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재조합 단백질 과발현을 위한 Bacillus snbtilis 포자형성 변이주의 개발 및 특성 분석
오민규,박승환김병기 한국생물공학회 1994 KSBB Journal Vol.9 No.1
Bacillus subtilis를 재조합 이종 단백질 생산 균주로 만들기 위하여 포자형성 변이주를 만들었다. 균주는 두 개의 프로테아제가 제거된 균주인 DB104로부터 spoOJ와 spoIIG 변이주를 유전자 조작법에 의해 만들고 두 개의 유전자가 모두 제거된 균주도 만들었다. 이에 목적 aprE 유전자를 삽입 벡터 형태로 만들어 integration시킨 뒤 변이주 각각의 형태적인 변화를 투과성 전자현미경으로 살펴 보았다. 각각 변이주의 모습은 이전에 보고된 것과 거의 일치하였으며 spoOJ spoIIG 이중포자변이주의 경우는 spoIIG 변이주와 더욱 닮은 것을 알 수 있었으며, 훨씬 주름진 것과 같은 투박한 세포벽 및 막을 가지고 있음을 관찰하였다. spoOJ 변이는 포자형성 빈도를 낮추고 aprE 활성을 감소시키는 반면, spoIIG 변이는 포자형성을 거의 하지 않으면서 aprE 활성에 상승효과를 가져왔다. spoOJ와 spoIIG 이중포자변이주는 spoOJ 변이의 효과는 거의 나타나지 않은채, spoIIG와 비슷한 aprE 활성을 보였다. Sporulation mutants of Bacillus subtilis were developed for overproduction of heterologous proteins. The strains spoOJ spoIIG, and spoOJ spoIIG double mutant were constructed from two pretense-delfted mutant (DB104). The vector containing aprE gene was integrated in the chromosome of each strain, then the morphology of each strain was observed by TEM (trasmission electron microscopy). The morphology of spoOJ mutant and spoIIG mutant coincides with the description of the previous reports, respectively. The sporulating cells of spoOJ SpoIIG double mutation resemble spoIIG mutant more similarly, but with a little rougher cell wall membrane. The spoOJ mutation in B. subtilis gives negative effect on aprE activity with only a decreased sporulation frequency. On the contrary spoIIG mutation increases the aprE activity twice with an undetectable sporulation frequency. In the case of spoOJ and spolIG, i. e. double mutation, the effect of spoOJ on aprE activity seems to be relieved and the double mutant shows more or less the same aprE activity compared to spoIIG mutant.
Metabolic Engineering for Utilizing Various Carbon Sources
오민규 한국공업화학회 2020 한국공업화학회 연구논문 초록집 Vol.2020 No.-
The development of industrial strains that can efficiently utilize various carbon sources has been emphasized in metabolic engineering because cheaper carbon source should be utilized in biorefinery. In case of glucose, overflow metabolism is the main problem in because high amounts of byproducts are produced. This can be significantly reduced if the glucose uptake rate is regulated in various strains. To utilize lignocellulosic biomass, the strain must be able to co-utilize glucose and xylose. This can be resolved by minimizing carbon catabolic repression (CCR), which is regulated by phosphotransferase system (PTS). Disruption of PTS was not enough so that further engineering to reduce metabolic burden was applied. Fourth generation carbon source, C1 gas, can also be utilized for PHB production. Useful metabolic engineering strategy for utilizing C1 gas in industrial strain will also presented.
Production of bio-based chemicals from Klebsiella pneumonia
오민규,정무영 한국공업화학회 2015 한국공업화학회 연구논문 초록집 Vol.2015 No.0
The fermentative production of 2,3-butanediol has been intensively studied because of the potential of these products as platform chemicals, such as polymers, fuels, and plastics. Klebsiella pneumoniae is a good host for production of 2,3-butanediol from a wide range of carbon sources. We metabolically engineered K. pneumoniae (△wabG △ldhA △pflB) to reduce the production of byproducts. In particular, the inactivation of pflB significantly improved the yield of 2,3-butanediol from glucose to 92.2% of the theoretical maximum. We also engineered Enterobacter aerogenes for efficient utilization of sugarcane molasses in 2,3-butanediol production by elimination of carbon catabolite repression. Deletion of scrR and cra as well as overexpression of scrAB improved the strain’s performance of utilizing three sugars in sugarcane molasses and producing 2,3-butanediol.
공랭형 NH3/H2O 흡수식 냉동기의 모사 및 실험적 연구
오민규,김현준,김성수,강용태 대한설비공학회 2005 설비공학 논문집 Vol.17 No.11
The objective of this paper is to study the effects of the cooling air mass flow rate and the heat input variation by the simulation and the experiment. An air-cooled NH3/H2O absorption chiller is tested in the present study. The nominal cooling capacity of the single effect machine is 17.6kW(5.0USRT). The overall conductance(UA) of each component, the cooling capacity, coefficient of performance and each state point are measured with the variation of the cooling air mass flow rate and the heat input. It is found that the COP and cooling capacity increase and then decreases with increasing the heat input. It is also found that the COP and the cooling capacity increase and keep constant with increasing the cooling air mass flow rate. The maximum COP is estimated as 0.51 and the optimum cooling air mass flow rate is 217m3/min from the present experiment.
LCC 분석에 의한 상온 에너지수송용 흡수식 냉동기(STA)의 경제성 평가
오민규,이봉진,홍희기,강용태 대한설비공학회 2004 설비공학 논문집 Vol.16 No.6
The objective of this paper is to evaluate the economic performance of latent and sensible energy transportation systems. LCC (Life-Cycle Cost) analysis is a practical method and a guideline for evaluating the economic performance of considered systems during the Life-Cycle Period. By comparing the LCC of alternatives, The most ideal alternative is determined which has the lowest LCC. It is concluded that the cost of STA (Solution Transportation Absorption system) can be reduced by 67% to that of sensible energy transportation for study period with 10 km transportation distance.
DNA Microarray Analysis of Immediate Response to EGF Treatment in Rat Schwannoma Cells
오민규,Daniel R. Scoles,Stefan-M. Pulst 한국생물공학회 2005 Biotechnology and Bioprocess Engineering Vol.10 No.5
Epidermal growth factor (EGF) activates many intracellular effector molecules, which subsequently in-fluence the expression levels of many genes involved in cell growth, apoptosis and signal transduction, etc. In this study, the early response of gene expressions due to EGF treatment was monitored using oligonucleotide DNA microarrays in rat schwannoma cell lines. An immunoblotting experiment showed the successful activation of EGF receptors and an effector protein, STAT5, due to EGF treatment. The microarray study showed that 35 genes were significantly induced and 2 were repressed within 60 min after the treatment. The list of induced genes included early growth response 1, suppressor of cytokine signaling 3, c-fos, interferon regulatory factor 1 and early growth response 2, etc. According to the microarray data, six of these were induced by more than 10-fold, and showed at least two different induc-tion patterns, indicating complicated regulatory mechanisms in the EGF signal transduction.