Homologous cloning, expression, and characterisation of a laccase from Streptomyces coelicolor and enzymatic decolourisation of an indigo dye.
The lack of a commercially available robust and inexpensive laccase is a major barrier to the widespread application of this enzyme in various industrial sectors. By using an efficient system developed in Streptomyces lividans, we have produced by homologous expression 350 mg L(-1) of a bacterial laccase with a high purity and without any extensive purification. This is the highest production yield reported in the literature for a bacterial laccase. The secreted enzyme achieved oxidation under a wide pH range depending on the substrate: 4.0 for 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonate) and 9.0 for 2,6-dimethoxyphenol. Furthermore, this bacterial laccase was found to be quite resistant under various conditions. It withstands pH from 3.0 to 9.0, shows a great thermostability at 70 degrees C and was highly resistant toward conventional inhibitors. For instance, while the laccase of Trametes versicolor was completely inhibited by 1 mM NaN(3), the laccase of Streptomyces coelicolor was fully active under the same conditions. To assess application potential of this laccase, we have investigated its ability to decolourise Indigo carmine. This enzyme was able to rapidly decolourise the dye in the presence of syringaldehyde as a redox mediator.
Dubé
Etienne
E
Département de Chimie-Biologie, Université du Québec à Trois-Rivières, Trois-Rivières, Quebec, Canada.
Shareck
François
F
Hurtubise
Yves
Y
Daneault
Claude
C
Beauregard
Marc
M
D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D003001:Cloning, Molecular; D004396:Coloring Agents / Q000378:metabolism*; D004795:Enzyme Stability; D015870:Gene Expression; D007203:Indigo Carmine; D007211:Indoles / Q000378:metabolism*; D007218:Industrial Microbiology; D042845:Laccase / Q000737:chemistry* / Q000235:genetics / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000201:enzymology*; D048170:Streptomyces lividans / Q000235:genetics / Q000378:metabolism; D013696:Temperature
D001426:Bacterial Proteins; D004396:Coloring Agents; D007211:Indoles; D007203:Indigo Carmine; D042845:Laccase