Cloning and Characterization of Alginate Lyase from a Marine Bacterium Streptomyces sp. ALG-5 |
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Authors: | Dong Eun Kim Eun Yeol Lee Hee Sook Kim |
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Affiliation: | (1) Department of Food Science and Biotechnology, Kyungsung University, Busan, 608-736, South Korea;(2) Department of Chemical Engineering, Kyung Hee University, Gyeonggi-do, 446-701, South Korea |
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Abstract: | A marine bacterium was isolated from seaweeds for its ability to degrade alginate. Analysis of 16S ribosomal DNA sequence and chemotaxonomic characterizations revealed that the strain belongs to Streptomyces. The alginate lyase gene of Streptomyces sp. ALG-5 was cloned by using PCR with the specific primer designed from homologous nucleotide sequences. The consensus sequences of N-terminal YXRSELREM and C-terminal YFKAGXYXQ were conserved in the ALG-5 alginate lyase gene. The recombinant alginate lyase was purified by using Ni-Sepharose affinity chromatography. The alginate lyase appears to be poly-guluronate lyase degrading poly-G block preferentially than poly-M block. The degraded products were determined to be di-, tri-, tetra- and pentasaccharides by using BioGel P-2 gel filtration chromatography and ionization mass spectroscopy method. |
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Keywords: | Alginate lyase Alginate oligosaccharide Streptomyces sp. ALG-5 Unsaturated uronate |
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