期刊文献+

DEAE-Sepharose Fast Flow分离纯化刺芹侧耳木质素降解酶 被引量:4

Purification of Ligninolytic Enzymes from Pleurotus eryngii by DEAE-Sepharose Fast Flow
下载PDF
导出
摘要 [目的]为Pleurotus eryngii—Co60-7木质素降解酶的分离纯化和综合利用提供试验依据。[方法]采用DEAE—Sepharose^TM Fast Flow离子交换介质,分别考察缓冲液pH值、流速和洗脱方式等对刺芹侧耳木质素降解酶分离纯化的影响,确定了最佳分离纯化层析条件。[结果]DEAE-Sephalose^TM Fast Flow分离纯化Pleurotus eryngii-Co60-7木质素降解酶的最佳层析条件为:选择20mmol/L,pH值为5.0醋酸钠一醋酸缓冲体系,3ml/min的流速,进行分步洗脱(100、200~300和1000mmoL/L NaCl的三步洗脱),可较好地实现刺芹侧耳发酵液木质素降解酶初分,该纯化操作目标蛋白回收率达85%,纯化分离因素为2.71。[结论]该技术在分离纯化刺芹侧耳木质素降解酶上可行,具有潜在的工业应用价值。 [ Objective] The aim of the research was to provide the experimental basis for the purification and the comprehensive utilization of the Ligninolytic enzymes from Pleurotus eryngii-Co60-7 strain. [ Method ] Purification of the Ligninolytic enzymes from Pleurotus eryngii-Co60- 7 strain was investigated by the anion-exchange media DEAE-SepharoseTM Fast Flow, by optimizing pH value of the buffer solution, flow rate and elution condition, and its optimal condition of the chromatography was confirmed. [ Results ] The optimal technical condition of DEAE- SepharoseTM Fast Flow for purification of the Ligninolytic enzymes from Pleurotus eryngii-Co60-7 strain was as follows: the purification factor was 2.71 and Ligninolytic enzyme activity recovery was 85% through 20 mmol/L sodium acetate optimal buffer solution, pH value 5.0, 3 ml/min and 100,200 -300 mmol/L, 1 000 mmol/L three-step NaCl elution. [ Conclusion] The technique of separating the Ligninolytic enzymes from Pleurotus eryngii-Co60-7 strain feasible and have potential applied value in dustry.
出处 《安徽农业科学》 CAS 北大核心 2009年第2期448-450,共3页 Journal of Anhui Agricultural Sciences
基金 浙江省科技厅新苗计划(2007R40G2050021) 浙江省自然科学基金资助(Y505334)
关键词 PLEUROTUS eryngii-Co60-7 木质素降解酶 离子交换层析 DEAE-SepharoseTM FAST FLOW Pleurotus eryngii -Co60-7 Ligninolytic enzymes Ion exchange chromatography DEAE-Sepharose^TM Fast Flow
  • 相关文献

