The activities and structural specificities of extracellular enzymes that initiate microbial remineralization of high-molecular-weight (MW) organic matter were investigated in surface waters and sediments of an Arctic fjord of Svalbard. Hydrolysis rates of a suite of fluorescently labeled macromolecular substrates, including seven commercially available polysaccharides and three high-carbohydrate-content plankton extracts ranged from rapid to not detectable, and differed markedly between seawater and sediments. Order (fastest to slowest) of hydrolysis in surface water was laminarin, Spirulina extract, xylan>chondroitin, alginic acid, Wakame extract>arabinogalactan, fucoidan>Isochrysis extract>>>pullulan, while in sediments the order was pullulan, laminarin, alginic acid, Wakame extract>chondroitin, xylan>arabinogalactan, Isochrysis extract>Spirulina extract>fucoidan. These differences cannot be explained by simple scaling factors such as differences in microbial cell numbers between seawater and sediments. Other investigations have shown that microbial community composition of Svalbard sediments and of polar bacterioplankton samples differ markedly. These results demonstrate that sedimentary and seawater microbial communities also differ fundamentally in their abilities to access specific high-MW substrates. Substrate bioavailability depends on the capabilities of a microbial community, as well as the chemical and structural features of the substrate itself.