Characterization of the cellulolytic complex (cellulosome) produced by Clostridium cellulolyticum
Characterization of the cellulolytic complex (cellulosome) produced by Clostridium cellulolyticum
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DOI：
10.1128/aem.63.3.903-909.1997
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发表时间：
1997-03-01
影响因子：
4.4
通讯作者：
Belaich, JP
中科院分区：
文献类型：
--
作者：
Gal, L;Pages, S;Belaich, JP
从生长在纤维素上的纤维素分解梭菌中分离出纤维素分解复合物。经凝胶过滤，发现复合物主要由600-kDa单元组成，沿着有16-MDa聚集体。测定了其降解各种底物的能力和其与结晶纤维素结合的能力。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳，N-末端测序和印迹分析的结果表明，所有已知的纤维素酶的这种生物体都存在于这个复合物。观察到三个主要成分：第一个成分，一个非催化，大（160 kDa）的蛋白质，被确定为基于它的能力，结合到含有锚定蛋白的纤维素酶作为支架蛋白CipC。另外两种组分的分子量分别为94和80.6 kDa，分别被鉴定为CelE和CelF。鉴定的纤维素酶和纤维素酶体的一些其他组分能够结合到miniCipC 1构建体。除了提供了一个广泛的描述系统，本研究的结果证实，锚定蛋白的粘附结构域的相互作用中起着至关重要的作用，在多纤维素酶体的宪法。
The cellulolytic complex was isolated from Clostridium cellulolyticum grown on cellulose. Upon gel filtration, the complex was found to consist mainly of 600-kDa units, along with a 16-MDa aggregate, Its ability to degrade various substrates and its capacity to bind to the crystalline cellulose were measured. The results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis, N-terminal sequencing, and blotting analysis showed that all of the known cellulases of this organism are present in this complex. Three major components were observed: the first component, a noncatalytic, large (160-kDa) protein, was identified based on its ability to bind to the dockerin-containing cellulases as scaffolding protein CipC. The other two components, which had molecular masses of 94 and 80.6 kDa, were identified as CelE and CelF, respectively. The identified cellulases and some other components of the cellulosome were able to bind to a miniCipC1 construct. In addition to providing an extensive description of the system, the results of the present study confirm that the dockerin-cohesin domain interaction plays an essential role in the constitution of the cellulosome.