• Journal of Microbiology and Biotechnology
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• 제3권2호
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• pp.115-122
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• 1993

Poly(3-hydroxybutyrate)(P(3HB)) accumulation under nutrient-rich condition with various amounts of $(NH_4)_2 SO_4$ was systematically investigated. The results of the electron-microscopy and the solvent extraction showed that the P(3HB) accumulation is unavoidable even under nutrient-rich condition. This indicates that in a two-step culture of Alcaligenes eutrophus H16, the researches should be careful in interpreting the data of polyhydroxyalkanoates(PHAs) accumulation in terms of the carbon-source fed in the second step because the two-step culture product contains the P(3HB) produced under nutrient-rich condition. The polyester production capability in a two-step batch culture of A. eutrophus H16(ATCC 17699) was also investigated using various saccharides and their derivatives such as glucose, fructose, gluconic acid, glucaric acid, sorbitol, lactose, galactose, and mannose. The polyesters synthesized were characterized by 500 MHz$^{1}H-NMR$ spectroscopy, intrinsic viscosity$[\eta]$ measurement in chloroform and differential scanning calorimetry(DSC). 500 MHz $^{1}H-NMR$ analysis showed that all polyesters synthesized generally contained 1~2 mol% of 3HV. Another finding is that the glucose utilization can be increased by changing the autoclaving procedure of the substrate to enhance the P(3HB) production yield up to 46 wt% of P(3HB) in dry cells.

• Journal of Microbiology and Biotechnology
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• 제3권2호
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• pp.123-128
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• 1993

The polyesters (polyhydroxyalkanoates; PHAs) production capability in a two-step cultivation of Alcaligenes eutrophus H16(ATCC 17699) was investigated by using various organic carbon sources. The carbon sources used included linear $C_2~C_10$ monocarboxylic acids, $C_3~C_10$ dicarboxylic acids, crotonic acid, and several linear vicinal and $\omega$-diols. The polyesters synthesized were characterized by 500 MHz $^1 H-NMR$ spectroscopy, intrinsic viscosity$[\eta]$ measurement in chloroform and differential scanning calorimetry (DSC). The PHAs synthesis data showed that the use of C-odd ($C_3, C_5, and C_7$) monocarboxylic acids resulted in poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(P(3HB-co-3HV) (3HV content ranging 40 to 70 mol%) while the use of $C_9$ substrate gave the copolyester containing only 4 mol% of 3HV. All culture products obtained on $C_3$~C$_{10}$ dicarboxylic acids gave exclusively P(3HB). 500 MHz $^1 H-NMR$ analysis showed that all polyesters synthesized generally contained 1~2 mol% 3HV even for the unrelated substrates such as the carboxylic acids with even number of carbon. When $\alpha, \omega$-diols with even number of carbon were used as substrates, 4-hydroxybutyrate(4HB) was inserted into the polyester chain composed of P(3HB-co-4HB). Vicinal diols were generally not utilized by the bacterium for polyester production.n.

• 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

• 한국미생물생명공학회:학술대회논문집
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• 한국미생물생명공학회 2000년도 Proceedings of 2000 KSAM International Symposium and Spring Meeting
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• pp.100-107
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• 2000

In a recently developed strategy for in-situ treatment of polychlorinated biphenyls (PCB), bioaugmentation was used in conjunction with a surfactant, sorbitan trioleate, as a carbon source for the degrader bacteria, along with the monoterpene, carvone, and salicylic acid as inducing substrates. Two bacteria were used for soil inoculants, including Arthrobacter sp. st. B1B and Ralstonia eutrophus H850. This methodology achieved 60% degradation of PCBs in Aroclor 1242 after 18 weeks in soils receiving 34 repeated applications of the degrader bacteria. However, an obvious limitation was the requirement for soil mixing after every soil inoculation. In the research reported here, bioaugmentation and biostimulation treatment strategies were modified by using the earthworm, Pheretima hawayana, as a vector for dispersal and mixing of surface-applied PCB-degrading bacteria and soil chemical amendments. Changes in microbial biomass and microbial community structure due to earthworm effects were examined using DNA extraction and PCR-DGGE of 16S rDNA. Results showed that earthworms effectively promoted biodegradation of PCBs in bioaugmented soils to the same extent previously achieved using physical soil mixing, and had a lesser, but significant effect in promoting PCB biodegradation in biostimulated soils treated with carvone and salicylic acid. The effects of earthworms were speculated to involve many interacting factors including increased bacterial transport to lower soil depths, improved soil aeration, and enhanced microbial activity and diversity.