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Protein and Peptide Letters 2021Class III plant peroxidases play important role in a number of physiological processes in plants such as lignin biosynthesis, suberization, cell wall biosynthesis,...
BACKGROUND
Class III plant peroxidases play important role in a number of physiological processes in plants such as lignin biosynthesis, suberization, cell wall biosynthesis, reactive oxygen species metabolism and plant defense against pathogens. Peroxidases are also of significance in several industrial applications. In view of this, the production and identification of novel peroxidases having resistance towards temperature, pH, salts is desirable.
OBJECTIVE
The objective of the present work was to clone and characterize a novel plant peroxidase suitable for industrial application.
METHODS
A full length cDNA clone of lemon peroxidase was isolated using PCR and RACE approaches, characterized and heterologously expressed in Escherichia coli using standard protocols. The expressed peroxidase was purified using Ni-NTA agarose column and biochemically characterized using standard protocols. The peroxidase was also in-silico characterized at nucleotide as well as protein levels using standard protocols.
RESULTS
A full length cDNA clone of lemon peroxidase was isolated and expressed heterologously in E. coli. The expressed recombinant lemon peroxidase (LPRX) was activated by in-vitro refolding and purified. The purified LPRX exhibited pH and temperature optima of pH 7.0 and 50°C, respectively. The LPRX was found to be activated by metal ions (Na+, Ca2+, Mg2+ and Mn2+) at lower concentration. The expressional analysis of the transcripts suggested involvement of lemon peroxidase in plant defense. The lemon peroxidase was in silico modelled and docked with the substrates guaiacol, and pyrogallol and shown the favourability of pyrogallol over guaiacol, which is in agreement with the in-vitro findings. The protein function annotation analyses suggested the involvement of lemon peroxidase in the phenylpropanoid biosynthesis pathway and plant defense mechanisms.
CONCLUSION
Based on the biochemical characterization, the purified peroxidase was found to be resistant towards the salts and thus, might be a good candidate for industrial exploitation. The in-silico protein function annotation and transcript analyses highlighted the possible involvement of the lemon peroxidase in plant defense response.
Topics: Citrus; Gene Expression; Peroxidase; Plant Proteins; Recombinant Proteins
PubMed: 32981494
DOI: 10.2174/0929866527666200925114054 -
Molecules (Basel, Switzerland) Jul 2022The peroxidase-like activity of vitamin B6 (VB6) was firstly demonstrated by catalyzing the peroxidase chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) at the...
The peroxidase-like activity of vitamin B6 (VB6) was firstly demonstrated by catalyzing the peroxidase chromogenic substrate 3,3',5,5'-tetramethylbenzidine (TMB) at the existence of HO. The influence of different factors on the catalytic property of VB6, including pH, temperature, VB6 concentration, and incubation time, were investigated. The steady-state kinetic study results indicate that VB6 possesses higher affinity to HO than natural horseradish peroxidase and some other peroxidase mimics. Besides, the radical quenching experiment results confirm that hydroxyl radical (•OH) accounts for the catalytic process. Based on the excellent peroxidase-like catalytic activity of VB6, the colorimetric methods for HO and gallic acid (GA) detection were developed by measuring the absorbance variance of the catalytic system. Under the optimal conditions, the linear ranges of the methods for HO and GA determination with good selectivity are 50.0-600.0 μM and 10.0-50.0 μM, respectively. In addition, the developed method was applied in the detection of HO in milk samples and evaluation of total antioxidant capacity of different tea infusions. This study may broaden the application prospect of VB6 in environmental and biomedical analysis fields, contribute to profound insight of the physiological functions of VB6, as well as lay foundation for further excavation of small-molecule peroxidase mimics.
Topics: Antioxidants; Biomimetic Materials; Colorimetry; Hydrogen Peroxide; Peroxidase; Peroxidases; Vitamin B 6
PubMed: 35807507
DOI: 10.3390/molecules27134262 -
Frontiers in Cellular and Infection... 2023The fungal G protein-coupled receptors Ste2 and Ste3 are vital in mediating directional hyphal growth of the agricultural pathogen towards wheat plants. This...
INTRODUCTION
The fungal G protein-coupled receptors Ste2 and Ste3 are vital in mediating directional hyphal growth of the agricultural pathogen towards wheat plants. This chemotropism is induced by a catalytic product of peroxidases secreted by the wheat. Currently, the identity of this product, and the substrate it is generated from, are not known.
METHODS AND RESULTS
We provide evidence that a peroxidase substrate is derived from conidia and report a simple method to extract and purify the Ste2-activating ligand for analyses by mass spectrometry. The mass spectra arising from t he ligand extract are characteristic of a 400 Da carbohydrate moiety. Consistent with this type of molecule, glycosidase treatment of conidia prior to peroxidase treatment significantly reduced the amount of ligand extracted. Interestingly, availability of the peroxidase substrate appears to depend on the presence of both Ste2 and Ste3, as knockout of one or the other reduces the chemotropism-inducing effect of the extracts.
