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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 -
Chemosphere Jan 2024Single-atom nanozymes (SANs) are nanomaterials-based nanozymes with atomically dispersed enzyme-like active sites. SANs offer improved as well as tunable catalytic... (Review)
Review
Single-atom nanozymes (SANs) are nanomaterials-based nanozymes with atomically dispersed enzyme-like active sites. SANs offer improved as well as tunable catalytic activity. The creation of extremely effective SANs and their potential uses have piqued researchers' curiosity due to their advantages of cheap cost, variable catalytic activity, high stability, and large-scale production. Furthermore, SANs with uniformly distributed active centers and definite coordination structures offer a distinctive opportunity to investigate the structure-activity correlation and control the geometric and electrical features of metal centers. SANs have been extensively explored in photo-, thermal-, and electro-catalysis. However, SANs suffer from the following disadvantages, such as efficiency, non-mimicking of the 3-D complexity of natural enzymes, limited and narrow range of artificial SANs, and biosafety aspects. Among a quite limited range of artificial SANs, the peroxidase action of SANs has attracted significant research attention in the last five years with the aim of producing reactive oxygen species for use in cancer therapy, and water treatment among many other applications. In this review, we explore the recent progress of different SANs as peroxidase mimics, the role of the metal center in enzymatic activity, possible prospects, and underlying limitations in real-time applications.
Topics: Biomimetic Materials; Nanostructures; Peroxidase; Catalysis; Peroxidases
PubMed: 38303399
DOI: 10.1016/j.chemosphere.2023.140557 -
Nature Communications May 2022Developing tunable and stable peroxidase mimetics with high catalytic efficiency provides a promising opportunity to improve and expand enzymatic catalysis in lignin...
Developing tunable and stable peroxidase mimetics with high catalytic efficiency provides a promising opportunity to improve and expand enzymatic catalysis in lignin depolymerization. A class of peptoid-based peroxidase mimetics with tunable catalytic activity and high stability is developed by constructing peptoids and hemins into self-assembled crystalline nanomaterials. By varying peptoid side chain chemistry to tailor the microenvironment of active sites, these self-assembled peptoid/hemin nanomaterials (Pep/hemin) exhibit highly modulable catalytic activities toward two lignin model substrates 2,2-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) and 3,3',5,5'-tetramethylbenzidine. Among them, a Pep/hemin complex containing the pyridyl side chain showed the best catalytic efficiency (V/K = 5.81 × 10 s). These Pep/hemin catalysts are highly stable; kinetics studies suggest that they follow a peroxidase-like mechanism. Moreover, they exhibit a high efficacy on depolymerization of a biorefinery lignin. Because Pep/hemin catalysts are highly robust and tunable, we expect that they offer tremendous opportunities for lignin valorization to high value products.
Topics: Biomimetics; Coloring Agents; Hemin; Lignin; Peptoids; Peroxidase; Peroxidases
PubMed: 35641490
DOI: 10.1038/s41467-022-30285-9 -
International Journal of Molecular... Apr 2023Phenols are widely used in industries despite their toxicity, which requires governments to limit their concentration in water to 5 mg/L before discharge to the city...
Phenols are widely used in industries despite their toxicity, which requires governments to limit their concentration in water to 5 mg/L before discharge to the city sewer. Thus, it is essential to develop a rapid, simple, and low-cost detection method for phenol. This study explored two pathways of peroxidase immobilization to develop a phenol detection system: peroxidase encapsulation into polyelectrolyte microcapsules and peroxidase captured by CaCO. The encapsulation of peroxidase decreased enzyme activity by 96%; thus, this method cannot be used for detection systems. The capturing process of peroxidase by CaCO microspherulites did not affect the maximum reaction rate and the Michaelis constant of peroxidase. The native peroxidase-Vmax = 109 µM/min, Km = 994 µM; CaCO-peroxidase-Vmax = 93.5 µM/min, Km = 956 µM. Ultimately, a reusable phenol detection system based on CaCO microparticles with immobilized peroxidase was developed, capable of detecting phenol in the range of 700 ng/mL to 14 µg/mL, with an error not exceeding 5%, and having a relatively low cost and production time. The efficiency of the system was confirmed by determining the content of phenol in a paintwork product.
