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PMID:16545947
| Citation |
Ochiai, A, Hashimoto, W and Murata, K (2006) A biosystem for alginate metabolism in Agrobacterium tumefaciens strain C58: molecular identification of Atu3025 as an exotype family PL-15 alginate lyase. Res. Microbiol. 157:642-9 |
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| Abstract |
The Gram-negative bacterium Sphingomonas sp. strain A1 (strain A1) has a peculiar biosystem to directly import and depolymerize a macromolecule, alginate, which is encoded by a cluster of genes on the genome. We identified five clustered ORFs homologous to some genes of the strain A1 cluster in the genome of Agrobacterium tumefaciens strain C58 (strain C58). These ORFs are Atu3021, Atu3022, Atu3023, and Atu3024, encoding a putative sugar ABC transporter system and Atu3025, which encodes a putative alginate lyase. We analyzed the involvement of this gene cluster in alginate metabolism. Strain C58 cells grew significantly on low-molecular-weight (LMW) alginate (average molecular weight, 1000), and we detected specific alginate-induced expression of Atu3024 and Atu3025. This strain does not grow on alginate (average molecular weight, 25,600), suggesting that the strain C58 gene cluster is involved in importing and degrading LMW alginate. One protein, Atu3025, purified from strain C58, was identified as an alginate lyase, and the enzyme overexpressed in Escherichia coli was further characterized. Atu3025 released monosaccharides specifically from alginate most efficiently at pH 7.3 and 30 degrees C through a beta-elimination reaction, indicating that Atu3025 is an exotype alginate lyase potentially involved in the assimilation of LMW alginate in strain C58. |
| Links |
PubMed Online version:10.1016/j.resmic.2006.02.006 |
| Keywords |
Agrobacterium tumefaciens/enzymology; Agrobacterium tumefaciens/genetics; Agrobacterium tumefaciens/growth & development; Alginates/metabolism; Bacterial Proteins/genetics; Bacterial Proteins/metabolism; Culture Media; Escherichia coli/enzymology; Escherichia coli/genetics; Genes, Bacterial; Glucuronic Acid/metabolism; Hexuronic Acids/metabolism; Lyases/genetics; Lyases/metabolism; Multigene Family; Plasmids; Recombinant Proteins/genetics; Recombinant Proteins/metabolism |
| edit table |
Significance
Annotations
| Gene product | Qualifier | GO Term | Evidence Code | with/from | Aspect | Extension | Notes | Status |
|---|---|---|---|---|---|---|---|---|
| GO:0052764: exo-oligoalginate lyase activity |
ECO:0000314: |
F |
According to Figure 2B from this paper, C58 cells only grew on LMW alginate, this suggests that these cells are involved in LMW alginate metabolism. According to Figure 4B, the reaction with Atu 3025 was detected on a TLC plate involving TBA, thus further characterizing Atu 3025 as an exotype oligoalginate lyase. |
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