|本期目录/Table of Contents|

[1]朱年青,夏文静,勇强.里氏木霉纤维素酶的分离纯化及酶学性质[J].生物加工过程,2010,8(03):40-43.[doi:10.3969/j.issn.1672-3678.2010.03.008]
　ZHU Nian-qing,XIA Wen-jing,YONG Qiang.Purification and characterization of cellulase from Trichoderma reesei[J].Chinese Journal of Bioprocess Engineering,2010,8(03):40-43.[doi:10.3969/j.issn.1672-3678.2010.03.008]
点击复制

里氏木霉纤维素酶的分离纯化及酶学性质
(/HTML)
分享到：

《生物加工过程》[ISSN:1672-3678/CN:32-1706/Q]

卷:
8
期数:
2010年03期
页码:
40-43
栏目:
出版日期:
2010-05-30

文章信息/Info

Title:
Purification and characterization of cellulase from Trichoderma reesei
文章编号:
1672-3678(2010)03-0040-04
作者:
朱年青; 夏文静; 勇强
南京林业大学 林木遗传与生物技术省部共建教育部重点实验室，南京210037
Author(s):
ZHU Nian-qing;  XIA Wen-jing; YONG Qiang
Key Laboratory of Forest Genetics ＆ Biotechnology,Nanjing Foresty University, Nanjing 210037, China
关键词:
里氏木霉; 纤维素酶; 酶学性质; 分离纯化
分类号:
Q814
DOI:
10.3969/j.issn.1672-3678.2010.03.008
文献标志码:
A
摘要:
通过(NH4)2SO4分级沉淀、HiPrep 26/10 Desalting凝胶色谱脱盐、Source 15 Q阴离子交换色谱技术，里氏木霉（Rut C-30）纤维素酶主要组分得以初步分开，再经过Source 15 S阳离子交换色谱、HiPrep Sephacryl S-100 HR凝胶过滤色谱、Superdex 75 PrepGrade凝胶过滤色谱进一步分离纯化，得到2个纯化的内切葡聚糖酶组分EGⅡ、EGⅠ和一个外切葡聚糖酶组分CBHⅠ；经过SDS-PAGE电泳鉴定为电泳纯，测得相对分子质量分别为5.22×104，5.62×104和6.90×104。EGⅡ的最适反应pH是5.6，最适反应温度为65 ℃；EGⅠ的最适反应pH是4.4，最适反应温度为55 ℃；以羧甲基纤维素(CMC)为底物时，EGⅠ、EGⅡ的米氏常数(Km )分别为2.20 mg/mL、3.38 mg/mL。CBHⅠ的最适反应pH是5.8，最适反应温度为60 ℃，以对硝基苯基β-D-纤维二糖苷(PNPC)为底物时，米氏常数(Km )为0.12 mg/mL。

参考文献/References:

