Microbial enzyme activities: polysaccharide hydrolase activities in bulk seawater samples from the RV\Sonne cruise SO248 in the South and North Pacific, along 180 W, May, 2016 (NCEI Accession 0294403)

Acquisition Description:
Water was collected via Niskin bottles mounted on a rosette, equipped with a CTD.

The potential of the seawater microbial community to hydrolyze six high-molecular-weight polysaccharides (arabinogalactan, chondroitin sulfate, fucoidan, laminarin, pullulan, and xylan) was investigated in surface and bottom water. For each substrate, three 50 mL falcon tubes were filled with seawater and one 50 mL falcon tube was filled with autoclaved seawater to serve as a killed control. Substrate was added at 3.5 uM monomer-equivalent concentrations, except for fucoidan, which was added at 5 uM concentrations (a higher concentration was necessary for sufficient fluorescence signal). Two 50 mL falcon tubes – one with seawater and one with autoclaved seawater – with no added substrate served as blank controls. Incubations were stored in the dark at as close to in situ temperature as possible. Subsamples of the incubations were collected at time zero, and at six subsequent timepoints (t1-t6): 2 days, 5 days, 10 days, 17 days, 30 days, and 42 days. At each timepoint, 2 mL of seawater was collected from the 50 mL falcon tube using a sterile syringe, filtered through a 0.2 um pore size syringe filter, and stored frozen until processing.

The hydrolysis of high molecular weight substrate to lower molecular weight hydrolysis products was measured using gel permeation chromatography with fluorescence detection, after the method of Arnosti [1996, 2003]. In short, the subsample was injected onto a series of columns consisting of a 21 cm column of G50 and a 19 cm column of G75 Sephadex gel. The fluorescence of the column effluent was measured at excitation and emission wavelengths of 490 and 530 nm, respectively. Hydrolysis rates were calculated from the change in molecular weight distribution of the substrate over time, as described in detail in Arnosti [2003].

ara = arabinogalactan
chn = chondroitin sulfate
fuc = fucoidan
lam = laminarin
pul = pullulan
xyl = xylan