期刊文献+

豆豉纤溶酶Subtilisin FS33的提取纯化与鉴定 被引量:7

Preparation and identification of a novel fibrinolytic enzyme FS33 from Bacillus subtilis DC_(33)
下载PDF
导出
摘要 探讨了凝胶层析法提取纯化新型纤溶酶Subtilisin FS33的方法。芽胞杆菌(Bacillus subtilis)DC33发酵液或固态豆豉抽提液在30%~65%硫酸铵饱和度下分段盐析沉淀,再经DEAE-Sepharose FF、Phenyl Sepharose 6FF、Sephadex G-50连续凝胶层析纯化,得到电泳纯纤溶酶Subtilisin FS33,其最终纯化倍数和回收率分别达到34.6倍和13.0%,酶蛋白比活力达到15495U/mg。该酶在还原和非还原条件下SDS-PAGE电泳分析均呈现单一条带,分子量为30kDa;活性电泳FS33凝胶条带在纤维蛋白平板上也表现出强烈纤溶活性。 The method of purifying a novel fibrinolytic enzyme subtilisin FS33 by gel chromography was investigated in this paper. The crude enzyme produced by Bacillus subtilis DC33 was precipitated by 30%-65% saturation of (NH4)2SO4, and the protein collected by centrifugation (10000xg, 15min) was applied to chromatographic procedures including hydrophobic chromatography, anion exchange and gel filtration chromatography. The finally eluted proteins by Sephadex G-50 gel filtration were subjected to SDS-PAGE. Under these conditions, the purified rate and the recovery of final enzyme were 36.6 fold and 13.0%, respectively. The specific activity of the fibrinolytic enzyme was 15495U/mg, and only one band was observed in the purified sample under reducing or non-reducing conditions. The molecular mass was estimated to be approximately 30kDa. The higher fibrinolytic activities of the purified enzyme band were observed on the plasminogen-fxee fibrin plate.
出处 《中国酿造》 CAS 北大核心 2009年第2期29-33,共5页 China Brewing
基金 国家重点基础研究发展计划973项目(2007CB707802)
关键词 纤溶酶 芽胞杆菌DC33 凝胶层析 电泳 纯化 纤溶活性 fibrinolytic subtilisin Bacillus sublilis chromatography electrophoresis purification fibrinolytic activity
  • 相关文献

参考文献18

  • 1赵友春,赵淑梅,王革,张长铠.第三代溶血栓药物研究进展[J].药学进展,2004,28(2):72-75. 被引量:25
  • 2NAKAMURA T, YAMAGATA Y, ICHISHIMA E. Nucleotide sequence of the subtilisin NAT gene, aprN, of Bacillus subtilis (natto) [J] . Biosei Biotech Bioch, 1992,56 (11) : 1869-1871.
  • 3QIU D, OSHIMA H, OHASHI Y. Construction of physical maps of Bacillus subtilis (natto) strains [J]. Nucleic Acids Res Suppl, 2003,3: 207 -208.
  • 4FUJITA M, NOMURRA K, HONG K, et al. Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, a popular soybean fermented food in Japan [J]. Bioehem Biophy Res Co, 1993, 197: 1340-1347.
  • 5FUJITA M, HOMG K, ITO Y, et al. Thrombolytic effect of nattokinase on a chemically induced thrombosis model in rat [J]. Biol Pharm Bull, 1995a, 18: 1387-1391.
  • 6段智变,江汉湖,张书霞,江晓,董名盛,赵晓燕.纳豆激酶粗制液对家兔溶血栓作用及其机制研究[J].营养学报,2003,25(1):46-51. 被引量:30
  • 7KIM W, CHOI KIM Y. Purification and characterization of a fibrinolytic enzyme produced from Bacillus sp. Strain CKll-4 screened from Chungkook-Jang [J]. Appl Environ Microb, 1996, 62: 2482-2488.
  • 8KO J H, YAN J P, ZHU L, et al. Identification of two novel fibrinolytic enzymes from Bacillus subtilis QK-2 [J]. Comp Biochem Phys C, 2004, 137: 65-74.
  • 9王敏,王骏,邵明远,王敏,王以光.链霉菌产生的新型纤溶酶的纤溶性质和溶栓作用[J].药学学报,1998,33(7):481-485. 被引量:18
  • 10WANG C T, JIB P, LI B, et al. Purification and characterization of a fibrinolytic enzyme of Bacillus subtilis DC33, isolated from Chinese traditional Douchi [J]. J Ind Mierobiol Blot, 2006,33: 750-758.

二级参考文献17

  • 1崔立斌 马清俊.vt-PA缺失突变体K2P t-PA的基因工程研究[A]..全军生物化学与分子生物学研究进展[C].,1998.p104.
  • 2刘永军.基因工程产品reteplase[OL].http://www.hx863.com.,2002-07-23.
  • 3上海新生源医药研究有限公司.组织型纤溶酶原激活物突变体的生产方法[P].中国发明专利,申请号:01145280.3.2002.
  • 4叶林柏 郜金荣.重建人t-PA及表达重建人t-PA蛋白的工程菌株[P].CP0015972.0.2001-01-24.
  • 5黄建英 牛瑞芳.细胞工程法制备t-PA及其应用[J].科学技术成果公报,1993,5:75-75.
  • 6国家药品监督管理局注册司.关于已批准临床研究的新药品种公告(第43号).药监注函[2002]113号[Z].,2002-4-22..
  • 7江苏吴中实业股份有限公司.2002年第3季度报告[N].上海证券报,2002-10-23.
  • 8刘城.[D].南京农业大学食品科技学院,2000.
  • 9周元聪,生物化学与生物物理学报,1988年,20卷,35页
  • 10路英华,生物化学杂志,1988年,4卷,166页

共引文献69

同被引文献132

引证文献7

二级引证文献23

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部