Characterization of yeast cell wall lytic enzyme from Fusarium moniliforme

Fusarium moniliforme이 생산하는 효모세포벽 분해효소의 특성

Yeast cell wall lytic enzyme was purified from Fusarium moniliforme by ammonium sulfate fractionation and gel column chromatography. The lytic activity was found to consist of three enzyme activities which were resolved on Sephadex G-100. The first peak on chromatogram exhibited proteolytic, lytic and laminarinase activities, and the second had both lytic and laminarinase activities, whereas the third peak was shown to contain lytic activity only. Three enzyme activities showed the synergistic effect and reducing agents accelerated the yeast roil wall lysis. This indicates that lytic, proteolytic and laminarinase activity acted cooperatively in the lysis of intact cells. Tannic acid precipitate of crude enzyme constituted of three enzyme activities had a high lytic activity on viable yeast cell and has proved useful in yeast protoplast formation.

Fusarium moniliforme으로부터 순모세포벽 분해효소를 생산하고 분리, 정제하여 효소특성 및 protoplast 제조실험을 하였다. Ammonium nitrate를 0.2％ 첨가한 Baker's yeast 배지에서 7일간 진탕배양으로 효소를 생산한 후 Ammonium sulfate로 분획하고 Sephadex(G-100) column chromatography하여 세개의 peak를 얻었다 첫 번째 peak는 proteolytic, lytic activity 및 laminarin 분해력가를 보였으며, 두 번째 Peak는 lytic activity와 laminarin 분해력가를 동시에 가지고 있었으며, 세 번째 peak는 lytic activity만을 가지고 있었다. 분리된 세개의 peak를 혼합하였을때 개개의 peak보다 훨씬 높은 역가을 나타내어 상승효과를 보였고 또한 환원제에 의한 효소력가의 상승효과도 있었다. protoplast 수율은 99.2％정도였다