嗜热栖热放线菌碳水化合物降解酶编码基因在大肠杆菌中的克隆与表达
Cloning and expression of genes coding for carbohydrate degrading enzymes of Anaerocellum thermophilum in E.coli.
作者信息
Bolshakova E V, Ponomariev A A, Novikov A A, Svetlichnyi V A, Velikodvorskaya G A
机构信息
Institute of Molecular Genetics, Russian Academy of Science, Moscow.
出版信息
Biochem Biophys Res Commun. 1994 Jul 29;202(2):1076-80. doi: 10.1006/bbrc.1994.2038.
A gene library of Anaerocellum thermophilum Z-1320 was constructed in Escherichia coli using plasmid pUC18. Clones were selected by screening for hydrolysis of CMC, MU-Cel, lichenan and xylan. Recombinant clones expressing two beta-1,4-glucanases and two different xylanases were obtained. Both beta-1,4-glucanases were capable of degrading Avicel. Clones with xylanase activity also cleft PNPXyl, MU-Glu and MU-Cel. The genes of one beta-1,4-glucanase and one xylanase are probably located in the same region of A.thermophilum chromosome.
利用质粒pUC18在大肠杆菌中构建嗜热厌氧纤维素菌Z-1320的基因文库。通过筛选羧甲基纤维素（CMC）、微晶纤维素（MU-Cel）、地衣多糖和木聚糖的水解情况来选择克隆。获得了表达两种β-1,4-葡聚糖酶和两种不同木聚糖酶的重组克隆。两种β-1,4-葡聚糖酶都能够降解微晶纤维素。具有木聚糖酶活性的克隆也能裂解对硝基苯木糖苷（PNPXyl）、4-甲基伞形酮基-β-D-葡萄糖苷（MU-Glu）和微晶纤维素。一种β-1,4-葡聚糖酶和一种木聚糖酶的基因可能位于嗜热厌氧纤维素菌染色体的同一区域。
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