Direct over-expression, characterization and H2O2 stability study of active Pleurotus eryngii versatile peroxidase in Escherichia coli

Biotechnology Letters
Xue BaoJian-Jun Li

Abstract

The vpl2 gene, encoding versatile peroxidase (VP) from Pleurotus eryngii, was synthesized with codon optimization and cloned into vector-pET-32a(+) and over-expressed in Escherichia coli BL21(DE3). An active peroxidase fused to the thioredoxin-hexahistidine tag was directly obtained by IPTG induction in the presence of hemin. Most of over-expressed protein was in the soluble form, and was purified on a nickel column with >85 % purity at a yield of 12.5 mg/l. The purified fusion protein, having an Rz value (A(407)/A(280), a measure of hemin content of the peroxidases) of 1.2, oxidized ABTS veratryl alcohol, Mn(2+), and Reactive Black 5. Activity of the enzyme increased after removing the tag. It lost only 5 % of its activity in 6.4 mM H(2)O(2). This is the first report on direct over-expression of active VP in E. coli.

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Citations

Sep 7, 2014·Biotechnology Advances·Katerina ZelenaRalf G Berger
Oct 2, 2014·Applied Microbiology and Biotechnology·Nancy Coconi-LinaresMiguel A Gómez-Lim
Feb 5, 2015·The FEBS Journal·Loredano PollegioniElena Rosini
Apr 24, 2016·Applied Biochemistry and Biotechnology·Christoph J BehrensRalf G Berger
Apr 30, 2015·PloS One·Verónica Sáez-JiménezFrancisco J Ruiz-Dueñas
Apr 25, 2017·Biotechnology and Bioengineering·Yongkun LvJian Chen
Oct 15, 2020·Enzyme and Microbial Technology·Odwa D V BikoWillem H van Zyl

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