Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study.
The main aim of this study was to prepare gelatine-based hydrogels containing entrapped substrate and to examine the applicability of these matrices for detection of enzymes with a specified catalytic activity. The general research concept assumed the use of a substrate that, in the presence of a pa...
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| Format: | Article |
| Language: | English |
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Public Library of Science (PLoS)
2018-01-01
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| Series: | PLoS ONE |
| Online Access: | http://europepmc.org/articles/PMC6181394?pdf=render |
| _version_ | 1818457177028820992 |
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| author | Karolina Labus |
| author_facet | Karolina Labus |
| author_sort | Karolina Labus |
| collection | DOAJ |
| description | The main aim of this study was to prepare gelatine-based hydrogels containing entrapped substrate and to examine the applicability of these matrices for detection of enzymes with a specified catalytic activity. The general research concept assumed the use of a substrate that, in the presence of a particular enzyme, will quickly undergo conversion to a coloured product. ortho-Nitrophenyl-β-D-galactopyranoside (ONPG) was used as the immobilized substrate and β-galactosidase from Kluyveromyces lactis as the biocatalyst to be determined. Among other factors, the range of detectable concentrations of galactosidase, the operational pH range, the time necessary to achieve a visible response and the preferred storage conditions for the test were determined. As a result, an effective colourimetric test for β-galactosidase detection was obtained. Its main advantages include (i) the effective detection of the enzyme at concentrations greater than or equal to 0.6 mg.L-1, (ii) the ability to perform initial quantification of the enzyme on the basis of the intensity of the obtained colour (iii) applicability in a wide pH range (from 4.0 to 9.0), (iv) a relatively short response time (from 1 to a maximum of 30 minutes) and (v) stability in long-term storage at 4°C (90 days without loss of specific properties). |
| first_indexed | 2024-12-14T22:38:25Z |
| format | Article |
| id | doaj.art-e89745933d9044b2b053558a229431e6 |
| institution | Directory Open Access Journal |
| issn | 1932-6203 |
| language | English |
| last_indexed | 2024-12-14T22:38:25Z |
| publishDate | 2018-01-01 |
| publisher | Public Library of Science (PLoS) |
| record_format | Article |
| series | PLoS ONE |
| spelling | doaj.art-e89745933d9044b2b053558a229431e62022-12-21T22:45:04ZengPublic Library of Science (PLoS)PLoS ONE1932-62032018-01-011310e020553210.1371/journal.pone.0205532Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study.Karolina LabusThe main aim of this study was to prepare gelatine-based hydrogels containing entrapped substrate and to examine the applicability of these matrices for detection of enzymes with a specified catalytic activity. The general research concept assumed the use of a substrate that, in the presence of a particular enzyme, will quickly undergo conversion to a coloured product. ortho-Nitrophenyl-β-D-galactopyranoside (ONPG) was used as the immobilized substrate and β-galactosidase from Kluyveromyces lactis as the biocatalyst to be determined. Among other factors, the range of detectable concentrations of galactosidase, the operational pH range, the time necessary to achieve a visible response and the preferred storage conditions for the test were determined. As a result, an effective colourimetric test for β-galactosidase detection was obtained. Its main advantages include (i) the effective detection of the enzyme at concentrations greater than or equal to 0.6 mg.L-1, (ii) the ability to perform initial quantification of the enzyme on the basis of the intensity of the obtained colour (iii) applicability in a wide pH range (from 4.0 to 9.0), (iv) a relatively short response time (from 1 to a maximum of 30 minutes) and (v) stability in long-term storage at 4°C (90 days without loss of specific properties).http://europepmc.org/articles/PMC6181394?pdf=render |
| spellingShingle | Karolina Labus Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study. PLoS ONE |
| title | Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study. |
| title_full | Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study. |
| title_fullStr | Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study. |
| title_full_unstemmed | Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study. |
| title_short | Effective detection of biocatalysts with specified activity by using a hydrogel-based colourimetric assay - β-galactosidase case study. |
| title_sort | effective detection of biocatalysts with specified activity by using a hydrogel based colourimetric assay β galactosidase case study |
| url | http://europepmc.org/articles/PMC6181394?pdf=render |
| work_keys_str_mv | AT karolinalabus effectivedetectionofbiocatalystswithspecifiedactivitybyusingahydrogelbasedcolourimetricassaybgalactosidasecasestudy |