Article
Cloning and expression of the algL gene, encoding the Azotobacter chroococcum alginate lyase: Purification and characterization of the enzyme
Author/s | Peciña, Ana
Pascual, Alberto Paneque Guerrero, Antonio |
Department | Universidad de Sevilla. Departamento de Bioquímica Vegetal y Biología Molecular |
Publication Date | 1999 |
Deposit Date | 2017-07-25 |
Published in |
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Abstract | The alginate lyase-encoding gene (algL) of Azotobacter chroococcum was localized to a 3.1-kb EcoRI DNA fragment that revealed an open reading frame of 1,116 bp. This open reading frame encodes a protein of 42.98 kDa, in ... The alginate lyase-encoding gene (algL) of Azotobacter chroococcum was localized to a 3.1-kb EcoRI DNA fragment that revealed an open reading frame of 1,116 bp. This open reading frame encodes a protein of 42.98 kDa, in agreement with the value previously reported by us for this protein. The deduced protein has a potential N-terminal signal peptide that is consistent with its proposed periplasmic location. The analysis of the deduced amino acid sequence indicated that the gene sequence has a high homology (90% identity) to the Azotobacter vinelandii gene sequence, which has very recently been deposited in the GenBank database, and that it has 64% identity to the Pseudomonas aeruginosa gene sequence but that it has rather low homology (15 to 22% identity) to the gene sequences encoding alginate lyase in other bacteria. The A. chroococcum AlgL protein was overproduced in Escherichia coli and purified to electrophoretic homogeneity in a two-step chromatography procedure on hydroxyapatite and phenyl-Sepharose. The kinetic and molecular parameters of the recombinant alginate lyase are similar to those found for the native enzyme. |
Citation | Peciña, A., Pascual, A. y Paneque, A. (1999). Cloning and expression of the algL gene, encoding the Azotobacter chroococcum alginate lyase: Purification and characterization of the enzyme. Journal of Bacteriology, 181 (5), 1409-1414. |
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