• 한국생물공학회:학술대회논문집
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• 한국생물공학회 2000년도 추계학술발표대회 및 bio-venture fair
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• pp.356-359
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• 2000

Xylitol의 생산성을 높이기 위해 two-stage fed-batch를 수행하였다. Glucose가 고갈되어 pH가 5.7에서 올라가면 glucose를 공급하는 방법에서 최종세포의 OD 185.0과 최종 ethanol 농도 1.0 g/L를 얻었다. 산소전달에 대한 xylitol의 생성 영향에서는 통기량 1 vvm에서 500 rpm의 교반속도일 때 xylitol 수율 55.2%와 생산성 $2.19\;g-xylitol/L\;{\cdot}\;h$를 얻었다.

• Journal of Microbiology and Biotechnology
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• 제19권5호
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• pp.474-481
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• 2009

A novel microorganism, designated as LG12, was isolated from soil based on its ability to use acrylic acid as the sole carbon source. An electron microscopic analysis of its morphological characteristics and phylogenetic classification by 16S rRNA homology showed that the LG12 strain belongs to Rhodococcus erythropolis. R. erythropolis LG12 was able to metabolize a high concentration of acrylic acid (up to 40 g/l). In addition, R. erythropolis LG12 exhibited the highest acrylic acid-degrading activity among the tested microorganisms, including R. rhodochrous, R. equi, R. rubber, Candida rugosa, and Bacillus cereus. The effect of the culture conditions of R. erythropo/is LG12 on the production of 3-hydroxypropionic acid (3HP) from acrylic acid was also examined. To enhance the production of 3HP, acrylic acid-assimilating activity was induced by adding 1 mM acrylic acid to the culture medium when the cell density reached an $OD_{600}$ of 5. Further cultivation of R. erythropo/is LG 12 with 40 g/l of acrylic acid resulted in the production of 17.5 g/l of 3HP with a molar conversion yield of 44% and productivity of 0.22 g/l/h at $30^{\circ}C$ after 72 h.

• Food Science and Biotechnology
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• 제18권4호
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• pp.954-958
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• 2009

Effects of controlled- and uncontrolled-pH operations on phenylalanine ammonia lyase (PAL) production by a recombinant Escherichia coli strain were investigated at uncontrolled-pH ($pH_{UC}$) and controlled-pH ($pH_C$) of 5.5, 6.0, 6.5, 7.0, 7.5, 8.0, and 8.5 in bioreactor systems. The results showed that the recombinant PAL activity was improved significantly by controlled pH strategy. Among the $pH_C$ operations, the highest PAL activities were obtained under $pH_C$ 7.5 strategy where cell mass ($OD_{600\;nm}$) and PAL activity was 1.3 and 1.8 fold higher than those of $pH_{UC}$, respectively. The maximum PAL activity reached 123 U/g. The $pH_C$ 7.5 strategy made recombinant plasmid more stable and therefore allowed easier expression of PAL recombinant plasmid, which increased PAL production. It was indicated that the new approach (controlled-pH strategy) obtained in this work possessed a high potential for the industrial production of PAL, especially in the biosynthesis of L-phenylalanine.

• 생명과학회지
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• 제24권10호
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• pp.1046-1054
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• 2014

