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베타-1,3-글루칸 합성효소 저해제의 스크리닝을 위한 효소원 제조법
박희문,이동원,송미령,김정윤,김성욱,복성해 한국산업미생물학회 1995 한국미생물·생명공학회지 Vol.23 No.3
Assay conditions for screening of β-1,3-glucan synthase inhibitor were evaluated. Cells in the beginning of mid-log phase showed the highest activity of the β-1,3-glucan synthase. Cells permeabilized with 1% digitonin treatment could be used as a good crude enzyme source for convenient screening of the β-1,3-glucan synthase inhibitors. Calcofluor white (0.125% in final) and papulacandin B (25 ㎍/㎖) inhibit 90% and more than 50% of the β-1,3-glucan synthase activity, respectively. Cells grown at 37℃ showed higher enzyme activity than those of 25℃. Catalytic factor of the β-1,3-glucan synthase was solubilized from particulated membrane preparations, holoenzyme, by extracting with 0.00938% CHAPS.