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利用pBluescripts构建巨大芽孢杆菌基因组文库,在ABP平板上测定酶活性,共有78株阳性克隆子具有内切葡聚糖酶活性。将酶活性表达较好的一株菌株进行序列测定。测定结果这个片段共941bp,含有一个开放阅读框架,共348个核苷酸,可编码116个氨基酸。序列比较结果表明,该基因片段同已发表的枯草芽孢杆菌glyB aprE之间的同源性为35%;同芽孢杆菌BP23celB、短小芽孢杆菌内切葡聚糖酶和多粘芽孢杆菌β 1,4 内切葡聚糖酶的编码基因的同源性只有27%。虽然同源性较低,但酶活性表达较强,认为该基因是编码内切葡聚糖酶的一个新基因片段。
The B. megaterium genomic library was constructed using pBluescripts and the enzyme activity was determined on ABP plates. A total of 78 positive clones showed endoglucanase activity. A strain with better enzyme activity was sequenced. The results of this fragment of 941bp, contains an open reading frame, a total of 348 nucleotides, can encode 116 amino acids. The results of sequence comparison showed that the homology between this gene fragment and published B. subtilis glyB aprE was 35%. The homology with Bacillus sp. BP23celB, B. pumilus endoglucanase and B. polymyxa beta 1,4 Endoglucanase encoding gene homology is only 27%. Although its homology is low, its activity is strong, which is considered as a new gene fragment encoding endoglucanase.