黄振瑞 柳颖 黎志德

摘  要：为提高烟叶中茄尼醇的提取率，通过纤维素酶-半纤维素酶联用的方法对烟叶进行预处理，研究了酶种类、添加量、温度、pH和反应时间对烟叶提取液中游离茄尼醇和总茄尼醇含量的影响，并以Box-Behnken响应面法优化酶解工艺。结果表明，在单因素试验中，最优工艺为纤维素酶-半纤维素酶联用（酶添加量为每克烟叶各1.00 U）、温度55 ℃、pH 5.2和反应时间60 min;与未经酶解预处理的空白组（CK）相比，纤维素酶-半纤维素酶联用处理后，游离和总茄尼醇提取率分别增加305.61%和113.55%，响应面法优化后，总茄尼醇提取率较CK提高158.78%。本研究探索出一种提高茄尼醇提取率的有效方法，对烟草综合利用具有指导意义。

关键词：烟叶;茄尼醇;酶解;响应面法

Abstract： In this study， both cellulase and hemicellulose were used in pretreatment process in order to improve the extraction rate of solanesol in tobacco leaves. The effects of enzyme types， dosage， temperature， pH and reaction time on the content of free solanesol and total solanesol in tobacco leaf extracts were studied. Optimization of enzymatic hydrolysis was carried out by the Box-Behnken response surface method. The results of single-factor experiment showed that the optimal enzymatic hydrolysis process was cellulase-hemicellulase combined use （1.00 U per gram of tobacco leaves）， temperature 55 ℃， pH 5.2 and reaction time 60 min; Compared with the control group without enzymatic hydrolysis pretreatment， the extraction rate of free solanesol of the cellulase-hemicellulase combined use group was increased by 305.61% and the total solanesol was increased by 113.55%. After response surface optimization， the total solanesol extraction rate was increased by 158.78% compared with the control group. This study has explored an effective way to improve the extraction rate of solanesol in tobacco leaves， which has guiding significance for the comprehensive utilization of tobacco.

Keywords： tobacco leaf; solanesol; enzymatic hydrolysis; response surface methodology

茄尼醇（Solanesol）是一种不饱和的聚异戊二烯醇，属四倍半萜醇和长链脂肪醇[1]，具有抗菌、消炎和止血作用，同时也是许多药物的重要中间体，可用于合成N-茄尼基谷酰氨、辅酶Q10[2]和维生素K2[3]等，广泛存在于烟草、马铃薯和桑树等植物中，以烟草中的含量最为突出，最高可达3%[4]。其在烟草中以游离态或化合态形式存在，新鲜烟叶中通常以化合态为主，随着贮存时间的延长逐渐转化为游离态[5]。茄尼醇易溶于正己烷等有机溶剂，微溶于乙醇[6]，在光照、加热环境中易发生衰变[7]。目前茄尼醇的提取方式以有机溶剂（如正己烷、乙醇）提取为主，存在提取时间长、溶剂使用量大，易出现乳化现象导致难以分层等问题[8]。其他提取方法如超临界二氧化碳萃取、分子蒸馏和微波辅助萃取等，存在前期设备投入大，工艺复杂不利于扩大等问题，在实际生产中鲜有应用[9]。……