Enhanced catalytic efficiency of CotA-laccase by DNA shuffling
نویسندگان
چکیده
منابع مشابه
CotA of Bacillus subtilis is a copper-dependent laccase.
The spore coat protein CotA of Bacillus subtilis displays similarities with multicopper oxidases, including manganese oxidases and laccases. B. subtilis is able to oxidize manganese, but neither CotA nor other sporulation proteins are involved. We demonstrate that CotA is a laccase. Syringaldazine, a specific substrate of laccases, reacted with wild-type spores but not with DeltacotA spores. Co...
متن کاملDirected evolution of CotA laccase for increased substrate specificity using Bacillus subtilis spores.
Directed evolution is an effective strategy to engineer and optimize protein properties, and microbial cell-surface display is a successful method to screen protein libraries. Protein surface display on Bacillus subtilis spores is demonstrated as a tool for screening protein libraries for the first time. Spore display offers advantages over more commonly utilized microbe cell-surface display sy...
متن کاملEnzymatic biotransformation of the azo dye Sudan Orange G with bacterial CotA-laccase.
In the present study we show that recombinant bacterial CotA-laccase from Bacillus subtilis is able to decolourise, at alkaline pH and in the absence of redox mediators, a variety of structurally different synthetic dyes. The enzymatic biotransformation of the azo dye Sudan Orange G (SOG) was addressed in more detail following a multidisciplinary approach. Biotransformation proceeds in a broad ...
متن کاملCharacterization of an Alkali- and Halide-Resistant Laccase Expressed in E. coli: CotA from Bacillus clausii
The limitations of fungal laccases at higher pH and salt concentrations have intensified the search for new extremophilic bacterial laccases. We report the cloning, expression, and characterization of the bacterial cotA from Bacillus clausii, a supposed alkalophilic ortholog of cotA from B. subtilis. Both laccases were expressed in E. coli strain BL21(DE3) and characterized fully in parallel fo...
متن کاملInsight into stability of CotA laccase from the spore coat of Bacillus subtilis.
The axial ligand of the catalytic mononuclear T1 copper site (Met(502)) of the CotA laccase was replaced by a leucine or phenylalanine residue to increase the redox potential of the enzyme. These mutations led to an increase in the redox potential by approx. 100 mV relative to the wild-type enzyme but the catalytic constant k(cat) in the mutant enzymes was severely compromised. This decrease in...
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ژورنال
عنوان ژورنال: Bioengineered
سال: 2019
ISSN: 2165-5979,2165-5987
DOI: 10.1080/21655979.2019.1621134