Cloning and expression of a new manganese peroxidase from Irpex lacteus F17 and its application in decolorization of reactive black 5

Chen, Wenting; Zheng, Leilei; Jia, Rong; Wang, Nan

doi:10.1016/j.procbio.2015.07.009

Cited by 42 publications

(37 citation statements)

References 64 publications

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“…Our previous studies indicated that the MnP preprotein from I. lacteus F17 is composed of 359 amino acid residues, including a 26-amino acid signal peptide and four disulfide bridges essential for the native conformation of the enzyme [22]. A cDNA encoding the mature 333-amino acid MnP protein without the signal sequence was subcloned into pET28a and expressed in E. coli Rosetta (DE3) cells.…”

Section: Discussionmentioning

confidence: 99%

“…Irpex lacteus F17 is a local, MnP-producing, white-rot basidiomycete fungus, and an MnP-encoding gene, imnp , from this species has been cloned and expressed in E. coli by our laboratory [22]. As expected, the expressed protein accumulated in an insoluble form as inclusion bodies.…”

Section: Introductionmentioning

confidence: 88%

“…The mature MnP cDNA sequence, named imnp , was amplified by polymerase chain reaction using the genomic DNA of I. lacteus F17 as a template [22]. The imnp cDNA was excised with Xho I and Nco I, and inserted into the expression vector pET28a(+).…”

Section: Methodsmentioning

confidence: 99%

“…In vitro refolding was performed according to the method of Chen et al [22]. The optimal refolding conditions were further studied in small-scale experiments.…”

Section: Methodsmentioning

confidence: 99%

“…As expected, the expressed protein accumulated in an insoluble form as inclusion bodies. Subsequent in vitro refolding and incorporating exogenous hemin into the recombinant protein led to the formation of biologically active MnP [22]. In the present study, we further optimized the in vitro refolding conditions to improve the efficiency of renaturation from inclusion bodies using a one-factor-at-a-time method and an orthogonal experimental design based on our previous results.…”

Section: Introductionmentioning

confidence: 99%

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Expression of a fungal manganese peroxidase in Escherichia coli: a comparison between the soluble and refolded enzymes

et al. 2016

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BackgroundManganese peroxidase (MnP) from Irpex lacteus F17 has been shown to have a strong ability to degrade recalcitrant aromatic pollutants. In this study, a recombinant MnP from I. lacteus F17 was expressed in Escherichia coli Rosetta (DE3) in the form of inclusion bodies, which were refolded to achieve an active enzyme. Further, we optimized the in vitro refolding conditions to increase the recovery yield of the recombinant protein production. Additionally, we attempted to express recombinant MnP in soluble form in E. coli, and compared its activity with that of refolded MnP.ResultsRefolded MnP was obtained by optimizing the in vitro refolding conditions, and soluble MnP was produced in the presence of four additives, TritonX-100, Tween-80, ethanol, and glycerol, through incubation at 16 °C. Hemin and Ca2+ supplementation was crucial for the activity of the recombinant protein. Compared with refolded MnP, soluble MnP showed low catalytic efficiencies for Mn2+ and H2O2 substrates, but the two enzymes had an identical, broad range substrate specificity, and the ability to decolorize azo dyes. Furthermore, their enzymatic spectral characteristics were analysed by circular dichroism (CD), electronic absorption spectrum (UV-VIS), fluorescence and Raman spectra, indicating the differences in protein conformation between soluble and refolded MnP. Subsequently, size exclusion chromatography (SEC) and dynamic light scattering (DLS) analyses demonstrated that refolded MnP was a good monomer in solution, while soluble MnP predominantly existed in the oligomeric status.ConclusionsOur results showed that two forms of recombinant MnP could be expressed in E. coli by varying the culture conditions during protein expression.Electronic supplementary materialThe online version of this article (doi:10.1186/s12896-016-0317-2) contains supplementary material, which is available to authorized users.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 88%

Section: Methodsmentioning

confidence: 99%

“…In vitro refolding was performed according to the method of Chen et al [22]. The optimal refolding conditions were further studied in small-scale experiments.…”

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Expression of a fungal manganese peroxidase in Escherichia coli: a comparison between the soluble and refolded enzymes

et al. 2016

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A robust and stable nano‐biocatalyst by co‐immobilization of chloroperoxidase and horseradish peroxidase for the decolorization of azo dyes

Jin

Long

et al. 2017

J of Chemical Tech & Biotech

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BACKGROUND Immobilization of more than one peroxidase on the same support is an ideal technology for application because such a multi‐enzyme catalytic system may show high activity over a very wide optimal range of pH, temperature and H2O2 concentration. In this work, chloroperoxidase (CPO) and horseradish peroxidase (HRP) were co‐immobilized on ZnO nanowires/macroporous SiO2 composite support through an in situ cross‐linking method. An anionic bi‐epoxy cross‐linker was used by adsorption on the surface of ZnO nanowires before cross‐linking. RESULTS The cross‐linking was carried out under suitable conditions: pH 6.5, reaction temperature 15°C and reaction time 15 h. Using a 1/1 mixture of CPO and HRP resulted in a co‐immobilized enzyme with loading 79.6 mgCPO g‐1support and 52.8 mgHRP g‐1support, and total specific activity up to 15.7 U per mgsupport. The co‐immobilized enzyme also showed good stability after 60 days of storage, and excellent reusability over 20 repeat uses. CONCLUSIONS For the decolorization of azo dyes the co‐immobilized CPO (60%)/HRP (40%) exhibited high catalytic activity over very wide ranges of pH, temperature and H2O2 concentration. Using this robust biocatalyst, complete decolorizations of azo dyes have been realized within 3 h of reaction. © 2017 Society of Chemical Industry

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Studies on the characteristics and mechanism of aerobic biodegradation of tetrabromobisphenol A by Irpex lacteus F17

Chang

Fan

et al. 2021

J Basic Microbiol

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The study investigated the characteristics of aerobic degradation of tetrabromobisphenol A (TBBPA) by Irpex lacteus F17 (I. lacteus F17) under four different cometabolic substrates (phenol, glucose, sodium pyruvate, and sodium citrate). The biodegradation of TBBPA by I. lacteus F17 could be enhanced via cometabolism, and glucose (8 g/L) was confirmed to be the optimum carbon source. For different initial solution pH ranging from 3.0 to 8.0, the results showed that I. lacteus F17 could be applied to biodegrade TBBPA in a wide pH range of 4.0-8.0, and the degradation rate could reach the maximum 75.31%, while the debromination rate reached the maximum 12.40% under pH 5.0. In addition, it has been confirmed that Mn 2+ (50 μmol/L) could promote the secretion of manganese peroxidase and TBBPA biodegradation efficiency. Seven intermediates were identified by gas chromatography-mass spectrometry analysis, and the possible degradation pathways were proposed, which indicated the biodegradation of TBBPA might be subjected to debromination, β-scission, hydroxylation, deprotonation, and oxidation reactions.

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Cloning and expression of a new manganese peroxidase from Irpex lacteus F17 and its application in decolorization of reactive black 5

Cited by 42 publications

References 64 publications

Expression of a fungal manganese peroxidase in Escherichia coli: a comparison between the soluble and refolded enzymes

Expression of a fungal manganese peroxidase in Escherichia coli: a comparison between the soluble and refolded enzymes

A robust and stable nano‐biocatalyst by co‐immobilization of chloroperoxidase and horseradish peroxidase for the decolorization of azo dyes

Studies on the characteristics and mechanism of aerobic biodegradation of tetrabromobisphenol A by Irpex lacteus F17

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