Premium
Purification and characterization of a recombinant laccase‐like multi‐copper oxidase from Paenibacillus glucanolyticus SLM 1
Author(s) -
Mathews S.L.,
Smithson C.E.,
Grunden A.M.
Publication year - 2016
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.13241
Subject(s) - abts , laccase , multicopper oxidase , escherichia coli , chemistry , recombinant dna , enzyme , biochemistry , guaiacol , antioxidant , gene , dpph
Abstract Aims The aim of this study was to evaluate the activity of a novel bacterial laccase‐like multi‐copper oxidase ( LMCO ) from Paenibacillus glucanolyticus SLM 1: a bacterium isolated from pulp and paper waste. Methods and Results A new bacterial LMCO gene ( CuOx ) from P. glucanolyticus SLM 1 was identified and cloned into pET 22b. The protein it encodes was recombinantly expressed in Escherichia coli . The recombinant P. glucanolyticus LMCO had a molecular weight of approximately 90 kD a and demonstrated oxidation of the LMCO substrates 2,2′‐Azino‐bis(3‐ethylbenzothiazoline‐6‐sulfonate) ( ABTS ), catechol, and 2,6‐Dimethoxyphenol (2,6‐ DMP ), with the oxidation of ABTS occurring to the greatest extent (776 U mg −1 under optimal conditions of pH 7 and 40°C). Furthermore, recombinant P. glucanolyticus CuOx retained activity against ABTS in the presence of 1 mol l −1 NaCl, 50% dimethyl sulfoxide and 5% Tween‐80 and can decolorize several types of dyes. Conclusions This enzyme has a neutral pH optimum, is capable of decolorizing dyes, and is active in the presence salt, detergents and surfactant. The characteristics of this enzyme suggest that it could be used for a variety of industrial applications. Significance and Impact of the Study This work characterizes a unique bacterial LMCO with activity higher than that of previously characterized fungal or bacterial LMCO s. This enzyme may have utility for industrial bleaching, treatment of dye effluent, and lignin removal.