Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5
Catechol 2,3-dioxygenases (C23Os, E.C.1.13.12.2) are two domain enzymes that catalyze degradation of monoaromatic hydrocarbons. The catalytically active C-domain of all known C23Os comprises ferrous ion ligands as well as residues forming active site pocket. The aim of this work was to examine and d...
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Wiley
2014-01-01
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| Series: | The Scientific World Journal |
| Online Access: | http://dx.doi.org/10.1155/2014/598518 |
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| author | Katarzyna Hupert-Kocurek Danuta Wojcieszyńska Urszula Guzik |
| author_facet | Katarzyna Hupert-Kocurek Danuta Wojcieszyńska Urszula Guzik |
| author_sort | Katarzyna Hupert-Kocurek |
| collection | DOAJ |
| description | Catechol 2,3-dioxygenases (C23Os, E.C.1.13.12.2) are two domain enzymes that catalyze degradation of monoaromatic hydrocarbons. The catalytically active C-domain of all known C23Os comprises ferrous ion ligands as well as residues forming active site pocket. The aim of this work was to examine and discuss the effect of nonsense mutation at position 289 on the activity of catechol 2,3-dioxygenase from Planococcus strain. Although the mutant C23O showed the same optimal temperature for activity as the wild-type protein (35°C), it exhibited activity slightly more tolerant to alkaline pH. Mutant enzyme exhibited also higher affinity to catechol as a substrate. Its Km (66.17 µM) was approximately 30% lower than that of wild-type enzyme. Interestingly, removal of the C-terminal residues resulted in 1.5- to 1.8-fold (P<0.05) increase in the activity of C23OB61 against 4-methylcatechol and 4-chlorocatechol, respectively, while towards catechol the activity of the protein dropped to about 80% of that of the wild-type enzyme. The results obtained may facilitate the engineering of the C23O for application in the bioremediation of polluted areas. |
| format | Article |
| id | doaj-art-767bd69d256d43b6834d334743310d99 |
| institution | OA Journals |
| issn | 2356-6140 1537-744X |
| language | English |
| publishDate | 2014-01-01 |
| publisher | Wiley |
| record_format | Article |
| series | The Scientific World Journal |
| spelling | doaj-art-767bd69d256d43b6834d334743310d992025-08-20T02:07:13ZengWileyThe Scientific World Journal2356-61401537-744X2014-01-01201410.1155/2014/598518598518Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5Katarzyna Hupert-Kocurek0Danuta Wojcieszyńska1Urszula Guzik2Department of Biochemistry, Faculty of Biology and Environment Protection, University of Silesia in Katowice, Jagiellonska 28, 40-032 Katowice, PolandDepartment of Biochemistry, Faculty of Biology and Environment Protection, University of Silesia in Katowice, Jagiellonska 28, 40-032 Katowice, PolandDepartment of Biochemistry, Faculty of Biology and Environment Protection, University of Silesia in Katowice, Jagiellonska 28, 40-032 Katowice, PolandCatechol 2,3-dioxygenases (C23Os, E.C.1.13.12.2) are two domain enzymes that catalyze degradation of monoaromatic hydrocarbons. The catalytically active C-domain of all known C23Os comprises ferrous ion ligands as well as residues forming active site pocket. The aim of this work was to examine and discuss the effect of nonsense mutation at position 289 on the activity of catechol 2,3-dioxygenase from Planococcus strain. Although the mutant C23O showed the same optimal temperature for activity as the wild-type protein (35°C), it exhibited activity slightly more tolerant to alkaline pH. Mutant enzyme exhibited also higher affinity to catechol as a substrate. Its Km (66.17 µM) was approximately 30% lower than that of wild-type enzyme. Interestingly, removal of the C-terminal residues resulted in 1.5- to 1.8-fold (P<0.05) increase in the activity of C23OB61 against 4-methylcatechol and 4-chlorocatechol, respectively, while towards catechol the activity of the protein dropped to about 80% of that of the wild-type enzyme. The results obtained may facilitate the engineering of the C23O for application in the bioremediation of polluted areas.http://dx.doi.org/10.1155/2014/598518 |
| spellingShingle | Katarzyna Hupert-Kocurek Danuta Wojcieszyńska Urszula Guzik Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5 The Scientific World Journal |
| title | Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5 |
| title_full | Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5 |
| title_fullStr | Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5 |
| title_full_unstemmed | Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5 |
| title_short | Activity of a Carboxyl-Terminal Truncated Form of Catechol 2,3-Dioxygenase from Planococcus sp. S5 |
| title_sort | activity of a carboxyl terminal truncated form of catechol 2 3 dioxygenase from planococcus sp s5 |
| url | http://dx.doi.org/10.1155/2014/598518 |
| work_keys_str_mv | AT katarzynahupertkocurek activityofacarboxylterminaltruncatedformofcatechol23dioxygenasefromplanococcussps5 AT danutawojcieszynska activityofacarboxylterminaltruncatedformofcatechol23dioxygenasefromplanococcussps5 AT urszulaguzik activityofacarboxylterminaltruncatedformofcatechol23dioxygenasefromplanococcussps5 |