• KSBB Journal
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• 제22권3호
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• pp.146-150
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• 2007

Poly (3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] 미립구를 용매증발법으로 제조하였다. 3.9 mol% 4HB 조성의 P(3HB-co-4HB)를 Ralstonia eutropha의 유가식 배양으로부터 합성하였다. 계면활성제의 농도 및 종류(Tween 80, sodium dodecylsulfate, polyvinyl alcohol), 분산안정제 (Acacia)의 첨가, 고분자 및 모델 약물 (bovine serum albumin)의 농도 등이 미립구 입자 크기에 미치는 효과 및 in vitro 약물 방출 특성을 조사하였다. 평균 입자크기는 분산 안정제 첨가시 감소하였으며, 계면활성제, 약물 및 고분자의 농도가 증가할수록 증가하였다. 약물 방출량은 입자 크기가 감소할수록 증가하였다.

• KSBB Journal
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• 제14권4호
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• pp.422-428
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• 1999

Alcaligenes latus 유래의 phbC 유전자를 A. eutrophus에 재도입시킨 형질전환균주를 이용하여 높은 4HB 몰분율을 갖는 P(3HB-4HB)의 고농도 생산을 시도하였다. 형질전환균주는 총균체량, P(3HB-4HB) 농도, 그리고 축적률에서 모균주에 비해 다소 증가한 반면 P(3HB-4HB)내의 4HB 몰분율은 23.5 mol%로 모균주의 12.3 mol%에 비해 현저히 증가하였다. 이는 phbC유전자의 증폭으로 인해 해당과정에서 생성된 3HB와 전구물질인 ${\gamma}$-butyrolacton에서 전환된 4HB의 중합반응이 촉진되기 때문으로 사료된다. 또한 ${\gamma}$-butyrolacton의 농도 $Mg^{2-}$ 이온, 그리고 citrate 첨가량이 P(3HB-4HB)의 농도, 축적률, 그리고 4HB 몰분율에 미치는 영향을 검토하였다. P(3HB-4HB)내의 4HB 몰분율을 증대시키기 위하여 일반적으로 사용되는 2단계 배양법을 변형시켜 ${\gamma}$-butyrolacton과 citrate의 첨가시기를 늦춘 2단계 배양법을 활용하여 P(3HB-4HB)내의 4HB 몰분율을 61.0 mol%로 증가시킬 수 있었다. 또한 ${\gamma}$-butyrolacton의 첨가량을 조절하여 P(3HB-4HB)내의 4HB 몰분율이 92.0 mol%에 이르는 homopolymeric P(4HB)를 생산할 수 있었으며, 그 구조를 $^1$H-NMR을 통해 확인하였다.

• Journal of Microbiology and Biotechnology
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• 제11권2호
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• pp.333-336
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• 2001

An isolated phbC gene from Alcaligenes latus was reintroduced into the parent A. latus through the transformation process, and the effect of the amplified phbC gene on the biosynthesis of P(3-hydroxybutyrate-3-hydroxyvalerate) [P(3HB-3HV)] and P(3-hydroxybutyrate-4-hydroxybutyrate) [P(3HB-4HB)] in the transformant A. latus was investigated. The biosynthesis rate and content of the above copolymers increased up to 1.3-fold after enforcing its own phbC gene, and the molar fractions of 3HV and 4HB in P(3HB-3HV) and P(3HB-4HB) also changed remarkably from 35.0 to 48.0% and from 34.0 to 56.0%, respectively, showing a critical role of PHB synthase which catalyzes the polymerizing reactions between eiher 3HV or 4HB from precursor compounds and 3HB.