${\beta}$-1,3-glucanase는 다양한 바이오공정에 널리 사용되어지는 효소로서 산업적 이용가치 증대를 위해 ${\beta}$-1,3-glucanase의 대량생산이 요구되어 지고 있다. 본 연구에서는 반복서열 ${\delta}$-sequence에 의한 integration를 통해 Aspergillus oryzae유래의 ${\beta}$-1,3-glucanase (EXGA)의 과발현 유도를 연구하였다. 먼저 효모내의 여러 염색체상에 EXGA 유전자를 integration하기 위해 $pRS{\delta}K$-exgA와 $pRS{\delta}K$-exgA 플라스미드를 구축하였다. 이 플라스미드는 유전자의 구성적 발현을 위한 ADH1 프로모터, 분비생산을 위한 signal sequence와 ${\beta}$-1,3-glucanase유전자의 integration을 위한 ${\delta}$-sequence를 포함하고 있다. 먼저 $pRS{\delta}K$-exgA 플라스미드를 $BY4742{\Delta}exg1$ 균주에 형질전환하고, 재조합 ${\beta}$-1.3-glucanase가 안정하게 과발현 및 분비생산됨을 확인하였다. 다음으로 geneticin 선별을 통한 integration 유도와 ${\beta}$-1,3-glucanase 활성과의 관계를 조사해보기 위해 $pRS{\delta}K$-exgA 플라스미드를 $BY4742{\Delta}exg1$ (YKY082) 균주에 도입한 결과, geneticin 농도 증가에 따라 ${\beta}$-1,3-glucanase 활성도 증가되었고, geneticin 농도 0.8 mg/ml가 ${\beta}$-1,3-glucanase과발현에 적합한 농도임을 확인 할 수 있었다. 이어서 $pRS{\delta}K$-exgA 플라스미드는 연속적 ${\delta}$-integration에 의해 효모세포 내에 도입되어, 한번, 두번, 세번 그리고 네번의 integration에 의해 ${\beta}$-1,3-glucanase의 비활성은 0.063, 0.095, 0.131 그리고 0.165 unit/ml/$OD_{600}$로 증가되었다. 또한 연속적 integration에 의해 ${\beta}$-1,3-glucanase의 활성이 증가됨에 따라 다양한 염색체에 도입된 EXGA 유전자의 복제수(integration 빈도)도 같이 증가되었음을 확인하였다. 따라서 본 연구 결과는 반복적 ${\delta}$-sequence integration방법을 통해 재조합 ${\beta}$-1,3-glucanase의 활성을 점진적으로 안정하게 증가시킬 수 있음을 제시하였다.

• Journal of Microbiology and Biotechnology
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• 제19권9호
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• pp.982-986
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• 2009

For the production of ghost bacteria vaccine to prevent the streptococcal disease in aquaculture fish species, a double cassettes vector was constructed and cloned in Escherichia coli DH5${\alpha}$. Ghost bacteria vaccine production from Escherichia coli DH5${\alpha}$/pHCE-InaN-GAPDH-Ghost 37 SDM (SIG) was maximized at a glucose concentration of 1 g/l, agitation of 300 rpm, and aeration of 1 vvm. The maximal efficiency of ghost bacteria formation was obtained at the mid-exponential phase ($OD_{600}=2.0$) with the concentration of 0.77 g/l for SIG. The molecular mass of GAPDH was detected at 67 kDa with the insoluble fraction, by SDS-PAGE and Western blot. The protective efficacy of ghost bacteria vaccine was evaluated by challenge test using olive flounder. The cumulative mortalities of the positive control, formalin-killed cell (FKC) vaccine, and SIG vaccine immunized groups were 91%, 74%, and 57%, respectively. These results suggest that SIG vaccine showed efficacy as a vaccine and had a higher potential to induce protective antibodies than did FKC vaccine.

• 한국미생물·생명공학회지
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• 제27권4호
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• pp.327-332
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• 1999

A highly productive fed-batch fermentation process was developed for the production of L-ornithine by using a new stabilized strain, Breviabcterium ketoglutamicum BK52. Fed-batch cultures with a continuous feeding of the complex medium were conducted on various operating conditions. The optimal concentration of phosphate in the complex medium was 2.1g/L. The optimal feeding rate of L-arginine was 0.028g/L/hr. The optimal feeding point of the complex medium was determined to be at 40 OD of the cell mass. The final L-ornithine concentrations within 64hrs of cultivation in 5 and 50 liter fermenters were 73g/L and 71g/L, respectively. The maximum overall L-ornithine productivity was 1.14g/L/hr which was about 2 times higher than that of the conventional fed-batch culture with intermittent feeding. The overall productivity of the fermentation system is remarkably improved by employing the optimized conditions, and it offers a significant potential for industrial application.

• KSBB Journal
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• 제26권3호
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• pp.223-228
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• 2011

The effects of pH, glycerol concentration and salt on cell growth and ethanol production using Enterobacter aerogenes KCTC 2190 were evaluated in the anaerobic culture condition. In condition of initial pH 5, cell growth and ethanol production were highest. An initial concentration of 10 g/L of pure glycerol gave the highest cell growth and ethanol production. However, in case of over 15 g/L of pure glycerol, they decreased. The cell growth and ethanol production decreased with the increase of salt concentration. When 10 g/L of crude glycerol was used as the carbon source, the cell growth and ethanol production were $1.32\;OD_{600}$ and 3.95 g/L, respectively, which were about 94.4% and 88.5% compared to those of pure glycerol. These result indicates that the crude glycerol produced in the biodiesel manufacturing process maybe useful as a potential carbon source for ethanol production form Enterobacter aerogenes KCTC 2190.

