López-Contreras Ana M, Gabor Krisztina, Martens Aernout A, Renckens Bernadet A M, Claassen Pieternel A M, Van Der Oost John, De Vos Willem M
Laboratory of Microbiology, Wageningen University and Research Centre, P.O. Box 17, 6700 AA Wageningen, The Netherlands.
Appl Environ Microbiol. 2004 Sep;70(9):5238-43. doi: 10.1128/AEM.70.9.5238-5243.2004.
Clostridium acetobutylicum ATCC 824 is a solventogenic bacterium that grows heterotrophically on a variety of carbohydrates, including glucose, cellobiose, xylose, and lichenan, a linear polymer of beta-1,3- and beta-1,4-linked beta-D-glucose units. C. acetobutylicum does not degrade cellulose, although its genome sequence contains several cellulase-encoding genes and a complete cellulosome cluster of cellulosome genes. In the present study, we demonstrate that a low but significant level of induction of cellulase activity occurs during growth on xylose or lichenan. The celF gene, located in the cellulosome-like gene cluster and coding for a unique cellulase that belongs to glycoside hydrolase family 48, was cloned in Escherichia coli, and antibodies were raised against the overproduced CelF protein. A Western blot analysis suggested a possible catabolite repression by glucose or cellobiose and an up-regulation by lichenan or xylose of the extracellular production of CelF by C. acetobutylicum. Possible reasons for the apparent inability of C. acetobutylicum to degrade cellulose are discussed.
丙酮丁醇梭菌ATCC 824是一种产溶剂细菌,它能在多种碳水化合物上进行异养生长,这些碳水化合物包括葡萄糖、纤维二糖、木糖和地衣多糖(一种由β-1,3-和β-1,4-连接的β-D-葡萄糖单元组成的线性聚合物)。丙酮丁醇梭菌不能降解纤维素,尽管其基因组序列包含几个纤维素酶编码基因和一个完整的纤维小体基因簇。在本研究中,我们证明在木糖或地衣多糖上生长期间会发生低水平但显著的纤维素酶活性诱导。位于类纤维小体基因簇中的celF基因编码一种属于糖苷水解酶家族48的独特纤维素酶,该基因在大肠杆菌中克隆,并针对过量产生的CelF蛋白产生抗体。蛋白质免疫印迹分析表明,葡萄糖或纤维二糖可能存在分解代谢物阻遏作用,而地衣多糖或木糖可上调丙酮丁醇梭菌胞外CelF的产生。本文讨论了丙酮丁醇梭菌明显无法降解纤维素的可能原因。