Please use this identifier to cite or link to this item: http://hdl.handle.net/11189/8332
Title: Production and characterisation of a novel actinobacterial DyP-type peroxidase and its application in coupling of phenolic monomers
Authors: Musengi, Amos 
Durrell, Kim 
Prins, Alaric 
Khan, Nuraan 
Agunbiade, Mayowa 
Kirby-McCullough, Bronwyn 
Burton, Stephanie G. 
Kudanga, Tukayi 
Pletschke, Brett I. 
Le Roes-Hill, Marilize 
Keywords: Actinobacteria;biocatalysis;DyP-type peroxidase;purification;streptomyces albidoflavus;genome
Issue Date: 2020
Publisher: Elsevier
Source: Musengi, A., Durrell, K., Prins, A. et al. 2020. Production and characterisation of a novel actinobacterial DyP-type peroxidase and its application in coupling of phenolic monomers. Enzyme and Microbial Technology. 141. 1-11. [https://doi.org/10.1016/j.enzmictec.2020.109654]
Journal: Enzyme and Microbial Technology 
Abstract: The extracellular peroxidase from Streptomyces albidoflavus BSII#1 was purified to near homogeneity using sequential steps of acid and acetone precipitation, followed by ultrafiltration. The purified peroxidase was characterised and tested for the ability to catalyse coupling reactions between selected phenolic monomer pairs. A 46-fold purification of the peroxidase was achieved, and it was shown to be a 46 kDa haem peroxidase. Unlike other actinobacteria-derived peroxidases, it was only inhibited (27 % inhibition) by relatively high concentrations of sodium azide (5 mM) and was capable of oxidising eleven (2,4-dichlorophenol, 2,6-dimethoxyphenol, 4-tert-butylcatechol, ABTS, caffeic acid, catechol, guaiacol, l-DOPA, o-aminophenol, phenol, pyrogallol) of the seventeen substrates tested. The peroxidase remained stable at temperatures of up to 80 °C for 60 min and retained >50 % activity after 24 h between pH 5.0–9.0, but was most sensitive to incubation with hydrogen peroxide (H2O2; 0.01 mM), l-cysteine (0.02 mM) and ascorbate (0.05 mM) for one hour. It was significantly inhibited by all organic solvents tested (p ≤ 0.05). The Km and Vmax values of the partially purified peroxidase with the substrate 2,4-DCP were 0.95 mM and 0.12 mmol min−1, respectively. The dyes reactive blue 4, reactive black 5, and Azure B, were all decolourised to a certain extent: approximately 30 % decolourisation was observed after 24 h (1 μM dye). The peroxidase successfully catalysed coupling reactions between several phenolic monomer pairs including catechin-caffeic acid, catechin-catechol, catechin-guaiacol and guaiacol-syringaldazine under the non-optimised conditions used in this study. Genome sequencing confirmed the identity of strain BSII#1 as a S. albidoflavus strain. In addition, the genome sequence revealed the presence of one peroxidase gene that includes the twin arginine translocation signal sequence of extracellular proteins. Functional studies confirmed that the peroxidase produced by S. albidoflavus BSII#1 is part of the dye-decolourising peroxidase (DyP-type) family.
URI: http://hdl.handle.net/11189/8332
ISSN: 0141-0229
DOI: https://doi.org/10.1016/j.enzmictec.2020.109654
Appears in Collections:Appsc - Journal Articles (DHET subsidised)

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