Cloning and characterization of a new laccase from Bacillus licheniformis catalyzing dimerization of phenolic acids

A new laccase gene (cotA) was cloned from Bacillus licheniformis and expressed in Escherichia coli. The recombinant protein CotA was purified and showed spectroscopic properties, typical for blue multi-copper oxidases. The enzyme has a molecular weight of ~65 kDa and demonstrates activity towards ca...

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Bibliographic Details
Published in:Applied microbiology and biotechnology Vol. 79; no. 2; pp. 217 - 224
Main Authors: Koschorreck, Katja, Richter, Sven M, Ene, Augusta B, Roduner, Emil, Schmid, Rolf D, Urlacher, Vlada B
Format: Journal Article
Language:English
Published: Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01.05.2008
Springer Berlin Heidelberg
Springer
Springer Nature B.V
Subjects:
Acids
analysis
Bacillus
Bacillus - enzymology
Bacillus - genetics
Bacillus - metabolism
Bacillus licheniformis
Bacterial Proteins
Bacterial Proteins - chemistry
Bacterial Proteins - genetics
Bacterial Proteins - metabolism
Benzoquinone
Benzothiazoles
Benzothiazoles - metabolism
Biological and medical sciences
Biology of microorganisms of confirmed or potential industrial interest
Biotechnologically Relevant Enzymes and Proteins
Biotechnology
Caffeic acid
chemistry
Cinnamic acid
Cloning
Cloning, Molecular
Copper
Coumaric acid
Dimerization
E coli
Enzymes
enzymology
Escherichia coli
Escherichia coli - genetics
Ferulic acid
Fundamental and applied biological sciences. Psychology
Fungi
genes
Genes, Bacterial
genetics
Hydrazones
Hydrazones - metabolism
Hydroxybenzoates
Hydroxybenzoates - metabolism
Laccase
Laccase - analysis
Laccase - metabolism
Laccase - physiology
Life Sciences
metabolism
Microbial Genetics and Genomics
Microbiology
Miscellaneous
Mission oriented research
Molecular weight
Oxidation
Phenol oxidation
Phenolic acids
Phenols
physiology
Potassium
Proteins
recombinant proteins
Sinapic acid
Sodium
spectral analysis
Studies
Substrate Specificity
Sulfonic acid
Sulfonic Acids
Sulfonic Acids - metabolism
Syringaldazine
syringic acid
Vanillic acid
Phenol oxidation
Dimerization
Laccase
Enzyme
Phenolic acid
Bacillus licheniformis
Laccase,Phenol oxidation,Dimerization
Bacillaceae
Characterization
Phenol
Bacillales
Bacteria
Oxidation
Oxidoreductases
ISSN:0175-7598, 1432-0614
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Summary:A new laccase gene (cotA) was cloned from Bacillus licheniformis and expressed in Escherichia coli. The recombinant protein CotA was purified and showed spectroscopic properties, typical for blue multi-copper oxidases. The enzyme has a molecular weight of ~65 kDa and demonstrates activity towards canonical laccase substrates 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS), syringaldazine (SGZ) and 2,6-dimethoxyphenol (2,6-DMP). Kinetic constants K M and k cat for ABTS were of 6.5 ± 0.2 μM and 83 s⁻¹, for SGZ of 4.3 ± 0.2 μM and 100 s⁻¹, and for 2,6-DMP of 56.7 ± 1.0 μM and 28 s⁻¹. Highest oxidizing activity towards ABTS was obtained at 85°C. However, after 1 h incubation of CotA at 70°C and 80°C, a residual activity of 43% and 8%, respectively, was measured. Furthermore, oxidation of several phenolic acids and one non-phenolic acid by CotA was investigated. CotA failed to oxidize coumaric acid, cinnamic acid, and vanillic acid, while syringic acid was oxidized to 2,6-dimethoxy-1,4-benzoquinone. Additionally, dimerization of sinapic acid, caffeic acid, and ferulic acid by CotA was observed, and highest activity of CotA was found towards sinapic acid.
Bibliography:http://dx.doi.org/10.1007/s00253-008-1417-2
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ISSN:0175-7598
1432-0614
DOI:10.1007/s00253-008-1417-2