P(3HB-co-3HV) 함량 33.4%, 3HV 함량 6.26 mol%를 얻을 수 있었다.

• Journal of Applied Biological Chemistry
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• 제42권1호
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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.

• 한국미생물·생명공학회지
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• 제19권1호
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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).

• 한국염색가공학회지
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• 제7권1호
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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.