Findings from recent studies have indicated that enzymes containing more than one catalytic domain may be particularly powerful in the degradation of recalcitrant polysaccharides such as chitin and cellulose. Some known multicatalytic enzymes contain several glycoside hydrolase domains and one or more carbohydrate-binding modules (CBMs). Here, using bioinformatics and biochemical analyses, we identified an enzyme, Jd1381 from the actinobacterium Jonesia denitrificans, that uniquely combines two different polysaccharide-degrading activities. We found that Jd1381 contains an N-terminal family AA10 lytic polysaccharide monooxygenase (LPMO), a family 5 chitin-binding domain (CBM5), and a family 18 chitinase (Chi18) domain. The full-length enzyme, which seems to be the only chitinase produced by J. denitrificans, degraded both a- and b-chitin. Both the chitinase and the LPMO activities of Jd1381 were similar to those of other individual chitinases and LPMOs, and the overall efficiency of chitin degradation by full-length Jd1381 depended on its chitinase and LPMO activities. Of note, the chitin-degrading activity of Jd1381 was comparable with or exceeded the activities of combinations of well-known chitinases and an LPMO from Serratia marcescens. Importantly, comparison of the chitinolytic efficiency of Jd1381 with the efficiencies of combinations of truncated variants—JdLPMO10 and JdCBM5-Chi18 or JdLPMO10-CBM5 and JdChi18—indicated that optimal Jd1381 activity requires close spatial proximity of the LPMO10 and the Chi18 domains. The demonstration of intramolecular synergy between LPMOs and hydrolytic enzymes reported here opens new avenues toward the development of efficient catalysts for biomass conversion.
Bibliographical noteFunding Information:
Funding and additional information—This work was supported by the Research Council of Norway through Project 221576 (to S. M., T. R. T., G. V.-K., and V. G. H. E.), called MarPol, and Project 247001 (to S. M., A. N., J. M., G. V.-K., and V. G. H. E.), which is Norway’s contribution to a project in the European Research Area Industrial Biotechnology (ERA-IB) called ChitoTex: Development and production of new insect chitosan and chitosan based functional coatings for yarns and textile fabrics.
© 2020 Mekasha et al.
PubMed: MeSH publication types
- Journal Article
- Research Support, Non-U.S. Gov't