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[18]许尧兴,李艳丽,柳永,等.黑曲霉变种RM480【一半乳学报:农业与生命科学版,2009,35(2):147—152糖苷酶的分离纯化及其酶学性质研究[J].浙江大学Purification andCharacterization ofIntracellular0【·galactosidase fromRh&opus sp.A01WangJian-fen91,ChenJin.zha02,LiJian91,RaoJunl1(Department ofBiology,EastChinaInstitute ofTechnology,Fuzhou,China)2(College ofLifeScience andTechnology,YangtzeNormalUniversity,Chongqing,China)ABSTRACTUsing three—phase partitioning followed by fihration chromatography withSephadexG一100,an intra. cellular仅-galactosidase fromRhizopus sp.A01 grown on soya bean dregs broth was purified to homogeneity with a54.8一fold increase in specific activity and27.3%recovery.The relative molecular weight of the enzyme was about302 ku through gel filtration and85.6 ku throughSDS—polyacrylamide gel electrophoresis,suggesting that the native enzyme was a tetramer.The仅一galactosidase showed high activity againstP—nitrophenyl—d—d-galactopyranoside(pNP-Gal)but had little activity for melibiose and raffinose,and the optimal activity was observed at pH4.5 and55 oC.The kinetic parameters ofK。and知cal/K。were0.2424-0.027 mmol/L and4.089×105L/(mol-S)with pNPGal, and the rates of hydrolysis for melibiose and raffinose were138.3斗m01./(h·mg)and19.7 p。mol/(h·mg),respec— tively.The enzyme activity was significantly activated byFe2+andFe3+,but strongly inhibited byMn2+,Cu2+,Hg+ andMg“at5.0 mmol/L.The0【一galactosidase was highly stable over pH range from4.0 to8.2 at25℃.and it re. tained approximately48%of the original activity after incubation for90min at50。C.Key words raffinose,melibiose,d—galactosidase,three—phase partitioning申画食岛麟重些蠕窥院猢匠康而蛋囟功能艨重程拔术研窥中心镞批准感查北京市科委正式公布的“2011年度认定北京市重点实验室和北京市S-程技术研究中心名单”中,以中国食品发酵工业研究院为依托单位申报的“北京市蛋白功能肽工程技术研究中心”名列其中。北京市蛋白资源与功能肽工程技术研究中心由中国食品发酵工业研究院与北京同仁堂健康药业股份有限公司、北京中食海氏生物技术有限公司共同建立,旨在构建面向农产品加工业、食品工业及传统医药等行业的天然蛋白资源的深度加工、高值利用技术研发平台,涵盖天然蛋白资源综合利用、食源性功能肽研制、体外功能评价、结构鉴定、终端产品开发及工业化生产技术研究,建立技术标准和体外功能评价体系,对于实现低值农林资源的高值化利用,促进我国传统医药、营养保健食品的升级换代具有重要作用。(来源:中国食品发酵工业研究院)垫!呈生差塑鲞差曼翅(璺蔓!!!塑引69
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