2012
DOI: 10.1007/s10529-012-0940-5
|View full text |Cite
|
Sign up to set email alerts
|

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

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(4… Show more

Help me understand this report

Search citation statements

Order By: Relevance

Paper Sections

Select...
3
1
1

Citation Types

4
12
0
3

Year Published

2014
2014
2017
2017

Publication Types

Select...
7

Relationship

1
6

Authors

Journals

citations
Cited by 23 publications
(19 citation statements)
references
References 16 publications
4
12
0
3
Order By: Relevance
“…Similar studies report that the half-lives of peroxidases in the presence of excess H 2 O 2 lie in the minute range [2,7,45], reaching up to 60 min, even after directed evolution approaches for the improvement of their properties. MtPerII appears relatively stable after 24 h in the presence of 5000-fold excess peroxide.…”
Section: Discussionsupporting
confidence: 61%
See 3 more Smart Citations
“…Similar studies report that the half-lives of peroxidases in the presence of excess H 2 O 2 lie in the minute range [2,7,45], reaching up to 60 min, even after directed evolution approaches for the improvement of their properties. MtPerII appears relatively stable after 24 h in the presence of 5000-fold excess peroxide.…”
Section: Discussionsupporting
confidence: 61%
“…The recombinant enzyme, named MtPerII was expressed and fully characterized. The expression yield obtained is almost 2.5-fold higher than levels reported in other studies involving heterologous expression of peroxidase genes in Aspergillus nidulans [41], P. pastoris [3,4] and E. coli [2]. Similar studies are sparse, probably due to the fact that heterologous expression of peroxidase genes poses many problems, such as the number of introns per ORF, the need for hemin addition, and the instability of the product, mainly due to protease degradation [42].…”
Section: Discussionmentioning
confidence: 79%
See 2 more Smart Citations
“…The model bacterium Escherichia coli has been extensively developed as an expression platform for fungal enzymes, and many recombinant enzymes have been isolated successfully from inclusion bodies and refolded with 1-28% efficiency [23][24][25][26][27][28][29][30] and maximum yields of 1.5-14 mg L ¡1 after in vitro activation and purification. 29 A soluble form of a VP was produced using a thioredoxin tag 31,32 or following intensive screening. 31 Few publications have described the activity of such recombinant enzymes against complex lignin model compounds or synthetic lignin.…”
Section: Recombinant Lignin-degrading Peroxidasesmentioning
confidence: 99%