以指状青霉(Penicillium digitatum) DSM62840为研究对象，采用高压细胞破碎法破碎菌体，对其转化柠檬烯生成α-松油醇过程中的相关酶的提取条件进行优化。以柠檬烯转化酶活性为指标，在单因素试验的基础上结合响应面试验法，确定最佳提取条件；同时，对柠檬烯转化酶的酶学性质进行了初步探索。结果显示，最佳提取条件为：破碎压力100 MPa、破碎6次、液料比15∶1 (mL/g)，在此条件下，α-松油醇的质量浓度为855.75 mg/L，酶活力达到71.31 U。柠檬烯转化酶最佳转化时间为4 h，在磷酸盐缓冲液中获得的酶活性最高；该酶可能为细胞色素P450，且不同的金属离子对酶活性的影响不同：Fe2+对酶有轻微的抑制作用，而Ca2+、Mg2+、Mn2+、Zn2+、Co2+、Ba2+、Na+、K+、Fe3+、Ni2+则对酶有不同程度的激活作用。粗酶液SDS-PAGE电泳结果表明优化后的提取工艺效果较好。
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.