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The cloning and expression of a family of five modular-type mannuronan C-5-epimerase genes fromAzotobacter vinelandii (algE1 to -5 ) has previously been reported. The corresponding proteins catalyze the Ca2+ -dependent polymer-level epimerization of β-d -mannuronic acid to α-l -guluronic acid (G) in the commercially important polysaccharide alginate. Here we report the identification of three additional structurally similar genes, designatedalgE6 ,algE7 , andalgY . All three genes were sequenced and expressed inEscherichia coli . AlgE6 introduced contiguous stretches of G residues into its substrate (G blocks), while AlgE7 acted as both an epimerase and a lyase. The epimerase activity of AlgE7 leads to formation of alginates with both single G residues and G blocks. AlgY did not display epimerase activity, but a hybrid gene in which the 5′-terminal part was exchanged with the corresponding region inalgE4 expressed an active epimerase. Southern blot analysis of genomicA. vinelandii DNA, using the 5′ part ofalgE2 as a probe, indicated that all hybridization signals originated fromalgE1 to -5 or the three new genes reported here.

journal of bacteriology

Cloning and Expression of Three New <i>Azotobacter vinelandii</i> Genes Closely Related to a Previously Described Gene Family Encoding Mannuronan C-5-Epimerases

Cloning and Expression of Three New Azotobacter vinelandii Genes Closely Related to a Previously Described Gene Family Encoding Mannuronan C-5-Epimerases