Zverlov Vladimir V, Schwarz Wolfgang H
Department of Microbiology, Technische Universität München, Am Hochanger 4, D-85350 Freising, Germany.
Ann N Y Acad Sci. 2008 Mar;1125:298-307. doi: 10.1196/annals.1419.008.
Cellulose degradation is a rare trait in bacteria. However, the truly cellulolytic bacteria are extremely efficient hydrolyzers of plant cell wall polysaccharides, especially those in thermophilic anaerobic ecosystems. Clostridium stercorarium, a thermophilic ubiquitous soil dweller, has a simple cellulose hydrolyzing enzyme system of only two cellulases. However, it seems to be better suited for the hydrolysis of a wide range of hemicelluloses. Clostridium thermocellum, an ubiquitous thermophilic gram-type positive bacterium, is one of the most successful cellulose degraders known. Its extracellular enzyme complex, the cellulosome, was prepared from C. thermocellum cultures grown on cellulose, cellobiose, barley beta-1,3-1,4-glucan, or a mixture of xylan and cellulose. The single proteins were identified by peptide chromatography and MALDI-TOF-TOF. Eight cellulosomal proteins could be found in all eight preparations, 32 proteins occur in at least one preparation. A number of enzymatic components had not been identified previously. The proportion of components changes if C. thermocellum is grown on different substrates. Mutants of C. thermocellum, devoid of scaffoldin CipA, that now allow new types of experiments with in vitro cellulosome reassembly and a role in cellulose hydrolysis are described. The characteristics of these mutants provide strong evidence of the positive effect of complex (cellulosome) formation on hydrolysis of crystalline cellulose.
纤维素降解在细菌中是一种罕见的特性。然而,真正的纤维素分解细菌是植物细胞壁多糖的极其高效的水解者,尤其是在嗜热厌氧生态系统中的那些细菌。嗜热栖热菌是一种广泛存在于土壤中的嗜热菌,它具有仅由两种纤维素酶组成的简单纤维素水解酶系统。然而,它似乎更适合水解多种半纤维素。嗜热栖热放线菌是一种广泛存在的嗜热革兰氏阳性细菌,是已知最成功的纤维素降解菌之一。其细胞外酶复合物——纤维小体,是从在纤维素、纤维二糖、大麦β-1,3-1,4-葡聚糖或木聚糖与纤维素的混合物上生长的嗜热栖热放线菌培养物中制备的。通过肽色谱法和基质辅助激光解吸电离飞行时间串联质谱法鉴定单个蛋白质。在所有八种制剂中都能发现八种纤维小体蛋白,至少有一种制剂中出现32种蛋白。许多酶组分以前尚未被鉴定。如果嗜热栖热放线菌在不同底物上生长,组分的比例会发生变化。描述了缺乏支架蛋白CipA的嗜热栖热放线菌突变体,这些突变体现在允许进行新型的体外纤维小体重组实验以及在纤维素水解中的作用。这些突变体的特性为复合物(纤维小体)形成对结晶纤维素水解的积极作用提供了有力证据。