漆树漆酶的提取分离及性质研究  

Extraction, Separation and Properties of Lacquer Laccase

作　　者：尤俊昊 黄晓华[1] 赵爱国 YOU Jun-hao;HUANG Xiao-hua;ZHAO Ai-guo(Northwest A&F University,Yangling,Shaanxi,712100)

机构地区：[1]西北农林科技大学林业科学研究院,陕西杨凌712100

出　　处：《陕西林业科技》2019年第6期1-5,共5页Shaanxi Forest Science and Technology

基　　金：国家重点研发计划“漆树丰产经营与精细产品开发研究技术”(2017YFD0600705)。

摘　　要：以生漆原液为原料,用冷丙酮和硫酸铵溶液成功提取漆树漆酶后对所提溶液蛋白浓度进行了测定和SDS-PAGE凝胶电泳检测,漆树漆酶蛋白分子量约为100 Kda。并以2,2′-连氮-双-3-乙基苯并噻唑啉-6-磺酸(简称ABTS)为底物,测定漆树漆酶活性,探讨了pH、温度、金属离子种类及浓度对漆树漆酶活性的影响。结果表明,以ABTS为底物的测定漆树漆酶酶活最佳条件为:pH值为2.6的柠檬酸和磷酸氢二钠缓冲溶液为反应溶液,反应温度35℃。铜、镍离子在低浓度时(小于10 mM)对漆树漆酶酶活起激活作用,其中铜离子激活作用最强;三价铁离子对漆树漆酶酶活起强抑制作用;低浓度(小于10 mM)钙、镁、锌、钾离子对漆树漆酶酶活影响不大;在高浓度时,各金属离子均不同程度抑制漆酶酶活,对漆树漆酶活性抑制作用近似饱和时,金属离子浓度对漆树漆酶活性的影响不再突出。The raw lacquer stock as raw material,the molecular weight and concentration of lacquer laccase protein were measured after the lacquer laccase was successfully extracted with cold acetone and ammonium sulfate solution.ABTS is used as the substrate to determine the lacquer laccase activity,and discuss the effects of pH,temperature,metal ion species and concentration on the lacquer laccase activity.The results show that the ABTS as the substrate of the determination of the best conditions for lacquer laccase activity:the citric acid and disodium hydrogen phosphate buffer solution with pH 2.6 is used as the reaction solution,reaction temperature is 35℃.At low concentration(<10 mM),copper and nickel ions activate the lacquer laccase activity,especially copper ions.Ferric ions strongly inhibit the lacquer laccase activity;Calcium,zinc and potassium ions(<10 mM)had no significant effect on lacquer laccase activity.At high concentration,all metal ions inhibit lacquer laccase activity to different extents,and the inhibitory effect on lacquer laccase activity is approximately saturated,and the concentration of metal ions is no longer significant for lacquer laccase activity.

关 键 词：漆树漆酶 酶活 ABTS 金属离子 

分 类 号：Q554.9[生物学—生物化学]