Preparation of 4-Deoxy-<scp>L</scp>-<i>erythro</i>-5-hexoseulose Uronic Acid (DEH) and Guluronic Acid Rich Alginate Using a Unique <i>exo</i>-Alginate Lyase from <i>Thalassotalea crassostreae</i>

Abstract

Marine multicellular algae are considered promising crops for the production of sustainable biofuels and commodity chemicals. However, their commercial exploitation is currently limited by a lack of appropriate and efficient enzymes for converting alginate into metabolizable building blocks, such as 4-deoxy-L-erythro-5-hexoseulose uronic acid (DEH). Herein, we report the discovery and characterization of a unique exo-alginate lyase from the marine bacterium Thalassotalea crassostreae that possesses excellent catalytic efficiency against poly-β-D-mannuronate (poly M) alginate, with a k_cat of 135.8 s^-1, and a 5-fold lower k_cat of 25 s^-1 against poly-α-L-guluronate (poly G alginate). We propose that this preference for poly M is due to a structural feature of the protein's active site. The mode of action and specificity of this enzyme has made it possible to design an effective and environmentally friendly process for the production of DEH and low molecular weight guluronate-enriched alginate.