A novel alginate lyase and its domain functions for the preparation of unsaturated monosaccharides

Brown algae are considered promising crops for the production of sustainable biofuels. However, the commercial application has been limited by lack of efficient methods for converting alginate into fermentable sugars. Herein, we cloned and characterized a novel alginate lyase AlyPL17 from Pedobacter hainanensis NJ-02. It possessed outstanding catalytic efficiency toward polymannuronic acid (polyM), polyguluronic acid (polyG), and alginate sodium, with k(cat) of 39.42 +/- 1.9 s(-1), 32.53 +/- 0.88 s(-1), and 38.30 +/- 2.12 s(-1), respectively. AlyPL17 showed maximum activity at 45 degrees C and pH 9.0. The domain truncation did not change the optimal temperature and optimal pH, but greatly reduced the activity. In addition, AlyPL17 degrades alginate through the cooperative action of two structural domains in an exolytic mode. The minimal degradation substrate of AlyPL17 is a disaccharide. Furthermore, AlyPL17 and AlyPL6 can synergistically degrade alginate to prepare unsaturated monosaccharides that can be converted to 4-deoxy-l-erythron-5-hexoseuloseuronate acid (DEH). DEH is reduced to KDG by DEH reductase (Sdr), which enters the Entner-Doudoroff (ED) pathway as a common metabolite and is converted to bioethanol.

Automated summary

A novel alginate lyase, AlyPL17, was cloned and characterized from Pedobacter hainanensis NJ-02. It exhibited efficient catalytic activity towards polymannuronic acid, polyguluronic acid, and alginate sodium. AlyPL17 showed maximum activity at 45°C and pH 9.0, and degraded alginate through the cooperative action of two structural domains in an exolytic mode.