• 한국미생물·생명공학회지
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• 제15권6호
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• pp.375-380
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• 1987

Alcaligenes eutrophus A52의 무세포 추출액으로부터 유안염석 및 DEAE-cellulose 이온교환 크로마토그래피로서 D-$\alpha$-Amino-$\varepsilon$-Caprolactam(DAC) racemase 와 L-$\alpha$-Amino-$\varepsilon$-Caprolactam(LAC) hydrolase를 분획하였다. A. eutrophus A52에 의한 DAC로부터 L-lysine으로의 전환은 DAC가 racemase에 의해 LAC로 전환된 후 hydrolase에 의해 L-lysine으로 가수분해됨을 확인하였다. DEAE-cellulose 이온교환 크로마토그래피에 의해 분리된 racemase의 분획은 최적온도가 55$^{\circ}C$, 최적 pH는 8.0이었으며 hydrolase의 분획은 최적온도가 $65^{\circ}C$ 최적 pH가 9.0이었다. 이 두 효소를 모두 함유하는 무세포 추출 액에 의한 DAC로부터 L-lysine으로의 전환은 6$0^{\circ}C$와 pH8.5에서 최대를 나타내었고 2% 이상의 DAC와 L-lysine에 의해 상당한 저해를 받았으나 0.5% DAC를 3.1mg의 단백질에 상당하는 무세포 추출액으로서 전환시킨 결과 55$^{\circ}C$에서 10시간 동안에 약 98%가 L-lysine으로 전환되었다.

• 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.

• 한국미생물·생명공학회지
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• 제19권2호
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• pp.186-192
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• 1991

It is very important to have a good kinetic model which considers the effects of both ammonium and glucose for the control and optimization of the poly-${\beta}$-hydroxybutyrate (PHB) fermentation. A kinetic model for the growth of Alcaligenes eutrophus and the biosynthesis of PHB under both ammonium and glucose limitation was proposed. Growth rate of residual biomass was expressed as a function of concentrations of residual biomass, glucose and ammonium having glucose inhibition. PHB production rate was expressed as a function of concentrations of residual biomass, glucose, ammonium and PHB content having ammonium and product inhibitions. Novel approaches were made to estimate the parameters in the model equations which considered two limiting substrates. Model parameters were evaluated by graphical and simplex methods. The proposed kinetic model fitted the data very well.

• 한국미생물·생명공학회지
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• 제21권3호
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• pp.221-228
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• 1993

In order to achieve poly-beta-hydroxybutyric acid (PHB) production using recombinant DNA in various host bacterial cells, the isolation of genes for PHB biosynthesis was attempted. As a result, a 5.2kb DNA fragment containing phbCAB operon of Alcaligenes eutrophus was isolated by colony hybridization using synthetic oligodeoxyribonucleotides as probes. The constructed recmbinant plasmid pSK(+)-phbCAB operon was transferred to Escherichia coli, and the obtained transformant accumulated considerable amount of PHB.

• KSBB Journal
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• 제13권3호
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• pp.272-278
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• 1998

Cell recovery from high cell density broths of Alcaligenes eutrophus by pretreatment with aluminum-based coagulants such as aluminum sulfate, polyaluminum hydrooxide chloride silicate (PACS), and polyaluminum hydrooxide chloride (Hi-PAX) was carried out. Cells coagulated with coagulants could be successfully recovered above 95-99% by centrifugation or filtration. The optimum initial pH of fermentation broths for cell recovery was in the range of 10 to 12. Optimum coagulants dosage for cell recovery increased with increasing of cell concentrations (21-160 g/L). The optimum coagulant dosages to recover cells with more than 95% cell recovery by centrifugation for the cell concentrations ranged 21-160 g/L were as follows: aluminum sulfate, 416-1708 mg Al/L; PACS, 211-826 mg Al/L; Hi-PAX, 320-960 mg Al/L. At optimum conditions for the coagulation of cells, centrifugal forces for 95% of cell recovery were dependent on the cell concentration. The centrifugal forces at 82 g/L and 160 g/L of cell concentration were only 45${\times}$g and 1600${\times}$g, respectively.

• 한국미생물·생명공학회지
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• 제15권6호
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• pp.369-374
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• 1987

하상 오니 및 토양으로부터 D-$\alpha$-Amino-$\varepsilon$-Caprolactam(DAC)의 자화능이 우수한 균을 분리하여 Alcaligenes eutrophus로 동정하였다. Alcaligenes eutrophus A52의 생육을 위한 배양조건은 배양 초기 최적 pH가 6.0, 최적온도는 3$0^{\circ}C$, 최적 DAC 농도는 0.2%로서 50시간 배양으로 0.2% DAC를 거의 다 자화하여 정지기에 도달하였다. 본 균은 탄소원으로 glucose와 fructose, 질소원으로는 NH$_4$Cl, NH$_4$NO$_3$, (NH$_4$)$_2$SO$_4$ 및 NaNO$_3$를 잘 자화하였다. 탄소원 및 질소원으로 Iysine, glutamate를 잘 이용하였으나 그 밖의 아미노산 자화능은 낮았다. 균체로부터 추출한 3.1mg의 단백질에 상당하는 무세포 추출 액으로 0.5% DAC를 55%에서 12시간 반응시킨 생성물은 약 98% L-Iysine으로 확인되었다.

