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This research was focused on investigation of the general condition for protoplast formation of Trichoderma speues. For protoplast formation, the mycelia cultured in YM medium were collected from each growth phase and were treated with the lytic enzymes. This procedure was carried out by all strains. The most optimal conditions of NOVOZYM 234 and DRISELASE were determined by T. saturnisporum IAM 12535 and T. longibruchiatum IBM 13107, respectively. The effect of osmotic stabilizers appeared KCl$gt;(NH₄)₂SO₄$gt;NaCl$gt;mannitol$gt;MgSO₄ and the optimal concentration of each osmotic stabilizer was determined by 0.6-0.9 M. The optimal condition of DRISELASE for protoplast formation ; optimal pH 5.0, optimal concentration, 2%, optimal reaction time, 4 hours, and optimal temperature, 30℃. The optimal condition of NOVOZYM 234 for protoplast formation ; optimal pH 5.5, optimal concentration 1%, optimal reaction Nme 3 hours, and optimal temperature 30℃. The optimal culture period of mycelia for protoplast formation was between the initial and the middle exponential phase. Generally, DRISELASE was more effective than NOVOZYM 234 on protoplast formation except for T. longibruchiatum IAM 13107 and T. viride IAM 5141.
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