• 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.

• Journal of Microbiology
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• 제39권1호
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• pp.79-82
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• 2001

The influence of levulinic acid (LA) on the production of copolyester consisting of 3-hydroxybutyrate (3HB) and 3-hydroxyvalerate (3HV) by Ralstonia eutropha was investigated. Addition of LA into the culture medium greatly increased the molar fraction of 3HV in the copolyester, indicating that LA can be utilized as a precursor of 3HV. In shake flask culture, the 3HV content in the copolyester increased from 7 to 75 mol% by adding 0.5 to 4.0 g/L LA to the medium containing fructose syrup as a main carbon source. A maximal copolyester concentration of 3.6 g/L (69% of dry cell weight) was achieved with a 3HV content of 40 mo1% in a jar fermentor culture containing 4.0 g/L of LA. When LA (total concentration, 4 g/L) was added repeatedly into a fermentor culture to maintain its concentration at a low level, the copolyester content and the 3HV yield from LA reached up to 85% of dry cell weight and 5.0 g/g, respectively, which were significantly higher than those when the same concentration of the LA was supplied al1 at once. The present results indicated that LA is more effective than propionate or valerate as a cosubstrate fur the production of copolyesters with varying molar fractions of 3HV by R. eutropha.

• KSBB Journal
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• 제25권3호
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• pp.239-243
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• 2010

식물성 오일 또는 glycerol로부터 Ralstonia eutropha 여섯 균주의 polyhydroxyalkanoates (PHA) 합성 특성을 조사하였다. 탄소원으로 soybean oil, olive oil, 또는 glycerol 만을 공급시 poly(3-hydroxybutyrate) homopolymer가 생성되었으며, $\gamma$-butyrolactone 또는 pentanoic acid를 함께 공급시 poly(3-hydroxybutyrate-co-4-hydroxybutyrate) 또는 poly(3-hydroxybutyrate-co-3-hydroxyvalerate) copolymer가 각각 합성되었다. 최종 균체농도 및 PHA 함량면에서 최적인 균주는 R. eutropha KCTC 2662로 결정되었으며, 최적 기질인 soybean oil 20 g/L로부터 72 h에 균체농도 1.7~9.2 g/L, PHA 함량 70~92 wt%를 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제32권1호
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• pp.110-116
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• 2022

Polyhydroxyalkanoates (PHAs) are emerging as alternatives to plastics by replacing fossil fuels with renewable raw substrates. Herein, we present the construction of engineered Escherichia coli strains to produce short-chain-length PHAs (scl-PHAs), including the monomers 4-hydroxyvalerate (4HV) and 3-hydroxyvalerate (3HV) produced from levulinic acid (LA). First, an E. coli strain expressing genes (lvaEDABC) from the LA metabolic pathway of Pseudomonas putida KT2440 was constructed to generate 4HV-CoA and 3HV-CoA. Second, both PhaAB enzymes from Cupriavidus necator H16 were expressed to supply 3-hydroxybutyrate (3HB)-CoA from acetyl-CoA. Finally, PHA synthase (PhaC_Cv) from Chromobacterium violaceum was introduced for the subsequent polymerization of these three monomers. The resulting E. coli strains produced four PHAs (w/w% of dry cell weight): 9.1 wt% P(4HV), 1.7 wt% P(3HV-co-4HV), 24.2 wt% P(3HB-co-4HV), and 35.6 wt% P(3HB-co-3HV-co-4HV).

• 생명과학회지
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• 제32권7호
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• pp.510-522
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• 2022

Poly (3-hydroxyalkanoates) (PHA)는 탄소원과 에너지원으로서 미생물이 과립형태로 생합성하는 열가소성 플라스틱이다. PHA 중 가장 연구가 많이 된 polyhydroxyburyrate (PHB)는 높은 결정성과 부서지기 쉬운 성질로 인해 사용 및 응용범위가 제한적이다. 따라서 본 연구에서는 polypropylene과 좀 더 유사한 성질을 지닌 PHA를 생합성하고자, 3-hydroxybutyrate와 3-hydroxyvalerat의 공중합체인 poly (3-hydroxybutyrate-co-3-hydroxyvalerat) (PHBV)를 생합성하는 Bacillus sp. EMK-5020 균주를 토양에서 분리하였다. 플라스크로 배양한 결과, Bacillus sp. EMK-5020 균주는 효소를 이용한 막걸리 주박 가수분해산물(enzymatic makgeolli less hydrolysate, MLEH)에 포함된 환원당(환원당_MLEH)을 단일 탄소원으로 이용하여 1.3%의 3HV가 함유된 PHBV를 생합성하였으며, 보조탄소원으로 첨가한 propionic acid의 양이 증가할수록 3HV의 함량이 증가하여 최대 48.6%의 3HV가 포함된 PHBV를 합성하였다. 이 결과를 바탕으로 환원당_MLEH (20 g/l)와 propionic acid (1 g/l)를 각각 주 탄소원 및 보조탄소원으로 이용하여 3 l 발효기에서 균주를 72시간 배양한 결과 6.4 g/l DCW와 8.9% 3HV를 함유하는 PHBV (MLEH-PHBV)를 50 wt% 생합성함을 확인하였다. 겔 투과 크로마토그래피 분석을 통해 MLEH-PHBV의 평균 분자량은 152 kDa으로 standard PHBV의 평균 분자량(314 kDa)에 비해 절반으로 감소한 것을 확인하였으며, 열중량을 분석한 결과 MLEH-PHBV의 분해 온도가 standard PHBV보다 20℃ 높은 273℃임을 확인하였다. 결론적으로 본 연구에서는 Bacillus sp. EMK- 5020균주를 이용하여 MLEH 및 propionic acid를 탄소원으로 사용하여 다양한 3HV 분획을 함유하는 PHBV생합성할 수 있었으며, 대량배양을 통해 생합성된 8.9% 3HV를 함유한 PHBV-MLEH는 standard PHBV (8% 3HV)에 비해 높은 열 안전성을 지닌 것을 확인하였다.