Topics: Phenol; Peroxidase; Phenols; Peroxidases; Enzymes, Immobilized; Horseradish Peroxidase
PubMed: 37047739
DOI: 10.3390/ijms24076766 -
The Journal of Physical Chemistry... Dec 2022As typical copper-based-manganese delafossites, CuMnO nanoflakes (CuMnO NFs) display excellent light absorption capacity, showing great promise for facile regulation of...
As typical copper-based-manganese delafossites, CuMnO nanoflakes (CuMnO NFs) display excellent light absorption capacity, showing great promise for facile regulation of catalysis reactions by light stimulation. Here, we report that the as-prepared CuMnO NFs possess intrinsic peroxidase- and oxidase-like activities, which are capable of catalyzing the oxidation of the enzymatic substrate 3,3',5,5'-tetramethylbenzidine (TMB) in the presence of HO or O, respectively. Additionally, CuMnO NFs exhibit significant light-enhanced dual enzyme-like performance (peroxidase and oxidase) for accelerating the oxidation of TMB, owing to a much greater light-induced reactive oxygen species (ROS) yield. The light-induced hydroxyl radicals (·OH) and superoxide radicals (O) hold the key to rapidly catalyzing the oxidation of TMB into the two-electron charge transfer complex diamine, subsequently improving the oxidation capacity and enzyme-like activity of CuMnO NFs as peroxidase and oxidase mimics. These findings confirm an efficient way to improve the enzyme-like performance of nanozymes and regulate the reactive oxygen species (ROS) level by simply employing irradiated light. This work also provides a general method for constructing external field-modulated smart nanozymes and an insightful approach to the corresponding enzyme-mimetic reaction mechanism.
Topics: Reactive Oxygen Species; Hydrogen Peroxide; Peroxidase; Oxidoreductases; Peroxidases; Coloring Agents
PubMed: 36516410
DOI: 10.1021/acs.jpclett.2c03204 -
Antioxidants & Redox Signaling May 2020Myeloperoxidase (MPO) is a heme peroxidase that is primarily expressed by neutrophils. It has the capacity to generate several reactive species, essential for its... (Review)
Review
Myeloperoxidase (MPO) is a heme peroxidase that is primarily expressed by neutrophils. It has the capacity to generate several reactive species, essential for its inherent antimicrobial activity and innate host defense. Dysregulated MPO release, however, can lead to tissue damage, as seen in several diseases. Increased MPO levels in circulation are therefore widely associated with conditions of increased oxidative stress and inflammation. Several studies have shown a strong correlation between MPO and cardiovascular disease (CVD), through which elevated levels of circulating MPO are linked to poor prognosis with increased risk of CVD-related mortality. Accordingly, circulating MPO is considered a "high-risk" biomarker for patients with acute coronary syndrome, atherosclerosis, heart failure, hypertension, and stroke, thereby implicating MPO as a multifaceted target for cardiovascular protection. Consistently, recent studies that target MPO in animal models of CVD have demonstrated favorable outcomes with regard to disease progression. Although most of these studies have established a critical link between circulating MPO and worsening cardiac outcomes, the mechanisms by which MPO exerts its detrimental effects in CVD remain unclear. Elucidating the mechanisms by which elevated MPO leads to poor prognosis and, conversely, investigating the beneficial effects of therapeutic MPO inhibition on alleviating disease phenotype will facilitate future MPO-targeted clinical trials for improving CVD-related outcomes.
Topics: Animals; Biomarkers; Cardiovascular Diseases; Humans; Peroxidase; Prognosis
PubMed: 31847538
DOI: 10.1089/ars.2019.7971 -
Journal of Biomaterials Applications Jul 2021Pectin has recently attracted increasing attention for biomedical and pharmaceutical applications. Due to the lack of adhesion molecules in polysaccharides, phenolic...
