Trans-二氢香芹酮广泛应用于香精香料、医药、农业等领域，具有高经济价值。为了利用克雷伯杆菌O852进行柠檬烯的生物转化生产trans-二氢香芹酮，该实验对克雷伯杆菌O852产trans-二氢香芹酮关键酶体系的提取工艺及酶学特性进行分析。研究通过单因素实验和响应面分析确定了细胞破碎的最佳条件，进一步分析金属离子、有机溶剂和抑制剂对酶活的影响，并测定酶的最适温度和pH值。研究发现，最佳的提取条件：超声时间21 min、料液比1:13、振幅36%。Fe2+、Mn2+、Zn2+显著提升酶的活性，而Cu2+和Fe3+则表现出强烈的抑制效果。使用1%（V/V）的二甲基亚砜（DMSO）和异丙醇时，酶活性分别提升至121.64%和142.62%，然而，当增至5%（V/V）时，有机溶剂对酶活性的抑制作用增强，尤其是氯仿，其抑制效果最为显著。对于抑制剂的影响，EDTA对酶活性的抑制最显著，其他抑制剂也表现出一定的抑制效果。此外，研究得出酶的最适温度20℃，最适pH值9.0。结果为该酶体系的工业化应用提供了理论依据和技术支持。

Trans-dihydrocarvone is widely used in flavor, medicine, agriculture and other fields, with high economic value. In order to produce trans-dihydrocarvone by limonene biotransformation using Klebsiella sp. O852, the extraction process and enzymatic characteristics of the key enzyme system of trans-dihydrocarvone production from Klebsiella sp. O852 were analyzed. The optimal conditions of cell fragmentation were determined by single factor experiment and response surface analysis. The effects of metal ions, organic solvents and inhibitors on enzyme activity were further analyzed, and the optimal temperature and pH value of the enzyme were determined. It was found that the optimal extraction conditions were as follows: ultrasonic time 21 min, solid-liquid ratio 1:13, amplitude 36%. Fe2+, Mn2+ and Zn2+ significantly increased the enzyme activity, while Cu2+ and Fe3+ showed strong inhibitory effects. With 1% (V/V) DMSO and isopropyl alcohol, the enzyme activity increased to 121.64% and 142.62%, respectively. However, when increased to 5% (V/V), the inhibition effect of organic solvents on the enzyme activity was enhanced, especially chloroform. As for the influence of inhibitors, EDTA has the most significant inhibition on enzyme activity, and other inhibitors also show certain inhibitory effects. In addition, the optimum temperature of the enzyme was 20℃ and the optimum pH value was 9.0. The results provided theoretical basis and technical support for the industrial application of the enzyme system.

基金项目：河南中烟工业有限责任公司科技项目计划（AW2024033），湖北省科技创新人才计划 (2023DJC116，2024DJC029)，云南省科技计划项目（202304BT090030）。