Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations
6 pages, 2 figures, 3 tables Measurements of potential enzyme activities are usually carried out to compare enzymatic activity between different systems and minimise the effect of product inhibition or substrate depletion during the assay. There is a general practice of estimating potential alkaline...
Published in: | Marine Ecology Progress Series |
---|---|
Main Authors: | , |
Format: | Article in Journal/Newspaper |
Language: | English |
Published: |
Inter Research
2004
|
Subjects: | |
Online Access: | http://hdl.handle.net/10261/342778 https://doi.org/10.3354/meps277285 |
id |
ftcsic:oai:digital.csic.es:10261/342778 |
---|---|
record_format |
openpolar |
spelling |
ftcsic:oai:digital.csic.es:10261/342778 2024-02-11T10:06:34+01:00 Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations Sebastián, Marta Niell, F. Xavier 2004-08 http://hdl.handle.net/10261/342778 https://doi.org/10.3354/meps277285 en eng Inter Research Publisher's version https://doi.org/10.3354/meps277285 Sí Marine Ecology Progress Series 277: 285-290 (2004) 0171-8630 http://hdl.handle.net/10261/342778 doi:10.3354/meps277285 1616-1599 open Alkaline phosphatase activity Exoenzyme Potential assays Marine environments artículo 2004 ftcsic https://doi.org/10.3354/meps277285 2024-01-24T00:42:59Z 6 pages, 2 figures, 3 tables Measurements of potential enzyme activities are usually carried out to compare enzymatic activity between different systems and minimise the effect of product inhibition or substrate depletion during the assay. There is a general practice of estimating potential alkaline phosphatase activity (APA) using single-substrate addition in excess of saturation, as complete kinetic experiments are time-consuming. Here, complete APA kinetic experiments were carried out in the Subtropical North Atlantic Ocean using from 0.1 to 100 µM concentration of the fluorogenic substrate MUF-P. There was a reduction of the velocity of reaction by excess of substrate in all experiments carried out (n = 7). This reduction could be related either to inhibition processes, to quenching effects, or to a loss of fluorescence of the product of reaction due to some kind of biological transformation. Moreover, the substrate concentration above which activity diminished was different between the cases studied. Therefore, there was only a narrow range of concentration in which single-substrate addition experiments would have given results comparable to the maximum enzyme activity, and this range differed between stations. Hence, in order to estimate APA potential activities in marine oligotrophic environments, complete kinetic experiments should be carried out, since the use of single additions in excess of substrate may give erroneous results. This conclusion may be extendable to other commonly performed assays of hydrolytic activities in aquatic environments that also use MUF-substrates Peer reviewed Article in Journal/Newspaper North Atlantic Digital.CSIC (Spanish National Research Council) Marine Ecology Progress Series 277 285 290 |
institution |
Open Polar |
collection |
Digital.CSIC (Spanish National Research Council) |
op_collection_id |
ftcsic |
language |
English |
topic |
Alkaline phosphatase activity Exoenzyme Potential assays Marine environments |
spellingShingle |
Alkaline phosphatase activity Exoenzyme Potential assays Marine environments Sebastián, Marta Niell, F. Xavier Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations |
topic_facet |
Alkaline phosphatase activity Exoenzyme Potential assays Marine environments |
description |
6 pages, 2 figures, 3 tables Measurements of potential enzyme activities are usually carried out to compare enzymatic activity between different systems and minimise the effect of product inhibition or substrate depletion during the assay. There is a general practice of estimating potential alkaline phosphatase activity (APA) using single-substrate addition in excess of saturation, as complete kinetic experiments are time-consuming. Here, complete APA kinetic experiments were carried out in the Subtropical North Atlantic Ocean using from 0.1 to 100 µM concentration of the fluorogenic substrate MUF-P. There was a reduction of the velocity of reaction by excess of substrate in all experiments carried out (n = 7). This reduction could be related either to inhibition processes, to quenching effects, or to a loss of fluorescence of the product of reaction due to some kind of biological transformation. Moreover, the substrate concentration above which activity diminished was different between the cases studied. Therefore, there was only a narrow range of concentration in which single-substrate addition experiments would have given results comparable to the maximum enzyme activity, and this range differed between stations. Hence, in order to estimate APA potential activities in marine oligotrophic environments, complete kinetic experiments should be carried out, since the use of single additions in excess of substrate may give erroneous results. This conclusion may be extendable to other commonly performed assays of hydrolytic activities in aquatic environments that also use MUF-substrates Peer reviewed |
format |
Article in Journal/Newspaper |
author |
Sebastián, Marta Niell, F. Xavier |
author_facet |
Sebastián, Marta Niell, F. Xavier |
author_sort |
Sebastián, Marta |
title |
Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations |
title_short |
Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations |
title_full |
Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations |
title_fullStr |
Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations |
title_full_unstemmed |
Alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations |
title_sort |
alkaline phosphatase activity in marine oligotrophic environments: implications of single-substrate addition assays for potential activity estimations |
publisher |
Inter Research |
publishDate |
2004 |
url |
http://hdl.handle.net/10261/342778 https://doi.org/10.3354/meps277285 |
genre |
North Atlantic |
genre_facet |
North Atlantic |
op_relation |
Publisher's version https://doi.org/10.3354/meps277285 Sí Marine Ecology Progress Series 277: 285-290 (2004) 0171-8630 http://hdl.handle.net/10261/342778 doi:10.3354/meps277285 1616-1599 |
op_rights |
open |
op_doi |
https://doi.org/10.3354/meps277285 |
container_title |
Marine Ecology Progress Series |
container_volume |
277 |
container_start_page |
285 |
op_container_end_page |
290 |
_version_ |
1790604363577688064 |