• 한국환경과학회:학술대회논문집
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• 한국환경과학회 2016년도 정기학술대회 발표논문집
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• pp.284-284
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• 2016

• 한국동물학회:학술대회논문집
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• 한국동물학회 2000년도 한국생물과학협회 학술발표대회
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• pp.228.2-228
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• 2000

No Abstract, See Full Text

• Carbon letters
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• 제21권
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• pp.116-121
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• 2017

• 한국미생물·생명공학회지
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• 제26권6호
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• pp.537-544
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• 1998

3-HV 몰분율이 증대된 P(3HB-3HV)의 고농도 생산을 위하여 재조합 phbC유전자를 모균주에 재도입시킨 형질전환 Alcaligenes eutrophus AER5의 배양공학적 연구를 수행하였다. 상기 형질전환균주를 전구물질인 valerate가 10.0g/L첨가된 최소배지에서 2단계 배양한 결과 P(3HB-3HV)내의 3-HV 몰분율이 52.2 mol%로 모균주 A. eutrophus H16의 30 mol%에 비해 현저히 증가하였다. 이와 같은 몰분율의 증가는 phbC 유전자의 산물인 PHB synthase의 증폭과 밀접한 관련이 있음을 확인하였다. 2단계 회분배양시 fructose의 보충첨가의 영향을 검토하였으며, 적정농도인 10.0g/L의 fructose를 valerate와 같이 첨가할 경우 총균체량, p(3HB-3HV)의 축적농도, 그리고 축적율은 현저히 증가한 반면, 3-HV 몰분율은 다소 감소하는 경향을 보였다. $Mg^{2+}$ 이온은 P(3HB-3HV) 축적 에 매우 민감한 영향을 미쳤으며, 적정 농도로 판단되는 0.1 g/L일 경우 P(3HB- 3HV) 축적농도는 6.1 g/L, 그리고 3-HV 몰분율은 71.3 %로 현저히 증가하였다. 또한 적정 pH 조절제의 선정을 위하여 NaOH, NaOH와 (NH$_4$)$_2$SO$_4$의 혼합액, 그리고 NH$_4$OH을 비교 검토하였다. 2단계 배양법의 번거로움을 극복하고저 1단계 간헐적 유가식배양법을 검토한 결과, 배양 72시간 후 총균체량 19.2g/L, P(3HB-3HV) 축적농도 10.4 g/L, 그리고 3-HV 몰분율 35 mol%로 2단계 회분 배양법보다 우수한 결과를 얻었다. 이는 형질전환균주를 이용한 3-HV 몰분율이 높은 P(3HB-3HV)의 고농도 생산을 위한 배양조건 확립을 위한 기초자료로 활용될 것이다.90배의 살충력을 보였다. SDS-PAGE와 Western blot 분석에서도 클론 pHLN2-72는 재조합 클론 pHLN2-80(-)보다 약간 높게 ICP가 생성이 되었었다. 이상의 결과는 과다발현에 Plac프로모터와 종결부위가 반드시 필요하며, -72 bp ICP 프로모터가 -80 bp 프로모터보다 과다발현률이 높았으며, ICP 유전자는 반드시 Plac프로모터의 전사 방향에 역방향으로 삽입이 되어야 하는 것으로 나타났다.사용함으로써 고품질의 전통주류 생산이 가능하다고 사료된다.noicacid, methylmalonic acid, 2-methyl-4-keto-pentan-2-ol 등과 지방산의 일종인 tetradecanoic acid, hexadecane, hept-adecanoic acid, octadecanoic acid 등이 확인되었다.er 7.6mm. Further rich management measure and investigation were recommended such as sapling protection, signboard construction, soil erosion controlling and regular monitoring within the community.ldom changed at level 1.will cause students to form the mathematical concepts correctly. 3. By visualizing the process of drawing the quadratic function graph, students understand the quadratic function graph structually. 4. Through the feedback, the recognition ability of the trigonometric function can be improved. 5. It is possible to change

• Macromolecular Research
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• 제10권3호
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• pp.150-157
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• 2002

Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) is a microbial storage polymer with biodegradable properties. In order to improve the cell compatibility of PHBV surfaces, the physicochemical treatments have been demonstrated. In this study, physical method was corona discharge treatment and chemical method was chloric acid mixture solution treatment. The physicochemically treated PHBV film surfaces were characterized by the measurement of water contact angle, electron spectroscopy for chemical analysis, and scanning electron microscopy (SEM). The water contact angle of the physicochemically treated PHBV surfaces decreased from 75 to 30～40 degree, increased hydrophilicity. due to the introduction of oxygen-based functional group onto the PHBV backbone chain. The mouse NIH/3T3 fibroblasts cultured onto the physicochemically treated PHBV film surfaces with different wettability. The effect of the PHBV surface with different wettability was determined by SEM as counts of cell number and [$^3$H]thymidine incorporation as measures of cell proliferation. As the surface wettability increased, the number of the cell adhered and proliferated on the surface was increased. The result seems closely related with the serum protein adsorption on the physicochemically treated PHBV surface. In conclusion, this study demonstrated that the surface wettabilily of biodegradable polymer as the PHBV plays an important role for cell adhesion and proliferation behavior for biomedical application.