Abstract:With Penicillium digitatum DSM62840 as the object of research,the extraction conditions of related enzymes in the process of converting limonene to α-terpenol were optimized to obtain high activity. And limonene converting-enzyme was extracted by high pressure cell crushing method. Taking the limonene converting-enzyme activity as evaluation indicator,the response surface methodology was used to optimize the extraction technology based on single factor test. At the same time,the properties of limonene converting-enzyme were preliminarily studied. The results showed that the optimum conditions for extracting were as follows: disruption pressure of 100 MPa,crushing times of 6,and liquid-to material ratio of 15∶1 (mL/g). Under the conditions,the yield of α-terpenol was 855.75 mg/L,and the enzyme activity was 71.31 U. The optimum conversion time of limonene converting-enzyme was 4 h,and the highest enzyme activity was obtained in phosphate buffer. Limonene converting-enzyme might be cytochrome P450,and different metal ions have different effects on the enzyme: Fe2+ had slight inhibition on the enzyme,while Ca2+,Mg2+,Mn2+,Zn2+,Co2+,Ba2+,Na+,K+,Fe3+ and Ni2+ had different activation effects on the enzyme. SDS-PAGE analysis of the crude enzyme solution showed that the extraction effect could be better under the optimum conditions.