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Title of Journal: Appl Microbiol Biotechnol

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Abbravation: Applied Microbiology and Biotechnology

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Springer-Verlag

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DOI

10.1007/11495628_7

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1432-0614

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Cloning and expression of a tyrosinase from Empha

Authors: Ashit Rao Priya Pimprikar Chandrika Bendigiri Ameeta Ravi Kumar Smita Zinjarde
Publish Date: 2011/06/18
Volume: 92, Issue: 5, Pages: 951-959
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Abstract

Amplification of the tyrosinase gene melO from the genomic DNA of Aspergillus oryzae NCIM 1212 yielded a 16kb product This gene was cloned into pYLEX1 and the resulting pTyroYLEX1 vector was transformed in Yarrowia lipolytica strain Po1g A clone displaying the highest specific activity for tyrosinase 1094 U/mg was used for obtaining the complementary DNA cDNA and for protein expression studies cDNA sequence analysis indicated the splicing of an intron present in the melO gene by Po1g Native polyacrylamide gel electrophoresis acidification at pH 30 followed by activity staining with lDOPA indicated the expression of an active tyrosinase The clone overexpressing the tyrosinase transformed ltyrosine to lDOPA On optimization of conditions for the biotransformation pH 40 temperature 60°C and with 35 mg of biomass 04 mg/ml of lDOPA was obtained

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