［1］Zhang Y H，Michael E H，Jonathan R M.Outlook for cellulase improvement:screening and selection strategies［J］.Biotechnology Advances，2006，24:452-481.
［2］Johan K，Matti S A,Maija T, et al.Enzymatic properties of the low molecular mass endoglucanases Cel12A(EGⅢ) and Cel45A (EGV) of Trichoderma reesei［J］. Biotech，2002，99:63-78.
［3］瞿丽莉，朱均均，刘敏,等.β葡萄糖苷酶的制备及在纤维素辅助水解上的应用研究［J］.林产化学与工业，2009，29(1):13-17.
Qu Lili，Zhu Junjun，Liu Min，et al.Study on preparation of β-glucosidase and its application in enzymatic hydrolysis of cellulose［J］.Chem Ind Forest Products，2009，29(1):13-17.
［4］曹健，郭德宪.里氏木霉纤维素酶的纯化和性质［J］.食品科学，2003,24(5):72-75.
Cao Jian，Guo Dexian.Study on purification and characteristics of Trichoderma reesei cellulase［J］.Food Chem，2003,24(5):72-75.
［5］江华.两级超滤部分纯化里氏木霉纤维素酶［J］.纤维素科学与技术，2007,15(4):9-15.
Jiang Hua.Partial purification of cellulases from Trichoderma reesei by a two-stage ultrafiltration［J］.J Sci Cellulose Tech，2007,15(4):9-15.
［6］Bardford M.A rapid and sensitive method for the quantitation of microgram quantities of protein uitilizing the principle of protein-dye binding［J］.Anal Biochem，1976，72(1/2):248-253.
［7］朱年青.里氏木霉纤维素酶的分离纯化及应用的研究［D］.南京：南京林业大学，2008.
［8］阎伯旭,高培基.外切葡聚糖纤维二糖水解酶的分离纯化和部分性质研究［J］.生物化学杂志，1997,13(3):362-364.
Yan Boxu，Gao Peiji.Purification of two cellobiohydrolases from Trichodema pseudokiningii S-38［J］.J Chem，1997,13(3):362-364.
［9］Medve J，Lee D，Tjerneldj F，et al.Ion-exchange chromatographic purification and quantitative analysis of Trichoderma reesei cellulases cellobiohydrolase I，II and endoglucanase II by fast protein liquid chromatography［J］.J Chromat A,1998，808(1/2):153-165.
［10］Gusakov A V，Salanovich T N，Antonov A I，et al.Design of highly efficient cellulase mixtures for enzymatic hydrolysis of cellulose［J］.Biotech Bioeng,2007，97:1028-1038.

相似文献/References:

