사상균 Naringin 분해 효소에 관한 연구-[제1보] 우량 균주의 분리 선별과 선별균의 조효소 성질에 관하여-

Studies on the Naringinase of Mold-[part 1] Screening test of Molds on the Production of Naringinase and some properties of Crude Enzyme of Selected strain-

밀감류 등의 고미 성분의 일종인 Naringin을 분해하는 효소를 생성하는 사상균을 분리 선별하여 실용화에 알맞는 S-1 균주 효소 Naringinase의 몇가지 효소학적 성질을 검토한 결과는 다음과 같다. 1. Naringinase S-1은 Pectinase의 생성이 비교적 약하였으며 최적 활성 pH의 범위는 pH $3.0{\sim}5.0$사이었다. 2. Naringinase의 생성은 배지에 Naringin첨가에 의하여 많이 증가되었다. 3. 적당한 효소농도 이상에서는 Naringin의 분해는 0.1% 기질 농도에서는 80% 까지는 거의 직선적이었으며 고농도의 기질농도에서는 활성도가 낮았다. 4. 최적 활성 pH는 $50^{\circ}C$이었으며 열안정성은 $50^{\circ}C$에서 2시간 열처리에 의하여서는 별 역가의 감소가 없었으나 60.C에서 30분에 40% $70^{\circ}C$에서 10분만에 70%의 불활성화를 나타내었다. 5. Glucose는 효소작용에 있어 경쟁적으로 저해하였으며 저해정수는 1.5 Mol이었고 Sucrose에 의한 저해는 l Mol 농도에서 56%의 저해를 가져왔다.

Fifty strains of mold which isolated from the various sources were screened for the production of Naringinase which hydrolyse naringin, the 7-rhamnoe-glucoside of 4'.5.7. - trihydroxyflavanonin, the main bitter principle of citrus fruits and grape fruits. Of the 4 strains yielded naringinase with significant activity, S-1 strain was selected on the criterion of industrial application, and some properties of crude naringinase of this S-1 was investigated. The results obtained were as follows. 1. Naringenase obtained from S-1 strain has optimum pH range from 3.0 to 5.0 for its activity. 2. Production of naringinase was increased on the addition of naringin to the medium. 3. Hydrolysis of naringin with approporiate concentration of naringinase was carried out linerly up to 80% on the 0.1% substrate solution. 4. The optimum temperature for its activity was $50^{\circ}C$, and this enzyme was inactivated 80% of its total activity at $70^{\circ}C$ for 10 minutes, 40% at $60^{\circ}C$ for 30 minutes. But signifiant decrease of activity were not occurred by heat treatment at $50^{\circ}C$ for 2 hours. 5. Crude enzyme of the naringinase obtained from S-1 strain was competitively inhibited by addition of glucose on the substrate, and inhibitor constant of the glucose on the this enzyme was 1.5 Mol, and inhibition rate were linearly increased according to the increase of sucrose concentration and 56% of its total activity was inhibited at 1 Mol sucrose solution.