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Article

Catechol 1,2-Dioxygenase is an Analogue of Homogentisate 1,2-Dioxygenase in Pseudomonas chlororaphis Strain UFB2

Discipline of Microbiology, School of Life Sciences, College of Agriculture, Engineering and Science, University of KwaZulu-Natal (Westville Campus), Private Bag X54001, Durban 4000, South Africa
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Int. J. Mol. Sci. 2019, 20(1), 61; https://doi.org/10.3390/ijms20010061
Received: 11 October 2018 / Revised: 27 November 2018 / Accepted: 29 November 2018 / Published: 24 December 2018
(This article belongs to the Section Biochemistry)
Catechol dioxygenases in microorganisms cleave catechol into cis-cis-muconic acid or 2-hydroxymuconic semialdehyde via the ortho- or meta-pathways, respectively. The aim of this study was to purify, characterize, and predict the template-based three-dimensional structure of catechol 1,2-dioxygenase (C12O) from indigenous Pseudomonas chlororaphis strain UFB2 (PcUFB2). Preliminary studies showed that PcUFB2 could degrade 40 ppm of 2,4-dichlorophenol (2,4-DCP). The crude cell extract showed 10.34 U/mL of C12O activity with a specific activity of 2.23 U/mg of protein. A 35 kDa protein was purified to 1.5-fold with total yield of 13.02% by applying anion exchange and gel filtration chromatography. The enzyme was optimally active at pH 7.5 and a temperature of 30 °C. The Lineweaver–Burk plot showed the vmax and Km values of 16.67 µM/min and 35.76 µM, respectively. ES-MS spectra of tryptic digested SDS-PAGE band and bioinformatics studies revealed that C12O shared 81% homology with homogentisate 1,2-dioxygenase reported in other Pseudomonas chlororaphis strains. The characterization and optimization of C12O activity can assist in understanding the 2,4-DCP metabolic pathway in PcUFB2 and its possible application in bioremediation strategies. View Full-Text
Keywords: catechol 1,2-dioxygenase; homogentisate 1,2-dioxygenase; Pseudomonas chlororaphis; Pseudomonas chlororaphis strain UFB2 catechol 1,2-dioxygenase; homogentisate 1,2-dioxygenase; Pseudomonas chlororaphis; Pseudomonas chlororaphis strain UFB2
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MDPI and ACS Style

Setlhare, B.; Kumar, A.; Mokoena, M.P.; Olaniran, A.O. Catechol 1,2-Dioxygenase is an Analogue of Homogentisate 1,2-Dioxygenase in Pseudomonas chlororaphis Strain UFB2. Int. J. Mol. Sci. 2019, 20, 61. https://doi.org/10.3390/ijms20010061

AMA Style

Setlhare B, Kumar A, Mokoena MP, Olaniran AO. Catechol 1,2-Dioxygenase is an Analogue of Homogentisate 1,2-Dioxygenase in Pseudomonas chlororaphis Strain UFB2. International Journal of Molecular Sciences. 2019; 20(1):61. https://doi.org/10.3390/ijms20010061

Chicago/Turabian Style

Setlhare, Boitumelo, Ajit Kumar, Mduduzi P. Mokoena, and Ademola O. Olaniran. 2019. "Catechol 1,2-Dioxygenase is an Analogue of Homogentisate 1,2-Dioxygenase in Pseudomonas chlororaphis Strain UFB2" International Journal of Molecular Sciences 20, no. 1: 61. https://doi.org/10.3390/ijms20010061

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