Hydrolysis rates from bulk samples, plate reader results from RV/Endeavor EN556, 2015 (Patterns of activities project) (NCEI Accession 0291494)
This dataset contains data collected on R/V Endeavor during cruise EN556 from 2015-04-27 to 2015-05-02. These data include depth. The instruments used to collect these data include Fluorometer, Niskin bottle, and plate reader. These data were collected by Carol Arnosti of University of North Carolina at Chapel Hill as part of the "Latitudinal and depth-related contrasts in enzymatic capabilities of pelagic microbial communities: Predictable patterns in the ocean? (Patterns of activities)" project. The Biological and Chemical Oceanography Data Management Office (BCO-DMO) submitted these data to NCEI on 2020-05-13.
The following is the text of the dataset description provided by BCO-DMO:
Bulk polysaccharide hydrolysis rates - EN556
Dataset Description:
This dataset includes polysaccharide hydrolysis rates to measure microbial enzyme activities and bacterial productivity.
See Niskin Bottle and Cast List EN556 to link specific casts and bottles to each experiment: https://www.bco-dmo.org/dataset/717427 .
The following is the text of the dataset description provided by BCO-DMO:
Bulk polysaccharide hydrolysis rates - EN556
Dataset Description:
This dataset includes polysaccharide hydrolysis rates to measure microbial enzyme activities and bacterial productivity.
See Niskin Bottle and Cast List EN556 to link specific casts and bottles to each experiment: https://www.bco-dmo.org/dataset/717427 .
Dataset Citation
- Cite as: Arnosti, Carol (2024). Hydrolysis rates from bulk samples, plate reader results from RV/Endeavor EN556, 2015 (Patterns of activities project) (NCEI Accession 0291494). [indicate subset used]. NOAA National Centers for Environmental Information. Dataset. https://www.ncei.noaa.gov/archive/accession/0291494. Accessed [date].
Dataset Identifiers
ISO 19115-2 Metadata
gov.noaa.nodc:0291494
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Ordering Instructions | Contact NCEI for other distribution options and instructions. |
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NOAA National Centers for Environmental Information +1-301-713-3277 NCEI.Info@noaa.gov |
Dataset Point of Contact |
NOAA National Centers for Environmental Information ncei.info@noaa.gov |
Time Period | 2015-04-27 to 2015-05-02 |
Spatial Bounding Box Coordinates |
West: -71.0086
East: -68.4078
South: 37.6675
North: 40.0725
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Dataset Progress Status | Complete - production of the data has been completed Historical archive - data has been stored in an offline storage facility |
Data Update Frequency | As needed |
Supplemental Information | Acquisition Description: Seawater was transferred to 20 L carboys that were rinsed three times with water from the sampling depth and then filled with seawater from a single Niskin bottle, using silicone tubing that had been acid washed then rinsed with distilled water prior to use. From each carboy, water was dispensed into smaller glass containers that were cleaned and pre-rinsed three times with water from the carboy prior to dispensing. This water was used to measure cell counts, bacterial productivity, and the activities of polysaccharide hydrolases, peptidases, and glucosidases. A separate glass Duran bottle was filled with seawater from the carboy and sterilized in an autoclave for 20-30 minutes to serve as a killed control for microbial activity measurements. Two substrates, -glucose and -glucose linked to a 4-methylumbelliferyl (MUF) fluorophore, were used to measure glucosidase activities. Five substrates linked to a 7-amido-4-methyl coumarin (MCA) fluorophore, one amino acid – leucine – and four oligopeptides – the chymotrypsin substrates alanine-alanine-phenylalanine (AAF) and alanine-alanine-proline-phenylalanine (AAPF), and the trypsin substrates glutamine-alanine-arginine (QAR) and phenylalanine-serine-arginine (FSR) – were used to measure exo- and endo-acting peptidase activities, respectively. Hydrolysis rates of the substrates were measured as an increase in fluorescence as the fluorophore was hydrolyzed from the substrate over time [as in Hoppe, 1993; Obayashi and Suzuki , 2005]. Incubations with the seven low molecular weight substrates were set up in a 96-well plate. For each substrate, triplicate wells were filled with a total volume of 200 L seawater for experimental incubations; triplicate wells were filled with 200 L autoclaved seawater for killed control incubations. Substrate was added at saturating concentrations. A saturation curve was determined with surface water from each station to determine saturating concentrations of substrate. The saturating concentration was identified as the lowest tested concentration of substrate at which additional substrate did not yield higher rates of hydrolysis. Fluorescence was measured over 24-48 hours incubation time with a plate reader (TECAN spectrafluor plus; 360 nm excitation, 460 emission), with time points taken every 4-6 hours. Hydrolysis rates were calculated from the rate of increase of fluorescence in the incubation over time relative to a set of standards of known concentration of fluorophore. Scripts to calculate hydrolysis rates and produce the figures shown here are available in the associated Github repository [ Hoarfrost , 2017]. 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 μM monomer-equivalent concentrations, except for fucoidan, which was added at 5 μM 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 time points (t1-t6): 2 days, 5 days, 10 days, 17 days, 30 days, and 42 days. At each time point, 2 mL of seawater was collected from the 50 mL falcon tube using a sterile syringe, filtered through a 0.2 μm pore size syringe filter, and stored frozen until processing. |
Purpose | This dataset is available to the public for a wide variety of uses including scientific research and analysis. |
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Platform keywords | NODC PLATFORM NAMES THESAURUS BCO-DMO Platform Names Global Change Master Directory (GCMD) Platform Keywords ICES/SeaDataNet Ship Codes |
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Last Modified: 2024-05-31T15:15:28Z
For questions about the information on this page, please email: ncei.info@noaa.gov
For questions about the information on this page, please email: ncei.info@noaa.gov