期刊文献+
任意字段
题名或关键词
题名
关键词
文摘
作者
第一作者
机构
刊名
分类号
参考文献
作者简介
基金资助
栏目信息

巨大芽孢杆菌产胞外核糖核酸酶的分离纯化及部分酶学性质研究 被引量：4

Purification and Partial Characterization of Extracellular Ribonuclease from Bacillus megaterium
在线阅读 下载PDF
导出
摘要 巨大芽孢杆菌发酵液经硫酸铵沉淀、CM-Sepharose离子交换层析和Superdex-200凝胶过滤层析,纯化得到核糖核酸酶电泳纯品,并对其性质进行研究。该酶比活力为54272.27U/mg,回收率为11.37%,纯化倍数为606.67倍。该酶分子质量约为33.3kD,最适温度为52.5℃,最适pH值为8.5,在20～40℃以及pH6.0～7.0范围内稳定性较好。Fe2+、Cu2+、SDS、抗坏血酸和草酸对该酶活性有抑制作用。 After amonanium sulfate precipitation followed by ion-exchange chromatography on CM-Sepharose column and Superdex-200 gel-filtration, ribonuclease from Bacillus megaterium was purified to electrophoretic homogeneity. Some of its enzymological properties were then explored. The results showed that the enzyme was 606.67-fold purified with specific activity of 54272.27 U/mg and recovery rate of 11.37%. The molecular weight was 33.3 kD. Optimum activity of the enzyme was achieved at 52.5 ℃ and pH 8.5. The enzyme displayed good stability at 20-40 ℃ and pH 6.0-7.0. The activity of this enzyme was inhibited by Fe^2＋, Cu^2＋, SDS, ascorbic acid and oxalic acid.
作者 赵芯 成丽丽 邓玉 敬海明 唐云明
机构地区 西南大学生命科学学院
出处 《食品科学》 EI CAS CSCD 北大核心 2012年第19期254-258,共5页 Food Science
基金 重庆市科学技术委员会重点攻关项目(CSTC,2011AB1027)
关键词 巨大芽孢杆菌 核糖核酸酶 分离纯化 性质 Bacillus megaterium ribonuclease isolation and purification characterization
分类号 Q556.1 [生物学—生物化学]
  • 相关文献

参考文献23

  • 1张颖,李伟军,苏志国.核糖核酸酶特殊生物学功能及药理作用研究新进展[J].生物医学工程学杂志,2001,18(3):456-460. 被引量：10
  • 2REDDY L G, HANKAR V. Immobilized nucleases[J]. Crit Rev Biotechnol, 1993, 13(3): 255-273.
  • 3LINDBLOM M, MOGREN H. Enzymatic RNA reduction in disintegrated cells of Saccharomyces cerevisiae[J]. Biotechnol Bioeng, 1974, 16(8): 1123-1133.
  • 4KOLPAKOV A I, KUPRIIANOVA-ASHINA F G, LESHCHINSKAIA I B. Dependence of the effect of RNAase from Bacillus intermedius on the growth of baker's yeast on the concentration of the exogenous enzyme[J]. Mikrobiologiia, 2000, 69(4): 478-482.
  • 5SKVORTSOVA M A, BOCHAROV A L, YAKOVLEV G I, et al. Novel extracellular ribonuclease from Bacillus intermedius-binase II : purification and some properties of the enzyme[J]. Biochemistry (Mose), 2002, 67(7): 802-806.
  • 6GUAN G P, WANG H X, NG T B. A novel ribonuclease with antiproliferative activity from fresh fruiting bodies of the edible mushroom Hypsizigus marmoreus[J]. Biochim Biophys Acta, 2007, 1770(12): 1593-1597.
  • 7LAM S K, NG T B. Isolation of a novel thermolabile heterodimeric ribonuclease with antifun gal and antiproliferative activities from roots of the sanchi ginseng (Panax notoginseng)[J]. Biochem Biophys Res Commun, 2001, 285(2): 419-423.
  • 8ILINSKAYA O, DECKER K, KOSCHINSKI A, et al. Bacillus intermedius ribonuclease as inhibitor of cell proliferation and membrane current[J]. Toxicology, 2001, 156(2/3): 101-107.
  • 9牟琳,王红宁,邹立扣.巨大芽孢杆菌表达外源蛋白的特点及其研究进展[J].中国生物工程杂志,2008,28(4):93-97. 被引量：13
  • 10ZNAMENSKAYA L V, VERSHININA O A, VERSHININA V I, et al. Expression of the genes for guanyl-specific ribonucleases fromBacillus intermedius and Bacillus pumilus is regulated by the two component signal transduction system PhoP-PhoR in B. subtilis[J]. FEMS Microbiol Lett, 1999. 173(1): 217-222.

