摘要
为分离纯化金银花过氧化物酶,对其酶学性质及抑制动力学进行研究,将三相法纯化所得金银花过氧化物酶,经DEAE纤维素DE-52离子交换层析分离,得到2种金银花过氧化物酶HSPⅠ和HSPⅡ,其比活力分别为3 312.3,564.8U/mg。HSPⅠ反应动力学分析表明金银花过氧化物酶的双底物酶促反应类型为乒乓机制。抑制动力学研究表明柠檬酸、偏重亚硫酸钠对金银花过氧化物酶的抑制类型为不可逆抑制。L-半胱氨酸对金银花过氧化物酶的抑制类型为可逆抑制,可逆抑制类型为竞争性可逆抑制。L-半胱氨酸抑制常数KI为0.053mmol/L。
In order to purify honeysuckle peroxides and study the enzymatic properties and inhibition kinetics,DEAE cellulose DE-52 ion exchange chromatography was used for purification of peroxidase from Lonicera Japonica Thunb prepared preliminarily by three-phase partitioning.Two kinds of peroxidase, HSP Ⅰ（honeysuckle peroxidaseⅠ）and HSP Ⅱ（honeysuckle peroxidase Ⅱ）, were purified,the specific activity were 3 312.3U/mg and 564.8U/mg respectively.The double substrate enzymatic reaction type was PingPong reaction analyzed by the reaction kinetics.Citric acid,Sodium metabisulfite showed irreversible inhibition to Lonicera Japonica Thunb peroxidase,L-cysteine showed reversible inhibition,and L-cysteine was a competitive inhibitor that the inhibition constant KI was 0.053mmol/L.
出处
《食品与机械》
CSCD
北大核心
2017年第7期111-116,共6页
Food and Machinery
基金
国家自然科学基金联合基金项目(编号:U1304330)
河南省高校科技创新团队计划(编号:17IRTSTHN016)
关键词
金银花
过氧化物酶
纯化
抑制动力学
Lonicera Japonica Thunb
peroxidase
purification
Inhibition kinetics
