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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Vydáno v:	Applied microbiology and biotechnology Ročník 79; číslo 2; s. 217 - 224
Hlavní autoři:	Koschorreck, Katja, Richter, Sven M, Ene, Augusta B, Roduner, Emil, Schmid, Rolf D, Urlacher, Vlada B
Médium:	Journal Article
Jazyk:	angličtina
Vydáno:	Berlin/Heidelberg Berlin/Heidelberg : Springer-Verlag 01.05.2008 Springer Berlin Heidelberg Springer Springer Nature B.V
Témata:	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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Shrnutí:	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.
Bibliografie:	http://dx.doi.org/10.1007/s00253-008-1417-2 ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 14 content type line 23
ISSN:	0175-7598 1432-0614
DOI:	10.1007/s00253-008-1417-2