Biocatalytic quantification of α-glucan in particulate marine organic matter

Abstract

Marine algae drive the marine carbon cycle, converting carbon dioxide into organic material. A major component of this produced biomass is a variety of glycans; and yet their chemical composition and individual involvement in production, sedimentation and bacterial uptake remain largely unknown due to a lack of analytical tools for glycan-specific quantification. Marine -glucans include a range of storage glycans from red and green algae, bacteria, fungi and animals. Although these compounds are likely to account for a high amount of the carbon stored in the oceans they have not been quantified in marine samples so far. Here we present a method to extract and quantify -glucans in particulate organic matter from algal cultures and environmental samples using a sequential physicochemical extraction and enzymes as -glucan-specific probes. This enzymatic assay is more specific and less susceptible to side reactions than chemical hydrolysis. Using HPAEC-PAD to detect the hydrolysis products allows for a glycan quantification in particulate marine samples even at low concentration of {approx} 2-7 {micro}g/L -glucans. We measured -glucans (and compared their concentration with the {beta}-glucan laminarin) in three microalgae laboratory cultures as well as in marine particulate organic matter from the North Sea and western North Atlantic Ocean. While laminarin from diatoms and brown algae is an essential component of marine carbon turnover, our results further indicate the significant contribution of starch-like -glucans to marine particulate organic matter. Henceforth, the combination of glycan-linkage-specific enzymes and chromatographic hydrolysis product detection can provide a powerful tool in the exploration of marine glycans and their role in the global carbon cycle.