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A Laccase of Fomes durissimus MTCC-1173 and Its Role in the Conversion of Methylbenzene to Benzaldehyde

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Abstract

A laccase has been purified from the liquid culture growth medium containing bagasse particles of Fomes durissimus. The method involved concentration of the culture filtrate by ultrafiltration and anion exchange chromatography on diethyl aminoethyl cellulose. The sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS-PAGE) and native polyacrylamide gel electrophoresis both gave single protein band indicating that the enzyme preparation was pure. The molecular mass of the purified laccase determined from SDS-PAGE analysis was 75 kDa. Using 2,6-dimethoxyphenol as the substrate, the determined K m and k cat values of the laccase are 182 μM and 0.35 s−1, respectively, giving a k cat/K m value of 1.92 × 103 M−1 s−1. The pH and temperature optimum were 4.0 and 35 °C, respectively. The purified laccase has yellow colour and does not show absorption band around 610 nm found in blue laccases. Moreover, it transformed methylbenzene to benzaldehyde in the absence of mediator molecules, property exhibited by yellow laccases.

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Acknowledgements

The authors acknowledge the financial support of the Department of Science Technology, Government India, New Delhi to Dr. R.S.S. Yadav in the form of DST Young Scientist Scheme, DO. No. SR/FTP/CS-90/2007, dated 22.04.2008 under which this work has been done. R. Sahay is thankful to University Grant Commission New Delhi for the award of Rajiv Gandhi National Fellowship (SRF), File. No.16-22/2006 (SA II) dated 05-03-2007 to him.

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  1. Department of Chemistry, D.D.U. Gorakhpur University, Gorakhpur, 273009, Uttar Pradesh, India

    R. Sahay, R. S. S. Yadav, Sudha Yadava & K. D. S. Yadav

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  1. R. Sahay
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  2. R. S. S. Yadav
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Sahay, R., Yadav, R.S.S., Yadava, S. et al. A Laccase of Fomes durissimus MTCC-1173 and Its Role in the Conversion of Methylbenzene to Benzaldehyde. Appl Biochem Biotechnol 166, 563–575 (2012). https://doi.org/10.1007/s12010-011-9448-z

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