以指狀青霉(Penicillium digitatum) DSM62840為研究對象，采用高壓細胞破碎法破碎菌體，對其轉化檸檬烯生成α-松油醇過程中的相關酶的提取條件進行優化。以檸檬烯轉化酶活性為指標，在單因素試驗的基礎上結合響應面試驗法，確定最佳提取條件；同時，對檸檬烯轉化酶的酶學性質進行了初步探索。結果顯示，最佳提取條件為：破碎壓力100 MPa、破碎6次、液料比15∶1 (mL/g)，在此條件下，α-松油醇的質量濃度為855.75 mg/L，酶活力達到71.31 U。檸檬烯轉化酶最佳轉化時間為4 h，在磷酸鹽緩沖液中獲得的酶活性最高；該酶可能為細胞色素P450，且不同的金屬離子對酶活性的影響不同：Fe2+對酶有輕微的抑制作用，而Ca2+、Mg2+、Mn2+、Zn2+、Co2+、Ba2+、Na+、K+、Fe3+、Ni2+則對酶有不同程度的激活作用。粗酶液SDS-PAGE電泳結果表明優化后的提取工藝效果較好。
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.