• 한국수산과학회지
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• 제50권2호
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• pp.169-174
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• 2017

Immunoglobulin M (IgM) was purified from olive flounder Paralichthys olivaceus sera using mannan-binding protein (MBP) and protein L affinity columns (designated as MBPIgM and ProLIgM, respectively). A monoclonal antibody (MAb) against olive flounder IgM was produced. The MBPIgM and ProLIgM had apparent molecular weights of 77, 73, and 28 kDa in SDS-PAGE. Nine hybridomas secreting MAbs against olive flounder IgM were established: five MAbs for MBPIgM (1, 2, 3, 4, and 5) and four for ProLIgM (6, 7, 8, and 9). Western blotting indicated that seven MAbs recognized heavy (H; MAbs 1, 2, 3, 4, 5, 6, and 7) chains and one recognized light (L; MAb 9) chains of IgM, while MAb 8 did not recognize IgM. The results of enzyme-linked immunosorbent assay (ELISA) with bovine serum albumin (BSA, antigen) and the nine MAbs revealed that the optical density (OD) values of sera differed significantly between BSA- and non-immunized fish, despite some sera from non-immunized fish with slight high OD values. These results suggest that the MAbs produced in this study reacted specifically with the IgM from olive flounder.

• 한국어병학회지
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• 제37권1호
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• pp.71-78
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• 2024

Streptococcosis, caused by Streptococcus iniae and Streptococcus parauberis is an important bacterial disease that affects in olive flounder in Korea. In Korea, multivalent bacterial vaccines are used to prevent streptococcal diseases in aquaculture. In this study, commercial vaccines containing formalin-inactivated bacterial cells of S. iniae and S. parauberis were administered at six fish farms and one unvaccinated fish farm were designated for investigation (Wando; 4 sites and Jeju; 3 sites). Blood was collected from vaccinated and unvaccinated olive flounders, and titers of antibodies against S. iniae and S. parauberis in serum were analyzed using ELISA. After a one shot vaccination in the farms at Jeju (farm A) and Wando (farm D), the proportion of individuals with specific antibodies against S. parauberis OD values of 0.4 or higher was 60% and 53.5%, respectively. But after booster vaccination, the proportion of individuals with serum OD values of 0.4 or higher was higher substantially increased to 96.6% (farm A) and 100% (farm D). The levels of S. parauberis specific antibodies of olive flounder were increased after vaccination in three fish farms (farm D, E, and F), but not S. iniae specific antibodies.

• 한국미생물·생명공학회지
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• 제44권4호
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• pp.497-503
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• 2016

전처리 폐글리세롤을 기질로 사용한 재조합 대장균 배양에서 트레할로스 생산성에 영향을 미치는 핵심 변수들을 도출하고 반응표면 분석법을 사용하여 트레할로스 생산을 극대화하기 위한 최적조건을 탐색하였다. $37^{\circ}C$에서 IPTG로 induction 하는 배양이 $27^{\circ}C$ 또는 induction 하지 않은 배양에 비하여 세포생장 및 트레할로스 생산성이 높았다. Box-Behnken design 실험계획법을 사용하여 배지중의 NaCl, 발리다마이신 및 IPTG의 농도를 최적화하였다. 통계분석결과 IPTG와 NaCl의 농도는 트레할로스 생산에 영향을 미쳤으나 발리다마신의 영향은 크지 않은 것으로 확인되었다. 등고선도 분석을 통해 298 mM NaCl이 첨가되고 0.1 mM의 IPTG로 induction되는 배양에서 가장 많은 양의 트레할로스가 생산되는 것으로 예측되었다. 최적화된 조건에서 생산 균주는 세포 밀도($OD_{600}$) $5.4{\pm}0.2$에서 $304{\pm}15mg/l$의 트레할로스를 생산하였다.