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Methylosinus trichosporium OB3b에서 세포내 메탄자화효소의 활성 측정
송효학,김경태,박성훈,강정미 釜山大學校生産技術硏究所 1996 生産技術硏究所論文集 Vol.50 No.-
메탄자화균을 이용한 지방족 염소화합물의 분해를 위해 메탄자화균의 일종인 Metlylosirus trichosporium OB3b를 배양하고 메탄자화효소의 활성을 5ml vial을 이용하여 여러조건에서 측정하였다. 기절로 propene을 사용하고 생성물인 propene oxide를 가스크로마토그래프(GC)로 분석하였다. 환원력 공급제로 formate가 첨가될 경우 그렇지 않은 경우에 비해 MMO의 활성이 4-5배 가량 높았고, 최적 pH와 온도는 각각 pH 7.0-7.5, 30℃였다. 기질에 대한 포화상수 Km의 값은 ?????mol/L였고 고농도 propene oxide는 활성을 저해하였다. 균체량이 너무 적으면 GC에서의 분석감도에 문제가 있었고 너무 많으면 산소전달이 제한되므로 MMO 활성 측정값이 저하되었다. 분석에 적합한 최적 균체량은 MMO 활성에 따라 달라지지만 대략 0.3∼0.8㎎/ml 정도였다. In order to determine the optimal assay conditions, the activity of the whole-cell methane monooxygenase(MMO) in Methylosinus trichosporium OB3b was measured under various conditions using gas chromatograph(GC). The enzymatic reaction monitored was propene oxidation to propene oxide and the reaction container was 5ml vial with a PTFE-faced rubber septum. When fermate was added as reducing agent, MMO activity was consistently 4-5 times higher compared to the case that fermate was missing. The optimal pH and temperature were 7.0-7.5 and 30℃, respectively. The dependence of enzymatic activity on propene concentration followed Michaelis-Menten kinetic model and the saturation constant(Km) was observed to be ?????mol/L The enzymatic activity was reduced with increasing incubation time or by high concentration of propene oxide. Considering the sensitivity of GC measurement and mass transfer resistance of gas substrates, proper cell density in the present assay system was estimated to be 0.3-0.8㎎/ml.
Bio platform of GS Caltex for Eco-friendly polymers
Hyohak Song(송효학) 한국고분자학회 2021 한국고분자학회 학술대회 연구논문 초록집 Vol.46 No.2
The bio-related R&D platform of GS Caltex is one of our strength, enabling us to create values with partners by using our resources and capabilities efficiently. Within lab., pilot, and demonstration plants and infrastructure, our expertise in the area of microorganisms, fermentation, and separation and purification makes possible to deliver new and existing products from one kilogram to a few tons. In particular, two demonstration plants located in Gusan and Yeosu cities are to shorten the time to market. We have developed a number of bio-originated monomers that can be used to synthesize eco-friendly polymers.