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http://dx.doi.org/10.5352/JLS.2015.25.2.121

Homology Modeling and Characterization of Oligoalginate Lyase from the Alginolytic Marine Bacterium Sphingomonas sp. Strain MJ-3  

Kim, Hee Sook (Department of Food Science and Biotechnology, Kyungsung University)
Publication Information
Journal of Life Science / v.25, no.2, 2015 , pp. 121-129 More about this Journal
Abstract
Alginates are found in marine brown seaweeds and in extracellular biofilms secreted by some bacteria. Previously, we reported an oligoalginate lyase from Sphingomonas sp. MJ-3 (MJ3-Oal) that had an exolytic activity and protein sequence homology with endolytic polymannuronate (polyM) lyase in the N-terminal region. In this study, the MJ3-Oal was tested for both exolytic and endolytic activity by homology modeling using the crystal structure of Alg17c from Saccharophagus degradans 2-40T. The tyrosine residue at the $426^{th}$ position, which possibly formed a hydrogen bond with the substrate, was mutated to phenylalanine. The FPLC profiles showed that MJ3-Oal degraded alginate quickly to monomers as a final product through the oligmers, whereas the Tyr426Phe mutant showed only exolytic alginate lyase activity. $^1H$-NMR spectra also showed that MJ3-Oal degraded the endoglycosidic bond of polyM and polyMG (polymannuronate-guluronate) blocks. These results indicate that oligoalginate lyase from Sphingomonas sp. MJ-3 probably catalyzes the degradation of both exo- and endo-glycosidic bonds of alginate.
Keywords
Endolytic activity; exolytic activity; homology modeling; oligoalginate lyase;
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