The Endo-α(1,3)-Fucoidanase Mef2 Releases Uniquely Branched Oligosaccharides from Saccharina latissima Fucoidans

Fucoidans are complex bioactive sulfated fucosyl-polysaccharides primarily found in brown macroalgae. Endo-fucoidanases catalyze the specific hydrolysis of alpha-L-fucosyl linkages in fucoidans and can be utilized to tailor-make fucoidan oligosaccharides and elucidate new structural details of fucoidans. In this study, an endo-alpha(1,3)-fucoidanase encoding gene, Mef2, from the marine bacterium Muricauda eckloniae, was cloned, and the Mef2 protein was functionally characterized. Based on the primary sequence, Mef2 was suggested to belong to the glycosyl hydrolase family 107 (GH107) in the Carbohydrate Active enZyme database (CAZy). The Mef2 fucoidanase showed maximal activity at pH 8 and 35 degrees C, although it could tolerate temperatures up to 50 degrees C. Ca2+ was shown to increase the melting temperature from 38 to 44 degrees C and was furthermore required for optimal activity of Mef2. The substrate specificity of Mef2 was investigated, and Fourier transform infrared spectroscopy (FTIR) was used to determine the enzymatic activity (Units per mu M enzyme: U-f/mu M) of Mef2 on two structurally different fucoidans, showing an activity of 1.2 x 10(-3) U-f/mu M and 3.6 x 10(-3) U-f/mu M on fucoidans from Fucus evanescens and Saccharina latissima, respectively. Interestingly, Mef2 was identified as the first described fucoidanase active on fucoidans from S. latissima. The fucoidan oligosaccharides released by Mef2 consisted of a backbone of alpha(1,3)-linked fucosyl residues with unique and novel alpha(1,4)-linked fucosyl branches, not previously identified in fucoidans from S. latissima.

Automated summary

This study cloned an endo-alpha(1,3)-fucoidanase encoding gene Mef2 from a marine bacterium and characterized its functional properties, showing its activity and specificity in fucoidan hydrolysis, as well as unveiling new structural details.