二级参考文献47

  • 1Beck A K, Pass H I, Carbone M, et al. Ranpimase as a potential antitumor ribonuclease treatment for mesothelioma and other malignancies. Future Oncol, 2008, 4(3) :341-349.
  • 2Matousek J. Ribonucleases and their antitumor activity. Corn PBiochem Physiol C Toxicol Pharmacol, 2001,129 ( 3 ) : 175- 191.
  • 3Dickson K A, Haigis M C, Raines R T. Ribonuclease inhibitor: structure and function. Prog Nucleic Acid Res Mol Biol, 2005, 80:349-374.
  • 4Ardelt W, Mikulski S M, Shogen K. Amino acid sequence of an anti-tumor protein from Rana pipiens oocytes and early embryos. Homology to pancreatic ribonucleases. J Biol Chem, 1991,266 ( 1 ) :245-251.
  • 5Rutkoski T J, Raines R T. Evasion of ribonuclease inhibitor as a determinant of ribonuclease cytotoxicity. Curr Pharm Biotechnol, 2008 ,9(3) :185-189.
  • 6Ardeh W, Shogen K, Darzynkiewicz Z. Onconase and amphinase, the antitumor ribonucleases from Rana pipiens oocytes. Curr Pharm Biotechnol, 2008, 9(3) :215-225.
  • 7Liao Y D. A pyrimidine-guanine sequence-specific ribonuclease from Rana catesbeiana (bullfrog) oocytes. Nucleic Acids Res, 1992, 20 (6) :1371-1377.
  • 8Kamiya Y, Oyama F, Oyama R, et al. Amino acid sequence of a lectin from Japanese frog ( Rana japonica ) eggs. J Biochem, 1990,108 (1) :139-143.
  • 9Bradford M M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of proteindye binding. Anal Biochem , 1976, 72: 248-254.
  • 10Rosenberg H F. Recombinant human eosinophil cationic protein. Ribonuclease activity is not essential for cytotoxicity. J Biol Chem, 1995, 270(14) :7876-7881.

共引文献33

同被引文献61

  • 1孟雅,李刚,崔焱,张龙.马铃薯多酚氧化酶的提取纯化条件对其活性影响的研究[J].化学与生物工程,2006,23(10):47-49. 被引量：20
  • 2李君兰,李怡华,赵秋玲,刘志芳,冯九海.鸡腿蘑多酚氧化酶特性研究[J].食品科学,2007,28(1):187-191. 被引量：23
  • 3布坎南RE 吉本斯NE.伯杰细菌鉴定手册[M].北京：科学技术出版社,1984.1228-1271.
  • 4Yu MY, Xu XY, Qing Y, et al. Isolation of an anti-tumor polysaccharide from Auricularia polytricha (jew's ear) and its effects on macrophage activation [ J ]. Eur Food Res Technol, 2009, 228(3 ) : 477-485.
  • 5Niu CP, Cai MH, Zhang YX, et al. Biosynthetic origin of the carbon skeleton of a novel anti-tumor compound, haloroqui- none, from a marine-derived fungus, Halorosellinia sp [J]. Biotechnol Lett, 2012, 34(11 ): 2119-2124.
  • 6Kamasuka T, Momoki Y, Sakai S. Antitumor, activity of polysaccharide fractions prepared from some strains of Basidiomycetes [ J ]. Gann, 1968, 59 (5) : 443-445.
  • 7Lee JS, Wittchen KD, Stahl C, et al. Cloning, expression, and carbon catabolite repression of the bamM gene encoding β- amylase of Bacillus megatefium DSM319 [J]. Appl Microbiol Biotechnol, 2001, 56(1-2) : 205-211.
  • 8Orhan E, Omay D, Gtivenilir Y. Partial purification and characterization of protease enzyme from Bacillus subtilis and Bacillus cereus [J]. Appl Biochem Biotechnol, 2005 (121-124): 183-194.
  • 9Kim SS, Kim YJ, Rhee IK. Purification and characterization of a novel extracellular protease from Bacillus cereus KCTC 3674 [J]. Arch Microbiol, 2001, 175(6): 458-461.
  • 10Sivapathasekaran C, Das P, Mukherjee S, et ol. Marine bacterium derived lipopeptides: Characterization and cytotoxic activity against cancer cell lines [J ]. Int J Pept Res Tber, 2010, 16 (4): 215-222.

引证文献4

二级引证文献5

食品科学
2012年 第19期

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