• 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 Microbiology and Biotechnology
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• 제5권3호
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• pp.167-171
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• 1995

A bacterial strain C-02 using methanol as a carbon source was isolated from Gumi Industrial Estate and selected based on its rapid growth and capability of poly-$\beta$-hydroxybutyrate accumulation. Characteristics of strain C-02 showed that it belongs to the Methylococcaceae family, Type II subgroup. Strain C-02 could incorporate valerate into the PHB chain to form 3-hydroxybutyrate and 3-hydroxyvalerate (P(3HB-co-3HV)). Among various nutrient limitation tests, the nitrogen limitation test resulted in the highest content of P(3HB-co-3HV) per dry cell weight, 50$%$. Under the nitrogen limited condition, the average molecular weight of P(3HB-co-3HV) obtained was determined to be approximately $2.8\times 10^5$ daltons.

• Biotechnology and Bioprocess Engineering:BBE
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• 제10권6호
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• pp.540-545
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• 2005

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate), poly(3HB-co-3HV), copolyesters, with 3-hydroxyvalerate (3HV) contents ranging from 17 to 60 mol%, were produced by Alcaligenes sp. MT-16, and their biocompatibility evaluated by the growth of Chinese hamster ovary (CHO) cells and the adsorption of blood proteins and platelets onto their film surfaces. The number of CHO cells that adhered to and grew on these films was higher with increasing 3HV content. In contrast, the tendency for blood proteins and platelets to adhere to the copolyester surfaces significantly decreased with increasing 3HV content. Examination of the surface morphology using atomic force microscopy revealed that the surface roughness was an important factor in determining the biocompatibility of theses copolyesters. The results obtained in this study suggest that poly(3HB-co-3HV) copolyesters, with >30 mol% 3HV, may be useful in biocompatible biomedical applications.

• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2001년도 추계학술발표대회
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• pp.321-324
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• 2001

R. eutropha 의 PHA 생합성 유전자를 포함하는 플라스미드 pSYLl07을 가진 재조합 대장균 GCSC6576 과 A. latus PHA 생합성 유전자를 포함하는 플라스미드 pJC4 한 가진 fadR atoC 돌연변이주 재조합 대장균 LS5218 의 유청으로부터 P(3HB- co -3HV) 합성을 비교하였다. 재조합 대장균 LS5218의 pH-stat 유가식 배양결괴 39 시간에 균체농도 31.8 g/L. P(3HB-co-3HV) 농도 10.6 g/L. P(3HB-co-3HV) 함량 33.4 wt%. 3HV 함량 6.26 mol%를 얻을 수 있었다.

• 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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• 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).

• 미생물학회지
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• 제36권2호
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• pp.155-160
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• 2000

Pseudomonas sp. HJ-2는 heptanoinc acid를 단일탄소원으로 이용하여 3-hydroxybutyrate (3HB),3-hydroxy-velerate (3HV) 및 3-hydroxybutyrate (3HHp)를 구성 단위체로 하는 고무탄성 polyhydroxyalkanoate (PHA)를 생합성한다. 이 미생물 고분자는 poly(3HB-co-3HV)공중합체와 poly(3HHp) 단일중합체로 이루어진 혼합물임이 밝혀졌다. 본 연구에서는 PHA가 고무탄성체로서의 성질을 유지하는데 필용한 단위체 조성과 HJ-2의 배양조건이 PHA의 생산 및 단위체 조성에 미치는 영향에 대하여 조사하였다. 생합성된 PHA의 탄성률은 poly(3HHp)의 존재로 크게 감소되었으나, 3HV의 함량이 높은 poly(3HB-co-3HV) 자체도 최대변형률 740%로서 고무탄성체로서의 성질을 보였다. HJ-2의 생장 및 PHA 생합성은 탄소원인 heptanoic acid의 초기농도가 40mMdlfEo 가장 높았으나, 50mM의 농도에서는 큰 저해를 받았다. PHA 생합성은 질소와 인이 결핍된 조건에서 크게 증가되었다. 배양액의 pH 및 통기는 HJ-2로부터 생합성되는 PHA의 단위체 조성에 큰 영향을 주는 것으로 나타났다. pH 7.5에서 생합성되는 고분자는 poly(3HB-co-38% 3HV)인 반면에 pH8.0에서의 고분자는 3HHp가 95%를 차지하였다. 발효조의 교반속도를 달리한 실험의 결과 고분자 내 3HHp의 함량은 산소전달 속도가 높아질수록 증가하였다.

