• Journal of Microbiology and Biotechnology
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• 제6권2호
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• pp.147-149
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• 1996

Poly(3-hydroxybutyrate) (PHB) was synthesized and accumulated intracellularly to a high concentration (7 g/l) by cultivating recombinant Escherichia coli XL1-Blue (pSYLl05) in a complex medium containing 20 g/l glucose. The morphology of PHB granules was examined by transmission electron microscopy. The PHB granules synthesized in recombinant E. coli were much larger than reported values for wild type microorganisms, and were often irregularly shaped. Some cells were apparently extruding PHB into the medium, which suggests that PHB granules maintain some fluidity and cells become fragile due to PHB accumulation.

• 한국미생물·생명공학회지
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• 제23권5호
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• pp.588-592
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• 1995

Fed-batch fermentation was used to produce the high concentrations of poly-$\beta $-hydroxybutyrate (PHB) and poly-$\beta $-(hydroxybutyrate-co-hydroxyvalerate) (PHB/V). Specific growth rate ($\mu $), yield of cell from glucose (Y$_{x/s}$) were calculated from the two samples in 3 to 5 hours of interval and they were reflected on the determination of glucose feeding rate to maintain the glucose concentration at around 10 g/l in the culture broth. PHB was accumulated after the nitrogen became limited at 60 g/l of dry cell weight by changing ammonia water to 4N-NaOH solution. As results, the final dry cell weight (DCW) of 170 g/l, PHB of 115 g/l were obtained in 50 hours and the overall productivity was 2.4 g/l$\cdot $h. After PHB accumulation, cosubstrate of glucose and propionic acid (PA) was fed to accumulate PHB/V. But, PA feeding rate was decreased from 3 g/l$\cdot $h to 1 g/l$\cdot $h to prevent PA from accumulating to high level in the broth, which is very inhibitory to the cells. As results, DCW, PHB and PHV were 147.5 g/l, 90 g/l and 8 mole % of hydroxyvalerate, respectively.

• 환경생물
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• 제17권3호
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• pp.249-255
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• 1999

Poly(3-hydroxybutyrate)의 표면 형태가 생분해성에 미치는 영향을 조사하였다. 용융 상태의 PHB를 서냉 ($-0.5^{\circ}C$/min)시켰을 때 PHB의 결정화도는 액체질소 속으로 급냉한 후 $90^{\circ}C$에서 2시간 동안 숙성한 PHB의 결정화도보다 높게 나타났으나 전자의 PHB 시료는 미세 균열의 존재로 인하여 후자의 PHB 시료보다 더 빠르게 생분해되었다. 거친 표면을 가진 평판 위에서 hot press하여 표면을 거칠게 만든 PHB film의 생분해는 표면이 평활한 film보다 훨씬 더 빠르게 진행되었다.

• 한국염색가공학회지
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• 제30권2호
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• pp.130-140
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• 2018

In the bio-synthesis of poly(3-hydroxybutyrate)(PHB), which is a kind of poly(3-hydroxyalkanoate)(PHA), aimed to control the low molecular weight of PHB and obtain a telechelic PHB. As a result of incubation of R. eutropha at $30^{\circ}C$ with ethylene glycol added as a chain transfer agent, PHB content on the dry cell weight increased up to 24h, however, it decreased after that, and the molecular weight of PHB increased from 9h to 12h, and then, decreased up to 72h. The decrease of the content and the molecular weight of PHB indicates that PHB was decomposed as an energy source in bacterial cells and was incorporated into metabolic pathways. $^1H-NMR$ of the obtained PHB after incubation for 72h was measured to determine the terminal groups of the PHB during incubation. As the results of $^1H-NMR$ measurement, the peaks derived from ethylene glycol in both terminals of PHB were observed. Which indicate that the terminal reaction was caused by the addition of ethylene glycol, and that telechelic PHB having hydroxyl group at the both terminals where molecular weight was controlled was successfully synthesized.

• Fibers and Polymers
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• 제4권4호
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• pp.195-198
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• 2003

Epoxidized polybutadiene (EPB) was prepared by polybutadiene (PB) with m-chloroperbenzoic acid (MCPBA) in homogeneous solution. EPB was blended with poly(3-hydroxybutyrate) (PHB) up to 30 wt% by solution-precipitation procedure. The thermal decomposition of PHB/EPB blends was studied by thermogravimetric analysis (TGA), differential scanning calorimetry (DSC) and differential thermal analysis (DTA). The thermograms of PHB/EPB blends contained a two-step degradation process, while that of pure PHB sample exhibited only one-step degradation process. This degradation behavior of PHB/EPB blends, which have a higher thermal stability as measured by maximum decomposition temperature and residual weight, is probably due to crosslinking reactions of the epoxide groups in the EPB component with the carboxyl chain ends of PHB fragments during the degradation process, and the occurrence of such reactions can be assigned to the exothermic peaks in their DTA thermograms.

• 한국균학회지
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• 제23권4호통권75호
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• pp.348-353
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• 1995

Poly (3-hydroxybutyrate)(PHB) 의 Penicillium pinophilum에 의한 생분해성을 변형 Sturm Test 법으로 조사하였다. 활성오니를 사용한 경우보다 PHB의 생분해성이 비교적 재현성있게 측정되었으며 PHB의 생분해를 가장 빠르게 진행시키는 최적 포자 접균량은 1 %(v/v)이었다. 생분해 속도는 시료의 표면적에 따라 증가하였으나 비례적으로 상승하지 않아 분해가 시료의 표면 뿐만 아니라 내부에서도 진행됨을 나타내었다. 반응 배지내의 질소원 함량에 따라 PHB의 생분해 속도는 증가하다가 점근값을 보여 질소원이 depolymerase 효소의 합성에 필요한 원소임을 보였다.

