摘要
为了研究表达在蛋清中的重组人组织激肽释放酶（rhK1）是否与天然组织激肽释放酶具有相似的理化特性，对鸡输卵管暂态生物反应器表达的rhK1与猪胰腺组织激肽释放酶（pK）的理化特性进行比较鉴定．Western blotting检测结果显示，表达在蛋清中的rhK1能被人组织激肽释放酶（hK1）特异抗体识别，与pK无交叉反应，rhK1酶原和成熟酶的分子量约为43kDa和37kDa．理化鉴定结果显示，rhK1在pH7～10的范围内活性稳定，而pK在pH8～9的范围内活性较稳定；rhK1和pK均能被Cu^2＋、Fe^3＋和Al^3＋完全抑制；rhK1和pK的热稳定性相似，但rhK1在37℃孵育20min后被显著激活、以上试验结果表明，表达在蛋清中的rhK1分子量正确，理化特性与猪的天然酶相似，具有开发利用价值．
To study the similarity between recombinant human tissue kallikrein （rhK1） and natural kallikrein, the physical and chemical properties of rhK1 expressed in chicken oviduct transient bioreactor and porcine pancreatic kallikrein （pK） were compared. Western blotting analysis showed the rhK1 was recognized specifically by hK1 antibody, and two bands with molecular weights of 37 kDa and 43 kDa were observed, corresponding to the mature- and pre-enzyme respectively, but pK failed to react with the antibody. Biochemical studies showed that the rhK1 was stable from pH 7 to pH 10, while pK was stable from pH 8 to pH 9, the activity of rhK1 and pK was inhibited completely by Cu^2＋ , Fe^3＋ and Al^3＋ . The rhK1 and pK showed similar thermo-stability, but rhK1 was activated significantly after 20 minutes with 37 ℃ incubation. These data indicated that the molecular weight of rhK1 was right, rhK1 and pK had similar thermo-stability, optimal pH value and sensitivity to ions, and the chicken oviduct-specific transient expression system can be used to produce commercial recombinant proteins.
出处
《浙江大学学报（农业与生命科学版）》
CAS
CSCD
北大核心
2008年第4期374-378,共5页
Journal of Zhejiang University:Agriculture and Life Sciences
基金
江苏省教育厅科研产业化资助项目(JH01-066)
关键词
鸡输卵管暂态生物反应器
重组人组织激肽释放酶
猪胰腺组织激肽释放酶
理化特性
chicken oviduct transient bioreactor
recombinant human tissue kallikrein
porcine pancreatic kallikrein
physical and chemical properties
