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KMID : 0880220190570100900
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Journal of Microbiology
2019 Volume.57 No. 10 p.900 ~ p.909
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Kinetic characterization of laccase from Bacillus atrophaeus, and its potential in juice clarification in free and immobilized forms
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Narnoliya Lokesh Kumar
Agarwal Neera
Patel Satya N.
Singh Sudhir P.
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Abstract
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In the present study, a laccase gene (BaLc) from a lignin degrading bacterium, Bacillus atrophaeus, has been cloned and expressed in Escherichia coli. The optimal catalytic activity of the protein was achieved at 5.5 pH and 35°C temperature, measured by oxidation of ABTS. The Km and Vmax values were determined as 1.42 mM and 4.16 μmole/min, respectively. To achieve the enzyme recovery, the biocatalyst (BaLc) was covalently attached onto the functionalized iron magnetic-nanoparticles. The nanoparticles were characterized by zeta-potential and FTIR analyses. The immobilized BaLc enzyme was physico-kinetically characterized, exhibiting retention of 60% of the residual activity after ten reaction cycles of ABTS oxidation. The immobilized biocatalyst system was tested for its biotechnological exploitability in plant juice processing, achieving 41?58% of phenol reduction, 41?58% decolorization, 50?59% turbidity reduction in the extracts of banana pseudo-stem and sweet sorghum stalk, and apple fruit juice. This is the first study to demonstrate the use of nanoparticle-laccase conjugate in juice clarification. The findings suggest that B. atrophaus laccase is a potential catalytic tool for plant juice bioprocessing activities.
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KEYWORD
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laccase, Bacillus atrophaeus, immobilization, magnetic nanoparticles, fruit juice clarification
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