Production, purification and characterization of a novel thermotolerant endoglucanase (CMCase) from Bacillus strain isolated from cow dung.
Production, purification and characterization of a novel thermotolerant endoglucanase (CMCase) from Bacillus strain isolated from cow dung.
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DOI：
10.1186/2193-1801-2-10
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发表时间：
2013-12
期刊：
影响因子：
--
通讯作者：
Maiti TK
中科院分区：
文献类型：
--
作者：
Sadhu S;Saha P;Sen SK;Mayilraj S;Maiti TK
为了从草食动物粪便中筛选出产纤维素酶的细菌，有可能从牛粪中分离出一种有效的细菌。利用16 S rDNA分子系统发育分析方法，将该菌鉴定为芽孢杆菌（Bacillus sp.）。测试了不同农业废弃物、造纸废弃物和羧甲基纤维素对内切葡聚糖酶产量的影响，发现8%的羧甲基纤维素产量最大。经硫酸铵饱和沉淀、DEAE-Sepharose柱层析纯化后，得到了高纯度的内切葡聚糖酶.粗提物纯化8.5倍，收率68.1%。SDS-PAGE测得该蛋白的分子量为97 kDa。洗涤剂（SDS、Tween-80）、金属离子（MnCl 2、ZnCl 2）和EDTA可适度降低酶活性。内切葡聚糖酶在pH 5.0 - 9.0和温度20 - 70°C之间稳定，在pH 7.0和温度50°C下具有最佳活性。酶对底物羧甲基纤维素的表观Km值记录为0.25 mg/ml。该内切葡聚糖酶在Ariel、Surf Excel和Tide等商品洗涤剂中稳定性较好，在洗涤剂工业中具有潜在的应用价值。该菌株的酶也可用于木质纤维素生物质转化为可发酵糖。
In an attempt to screen out cellulase producing bacteria from herbivorous animal fecal matter it was possible to isolate a potent bacterium from cow dung. The bacterium was identified as Bacillus sp. using 16S rDNA based molecular phylogenetic approach. The effect of different agricultural wastes, paper wastes and carboxymethyl cellulose on endoglucanase production was tested and was found to produce maximally at 8% carboxymethyl cellulose. The endoglucanase was precipitated by ammonium sulfate saturation and purified by DEAE- Sepharose column. The purification was achieved 8.5 fold from the crude extract with a yield of 68.1%. The molecular weight of the protein was determined to be 97 kDa by SDS-PAGE. The enzymatic activity was moderately reduced by detergents (SDS, Tween-80), metal ions (MnCl2, ZnCl2) and EDTA. The endoglucanase was stable between pH 5.0 – 9.0 and temperature between 20−70°C with optimal activity at pH 7.0 and temperature 50°C. The apparent Km value of the enzyme for the substrate carboxymethyl cellulose was recorded to be 0.25 mg/ml. The endoglucanase was stable in the presence of commercial detergents such as Ariel, Surf Excel and Tide, indicated might be of potential applications in detergent industry. The enzyme from this strain could also be applied in bioconversion of lignocellulosic biomass into fermentable sugars.
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DOI：
10.1080/00021369.1990.10869922
发表时间：
1990-01-01
期刊：
AGRICULTURAL AND BIOLOGICAL CHEMISTRY
影响因子：
--
作者：
SHIKATA, S;SAEKI, K;ITO, S
通讯作者：
ITO, S
影响因子：
11.4
作者：
Kaur, Jatinder;Chadha, Bhupinder S.;Saini, Harvinder S.
通讯作者：
Saini, Harvinder S.
影响因子：
3.6
作者：
Saratale, Ganesh D.;Oh, Sang Eun
通讯作者：
Oh, Sang Eun
影响因子：
4.4
作者：
TEATHER, RM;WOOD, PJ
通讯作者：
WOOD, PJ
影响因子：
15
作者：
Lineweaver, H;Burk, D
通讯作者：
Burk, D