黑曲霉TC-01产柚苷酶对柚皮苷酶解作用的研究

Study on enzymatic hydrolysis of naringin with naringinase from Aspergillus.niger TC-01

柚皮苷是一类主要的类黄酮物质,它是引起柚类和葡萄果实苦味的主要来源,去除柚皮苷会使果汁苦味减轻,而柚皮苷酶能在不影响柑橘果汁品质的前提下较好地去除苦味。但是由于不同来源的柚苷酶的结构不同,其催化特性也不尽相同,本文通过利用前期筛选的黑曲霉(Aspergillus.niger TC-01)产柚苷酶对柚皮苷酶解作用进行研究,实验采用高效液相色谱法测定柚皮苷、普鲁宁和柚皮素的三者之间含量变化,确定酶解最适温度、时间、pH及酶用量,并测定TC-01产柚苷酶的米氏常数以确定酶解反应的最佳条件。结果表明,TC-01产柚苷酶酶解最适温度为60℃、酶解最适时间为30-40min,最适pH4.0,酶最佳用量为24u/mL。米氏方程为Y=0.0018X-3.0×10-6,其中Km值为598.8ug/mL,Vmax=2.06g/(min.mL)。此研究结果为柚苷酶的应用提供了理论依据。

Naringin is a major class of flavonoids, It is the substance which causes pomelo and grape fruit bitter. Removing naringin can make juice less bitter. Naringinase can effectively removing bitterness without affect the citrus juice quality. But due to different sources of naringinase, their structure is different and their catalytic properties are not the same. In this paper, naringinase screened from Aspergillus niger TC -01 was used in studying the bitter removal effect. HPLC was used for the determination the content of naringin, naringenin, and pruning. The optimum temperature, time, pH, enzyme dosage, and Michaelis constant were also studied. The results show that optimum temperature for Aspergillus. niger TC -01 producing naringinase is 60℃, the suitable time is 30 -40min, the optimum pH4. 0, enzyme optimum dosage of 24u/mL. The Michaelis - Menten equation is Y = 0. 0018X - 3.0 × 10^-6, Km = 598.8ug/ mL, Vmax =2. 066g/ （min. mL）. The results of this research for naringinase provides a theoretical basis of the application.