【目的】本工作对棘孢曲霉固体发酵抽提酶液转化甜菊糖进行了研究,并对转化产物进行鉴定及纯化分析。【方法】用高效液相色谱、液质联用及红外光谱等方法对转化新产物进行鉴定,对上清液中莱鲍迪苷A(RA)成分进行纯化。【结果】棘孢曲霉酶液在10 h内对甜菊糖中的甜菊苷(SS)、莱鲍迪苷C(RC)进行高效特异性转化,以沉淀的形式析出的转化产物经鉴定为甜菊醇,转化率高达98.0%,分离提纯后纯度为95.2%,回收率达84.0%.由于甜菊醇的沉淀分离,留在溶液中的RA更易被纯化。RA通过树脂吸附分离的回收率为80.5%.【结论】棘孢曲霉酶液对甜菊糖的一次转化可以同时得到甜菊醇和莱鲍迪苷A两种产品,是一种经济高效的工艺。
[Objective] The purpose of this research was to apply Aspergillus aculeatus solid fermentation extracts to convert stevioside and rebaudioside C,followed by identifying and purifying the new conversion product.[Methods]The product was identified by high performance liquid chromatography,chromatography-mass spectrometry and Infrared spectrum.The new product and rebaudioside A exited in the supernatant were purified by alcohol and macroporous resin.[Results]The Aspergillus aculeatus enzyme extracts could convert the stevioside and rebaudioside C to the new product within 10 hours. The conversion rate was 98.0% in 24 hours.The conversion product existed in deposit was identified as steviol.The purity and recovery percent of steviol were 95.2% and 84.0% respectively.Because stevioside could occur to deposit,the rebaudioside A existed in supernatant was purified easily. We used the resin chromatography to purify RA and the recovery could reach 80.5%.[Conclusion]Aspergillus aculeatus enzyme extracts could convert stevioside efficiently and specifically,and we could obtain rebaudioside A and steviol at the same time.