参考文献22

  • 1M·霍夫利特,A·斯泰因比谢尔.生物高分子第1卷:木质素、腐殖质和煤[M].北京:化学工业出版社,2004:141-225.
  • 2MUNOZ C,GUILLEN F,MARTINEZ A T,et al. Laccase isoenzymes of Pleurotus eryngii:characterization, catalytic properties, and participation in activation of molecular oxygen and Mn^2+ oxidation[J]. Applied and Environmental Microbiology, 1997,6:2166 - 2174.
  • 3PEREZ - BOADA M, DOYLE W A, RUIZ-DUENAS F J, et al. Expression of Pleurotus eryngii versatile peroxidase in Escherichia coli and optimization of in vitro folding[J]. Enzyme and Microbial Technology. ,2002,30:518 -524.
  • 4LUCILIA CARAMELO,MARIA JESU S MARTINEZ,ANGEL T MARTINEZ. A search for ligninolytic peroxidases in the fungus Pleurotus eryngii involving a-keto-g-thiomethylbutyric acid and lignin model dimers[ J]. American Society for Microbiology,1999,65(3) :916-922.
  • 5HAKUHNEN N,ANDBERG M,KALLIO J,et al. A near atomic resolution structure of a Melanocarpus albomyces laccase[J]. Journal of Structural Biology ,2008,162:39.
  • 6VARELA E ,JESUS MARTINEZ M ,MARTINEZ A T. Martinez. Aryl-alcohol oxidase protein sequence:a comparison with glucose oxidase and other FAD oxidoreductases [ J ]. Biochimica et Biophysica Acta, 2000,1481 ( 1 ) : 202-208.
  • 7ANGEL T MARTINEZ. Molecular biology and structure-function of lignindegrading heine peroxidases[J]. Enzyme and Microbial Technology,2002, 30:425 -444.
  • 8SUSANA CAMARERO,GUIDO C GALLETTI, ANGEL T MARTINEZI. Preferential degradation of phenolic lignin units by two white rot fungi [ J]. American Society for Microbiology,1994,60(12) :4509 -4516.
  • 9TUNDE MESTER,KATIA AMBERT-BALAY,SIMONE CIOFI-BAFFONI ,et al. Oxidation of a tetrameric nonphenolic lignin model compound by lignin peroxidase [ J ]. American Society for Biochemistry and Molecular Biology ,2001,276 (25) :22985 - 22990.
  • 10RUIZ-DUENAS F J, MARTINEZ M J, MARTINEZ A T. Heterologous expression of Pleurotus eryngii peroxidase confirms its ability to oxidize Mn2. and different aromatic substrates[J]. American Society for Microbiology,1999,65 (10) :4705 -4707.

二级参考文献27

  • 1李红梅,梅乐和,URLACHER VLADA,SCHMID ROLF D.催化吲哚生成靛蓝的细胞色素P450BM-3定向进化研究[J].生物化学与生物物理进展,2005,32(7):630-635. 被引量:14
  • 2黄俊,梅乐和,胡升,盛清,许静,吴晖,姚善泾.纳豆中纳豆枯草杆菌的筛选和纳豆激酶的分离过程研究[J].高校化学工程学报,2005,19(4):518-522. 被引量:20
  • 3付利,杨志兴.纳豆激酶的研究与应用[J].生物工程进展,1995,15(5):46-49. 被引量:83
  • 4Sherry S. Recombinant tissue plasminogen activator(rt-PA): is it the thrombolytic agent of choice for an evolving acute myocardial infarction [J]. Am J Cardiol, 1987, 59(9): 984-989.
  • 5Sumi H, Hamada H, Tsushima H, et al. A novel fibrinolytic enzyme (nattokinase) in the vegetable cheese Natto; a typical and popular soybean food in the Japanese diet [J]. Experientia, 1987, 43(10): 1110-1111.
  • 6Sumi H, Hamada H, Nakanishi K, et al. Enhancement of the fibrinolytic activity in plasma by oral administration of nattokinase [J],Acta Haematol, 1990, 84(3): 139-143.
  • 7Bradford M M, A rapid and sensitive method for the quantitation of microgramquantities of protein utilizing the principle of protein-dye binding [J]. Aaml Biochem, 1976, 72(1-2): 248-254,.
  • 8Asturp T, Mttllertz S, The fibrin plate method for estimating fibrinolytic activity [J], Biochem Biophys, 1952, 40(2): 346-351.
  • 9Fujita M, Nomura K, Hong K, et al. Purification and characterization of a strong fibrinolytic enzyme (nattokinase) in the vegetable cheese natto, apopular soybean fermented food in Japan [J]. Biochem Biolahys Res Commun, 1993, 197(3): 1340-1347.
  • 10Omura T, Ishimura Y, Fujii K Y. CytochromeP450[M]. Kodanasha, VCH Tokyo,1993.

共引文献26

同被引文献54

引证文献4

二级引证文献16

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