Production of Methanol by Resting Cells of Methylosinus trichosporium OB3b

Methylosinus trichosporium OB3b를 이용한 메탄올의 생산

As an effort to develop an alternative transportation fuel, the production of methanol from methane gas was studied using the resting cells of an obligatory methanotroph, Methylosinus trichosporium OB3b. The reaction was carried out in high concentration phosphate buffer solutions with the flask-grown cells containing the exclusively cytoplasmic methane monooxygenase (sMMO) activity. The methanol accumulation rate was observed to be 79nmo1/mg·min during the initial 4.5hr. Phosphate-dependent inhibition was found for both sMMO and methanol dehydrogenase (MDH) activities, and the inhibition constants were 185mM and 42mM, respectively. The inhibition mode was noncompetitive. Methanol was found to be very inhibitory to the sMMO activity and the inhibition constant (noncompetitive) was 21mM when propylene was used as substrate. The sMO activity in the resting cells was declined very fast and the rate became very high during the methanol production. These results indicate that the use of M. trichosporium OB3b as a biocatalyst for the methanol production is heavily dependent on the stable maintenance of the whole-cell SMO activity as well as the effective alleviation of product inhibition.

본 연구에서는 sMMO를 갖는 메탄 자화균인 M. triclwsporium OB3b를 이용하여 메탄올 생산을 위한 기초실험을 수행하였다. 중요한 결과를 요약하면 다음과 같다(Table 2). 1. 세포 내 NADH의 재생을 위해 개미산을 첨가 할 때 whole-cell의 sMMO 활성은 pH 7.0 및 $30^{\circ}C$ 에서 최대값을 보이며 propylene을 기질로 할 경우 약 130nmol/mg cell min 정도이다. 2. 인산은 MMO와 MDH 활성을 모두 저해하나 M MDH에 대한 저해 정도가 훨씬 크므로 메탄올 합성 에 사용이 가능하다. Noncompetitive mode를 가정 할 때 저해상수는 각각 185mM(MMO) 및 42mM ( (MDH)이었다. 3. 메탄올은 MMO 활성을 저해하며 noncompeti­t tive mode를 가정할 때 propylene기질의 경우 2 21mM 이었다. 4. 균체 내 sMMO 활성은 성장이 멈춰진 상태에 셔 비교적 때}른 속도로 감소하며 고농도 인산용액에 서 그 속도가 더 빨라진다. 5. 인산농도 91mM에서 메탄은 메탄올로 산화되 어 축적되며 4.5시간 동안 에탄올의 생성속도는 평 균 79nmol/mg min이었다.