超声波辅助双水相萃取鲍鱼内脏的β-葡萄糖苷酶
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摘要
以皱纹盘鲍内脏为原料,通过超声波破碎辅助提取其中的β-葡萄糖苷酶,探讨一定量的预处理液体积下,不同超声波功率,超声时间和超声后的浸提时间对鲍鱼内脏中β-葡萄糖苷酶酶活力的影响,并通过单因素试验和正交试验最终确定超声波辅助提取的最佳工艺条件。在此基础上,采用聚乙二醇(polyethylene glycol,PEG)/硫酸铵([NH_4)_2SO_4]双水相系统对最优超声条件下提取的粗酶液进行萃取分离,确定最佳的双水相系统。结果表明超声功率对提取有显著性影响,超声时间则影响不显著,而超声后的浸提时间的影响可以忽略不计,所以当预处理液为10 mL时,超声功率为180 W,超声时间为20 min,超声后浸提时间为1 h为最优超声提取的工艺条件;当双水相体系的质量组成为10%的聚乙二醇1000、16%的(NH_4)_2SO_4、pH值为自然(pH≈5)的状态下萃取效果最好。
The abalone viscera was used as raw materials,which was extraction of β-glucosidase by ultrasonic assisted,and the different ultrasonic power,ultrasonic time and extraction time of a certain amount of the pretreatment liquid product in the influence of enzyme activity was discussed. The optimum process conditions were obtained by single factor test and orthogonal test. On this basis,polyethylene glycol(PEG)(/NH_4)_2 SO_4 aqueous two-phase system was applied to extract β-glucosidase from crude enzyme fluid and optimize the twophase system.The experimental results showed that ultrasonic power had a significant effect on the extraction,ultrasonic time had no significant influence,and the extraction time after ultrasonic could be neglected,so when the pretreatment liquid was 10 mL,ultrasonic power was 180 W,ultrasonic time was 20 min,extraction time after ultrasonic of 1 h were the optimal process conditions. The optimal two-phase system was the mass fraction of PEG1000 at 10 %,and the mass fraction of(NH_4)_2 SO_4 at 16 %,pH at nature(pH≈5).
The abalone viscera was used as raw materials,which was extraction of β-glucosidase by ultrasonic assisted,and the different ultrasonic power,ultrasonic time and extraction time of a certain amount of the pretreatment liquid product in the influence of enzyme activity was discussed. The optimum process conditions were obtained by single factor test and orthogonal test. On this basis,polyethylene glycol(PEG)(/NH_4)_2 SO_4 aqueous two-phase system was applied to extract β-glucosidase from crude enzyme fluid and optimize the twophase system.The experimental results showed that ultrasonic power had a significant effect on the extraction,ultrasonic time had no significant influence,and the extraction time after ultrasonic could be neglected,so when the pretreatment liquid was 10 mL,ultrasonic power was 180 W,ultrasonic time was 20 min,extraction time after ultrasonic of 1 h were the optimal process conditions. The optimal two-phase system was the mass fraction of PEG1000 at 10 %,and the mass fraction of(NH_4)_2 SO_4 at 16 %,pH at nature(pH≈5).
引文
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[3] Sinnott M L. Catalytic mechanisms of enzymic glycosyl transfer[J].Chemical Reviews, 1990, 90:1171-1202
[4]张儒,张变玲,谢涛,等.聚乙二醇/硫酸铵双水相萃取猪胃蛋白酶工艺研究[J].食品工业科技,2012,33(15):245-246
[5]张喜峰,孙亚莉,李鑫文,等.双水相萃取分离葡萄酒发酵下脚料中葡萄籽蛋白质[J].云南大学学报,2013,35(6):824-832
[6]单佳莉,黄帮裕,麦霖霞.双水相萃取技术在生物技术上的应用[J].广东化工,2014, 41(16):131-132
[7]石贤爱,张利,刘震,等.南日鲍β-葡萄糖苷酶分离纯化及表征[J].福州大学学报(自然科学版),2014,42(3):480-485
[8]罗小娟,杨娟,陈锦权,等.鲍鱼脏器β-葡萄糖苷酶提取工艺的优化[J].安徽农学通报,2016,22(18):16-19
[9]陈磊.西梅籽β-葡萄糖苷酶的分离纯化、性质及其固定化研究[D].广州:华南理工大学,2011
[10]董娜,董文宾,田颖.双水相萃取和壳聚糖沉淀相结合分离猪胃蛋白酶[J].食品科技,2011,36(1):26-29
[11]杨利民,吕金萍,冯研.蒲公英总黄酮在聚乙二醇-硫酸铵双水相体系中的分配与提取[J].化工进展, 2014, 33(8):1992-2011