Discovery of a catalytic domain defines a new glycoside hydrolase family containing endo-1,3-fucanase

Abstract

Sulfated fucans are important marine polysaccharides with various bioactivities. Fucanases are desirable tools for the structural elucidations and oligosaccharides preparation of sulfated fucans. Herein, a gene with unknown function was screened from a sulfated fucan utilization locus in genome of marine bacterium Wenyingzhuangia aestuarii OF219 with the assistance of a machine learning approach on the structural biology. An undefined catalytic domain that presented in this gene was further cloned and expressed in Escherichia coli. Utilizing a sulfated fucan tetrasaccharide with definite structure as substrate, the endo-acting cleavage point of expressed protein (named Fun187A) was identified as the α-l-1,3-glycosidic bond between Fucp and Fucp(2OSO3−). Fun187A demonstrated a novel cleavage specificity, that is the subsite −1 could tolerate α-l-Fucp, and the subsite +1 could tolerate α-l-Fucp(2OSO3−). A homologue of Fun187A was also validated to display the endo-1,3-fucanse activity. The sequence novelty of Fun187A and its homologue defines a new glycoside hydrolase family, GH187.