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单宁酶的固定化及其在酯型儿茶素水解反应中的应用

Immobilization of Tannase and Its Applicationin Hydrolysis of Estern-catechines

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【作者】 程琦程启坤李名君

【Author】 Cheng Qi;Cheng Qi-Kun;Li Ming-Jun(Department of Biochemistry,Tea Research Institute of Chinese Academy of Agricultural Science.Hangzhou 310008)

【机构】 中国农业科学院茶叶研究所生化室

【摘要】 采用自制的壳聚糖为载体对单宁酶(TA)固定化,TA与壳聚糖配比1:2.5,30℃固定2h,活力回收达23.6%~33.1%;偶联效率为84.9%~88.0%。固定化单宁酶(ITA)的表观Km值(以没食子酸丙酯为底物)为22.2×10-6mol/L,TA的Km值(以没食子酸丙酯为底物)为10×10-6mol/L,TA和ITA的最适反应温度分别为40℃和50℃;60℃处理15min,残存活性分别为13.6%和60.3%。TA和ITA的最适pH值分别为5.8和6.4;TA在pH4.8~7.8活力稳定,而ITA活力稳定范围在pH4.8~6.8.ITA作用于EGCG的半衰期为78.7h,EGCG水解率达90.3%。对茶多酚提取物进行水解,其所含的酯型儿茶素EGCG和ECG水解率分别为96.4%和96.8%,非酯型儿茶素EGC和EC的含量显著增加。

【Abstract】 Tannase(TA) was immobilized with self-made chitosan.The optimum conditions for immobilization were as follows:250 mg chitosan was dissolved in 1% solution of acetate acid,then 100 mg crude TA was added and the mixture was incubated at 30℃ for 2 hours.Enzyme activity recovery was 23.6%-33.1% and the efficiency of enzyme protein linked on chitosan was 84.9%-88.0%.The apparant Km of immobilized tannase(ITA) was 22.2×10-6 mol/L,while Km of TA was 10×10-e mol/L.The optimum reaction temperatures for TA and ITA were 40℃ and 50℃,respectively.Treated at 60℃ for 15 min,TA only retained 13.6% of the original activity.while ITA was more stable and retained 60.3% of the original activity. The optimum pHs for TA and ITA were 5. 8 and 6.4,respectively.TA was stable in the PH range of 4.8-7.8,while ITA was in pH 4.86.8 range.The half-life time of ITA(EGCG was used as substrate) was 78.7 hours.In the mean time the percentage of hydrolysis of EGCG was 90.3%.When treated by ITA,EGCG and ECG contained in tea polyphenol extracts were hydrolyzed up to 96.4% and 96.8%,respectively.The contents of EGC and EC in hydrolysate were increased greatly.

【基金】 农业部重点研究项目
  • 【分类号】Q550.2
  • 【被引频次】46
  • 【下载频次】361
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