泥蚶血红蛋白的制备及其抗菌活性研究  

Purification and antibacterial activity of hemoglobin from Tegillarca granosa

作　　者：王素芳[1,2] 包永波[2] 施淼江 郑丹妮[2] 杨婷婷[1,2] 林志华[2] 

机构地区：[1]宁波大学海洋学院,浙江宁波315211 [2]浙江万里学院浙江省水产种质资源高效利用技术研究重点实验室,浙江宁波315100

出　　处：《海洋学报》2014年第12期67-73,共7页

基　　金：国家863计划项目(2012AA10A410);宁波市自然基金项目(2013A610161);浙江省教育厅高校科研项目(Y201225951);国家级大学生创新创业训练计划项目(201310876018);宁波市海洋贝类科技创新团队(2011B82017);水产科学浙江省重中之重学科开放基金项目

摘　　要：利用凝胶层析技术从泥蚶血细胞裂解物中分离纯化得到两种血红蛋白,回收率为74.3%,并通过Tricine-SDS-PAGE和质谱技术鉴定为Tg-HbⅠ(同源二聚体)和Tg-HbⅡ(异源四聚体)。分别通过琼脂扩散法和分光光度法测其抗菌活性和过氧化物酶活性,结果发现,两种Tg-Hb对大肠杆菌和恶臭假单胞杆菌均有抗菌活性,Tg-HbⅡ对于恶臭假单胞菌和大肠杆菌的MIC值分别为0.063mg/mL和0.13mg/mL,Tg-HbⅠ对两种菌的MIC均为0.048mg/mL;两种Tg-Hb均具有过氧化物酶活性,能催化愈创木酚、邻苯二酚、对苯二酚、苯酚、左旋多巴等多种酚类物质。在Tg-Hb中添加GSH后,两种Tg-Hb均不再对恶臭假单胞杆菌具有抗菌活性。可见,两种Tg-Hb对恶臭假单胞杆菌等的抗菌活性均是通过其过氧化物酶活性发挥作用的。Two kinds of hemoglobins were purified by gel chromatography with a recovery 74. 3 %. They were iden- tified as Tg-HbI （homodimer） and Tg-HbⅡ （heterogeneous tetramer） by Tricine-SDS-PAGE and mass spectrome- ter. The agar diffusion method and spectrophotometric method were used to detect their antibacterial activity and peroxidase activity,respectively. Results showed that both hemoglobins exhibited antibacterial activities against Es-chetichia coli and Pseudomonas putida. The MIC values of Tg-HbⅡ towards E. coli and P. putida were 0. 063 mg/mL and 0. 13 mg/mL, while corresponding data of Tg-HbI were 0. 048 mg/mL. Both hemoglobins showed peroxidase activities,which could catalyze various of phenolic substances, such as guaiacol, pyrocatechol, hydroqui- none, phenol,L-DOPA, etc. Both hemoglobins lost their antibacterial activities against P. putida in the presence of GSH,indicating that they played the antibacterial abilities through their peroxidase activities.

关 键 词：泥蚶 血红蛋白 抗菌活性 过氧化物酶活性 

分 类 号：S944.44[农业科学—水产养殖]