Isolation and characterization of novel pI 4.8 MnP isoenzyme from white-rot fungus Irpex lacteus

Sklenář, Jan; Niku‐Paavola, Marja‐Leena; Santos, Sílvia; Man, Petr; Kruus, Kristiina; Novotný, Čeněk

doi:10.1016/j.enzmictec.2010.03.001

Cited by 19 publications

(18 citation statements)

References 47 publications

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“…The optimal pH of recombinant Il-MnP was 6.5, which is similar to that of native MnP from I. lacteus F17, but was different from most of the reported fungi MnPs that had optima between 4.0 and 5.5 [58,61,25]. Interestingly, the recombinant enzyme showed remarkable stability at alkaline pH (9.0), and no distinct inactivation was observed during 24 h at a pH range from 3.5 to 9.0.…”

Section: Discussionmentioning

confidence: 70%

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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

Chen

Zheng

Jia

et al. 2015

Process Biochemistry

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Section: Discussionmentioning

confidence: 70%

“…lacteus. Although many strains of I. lacteus have been reported for their good capacity for degradation of organopollutants, showing a crucial role of MnP in the bioremediation process [22][23][24][25], no information about MnP-encoding genes from the fungus was disclosed.…”

Section: Introductionmentioning

confidence: 99%

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

Chen

Zheng

Jia

et al. 2015

Process Biochemistry

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“…The production of lignolytic enzymes and its regulation has been intensively studied in various lignin degrading fungi (Bonnarme &Jeffries 1990;Hakala et al 2006; Jiménez-Tobon et al 2003;Kamitsuji et al 2004;Lankinen et al 2005; Martínez et al 1996;Moilanen et al 1996;Nuske et al 2002; Perie &Gold 1991;Palma et al 2000;Petruccioli et al 2009;Schneegab et al 1997; Swamy &Ramsay 1999;Steffen et al 2002;Susla et al 2008;Silva et al 2008;Sklenar et al 2010;Singh et al 2011; Taboada-Puig et al 2011;Vares et al 1995;Wang et al 2001;Wang et al 2008) and novel bacterial strains (Bharagava et al 2009; Mishra &Thakur 2010;Yadav et al 2011). Recombinant production has been studied in filamentous fungi (Conesa et al 2000;Irie et al 2001;Li et al 2001;Mayfield et al 1994;Stewart et al 1996), yeasts (Jiang et al 2008a), bacterial (Whitwam &Tien 1996) and insect (Johnson et al 1992 ; Pease et al 1991) hosts with successful production but modest yields of active enzyme.…”

Section: Introductionmentioning

confidence: 99%

Screening of white-rot fungi manganese peroxidases: a comparison between the specific activities of the enzyme from different native producers

et al. 2012

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In this study manganese peroxidase (MnP) enzymes from selected white-rot fungi were isolated and compared for potential future recombinant production. White-rot fungi were cultivated in small-scale in liquid media and a simplified process was established for the purification of extracellular enzymes.Five lignin degrading organisms were selected (Bjerkandera sp., Phanerochaete (P.) chrysosporium, Physisporinus (P.) rivulosus, Phlebia (P.) radiata and Phlebia sp. Nf b19) and studied for MnP production in small-scale. Extracellular MnP activity was followed and cultivations were harvested at proximity of the peak activity. The production of MnPs varied in different organisms but was clearly regulated by inducing liquid media components (Mn2+, veratryl alcohol and malonate). In total 8 different MnP isoforms were purified.Results of this study reinforce the conception that MnPs from distinct organisms differ substantially in their properties. Production of the extracellular enzyme in general did not reach a substantial level. This further suggests that these native producers are not suitable for industrial scale production of the enzyme. The highest specific activities were observed with MnPs from P. chrysosporium (200 U mg-1), Phlebia sp. Nf b19 (55 U mg-1) and P. rivulosus (89 U mg-1) and these MnPs are considered as the most potential candidates for further studies. The molecular weight of the purified MnPs was estimated to be between 45–50 kDa.

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“…Additionally, both of the soluble and refolded MnP exhibited a broad range of substrate specificities in the presence of Mn 2+ . Interestingly, soluble MnP had an optimum pH of 8.0 and an optimum temperature of 45 °C, which are obviously different from those of refolded MnP (pH 6.5 and 15 °C, respectively) and most of the reported fungal MnPs [47–49]. Furthermore, the two enzymes had the ability to decolorize the azo dyes Acid Red 18 and Orange G, but the decolorization efficiency of soluble MnP was lower than that of refolded MnP.…”

Section: Discussionmentioning

confidence: 99%

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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Isolation and characterization of novel pI 4.8 MnP isoenzyme from white-rot fungus Irpex lacteus

Cited by 19 publications

References 47 publications

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

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

Screening of white-rot fungi manganese peroxidases: a comparison between the specific activities of the enzyme from different native producers

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

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