• 분석과학
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• 제10권1호
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• pp.75-81
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• 1997

Poly-${\beta}$-hydroxybutyrates(PEB)는 토양에서 완전히 이산화탄소와 물로 분해되는 세포 내에 생성되는 고분자 물질로 잘 알려져 있다. Alcaligenes eutrophus의 세포 성장과 PHB의 생산성을 증가시키기 위해서 가압배양법이 사용되었다. 실험 데이타를 다양한 가압과 온도하에서 탄소 공급원과 기체성분의 영향에 관해서 분석한 결과 0.0075vvm의 기체수소를 공급한 배양이 수소를 공급하지 않은 배양에 비해 더 나은 PEB 생산성을 나타내었고, 6atm, $30^{\circ}C$에서 산소의 성분비를 2%에서 8%로 변화시켰을 때 더 높은 수율과 PEB의 생산성을 얻을 수 있었다.

• Journal of Microbiology and Biotechnology
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• 제6권5호
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• pp.309-314
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• 1996

Alcaligenes sp. SH-69 can synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from a single carbon source such as glucose. To clone the phaC gene from Alcaligenes sp. SH-69, a polymerase chain reaction was performed using the oligomers synthesized based on the conserved regions of the phaC genes from other bacteria. A PCR product (550 bp) was partially sequenced and the deduced amino acid sequence was found to be homologous to that of the phaC gene from Alcaligenes eutrophus. Using the PCR fragment Southern blotting of Alcaligenes sp. SH-69 genomic DNA digested with several restriction enzymes was carried out. To prepare a partial genomic library, about 5-Kb genomic DNA fragments digested with EcoRI, which showed a positive signal in the Southern blotting, were eluted from an agarose gel, ligated with pUC19 cleaved with EcoRI, and transformed into Escherichia coli. The partial library was screened using the PCR fragment as a probe and a plasmid, named pPHA11, showing a strong hybridization signal was selected. Restriction mapping of the insert DNA in pPHA11 was performed. Cotransformation into E. coli of the plasmid pPHA11 and the plasmid pPHA21 which has phaA and phaB from A. eutrophus resulted in turbid E. coli colonies which are indicative of PHA accumulation. This result tells us that the Alcaligenes sp. SH-69 phaC gene in the pPHA11 is functionally active in E. coli and can synthesize PHA in the presence of the A. eutrophus phaA and phaB genes.

• 한국식품과학회지
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• 제18권3호
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• pp.210-214
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• 1986

수소세균을 단세포단백질자원으로 이용하기 위한 기초연구로서 수소를 에너지원으로 이산화탄소를 탄소원으로 하는 autotrophic condition에서 수소세균 Alcaligenes eutrophus ATCC 17697의 배양조건 및 균체성분을 조사하였다. 균체성장을 위한 최적 pH와 온도는 각각 7.0과 $30^{\circ}C$ 이었으며 0.059atm부터 0.27atm까지의 산소분압범위에서 균체성장에 대한 산소분압의 영향을 본 결과 산소분압 0.11atm$(H_2:O_2:CO_2=7\;:\;1\;:\;1)$에서 최대비증식속도 ${\mu}max\;=0.31hr^{-1}$를 보였다. 건조균체의 조단백, 핵산, 회분함량은 각각 69.2%, 17.6%, 3.62%이었다.

• Journal of Microbiology and Biotechnology
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• 제5권5호
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• pp.292-296
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• 1995

Production of poly($\beta$-hydroxybutyrate-co-$\beta$-hydroxyvalerate)[poly(HB-co-HV) from glucose and propionic acid was studied in a two-stage fed-batch fermentation using Alcaligenes eutrophus NCIMB 11599. When either glucose became sufficient or the feeding rate of propionic acid decreased, production of poly(HB-co-HV) increased but concomitantly resulted in a reduced fraction of HV. During the copolymer accumulation stage, the specific production rate of hydroxyvalerate (HV) increased up to 0.013 (g-HV/g-RCM/h) but it decreased as propionic acid was accumulated. Control of the propionic acid concentration in the medium, therefore, is considered to be one of the most important operating parameters for production of poly(HB-co-HV) with a higher HV fraction. A high titre of poly(HB-co-HV) (85.6 g/I) with HV fraction of 11.4 mol$%$ could be obtained in 50 h by controlling the propionic acid concentration at 1 to 4 g/I.