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KMID : 0903520070500020107
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Journal of the Korean Society of Agricultural Chemistry and Biotechnology
2007 Volume.50 No. 2 p.107 ~ p.110
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Purification and Characterization of Overproduced E. coli Laccase
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Hong June-Hyuk
Kim Hyun-Jung
Kim Woo-Yeon
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Abstract
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The putative laccase gene, yacK of Escherichia coli, K-12 is not expressed in lab culture conditions. The laccase gene was amplified by PCR and subcloned into pET28a vector. The laccase overproduced in E. coli harboring pET28a was purified by His-affinity column chromatography. The purified laccase, which has the apparent molecular weight of 55,000 on the SDS-polyacrylamide gel showed enzyme activity on the guaiacol solution and agar plate. Optimun temperature and pH were around 65oC and 5.0, respectively.
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KEYWORD
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E. coli laccase expression, His-affinity column purification
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