CONCLUSIONS
While further characterization is necessary, identification of the -derived peroxidase substrate and the Ste2-activating ligand will unearth deeper insights into the intricate mechanisms that underlie fungal pathogenesis in cereal crops, unveiling novel avenues for inhibitory interventions.
Topics: Peroxidase; Ligands; Peroxidases; Fusarium; Plant Diseases
PubMed: 38239502
DOI: 10.3389/fcimb.2023.1287418 -
Nano Letters Jul 2023Pursuing effective and generalized strategies for modulating the electronic structures of atomically dispersed nanozymes with remarkable catalytic performance is...
Pursuing effective and generalized strategies for modulating the electronic structures of atomically dispersed nanozymes with remarkable catalytic performance is exceptionally attractive yet challenging. Herein, we developed a facile "formamide condensation and carbonization" strategy to fabricate a library of single-atom (M-NC; 6 types) and dual-atom (M/M-NC; 13 types) metal-nitrogen-carbon nanozymes (M = Fe, Co, Ni, Mn, Ru, Cu) to reveal peroxidase- (POD-) like activities. The FeCo-NC dual-atom nanozyme with Fe-N/Co-N coordination displayed the highest POD-like activity. Density functional theory (DFT) calculations revealed that the Co atom site synergistically affects the d-band center position of the Fe atom site and served as the second reaction center, which contributes to better POD-like activity. Finally, FeCo NC was shown to be effective in inhibiting tumor growth both and , suggesting that diatomic synergy is an effective strategy for developing artificial nanozymes as novel nanocatalytic therapeutics.
Topics: Peroxidase; Peroxidases; Carbon; Catalysis; Coloring Agents
PubMed: 37358396
DOI: 10.1021/acs.nanolett.3c01454 -
Applied Microbiology and Biotechnology Apr 2022Fungal dye-decolorizing peroxidases (DyPs) have found applications in the treatment of dye-contaminated industrial wastes or to improve biomass digestibility. Their...
Fungal dye-decolorizing peroxidases (DyPs) have found applications in the treatment of dye-contaminated industrial wastes or to improve biomass digestibility. Their roles in fungal biology are uncertain, although it has been repeatedly suggested that they could participate in lignin degradation and/or modification. Using a comprehensive set of 162 fully sequenced fungal species, we defined seven distinct fungal DyP clades on basis of a sequence similarity network. Sequences from one of these clades clearly diverged from all others, having on average the lower isoelectric points and hydropathy indices, the highest number of N-glycosylation sites, and N-terminal sequence peptides for secretion. Putative proteins from this clade are absent from brown-rot and ectomycorrhizal species that have lost the capability of degrading lignin enzymatically. They are almost exclusively present in white-rot and other saprotrophic Basidiomycota that digest lignin enzymatically, thus lending support for a specific role of DyPs from this clade in biochemical lignin modification. Additional nearly full-length fungal DyP genes were isolated from the environment by sequence capture by hybridization; they all belonged to the clade of the presumably secreted DyPs and to another related clade. We suggest focusing our attention on the presumably intracellular DyPs from the other clades, which have not been characterized thus far and could represent enzyme proteins with novel catalytic properties. KEY POINTS: • A fungal DyP phylogeny delineates seven main sequence clades. • Putative extracellular DyPs form a single clade of Basidiomycota sequences. • Extracellular DyPs are associated to white-rot fungi.
Topics: Basidiomycota; Coloring Agents; Fungal Proteins; Lignin; Peroxidase; Peroxidases
PubMed: 35435459
DOI: 10.1007/s00253-022-11923-0 -
Langmuir : the ACS Journal of Surfaces... Jul 2020The adsorption behavior and enzyme activity of horseradish peroxidase (HRP) was examined on a synthetic clay nanosheet, whose surface is flat at the atomic level and is...
The adsorption behavior and enzyme activity of horseradish peroxidase (HRP) was examined on a synthetic clay nanosheet, whose surface is flat at the atomic level and is negatively charged. The results showed that HRP is adsorbed effectively (adsorption equilibrium constant, = 1.61 × 10 L mol) and that the structure of HRP was altered on the clay surface. The enzyme activity of HRP on the clay surface was evaluated by using HO and -BuOOH as a substrate. As a result, HRP on the clay surface was able to work for -BuOOH, while HRP in solution did not show any activity. In addition, HRP on SSA showed reactivity even under the high-temperature conditions. These results indicate that the clay nanosheet can be a unique modifier for enzyme activity of HRP.
Topics: Clay; Horseradish Peroxidase; Hydrogen Peroxide; Peroxidase; Peroxidases
PubMed: 32407124
DOI: 10.1021/acs.langmuir.0c00607 -
Bioresource Technology Nov 2023Pseudomonas putida is a promising strain for lignin valorisation. However, there is a dearth of stable and efficient systems for secreting enzymes to enhance the...
