黄孢原毛平革菌茶皂素脱色酶的粗分离及酶学性质  

Separation and properties of Phanerochaete chrysosporium TS-07 tea saponin decoloring enzyme

作　　者：李艳丽[1,2] 陈列忠[1,2] 陶瑛妮 

机构地区：[1]浙江省农业科学院植物保护与微生物研究所,浙江杭州310021 [2]浙江省植物有害生物防控重点实验室-省部共建国家重点实验室培育基地,浙江杭州310021 [3]浙江中医药大学,浙江杭州310053

出　　处：《浙江农业学报》2014年第4期976-980,共5页Acta Agriculturae Zhejiangensis

基　　金：杭州市农业科研攻关项目(20120232B61);杭州市科技发展项目(20110232B11);浙江省重点科技创新团队项目(2010R50028)

摘　　要：对黄孢原毛平革菌(Phanerochaete chrysosporium)TS-07发酵液中的茶皂素脱色酶进行粗分离,并对其酶学性质进行了研究。结果表明:用硫酸铵沉降法所得粗酶粉的酶活力高达52 971 U·g-1;该酶的最适反应温度为40℃,最适作用pH值为4.5;该酶在常温下具有良好的热稳定性,在pH值3.0～7.5环境下都比较稳定;Fe3+、Ca2+和Ni2+对酶活影响不大,Hg2+能强烈抑制茶皂素脱色酶的活性;10%(V/V)有机溶剂丙酮、异丙醇、乙醇和乙腈能保留茶皂素脱色酶大部分的活力;反应体系中H2O2达到5%,茶皂素脱色酶仍能保持较高的活力;而在反应底物不存在的情况下,茶皂素脱色酶与0.25%的H2O2保温1 h后,酶活残余量仍能达到80%以上。A tea saponin decoloring enzyme （ TSDE） was separated from Phanerochaete chrysosporium TS-07 by using ammonium sulfate precipitation .The separated TSDE activity was 52 971 U· g^-1 .The effect of pH value on TSDE activity and stability was examined at pH values ranging from 3.0 to 10.0 .TSDE was completely stable in a wide pH range （3.0-7.5） and presented an optimum pH value of 4.5.The optimum temperature for TSDE was 40℃and the enzyme was stable at room temperature .It was also found that TSDE was strongly inhibited by Hg^ 2＋, while Fe^3＋, Ca^2＋and Ni^2＋did not cause any alteration in the activity .TSDE was also resistant to presence of up to 5%H2 O2 , but it was inhibited in presence of 10% H2 O2 .The TSDE showed a high percentage of activity in reaction mixtures containing 10% （ V/V ） of different organic solvents （ acetone , isopropanol , ethanol and acetonitrile）.The susceptibility of TSDE to inactivation by H2O2 in absence of substrate was also studied and it was＆amp;nbsp;found that the enzyme was very stable in presence of up to 0.25%H2 O2 .

关 键 词：黄孢原毛平革菌 茶皂素脱色酶 酶学性质 

分 类 号：S814.9[农业科学—畜牧学]