An enzymatic method for preparation of homopolymannuronate blocks and strictly alternating sequences of mannuronic and guluronic units

Two Pseudomonas aeruginosa alginates were lysed by an overexpressed polymannuronate lyase AlxMB (only acting on two or more consecutive, nonacetylated mannuronate units) to prepare either mannuronate blocks (poly-M blocks) with dp∼30, or strictly alternating sequences of mannuronic and guluronic acid (poly-MG blocks) with dp>20. The poly-M blocks were obtained by lysis of a P. aeruginosa polymannuronate that has 50% O-acetylation at C-2 and C-3. The poly-MG blocks were obtained from a P. aeruginosa alginate that contained both mannuronate and guluronate residues. The polysaccharide was first deacetylated and then treated with the lyase to excise the mannuronate units from the alternating-MG blocks. Both types of blocks should have potent biological effects and should provide useful specific substrates for characterisation of other alginate lyases.