Pseudomonas putida is a promising strain for lignin valorisation. However, there is a dearth of stable and efficient systems for secreting enzymes to enhance the process. Therefore, a novel secretion system for recombinant lignin-depolymerising peroxidase was developed. By adopting a flagellar type III secretion system, P. putida KT-M2, a secretory host strain, was constructed and an optimal secretion signal fusion partner was identified. Application of the dye-decolourising peroxidase of P. putida to this system resulted in efficient oxidation activity of the cell-free supernatant against various chemicals, including lignin model compounds. This peroxidase-secreting strain was examined to confirm its lignin utilisation capability, resulting in the efficient assimilation of various lignin substrates with 2.6-fold higher growth than that of the wild-type strain after 72 h of cultivation. Finally, this novel system will lead efficient bacterial lignin breakdown and utilization through enzyme secretion, paving the way for sustainable lignin-consolidated bioprocessing.
Topics: Lignin; Pseudomonas putida; Peroxidase; Peroxidases; Oxidoreductases; Coloring Agents
PubMed: 37739186
DOI: 10.1016/j.biortech.2023.129779 -
Chemistry (Weinheim An Der Bergstrasse,... Nov 2021The morphology of nanomaterials (geometric shape and dimension) play a significant role in its various physical and chemical properties. Thus, it is essential to link...
The morphology of nanomaterials (geometric shape and dimension) play a significant role in its various physical and chemical properties. Thus, it is essential to link morphology with performance in specific applications. For this purpose, the morphology of copper metal-organic polyhedra (Cu-MOP) can be modulated through distinct assembly process, which facilitates the exploration of the relationship between morphology and catalytic performance. In this work, the assemblies of Cu-MOP with three different morphologies (nanorods, nanofibers and nanosheets) were facilely prepared by the variation of solvent mixture of N, N-dimethylformamide (DMF) and methanol, revealed the important role of the interaction between the surface group and the solvent on the morphology of these assemblies. Cu-MOP nanofibers exhibited the highest mimetic peroxidase enzyme activity over the Cu-MOP nanosheets and nanorods, which have been utilized in the detection of glucose. Cu-MOPs assemblies with tunable morphology accompanied with adjustable mimic peroxidase activity, had great potential applications in the field of bioanalytical chemistry and biomedicals.
Topics: Catalysis; Copper; Oxidation-Reduction; Peroxidase; Peroxidases
PubMed: 34505733
DOI: 10.1002/chem.202102631 -
Environmental Microbiology Feb 2023Catalase-peroxidase is a heme oxidoreductase widely distributed in bacteria and lower eukaryotes. In this study, we identified a catalase-peroxidase PiCP1 (PITG_05579)...
Catalase-peroxidase is a heme oxidoreductase widely distributed in bacteria and lower eukaryotes. In this study, we identified a catalase-peroxidase PiCP1 (PITG_05579) in Phytophthora infestans. PiCP1 had catalase/peroxidase and secretion activities and was highly expressed in sporangia and upregulated in response to oxidative and heat stresses. Compared with wild type, PiCP1-silenced transformants (STs) had decreased catalase activity, reduced oxidant stress resistance and damped cell wall integrity. In contrast, PiCP1-overexpression transformants (OTs) demonstrated increased tolerance to abiotic stresses and induced the upregulation of PR genes in the host salicylic acid pathway. The high concentration of PiCP1 can also induced callose deposition in plant tissue. Importantly, both STs and OTs have severely reduced sporangia formation and zoospore releasing rate, but the sporangia germination rate and type varied depending on environmental conditions. Comparative sequence analyses show that catalase-peroxidases are broadly distributed and highly conserved among soil-borne plant parasitic oomycetes, but not in freshwater-inhabiting or strictly plants-inhabiting oomycetes. In addition, we found that silencing PiCP1 downregulated the expression of PiCAT2. These results revealed the important roles of PiCP1 in abiotic stress resistance, pathogenicity and in regulating asexual structure development in response to environmental change. Our findings provide new insights into catalase-peroxidase functions in eukaryotic pathogens.
Topics: Phytophthora infestans; Peroxidase; Catalase; Virulence; Stress, Physiological; Plant Diseases
PubMed: 36495132
DOI: 10.1111/1462-2920.16305 -
International Journal of Molecular... Apr 2023The development of artificial enzymes for application in sustainable technologies, such as the transformation of environmental pollutants or biomass, is one of the most...
The development of artificial enzymes for application in sustainable technologies, such as the transformation of environmental pollutants or biomass, is one of the most challenging goals in metalloenzyme design. In this work, we describe the oxidation of mono-, di-, tri- and penta-halogenated phenols catalyzed by the artificial metalloenzyme Fe-MC6*a. It promoted the dehalogenation of 4-fluorophenol into the corresponding 1,4-benzoquinone, while under the same experimental conditions, 4-chloro, 4-bromo and 4-iodophenol were selectively converted into higher molecular weight compounds. Analysis of the 4-chlorophenol oxidation products clarified that oligomers based on C-O bonds were exclusively formed in this case. All results show that Fe-MC6*a holds intriguing enzymatic properties, as it catalyzes halophenol oxidation with substrate-dependent chemoselectivity.
Topics: Peroxidase; Peroxidases; Oxidation-Reduction; Catalysis
PubMed: 37175773
DOI: 10.3390/ijms24098058