[1]叶锦,张静茹,覃益民,等.一种康氏木霉内切β-葡聚糖苷酶的分离纯化及酶学性质[J].江苏农业学报,2015,(05):995-1000.[doi:doi:10.3969/j.issn.1000-4440.2015.05.008]
　YE Jin,ZHANG Jing-ru,QIN Yi-min,et al.Purification of a new endo-β-glucanase from Trichoderma koningii and its characterization[J].,2015,(05):995-1000.[doi:doi:10.3969/j.issn.1000-4440.2015.05.008]
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一种康氏木霉内切β-葡聚糖苷酶的分离纯化及酶学性质()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年05期
页码:
995-1000
栏目:
遗传育种·生理生化
出版日期:
2015-10-31

文章信息/Info

Title:
Purification of a new endo-β-glucanase from Trichoderma koningii and its characterization
作者:
叶锦; 张静茹; 覃益民; 刘幽燕
（广西大学化学化工学院,广西南宁530004）
Author(s):
YE Jin; ZHANG Jing-ru; QIN Yi-min; LIU You-yan
(School of Chemistry﹠Chemical Engineering，Guangxi University,Nanning 530004，China)
关键词:
康氏木霉; 内切β-葡聚糖苷酶; 分离纯化; 酶学性质
Keywords:
Trichodermakoningii; endo-β-glucanase; isolation and purification; enzymatic property
分类号:
Q55
DOI:
doi:10.3969/j.issn.1000-4440.2015.05.008
文献标志码:
A
摘要:
为获得康氏木霉（Trichodermakoningii）内切β-葡聚糖苷酶组分，并研究其酶学性质，通过硫酸铵盐析、DEAE Sepharose FF阴离子交换层析和Sephadex G-75凝胶过滤层析对康氏木霉GIMP3.444纤维素酶主要组分进行分离纯化，SDS-PAGE电泳检测蛋白质纯度和分子量，MALDI-TOFMS鉴定蛋白质种类。结果显示，从康氏木霉所产的纤维素酶系中分离纯化获得一种内切β-葡聚糖苷酶组分，相对分子质量为44 600，酶的比活力为19.65 U/mg。该内切酶的最适反应pH值为4.5，最适反应温度为55 ℃。以羧甲基纤维素钠(CMC-Na)为底物时, 酶的米氏常数(Km)为7.22 mg/ml。不同金属离子对酶活性影响不同，其中Ca2+（0.3 mmol/L）对酶的抑制作用较强，抑制了近35%的活力。通过MALDI-TOFMS鉴定及数据库分析，确定该酶为一种新的内切β-葡聚糖苷酶组分。
Abstract:
To identify the composition of cellulase produced by Trichodermakoningii, ammonium sulfate precipitation, DEAE-sepharose FF anion exchange chromatography and sephadex G-75 gel filtration chromatography were used for purification of the cellulase. The molecular weight and purity were determined by SDS-PAGE, and the protein was identified by MALDI-TOFMS. A new endo-β-glucanase was identified with its molecular weight of 44 600 and the specific activity of 19.65 U/mg. The optimum pH and temperature for the endo-β-glucanase were 4.5 and 55 ℃. The Km value was 7.22 mg/ml (CMC-Na). Ca2+(0.3 mmol/L) could inhibit the enzymatic activity significantly by 35%. The enzymatic properties of the new endo-β-glucanase was distinct from other endo-β-glucanase. It could be used to was analyze the synergetic mechanism of cellulases.

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备注/Memo

备注/Memo:
收稿日期：2015-02-07 基金项目：国家自然科学基金项目（21276053）；广西生物炼制重点实验室培育基地开放课题（GXBF11-04） 作者简介：叶锦（1990-），女，河南信阳人，硕士，研究方向为生物化工。（Tel）15578445453；（E-mail）469157531@qq.com 通讯作者：覃益民,(E-mail) qym6289@sina.com
更新日期/Last Update: 2015-10-31