능이버섯(Sarcodon aspratus) 무름병소에서 분리한 MK1 균주의 최적 성장조건과 정제된 균체외다당류의 특성

Optimal Culture Conditions for MK1 Strain Isolated from Soft-Rotten Tissue of Neungee Mushroom (Sarcodon aspratus) and the Physico-Chemical Properties of the Purified Exopolysaccharide of MK1

능이버섯(Sarcodon aspratus)의 무름병소에서 분리한 MK1 균주는 절대호기성 타가영양 세균으로 거식세포의 활성을 유도하는 균체외다당류를 다량 생산한다. MK1의 최적 성장 조건과 MK1이 생산하는 균체외다당류의 몇가지 물리적 성질을 알아보았다. MK1은 glucose, galactose, fructose, sucrose를 탄소원으로 잘 이용하나 MK1의 lactose 이용은 다소 저조하였다. 한편 potato starch와 dextrin을 이용하지 못하였다. MK1은 pH 7.0, $30^{\circ}C$, 200 rpm, 2% glucose, 0.05~0.2% $(NH_4)_2SO_4$에서 최적 성장을 보였다. 최적 성장조건에서 얻은 균체외다당류($EPS_{opt}$)는 탄소(37.1%), 질소(2.2%), 산소(49.3%), 수소(6.4%)로 구성되었으며, 황(S)은 검출되지 않았다. 그리고 아미노당과 산성 당을 함유한 이종다당류로 여겨지며, SDS-polyacrylamide gel elctrophoresis로 분석한 $EPS_{opt}$의 분자량은 14.8~47.9 kDa였다. Glucose 배지에서 생산된 균체외다당류($EPS_{glu}$)의 점도를 측정한 바, pH 6.0과 7.5에서 얻은 것들(0.1 g/ml)의 상대 점도($\acute{\eta}_{rel}$)는 각각 1.23과 1.39이었다. 또한 형태 또는 질감에도 차이가 있었는데, 배지의 pH가 낮을수록 미세결정형을, pH가 높아질수록 끈끈한 질감을 보였다. 따라서 배지 pH가 균체외다당류의 물리적 성질에 영향을 주는 것으로 사료된다.

MK1 strain, an obligate aerobic heterotrophic bacterium isolated from the rotten tissue of Neungee mushroom (Sarcodon aspratus), produces a copious amount of exopolysaccharide (EPS), which could evoke macrophage activation. Investigations on optimal culture conditions of MK1 and physical properties of MK1 EPS were made. Glucose, galactose, fructose, and sucrose supported well growth of MK1, but potato starch and dextrin did not. However, lactose seemed to be a less favorable carbon source. Optimal growth of MK1 was obtained at pH 7.0, $30^{\circ}C$, and 200 rpm with 2% glucose, and 0.2~0.05% $(NH_4)_2SO_4$. $EPS_{opt}$ obtained from an optimal growth condition constituted of carbon (37.1%), nitrogen (2.2%), oxygen (49.3%), and hydrogen (6.4%), but no sulfur. Paper chrogromatogram of the acid-hydrolysate of $EPS_{opt}$ suggested that MK1 EPS seemed to be hetropolysaccharide composed of a few number of monosaccharides including amino- and acidic-sugars. Its molecular mass determined by SDS-polyacrylamide gel electrophoresis varied from 14.8 to 47.9 kDa. Physical properties of $EPS_{glu}$ obtained from cell grown in glucose medium, such as relative viscosity ($_{rel}$) and crystalline morphology were rather affected by pH of the growth medium. Relative viscosity ($_{rel}$) of exopolysaccaride (0.1 g/ml) harvested from cells grown at medium pH ranging from 6.0 and 7.5 was 1.23 and 1.39, respectively. The freeze-dried exopolysaccharide obtained at low pH (6.0 and 6.5) was fine crystaloid and water-soluble, whereas those obtained at high pH (7.0 and 7.5) was rather gluey and less water-soluble.