• Journal of Microbiology
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• 제42권4호
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• pp.346-352
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• 2004

Poly(3-Hydroxybutyrate-co­3-Hydroxyvalerate), poly(3HB-co-3HV), copolyesters with a variety of 3HV contents (ranging from 17 to $60\;mol\%$) were produced by Alcaligenes sp. MT-16 grown on a medium containing glucose and levulinic acid in various ratios, and the effects of hydrophilicity and crystallinity on the degradability of the copolyesters were evaluated. Measurements of thermo-mechanical pro­perties and Fourier-transform infrared spectroscopy in the attenuated total reflectance revealed that the hydrophilicity and crystallinity of poly(3HB-co-3HV) copolyesters decreased as 3HV content in the copolyester increased. When the prepared copolyester film samples were non-enzymatically hydrolysed in 0.01 N NaOH solution, the weights of all samples were found to have undergone no changes over a period of 20 weeks. In contrast, the copolyester film samples were degraded by the action of extra­cellular polyhydroxybutyrate depolymerase from Emericellopsis minima W2. The overall rate of weight loss was higher in the films containing higher amounts of 3HV, suggesting that the enzymatic degra­dation of the copolyester is more dependent on the crystallinity of the copolyester than on its hydro­philicity. Our results suggest that the degradability characteristics of poly(3HB-co-3HV) copolyesters, as well as their thermo-mechanical properties, are greatly influenced by the 3HV content in the copoly­esters.

• Journal of Microbiology
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• 제35권2호
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• pp.127-133
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• 1997

Biosynthesis of polyhydroxybutyrate and copolymer consisting of 3-hydroxybutyrate and 3-hydroxyvalerate [poly(3HB-co-3HV)] by Bacillus thuringiensis R-510 grown with glucose or with mixtures of glucose and propionate was investigated. n-Alkanoic acids other than propionate were not precursors of 3HV units. The fraction of 3HV unit in the copolymer increased from 0 to 84 mol% of 3HV. Polymer yield decreased as the fraction of propionate was increased but the molecular weight distribution was not affected by the composition of carbon substrate. The minimum melting temperature (around 65.deg.C) of poly (3HB-co-3HV) copolymers was observed for the polymer bearing approximately 35 mol% of 3HV. Polyhydroxyalkanoates production by this organism was not dependent on nutritional limitation, but remarkably influenced by dissolved oxygen concentration in the culture medium. Low level of dissolved oxygen concentration prevented spore formation in the cells and stimulated the synthesis of polyhydroxyalkanoate. The composition of poly (3HB-co-3HV) produced by B. thuringiensis R-510 lyhydroxyalkanoate. The composition of poly(3HB-co-3HV) propduced by B. thuringiensis R-510 varied according to the growth time. However, there was no evidence that polymers isolated from cells were mixtures of immiscible polymers.

• Biotechnology and Bioprocess Engineering:BBE
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• 제7권6호
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• pp.371-374
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• 2002

Production of poly(3-hydroxybutyrate-co-3-hydroxyvalerate)［P(3HB/V)］, by fed-batch culture of recombinant Escherichia coli harboring a plasmid containing the Alcaligenes latus polyhy-droxyalkanoate (PHA) biosynthesis genes, was examined in two pilot-scale fermentors with air supply only, In a 30 L fermentor having a XLa value of 0.11 S­$^1$, the final P(3HB/V) concentration and the P(3HB/V) content obtained were 29.6 g/L and 70.1 wt%, respectively giving a productivity of 1.37 g P(3HB/V)/L-h. In a 300 L fermentor having a XLa of 0.03 S­$^1$, the P(3HB/V) concentration and the P(3HB/V) content were 20.4 g/L and 69 wt%, respectively giving a productivity of 1.06g P(3HB/V)/L-h. These results suggest that economical production of P(3HB/V) is possible by fed-batch culture of recombinant E. coli in a large-scale fermentor having low KLa value.