• 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.

• 한국환경과학회지
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• 제6권4호
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• pp.379-384
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• 1997

The biodegradable characteristics of poly-$\beta$-hydroxybutyrate(PHB) film by fun맥 and soil burial are Investigated. As the results of the American Standards for Testing and Materials(ASTM) method, the you of Aspergillus niger was apparent on the PHB containing plate. This suggests that PHB was utilized as the sole carbon source by Aspergillus niger and ASTM method may have applications as measuring means of biome gradability of polyhydroxyalkanoic acid(PHA). PHB film was studied by monitoring the time-dependant changes in weight loss of PHB film under 30% and relative humidity 80 % during pot-test. As the results of pot-test, PHB film was decomposed about 87 % in 30 days by soul microorganisms. PHB film was more slowly degraded than PHB/HV film.

• Journal of Microbiology and Biotechnology
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• 제6권6호
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• pp.425-431
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• 1996

The enzymatic characteristics of Alcaligenes latus were investigated by measuring the variations of various enzyme activities related to biosynthesis and degradation of poly-${\beta}$-hydroxybutyrate (PHB) during cultivation. All PHB biosynthetic enzymes, ${\beta}$-ketothiolase, acetoacetyl-CoA reductase, and PHB synthase, were activated gradually at the PHB accumulation stage, and the PHB synthase showed the highest value among three enzymes. This indicates that the rate of PHB biosynthesis is mainly controlled by either ${\beta}$-ketothiolase or acetoacetyl-CoA reductase rather than PHB synthase. The enzymatic activities related to the degradation of PHB were also measured, and the degradation of PHB was controlled by the activity of PHB depolymerase. The effect of supplements of metabolic regulators, citrate and tyrosine, was also investigated, and the activity of glucose-6-phosphate dehydrogenase was increased by metabolic regulators, especially by tyrosine. The activities of ${\beta}$-ketothiolase and acetoacetyl-CoA reductase were also activated by citrate and tyrosine, while the activity of PHB depolymerase was depressed. The increased rate and yield of PHB biosynthesis by metabolic regulators may be due to the increment of acetyl-CoA concentration either by the repression of the TCA cycle by citrate through product inhibition or by the activation of sucrose metabolism by the supplemented tyrosine.

• 자연과학논문집
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• 제9권1호
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• pp.53-56
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• 1997

Cyanobacteria Chlorogloea fritschii를 여러 생장조건(인산 충분 혹은 결핍)에서 배양한 후 poly-$\beta$-hydroxybutyrate(PHB)를 추출하여 그의 구조를 적외선 분광법에 의해 분석하였다. 이들은 공통적으로 C=O의 신축 진동에 의한 1700-1800 $cm^-1$영역에서 대단히 강한 흡수 피이크를 나타내고 또한 2900$cm^-1$에서 C-H의 비대칭 및 대칭 신축 진동흡수 피이크를 나타냄으로써 PHB의 특징을 잘 나타내고 있었다. 그러나 생장조건에 따라 C-H의 신축 진동흡수 피이크 세기에 변화를 관측할 수 있었으며 인산 결핍 생장조건에서 C-H의 신축 진동흡수 피이크의 세기는 나머지 피이크에 비해 증가된 양상을 나타냄으로 인산의 공급여하에 따른 PHB구조의 변화를 시사하였다.

• 한국미생물·생명공학회지
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• 제20권5호
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• pp.597-606
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• 1992

Alcaligenes eutrophus 균주로 poly-Beta-hydroxybutyrate(PHB) 생산을 위한 연속배양 공정의 성능에 미치는 희석비율, 주입 포도당 및 염화암모늄농도의 영향에 대하여 연구하였다. 주입 기질농도가 일정할때( 주입 포도당농도=20g/l, 주입 염화암모늄농도=2g/l), 생체성장속도와 PHB생성속도는 희석비율이 각각 0.1, $0.06h^{-1}$에서 최고 값을 나타냈고, $0.13h^{-1}$에서 세포가 전부 배출되었다. 희석비율이 증가함에 따라 비PHB 생성속도는 계속 증가하였지만 PHB 축적비는 50%에서 25%로 감소하였다. 세포농도는 주입 염화암모늄농도가 2g/l일 때 최고값을 나타내었고, 그 이상의 농도에서는 감소하였다. 이 실험결과로 암모늄에 의한 기질저해가 있음을 알 수 있었다. 주입 포도당농도가 30g/l에서 세포농도는 최고값을 나타냈지만 PHB 농도는 계속 증가하였따. 모델속도식에 대한 매개변수는 도식적 방법과 매개변수 추정으로 구하였고 희석비율, 주입 포도당농도, 주입 염화암모늄농도의영향에 대하여 모사한 결과 실험데이타와 잘 일치하였다.

