Heterologous expression and characterization of functional mushroom tyrosinase (AbPPO4)

Scientific Reports
Matthias PretzlerAnnette Rompel

Abstract

Tyrosinases are an ubiquitous group of copper containing metalloenzymes that hydroxylate and oxidize phenolic molecules. In an application context the term 'tyrosinase' usually refers to 'mushroom tyrosinase' consisting of a mixture of isoenzymes and containing a number of enzymatic side-activities. We describe a protocol for the efficient heterologous production of tyrosinase 4 from Agaricus bisporus in Escherichia coli. Applying this procedure a pure preparation of a single isoform of latent tyrosinase can be achieved at a yield of 140 mg per liter of autoinducing culture medium. This recombinant protein possesses the same fold as the enzyme purified from the natural source as evidenced by single crystal X-ray diffraction. The latent enzyme can be activated by limited proteolysis with proteinase K which cleaves the polypeptide chain after K382, only one The latent enzyme can amino acid before the main in-vivo activation site. Latent tyrosinase can be used as obtained and enzymatic activity may be induced in the reaction mixture by the addition of an ionic detergent (e.g. 2 mM SDS). The proteolytically activated mushroom tyrosinase shows >50% of its maximal activity in the range of pH 5 to 10 and accepts a wide range of substr...Continue Reading

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Citations

Jun 1, 2017·Accounts of Chemical Research·Aleksandar Bijelic, Annette Rompel
Dec 10, 2017·Scientific Reports·Hyunsu DoYoo Seong Choi
Sep 30, 2018·Applied and Environmental Microbiology·Efstratios NikolaivitsEvangelos Topakas
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Aug 5, 2017·Acta Crystallographica. Section F, Structural Biology Communications·Ioannis KampatsikasAnnette Rompel
May 23, 2020·Critical Reviews in Biochemistry and Molecular Biology·Mark-Anthony McLarin, Ivanhoe K H Leung
Sep 30, 2020·Chemical Society Reviews·Nadiia I Gumerova, Annette Rompel
Mar 3, 2019·Angewandte Chemie·Ioannis KampatsikasAnnette Rompel
Mar 28, 2019·Scientific Reports·Joscha BreibeckAnnette Rompel
Jul 30, 2019·Journal of Food Biochemistry·Zohreh JamaliNematollah Gheibi
Jan 1, 2019·Chemtexts·Aleksandar Bijelic, Annette Rompel
Jun 28, 2018·Journal of Materials Chemistry. B, Materials for Biology and Medicine·Jagpreet Singh Sidhu, Narinder Singh
Feb 6, 2020·Scientific Reports·Felix PanisAnnette Rompel
Oct 28, 2020·Chembiochem : a European Journal of Chemical Biology·Ioannis Kampatsikas, Annette Rompel
Feb 21, 2021·Biochemical and Biophysical Research Communications·Ayu MasyitaTakayuki Kuraishi
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Aug 14, 2020·Journal of Medicinal Chemistry·Brayan RoulierRomain Haudecoeur
Oct 1, 2021·Scientific Reports·Raphael LamplAnnette Rompel

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Methods Mentioned

BETA
PCR
affinity chromatography
size exclusion chromatography
X-ray

Software Mentioned

Dataplot
TEW
XDS
SIFT
MolProbity

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