Composition of the Holdfast Polysaccharide from Caulobacter crescentus

Surface colonization is central to the lifestyles of many bacteria. Exploiting surface niches requires sophisticated systems for sensing and attaching to solid materials. Caulobacter crescentus synthesizes a polysaccharide-based adhesin known as the holdfast at one of its cell poles, which enables tight attachment to exogenous surfaces. The genes required for holdfast biosynthesis have been analyzed in detail, but difficulties in isolating analytical quantities of the adhesin have limited efforts to characterize its chemical structure. In this report, we describe a method to extract the holdfast from C. crescentus cultures and present a survey of its carbohydrate content. Glucose, 3-O-methylglucose, mannose, N-acetylglucosamine, and xylose were detected in our extracts. Our results provide evidence that the holdfast contains a 1,4-linked backbone of glucose, mannose, N-acetylglucosamine, and xylose that is decorated with branches at the C-6 positions of glucose and mannose. By defining the monosaccharide components in the polysaccharide, our work establishes a framework for characterizing enzymes in the holdfast pathway and provides a broader understanding of how polysaccharide adhesins are built.IMPORTANCE To colonize solid substrates, bacteria often deploy dedicated adhesins that facilitate attachment to surfaces. Caulobacter crescentus initiates surface colonization by secreting a carbohydrate-based adhesin called the holdfast. Because little is known about the chemical makeup of the holdfast, the pathway for its biosynthesis and the physical basis for its unique adhesive properties are poorly understood. This study outlines a method to extract the C. crescentus holdfast and describes the monosaccharide components contained within the adhesive matrix. The composition analysis adds to our understanding of the chemical basis for holdfast attachment and provides missing information needed to characterize enzymes in the biosynthetic pathway.