[1]宋娜娜,宋向阳,欧阳嘉,等.里氏木霉与黑曲霉混合发酵产纤维素酶及其水解特性[J].生物加工过程,2010,8(05):5.[doi:doi:10.3969/j.issn.1672-3678.2010.05.002]
　SONG Na-na,SONG Xiang-yang,OUYANG Jia,et al.Cellulose production from fungal mixed culture of Trichoderma reesei RUT C-30 and Apsergillus niger NL02 on steam-exploded corn stover and characteristics of hydrolysis[J].Chinese Journal of Bioprocess Engineering,2010,8(03):5.[doi:doi:10.3969/j.issn.1672-3678.2010.05.002]
[2]韩立荣,祝传书,孙平平,等.青霉F10-2固态发酵植物农药残渣产纤维素酶[J].生物加工过程,2010,8(05):11.[doi:doi:10.3969/j.issn.1672-3678.2010.05.003]
　HAN Li-rong,ZHU Chuan-shu,SUN Ping-ping,et al.Solid state fermentation for producing cellulase by Penicillum waksmanii F10-2 by using plant pestcide residue[J].Chinese Journal of Bioprocess Engineering,2010,8(03):11.[doi:doi:10.3969/j.issn.1672-3678.2010.05.003]
[3]于莹,姚日生,齐本坤,等.绿色木霉ZY-1固态发酵产纤维素酶[J].生物加工过程,2009,7(01):54.
　YU Ying,YAO Ri-sheng,QI Ben-kun,et al.Cellulase production by Trichoderma viride ZY-1 under solid-state fermentation[J].Chinese Journal of Bioprocess Engineering,2009,7(03):54.
[4]许云峰,周建芹,陈晶磊,等.内生青霉菌纤维素酶辅助提取槐米总黄酮[J].生物加工过程,2009,7(02):18.
　XU Yun-feng,ZHOU Jian-qin,CHEN Jing-lei,et al.Application of cellulase from endophytic Penicillium sp.B-4 on total flavonoid extraction from flower buds of Sophara japonica[J].Chinese Journal of Bioprocess Engineering,2009,7(03):18.
[5]乐易林,邵蔚蓝.极端耐热纤维素酶Cel74的表达、纯化与特性[J].生物加工过程,2009,7(03):64.
　LE Yi-lin,SHAO Wei-lan,Expression and characterization of Thermotoga maritima cellulase Cel74[J].Chinese Journal of Bioprocess Engineering,2009,7(03):64.
[6]梁娟然,戴菲辉,苏思莹,等.纤维素酶在二甲基亚砜微扰条件下的动力学行为和光谱学变化[J].生物加工过程,2011,9(02):29.[doi:doi:10.3969/j.issn.1672-3678.2011.02.006]
　LIANG Juanran,DAI Feihui,SU Siying,et al.Kinetics and spectrum changes of cellulase in dimethyl sulfoxide solution[J].Chinese Journal of Bioprocess Engineering,2011,9(03):29.[doi:doi:10.3969/j.issn.1672-3678.2011.02.006]
[7]李辉,王义强,陈介南,等.工业纤维素诱导里氏木霉RUT C-30产葡聚糖酶的条件[J].生物加工过程,2011,9(03):22.[doi:10.3969/j.issn.1672-3678.2011.03.005]
　LI Hui,WANG Yiqiang,CHEN Jienan,et al.Conditions of glucanase production for Trichoderma reesei RUT C-30[J].Chinese Journal of Bioprocess Engineering,2011,9(03):22.[doi:10.3969/j.issn.1672-3678.2011.03.005]
[8]钟耀华,钱远超,任美斌,等.丝状真菌降解转化纤维素的机制与遗传改良前景[J].生物加工过程,2014,12(01):46.[doi:10.3969/j.issn.1672-3678.2014.01.008]
　ZHONG Yaohua,QIAN Yuanchao,REN Meibin,et al.Progress in bioconversion of cellulose by filamentous fungi and related genetic engineering strategies[J].Chinese Journal of Bioprocess Engineering,2014,12(03):46.[doi:10.3969/j.issn.1672-3678.2014.01.008]
[9]欧阳嘉,李鑫,王向明,等.纤维素结合域的研究进展[J].生物加工过程,2008,6(02):10.[doi:10.3969/j.issn.1672-3678.2008.02.002]
　OUYANG Jia,LI Xin,WANG Xiang-ming,et al.Advances in cellulose-binding module[J].Chinese Journal of Bioprocess Engineering,2008,6(03):10.[doi:10.3969/j.issn.1672-3678.2008.02.002]
[10]杨晓辉,余晓斌.碱与微波联合处理蔗渣对里氏木霉发酵产纤维素酶的影响[J].生物加工过程,2008,6(02):61.[doi:10.3969/j.issn.1672-3678.2008.02.012]
　YANG Xiao-hui,YU Xiao-bin.Cellulase production by Trichoderma reesei from bagasse pretreated by alkali and microwave[J].Chinese Journal of Bioprocess Engineering,2008,6(03):61.[doi:10.3969/j.issn.1672-3678.2008.02.012]
[11]连之娜,李鑫,欧阳嘉,等.2种纤维素酶水解效果比较及酶蛋白组分分析[J].生物加工过程,2010,8(06):57.[doi:10.3969/j.issn.1672-3678.2010.06.011]
　LIAN Zhi-na,LI Xin,OUYANG Jia,et al.Comparison between two kinds of cellulases in enzymatic hydrolysis and their secreted proteins analysis[J].Chinese Journal of Bioprocess Engineering,2010,8(03):57.[doi:10.3969/j.issn.1672-3678.2010.06.011]
[12]祖彩霞,李志敏,叶勤.里氏木霉Trichoderma reesei产纤维素酶的发酵培养基碳氮源优化[J].生物加工过程,2011,9(04):6.[doi:10.3969/j.issn.1672-3678.2011.04.002]
　ZU Caixia,LI Zhimin,YE Qin.Culture medium optimization for producing cellulase by Trichoderma reesei[J].Chinese Journal of Bioprocess Engineering,2011,9(03):6.[doi:10.3969/j.issn.1672-3678.2011.04.002]

备注/Memo

备注/Memo:
收稿日期：2009-04-16
基金项目：国家自然科学基金资助项目(30471361)；江苏省高校自然科学重大基础研究项目(06KJ22015)
作者简介：朱年青（1983—），男， 江苏泰州人， 博士研究生，研究方向：蛋白质分离纯化；勇强（联系人），教授，E-mail：swhx@njfu.edu.cn
更新日期/Last Update: