Studies on the Production of Protease by Aspergillus oryzae KC-15 and Characteristics of the Enzymes

Aspergillus oryzae KC-15에 의한 protease의 생산 및 그 효소의 특징에 관한 연구

This experiment was conducted to investigate the conditions for production and the characteristics of pretenses. Aspergillus oryzae KC-15, which is selected as a superior strain for the production of the protease, was used in this study. The results obtained were as follows: 1. The optimum culture time for the production of acid, neutral and alkaline protease on wheat bran medium were about 48, 48 and 72hr, respectively. The protease-produced by the strain were mainly alkaline and neutral one, but the production of acid protease was feeble extremely. 2. The addition of NaH$_2$PO$_4$, Na$_2$HPO$_4$, glucose, rice powder and Na-glutamate respectively to wheat bran media were effective for the production of alkaline and neutral protease, and the addition of (NH$_4$)$_2$HPO$_4$, glucose and rice powder respectively were effective for the production of acid protease. 3. Characteristics of professes(equation omitted) 4. As a heat resistance agent, NaH$_2$PO$_4$was the most effective one. The optimum amount of NaH$_2$PO$_4$was 10mg for alkaline and neutral protease, and 5mg for acid protease. 5. The heat resistance of the Protease by NaH$_2$PO$_4$was not recognized mostly above 6$0^{\circ}C$. 6. After the treatment of enzyme solution with 10mg of NaH$_2$PO$_4$for 30 minutes at 55$^{\circ}C$, the residual activities measured for alkaline, neutral and acid protease were 58, 57 and 55％ respectively.

Protease의 생산능이 우수한 Asp. oryzae KC-15를 선정하고 다음과 같은 결과를 얻었다. 1. Wheat bran medium에서의 최적배양시간은 acid protease와 neutral protease는 약 48시간, alkaline protease는 약 72시간이었고 본 균주가 생산하는 protease는 alkaline protease와 neutral pro-tease가 주체이며 acid protease는 극히 미약하였다. 2. Wheat bran medium 에 $Na_2$HPO$_4$, NaH$_2$PO$_4$, Glucose, rice powder 및 Na-glutamate의 첨가는 alkaline protease와 neutral psotease의 생산에, (NH$_4$)$_2$HPO$_4$, glucose 및 rice powder의 첨가는 acid protease의 생산에 효과적이었다. 3. 조효소의 특징(equation omitted) 4. 내열제로서 NaH$_2$PO$_4$가장 효과적이었으며 최적첨가량은 alkaline protease와 neutral protease 에 대하여는 10mg, acid protease에 대하여는 5mg 이었다. 5. 6$0^{\circ}C$ 이상에서는 NaH$_2$PO$_4$의 내열효과는 거의 인정할 수 없었다. 6. NaH$_2$PO$_4$10mg을 첨가하고 55$^{\circ}C$에서 30분간 처리하였을 때의 잔존활성은 alkaline protease는 약 58%, neutral protease는 약 57%, acid protease는 약 55 %이었다.