• KSBB Journal
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• 제19권5호
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• pp.358-362
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• 2004

Characteristics of cell growth and poly(3-hydroxybutyrate-co-4-hydroxybutyrate) [P(3HB-co-4HB)] synthesis was investigated through flask and batch cultures of Alcaligenes latus and Comamonas acidovorans. The specific growth rate of C. acidovorans increased with yeast extract concentration and decreased with 1,4-butanediol concentration. Optimum glucose concentration for growth of C. acidovorans was 20 g/L. In one-step flask cultures of C. acidovorans, final dry cell weight and PHA content decreased with the ratio of 1,4-butanediol to glucose, while the 4HB fraction in copolymers gradually increased to 100 $mol\%$ with an initial 1,4-butanediol concentration of 20 g/L as single carbon source. The specific growth rate of A. latus decreased with v-butyrolactone concentration and optimum sucrose concentration for growth was 10 g/L. In batch cultures of A. latus, 4HB fraction increased with initial v-butyrolactone concentration. P(3HB-co-4HB) with 19 $mol\%$ 4HB was obtained when the initial ratio of v-butyloractone (g/L) to sucrose (g/L) was 10 : 10.

• 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.

• Journal of Microbiology and Biotechnology
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• 제9권6호
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• pp.722-725
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• 1999

Filamentation-suppressed recombinant Escherichia coli strain harboring the Alcaligenes latus polyhydroxyalkanoate (PHA) biosynthesis genes and the E. coli ftsZ gene was constructed and cultivated for the production of poly(3-hydroxybutyrate) [P(3HB)] with high concentration and high content. By the pH-stat fed-batch culture of this recombinant E. coli strain XL1-Blue(pJC5), the final cell concentration and P(3HB) concentration obtained in 44.25h were 172.2g cell dry weight/l and 141.9g P(3HB)/l, respectively, resulting in productivity of 3.21g P(3HB)/l-h. More importantly, the P(3HB) content obtained was 82.4 wt %, which was significantly higher than that obtained with the recombinant E. coli harboring only the PHA biosynthesis genes.

• Journal of Microbiology and Biotechnology
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• 제8권5호
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• pp.540-542
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• 1998

Five microorganisms capable of degrading an aromatic medium-chain-length polyhydroxyalkanoate ($PHA_{MCL}$), poly(3-hydroxy-5-phenylvalerate) (PHPV), were isolated from wastewater-treatment sludge. Among the isolates, JS02 showed degrading activity consistantly during several transfers. The isolate JS02 could hydrolyze another aromatic MCL copolyester, poly(3-hydroxy-5-phenoxyvalerate-co-3-hydroxy-7-phenoxyheptanoate), ［P(5POHV-co-7POHH)］, and other short-chain-length PHAs ($PHA_{SCL}) such as poly(3-hydroxybutyrate) ［P3(HB)］, poly(3-hydroxybutyrate-co-4-hydroxybutyrate) ［P(3 HB-co-4 HB)］, and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) ［P(3HB-co-3HV)］ with relatively low activity. The culture supernatant of JS02 showed hydrolyzing activity for the p-nitrophenyl esters of fatty acids.

• 폴리머
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• 제29권1호
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• pp.54-58
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• 2005

미생물 합성 poly[(R)3-hydroxybutyrate]](P(3HB))의 알칼리 및 효소 분해거동을 단결정과 Langmuir 단분자막을 모델시스템으로 하여 연구하였다. 단결정의 초기효소 및 알칼리 분해거동은 단결정의 장축에 대해 수직방향(b축)으로 분해가 일어났고 용융점 이하의 온도에서 열처리 또한 단결정의 b축을 따라 봉우리 형태의 형태학적 변화를 관찰하였다. 이러한 결과는 라멜라 단결정은 b축을 따라 불규칙한 영역을 가지고 있음을 의미하고 효소분해가 불규칙한 영역에서 선호적으로 일어난다고 설명할 수 있다. 한편, P(3HB), 단분자막의 효소 및 알칼리 분해경향은 분해매체와 표면압력에 크게 의존하였다. 알칼리 분해의 경우 낮은 표면압력에서도 분해를 나타내는 반면 효소 분해는 높은 표면 압력 하에서 분해거동을 나타내었다. 이러한 현상은 분자수준의 크기인 알칼리 분해매체는 P(3HB) 단분자막과 좁은 접촉면적(낮은 표면압력)에서도 활성을 보이는 반면 크기가 큰 분해효소는 보다 큰 활성 접촉면적(높은 표면압력)을 필요로 하는 것으로 판단된다.

• 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.

• 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.