• 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%.

• Journal of Microbiology
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• 제40권2호
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• pp.129-133
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• 2002

The fungus, Emericellopsis minima W2, capable of degrading poly(3-hydroxybutyrate) (PHB) was isolated from a waste water sample. Production of the PHB depolymerase from E. minima W2 (PhaZ/ sub Emi/) was significantly repressed in the presence of glucose. PhaZ/ sub Emi/ was purified by column chromatography on Octyl-Sepharose CL-4B and Sephadex G-100. The molecular mass of the PhaZ/ sub Emi/), which consisted of a single polypeptide chain, was estimated to be 48.0 kDa by SDS-PAGE and its pI vague was 4.4. The maximum activity of the PhaZ/ sub Emi/ was observed at pH 9.0 and 55$\^{C}$. It was significantly inactivated by 1mM dithiothreitol, 2mM diisopropyl fluorphosphate, 0.1mM Tween 80, and 0.1 mM Triton X-l00, but insensitive to phenylmethylsulfonyl fluoride and N-ethylmaleimide. The PhaZ/ sub Emi/ efficiently hydrolyzed PHB and its copolyester with 30 mol% 3-hydroxyvalerate, but did not act on poly(3-hydroxyoctanoate). It also hydrolyzed p-nitrophenylacetate and p-nitrophenylbutyrate but hardly affected the longer-chain forms. The main hydrolysis product of PHB was identified as a dimer of 3-hydroxybutyrate.

• 미생물학회지
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• 제37권1호
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• pp.92-99
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• 2001

Pi1ot plant (200 l)에서의 유가식 배양에 의한 Ralstonia eutopha KHB 8862의 고농도 배 양을 통하여 Poly(3-hydroxybutyrate) (PHB)의 대량생산을 모색하였다. 그 결과 배양 80시간 후 168 g/l의 건체량과 건체량의 74%에 달하는 PHB를 생산할 수 있었으며, 이때의 고과당 시럽으로부터의 PHB 전환수율 및 PHB 생산성은 각각 0.27 (w/w) 및 1.6 $gl^{-1}$ $h^{-1}$ /이었다. 이를 토대로 본 연구에서는 미생물 발효에 의한 PHB 생산 cost 및 그 경제성을 분석하였다. 신규설비투자를 고려하지 않은 경우의 PHB의 생산 cost는 US$2.41/kg으로 산출된 반면에 신규 설비투자를 고려한 경우에는 US$3.15/kg으로 상승되었다. 탄소원의 PHB로의 전환수율과 발효 생산성 모두 PHB 생산비를 결정하는 중요요인이지만 전체 생산비의 37%를 차지하는 탄소원 원료비의 비중이 설비투자의 감가상각비 비중 (17%)에 비해 높기 때문에 생산성을 높이는 노력보다는 전환수율을 개선하는 것이 PHB 생산비용 절감의 핵심이 되는 것으로 나타났다. PHB chip으로의 제조시 PHB 생산 cost는 US$4.0/kg의 수준으로 현재로서는 범용 합성플라스틱에 비하여 경쟁력을 확보하지 못한다. 따라서 생산비 절감을 통한 범용수지로써 경쟁력 제고와 함께 바이오의약 분야 등의 고부가가치 영역에서의 새로운 용도 개발 등이 적극 요구된다.

• 한국미생물·생명공학회지
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• 제26권3호
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• pp.206-212
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• 1998

생분해성 고분자인 poly(3-hydroxybutyrate, PHB)를 생산하는 Alcaligenes eutrophus의 배양액에 응집제를 첨가하여 회수한 균체로부터 용매추출법 에 의한 PHB 회수와 순도에 미치는 응집제의 영향을 조사하였다. 응집제는 Al계 3종류와 Fe계 2종류를 사용하였다. 균체로부터 PHB는 30%로 희석한 hypochlorite 용액과 chloloform 혼합용액을 이용하여 추출 회수하였다. Al계 응집제들의 경우, 응집제의 첨가량이 1,000 mg-Al/L 이하의 농도에서는 PHB 회수율이 응집제를 첨가하지 않은 경우의 회수율(82%)과 거의 비슷하였고, 1000 mg-Al/L 이상의 농도에서는 응집제 첨가량이 증가함에 따라 PHB 회수율이 감소하였다. PHB 추출에 사용한 hypochlorite 용액의 농도를 50%로 증가시키면, 응집제를 2000 mg-Al/L까지 높은 농도로 첨가하여도 90% 이상의 PHB 회수율을 얻을 수 있었다. 회수된 PHB의 순도는 응집제의 종류와 용량에 관계없이 98% 이상으로 매우 높았다. Fe계 응집제의 경우, 응집제의 용량은 PHB의 회수율에 거의 영향을 미치지 않았으나, 회수된 PHB중의 잔류 Fe의 농도가 높았고, 잔류 Fe에 의해 정제한 PHB가 엷은 적색으로 착색되었다. 세포의 회수와 PHB의 정제 공정에 적합한 응집제로는 응집효율, 잔류 Al과 Fe 및 정제 PHB의 색상 등을 고려할 때 Fe계 보다는 Al계 응집제가 적합하였다.