Masson J Y, Denis F, Brzezinski R
Groupe de Recherche en Biologie des Actinomycètes, Département de Biologie, Faculté des Sciences, Université de Sherbrooke, Québec, Canada.
Gene. 1994 Mar 11;140(1):103-7. doi: 10.1016/0378-1119(94)90738-2.
A 1.6-kb DNA fragment from the soil actinomycete, Streptomyces sp. strain N174, containing the gene (csn) encoding an extracellular chitosanase (CSN), has been isolated and its complete nucleotide sequence determined. The gene was expressed in Escherichia coli and Streptomyces lividans using appropriate vectors. The sequence was found to contain one large ORF which encodes a protein of 238 amino acids (aa). The deduced aa sequence begins with a signal peptide which has an unusual C-terminal segment, well recognized by the Streptomyces secretion system, but poorly in Escherichia coli. The strain N174 CSN aa sequence was compared with those of other proteins and significant homology was found only with the Bacillus circulans MH-K1 CSN but not with known chitinases or lysozymes. This suggests that CSN form a new enzyme family distinct from other endoglycosidases.
从土壤放线菌链霉菌属菌株N174中分离出一个1.6 kb的DNA片段,该片段含有编码胞外壳聚糖酶(CSN)的基因(csn),并测定了其完整的核苷酸序列。使用合适的载体在大肠杆菌和变铅青链霉菌中表达该基因。发现该序列包含一个大的开放阅读框,其编码一个238个氨基酸(aa)的蛋白质。推导的氨基酸序列起始于一个信号肽,该信号肽具有一个不寻常的C末端片段,能被链霉菌分泌系统很好地识别,但在大肠杆菌中识别效果较差。将菌株N174的CSN氨基酸序列与其他蛋白质的序列进行比较,发现仅与环状芽孢杆菌MH-K1的CSN有显著同源性,而与已知的几丁质酶或溶菌酶没有同源性。这表明CSN形成了一个不同于其他内切糖苷酶的新酶家族。