• KSBB Journal
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• 제10권2호
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• pp.176-182
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• 1995

Methylobacterium organophilum을 이용하여 다당류(메틸란)와 poly-${\beta}$-hydroxybutyrate(PHB)의 생산에 영향을 주는 환경 및 생리적 인자들을 조사하였다. PHB 축적을 위하여는 $38^{\circ}C$, 다당류의 생산을 위하여는 $30^{\circ}C$가 최적이었다. pH의 경우는 P PHB의 축적은 pH 7 ~ 8, 다당류의 생산은 pH 6~7 이 최적이였다. 질소원 제한 상태하에서의 $Mo^{2+}, Mg^{2+} 또는 Mn^{2+}$ 등의 결핍조건에셔는 질소원만의 제한조건보다 P PHB 축적은 증가하였으나, 다당류 생산은 감소하였 다. 질소원의 제한없이 $K^+$만의 결핍조건에서는 균 체증식도 억제되었고 다당류도 생성되지 않았지만 건조균체당 PHB 함량은 60%까지 증가하였다. PHB와 다당류의 동시 생산을 위한 C/N 비율의 효 는 C/N 비율이 높을수록 질소원의 제한효과 때문에 균체증식은 감소되었지만 건조균체량당 PHB 함량과 다당류의 생성은 현저히 증가하였다.

• 한국미생물·생명공학회지
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• 제18권3호
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• pp.273-279
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• 1990

PHB 생산을 위하여 메탄올을 기질로 한 선별배지에서 토양, 하천수, 퇴비 등으로부터 분홍색 색소를 가지는 PHB 축적 facultative methylotroph를 분리하여, 균주의 특성을 검토하였다. 분리균주의 최적 생육조건과 PHB 축적을 위한 배양조건을 조사한 결과 균체의 생육은 메탄올 농도 1.0(v/v), 질소원인$ NH_4C$ 농도 1.0g/l, 즉 C/N ratio 13.2 일때 그리고 pH 7.0과'$30^{\circ}C$에서 가장 좋았으며, PHB는 C/N ratio가 50.8, 즉 메탄올 농도 1.0(v/v )$NH_4CL$ 0.26g/l 일때, 그리고 pH 6.0일 때 건조중량의 약 40까지 축적되었다. 고농도 메탄올에 의한 생육저해를 극복하기 위하여 기질을 간헐적으로 계속 공급해주는 fed-batch 배양을 시도한 결과 균체량은 14g/l, PHB 축척량은 5.5g/l까지 증가시킬 수 있었다. 생산된 PHB를 분리.정제하여 IR과 $^I H-NMR$로 구조를 분석한 결과 3-hydroxybutyric acid 의 homopolymer임이 확인되었다. 또한 균주의 pink-pigment를 추출하여 absorption spectrum를 조사하여 그 특성을 규명하였다.

• 한국미생물·생명공학회지
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• 제22권4호
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• pp.401-406
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• 1994

The medium compositions of Alcaligenes eutrophus were optimized for increasing PHB productivity. It is very important to optimize the concentrations of inorganic salts and trace eleme- nts as well as carbon and nitrogen sources to maximize cell growth rate and productivity. The fed-batch culture of Alcaligenes eutrophus by dual feeding of ammonia water and glucose under optimized initial medium concentrations was carried out. Glucose was fed manually according to glucose consumption rate and ammonia water by pH-stat. The final cell concentrations and PHB content in 30 hours were 122 g/l and 65% of dry cell weight(yielding 79 g of PHB/l), respectively and 2.64 g/l/hr of PHB production rate was obtained.

• 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.

• KSBB Journal
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• 제6권1호
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• pp.35-43
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• 1991

The production of poly-$\beta$-hydroxybutyrate(PHB) from methanol by batch and fed-batch cultivations of Methylobacterium sp. GL-10 was studied. PHB accumulation was stimulated by the nutrients deficiency including, NH4+, SO42-, and K+. The nitrogen deficiency was the most critical factor for PHB accumulation. In batch cultivation, the maximum cell concentration and PHB content were 1.86g/l and 0.62g/l, respectively, with 1.0%(v/v) of methanol and 0.5g/1 of ammonium sulfate. The mass doubling time of Methylobacterum sp. GL-10 was in the range of 4-5 hrs. The cell growth and PHB accumulation were severely inhibited at the methanol concentration over than 2% (v/v). To overcome methanol Inhibition, constant feeding and intermittent feedillg fed-batch cultivations were adopted, using C/N molar ratio as a control factor. In constant feeding fed-batch process, cell concentration was increased up to 2.67g/1, and PHB yield was enhanced from 0.33 of batch culture to 0.53. The relatively low cell concentration was caused by methanol accumulated in culture broth at late growth phase. To prevent methanol accumulation and to maximize PHB production, DO-state intermittent fed-batch cultivation was attempted. The cell and PHB concentration was reached up to 4.55g/1 and 1.80g/1, respectively. It was possible to maintain methanol concentration low and also to feed nutrient of desired C/N molar ratio.