PMID:22805784

Citation	Lee, SI, Choi, SH, Lee, EY and Kim, HS (2012) Molecular cloning, purification, and characterization of a novel polyMG-specific alginate lyase responsible for alginate MG block degradation in Stenotrophomas maltophilia KJ-2. Appl. Microbiol. Biotechnol. 95:1643-53
Abstract	A gene for a polyMG-specific alginate lyase possessing a novel structure was identified and cloned from Stenotrophomas maltophilia KJ-2 by using PCR with homologous nucleotide sequences-based primers. The recombinant alginate lyase consisting of 475 amino acids was purified on Ni-Sepharose column and exhibited the highest activity at pH 8 and 40 °C. Interestingly, the recombinant alginate lyase was expected to have a similar catalytic active site of chondroitin B lyase but did not show chondroitin lyase activity. In the test of substrate specificity, the recombinant alginate lyase preferentially degraded the glycosidic bond of polyMG-block than polyM-block and polyG-block. The chemical structures of the degraded alginate oligosaccharides were elucidated to have mannuronate (M) at the reducing end on the basis of NMR analysis, supporting that KJ-2 polyMG-specific alginate lyase preferably degraded the glycosidic bond in M-G linkage than that in G-M linkage. The KJ-2 polyMG-specific alginate lyase can be used in combination with other alginate lyases for a synergistic saccharification of alginate.
Links	PubMed Online version:10.1007/s00253-012-4266-y
Keywords	Alginates/chemistry; Alginates/metabolism; Amino Acid Sequence; Bacterial Proteins/chemistry; Bacterial Proteins/genetics; Bacterial Proteins/isolation & purification; Bacterial Proteins/metabolism; Cloning, Molecular; Enzyme Stability; Glucuronic Acid/chemistry; Glucuronic Acid/metabolism; Hexuronic Acids/chemistry; Hexuronic Acids/metabolism; Hydrogen-Ion Concentration; Molecular Sequence Data; Polysaccharide-Lyases/chemistry; Polysaccharide-Lyases/genetics; Polysaccharide-Lyases/isolation & purification; Polysaccharide-Lyases/metabolism; Sequence Alignment; Stenotrophomonas maltophilia/chemistry; Stenotrophomonas maltophilia/enzymology; Stenotrophomonas maltophilia/genetics; Stenotrophomonas maltophilia/metabolism; Substrate Specificity
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Annotations

Gene product	Qualifier	GO Term	Evidence Code	with/from	Aspect	Extension	Notes	Status
STEMA:I6P4V6		GO:0016837: carbon-oxygen lyase activity, acting on polysaccharides	ECO:0000314:		F		Figure 4	complete CACAO 9103
STEMA:I6P4V6	enables	GO:0016837: carbon-oxygen lyase activity, acting on polysaccharides	ECO:0000314: direct assay evidence used in manual assertion		F		Seeded From UniProt	complete

References

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