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1、第31卷第1期水产学报Vol.31,No.12007年1月JOURNALOFFISHERIESOFCHINAJan.,2007文章编号:1000-0615(2007)01-0015-08皱纹盘鲍内脏酶的酶学性质及褐藻胶裂解酶的分离纯化康平,汪秋宽,宋琳琳,徐玲(大连水产学院食品工程系辽宁省水产品加工及综合利用重点开放实验室,辽宁大连116023)摘要:采用(NH4)2SO4分段盐析、透析、阴离子(DEAE252)交换柱层析、SephadexG2200凝胶柱层析等分离纯化技术,通过SDS2PAGE电泳分析了皱纹盘鲍
2、内脏酶的组成,结果表明鲍内脏酶主要含有两种褐藻胶裂解酶Ⅰ,Ⅱ,一种纤维素酶,一种琼脂酶。对酶的酶学性质分析结果表明两种褐藻胶裂解酶Ⅰ,Ⅱ的最适pH分别为8.6,7.2,最适温度为35℃,分子量分别为35.2ku,67ku;两种褐藻胶裂解酶的热稳定性比较差,且易受金属离子影响;纤维素酶的最适pH为5.0,最适温度为40℃。并确定了皱纹盘鲍内脏酶分离纯化的方法及参数,为进一步研究鲍内脏复合酶的性能提供了基础参数。关键词:皱纹盘鲍;褐藻胶裂解酶;纤维素酶;纯化中图分类号:Q814;S917文献标识码:ACharacte
3、risticsofenzymesextractedfromabalone(HaliotisdiscushannaiIno)intestineandpurificationofthealginatelyaseKANGPing,WANGQiu2kuan,SONGLin2lin,XULing(KeyandOpenLaboratoryofAquaticProductProcessingandUtilizationofLiaoningProvince,DepartmentofFoodEngineering,DalianFi
4、sheriesUniversity,Dalian116023,China)Abstract:Theenzymesextractedfromabalone(HaliotisdiscushannaiIno)intestinewerestudiedforfindingthealginateandcellulosedegradationenzymes.Theenzymeswereseparatedandpartiallypurifiedfromabaloneintestinebyusingammoniumsulfatep
5、recipitations,andsuccessivefractionationonDEAE252,SephadexG2200columnchromatography.Theresultsshowedthattheirwerefourtypesofenzymeseparated,whichwerealginatelyaseⅠ,lyaseⅡ,cellulaseandagarase.ThecharacteristicsofalginatelyaseⅠ,lyaseⅡandcellulaserelatedwithpH,t
6、emperatureandionswerealsodeterminedandtheresultsindicatedthatalginatelyaseⅠhasthemaximumactivityatpH8.6and35℃.AlginatelyaseⅡhasmaximumactivityatpH7.2and35℃.CellulasehasmaximumactivityatpH5.0and40℃.ThealginatelyaseⅠandlyaseⅡwerenotstableathightemperature.Thecr
7、udeenzymesexstractedcouldbeobviously2+3+2+2++activatedbyMn,Al,Ca,CuandinhibitedbyNa.SDS2PAGEwasusedforestimatingthemolecularweightsofenzymesandthefindingsshowedthatthemolecularweightsofalginatelyaseⅠ,Ⅱweretobe35.2ku,67ku.Thefindingsindicatedthatthereweretwoty
8、pesofalginateenzymecouldbeobtainedfromabalone(HaliotisdiscushannaiIno)intestineandthestudiesestablishedthepurificationsteps收稿日期:2006206222资助项目:国家“九四八”项目(2005-Z23);辽宁省科技厅攻关项目(2004209001);大
