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KMID : 0545119940040020102
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Journal of Microbiology and Biotechnology
1994 Volume.4 No. 2 p.102 ~ p.107
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Cloning of the Endoglucanase Gene from Actinomyces sp. 40 in Escherichia coli and Some Properties of the Gene Products
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Min, Hae Ki
Choi, Yun Jaie/Cho, Kwang Keun/Ha, Jong Kyu/Woo, Jung Hee
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Abstract
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The β-1,4-endoglucanase gene from Actinomyces sp. 40 was cloned into Escherichia coli DH5α with pUC19. Chromosomal DNA from Actinomyces sp. 40 was cleaved with the restriction enzyme Sau3Al and ligated into pUC19 for the transformation of Escherichia coli DH5α. Positive clones of β-1,4-endoglucanase gene were detected as the clear zones on a medium supplemented with carboxymethylcellulose (CMC). This transformant possessed a single plasmid, designated pDS1, which contained the vector DNA and a 3.5 kilobase (kb) Sau3Al insertion fragment encoding endoglucanase. The size of the cloned fragment was reduced to 2.0 kb. The endoglucanase activity produced by the E. coli DH5a (pDS6) was higher than that of Actinomyces sp. 40 strain. The optimum pH and temperature of the cloned enzyme were pH 4.0∼5.0 and 55℃, respectively. The cloned enzyme was stable at 55℃ or below and in buffer ranging from pH 4.0 to 7.0. The enzyme degraded CMC but did not degrade xylan, cellobiose, and methyl-umbelliferylcellobiopyranoside (MUC).
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