Challenges and perspectives in combinatorial assembly of novel exopolysaccharide biosynthesis pathways.

Because of their rheological properties various microbial polysaccharides are applied as thickeners and viscosifiers both in food and non-food industries. A broad variety of microorganisms secrete structurally diverse exopolysaccharides (EPS) that contribute to their surface attachment, protection against abiotic or biotic stress factors, and nutrient gathering. Theoretically, a massive number of EPS structures are possible through variations in monosaccharide sequences, condensation linkages and non-sugar decorations. Given the already-high diversity of EPS structures, taken together with the principal of combinatorial biosynthetic pathways, microbial polysaccharides are an attractive class of macromolecules with which to generate novel structures via synthetic biology approaches. However, previous manipulations primarily focused on increasing polysaccharide yield, with structural modifications restricted to removal of side chains or non-sugar decorations. This article outlines the biosynthetic pathways of the bacterial heteroexopolysaccharides xanthan and succinoglycan, which are used as thickening and stabilizing agents in food and non-food industries. Challenges and perspectives of combining synthetic biology approaches with directed evolution to overcome obstacles in assembly of novel EPS biosynthesis pathways are discussed.