Pectin has recently attracted increasing attention for biomedical and pharmaceutical applications. Due to the lack of adhesion molecules in polysaccharides, phenolic hydroxyl conjugated gelatin was added to enzymatically-gellable peroxidase-modified pectin derivative and compared with phenolic hydroxyl -pectin/collagen. Both pectin and gelatin were modified by tyramine hydrochloride in the presence of EDC/NHS. The phenolic hydroxyl -pectin/phenolic hydroxyl -gelatin, phenolic hydroxyl-pectin/collagen, and phenolic hydroxyl -pectin hydrogels were prepared using horseradish peroxidase and hydrogen peroxide The hydrogels were characterized by gelation time analysis. Morphology, enzymatic biodegradation, mechanical and swelling properties as well as water vapor transmission rate were also evaluated. Fibroblasts were cultured for 7 days, and the survival rate was evaluated using conventional MTT assay. Hydrogels composed of Ph-pectin/Ph-gelatin showed decreased biodegradation rate, and WVTR and further improved mechanical performance in comparison with other groups. Both phenolic hydroxyl -pectin/collagen and phenolic hydroxyl -pectin/phenolic hydroxyl -gelatin hydrogels exhibited porous structures. The hydrogels composed of collagen promoted cell survival rate 1.4 and 3.5 times compared to phenolic hydroxyl -gelatin and phenolic hydroxyl -pectin based hydrogels at the end of 7 days, respectively (p < 0.001). The study demonstrated the potential of enzymatically-gellable pectin-based hydrogels as cost-effective frameworks for use in tissue engineering applications.
Topics: Cell Survival; Collagen; Fibroblasts; Gelatin; Horseradish Peroxidase; Hydrogels; Pectins; Peroxidase; Peroxidases; Succinimides; Tissue Engineering
PubMed: 33302758
DOI: 10.1177/0885328220977601 -
The Journal of Biological Chemistry Aug 2022The protozoan parasite Trypanosoma cruzi is the causative agent of American trypanosomiasis, otherwise known as Chagas disease. To survive in the host, the T. cruzi...
The protozoan parasite Trypanosoma cruzi is the causative agent of American trypanosomiasis, otherwise known as Chagas disease. To survive in the host, the T. cruzi parasite needs antioxidant defense systems. One of these is a hybrid heme peroxidase, the T. cruzi ascorbate peroxidase-cytochrome c peroxidase enzyme (TcAPx-CcP). TcAPx-CcP has high sequence identity to members of the class I peroxidase family, notably ascorbate peroxidase (APX) and cytochrome c peroxidase (CcP), as well as a mitochondrial peroxidase from Leishmania major (LmP). The aim of this work was to solve the structure and examine the reactivity of the TcAPx-CcP enzyme. Low temperature electron paramagnetic resonance spectra support the formation of an exchange-coupled [Fe(IV)=O Trp] compound I radical species, analogous to that used in CcP and LmP. We demonstrate that TcAPx-CcP is similar in overall structure to APX and CcP, but there are differences in the substrate-binding regions. Furthermore, the electron transfer pathway from cytochrome c to the heme in CcP and LmP is preserved in the TcAPx-CcP structure. Integration of steady state kinetic experiments, molecular dynamic simulations, and bioinformatic analyses indicates that TcAPx-CcP preferentially oxidizes cytochrome c but is still competent for oxidization of ascorbate. The results reveal that TcAPx-CcP is a credible cytochrome c peroxidase, which can also bind and use ascorbate in host cells, where concentrations are in the millimolar range. Thus, kinetically and functionally TcAPx-CcP can be considered a hybrid peroxidase.
Topics: Antioxidants; Ascorbate Peroxidases; Ascorbic Acid; Chagas Disease; Cytochrome-c Peroxidase; Cytochromes c; Heme; Humans; Peroxidase; Peroxidases; Substrate Specificity; Trypanosoma cruzi
PubMed: 35772495
DOI: 10.1016/j.jbc.2022.102204 -
International Journal of Molecular... Mar 2021We aim to clarify the ligninolytic capabilities of dye-decolorizing peroxidases (DyPs) from bacteria and fungi, compared to fungal lignin peroxidase (LiP) and versatile...
We aim to clarify the ligninolytic capabilities of dye-decolorizing peroxidases (DyPs) from bacteria and fungi, compared to fungal lignin peroxidase (LiP) and versatile peroxidase (VP). With this purpose, DyPs from sp., , and , VP from , and LiP from were produced, and their kinetic constants and reduction potentials determined. Sharp differences were found in the oxidation of nonphenolic simple (veratryl alcohol, VA) and dimeric (veratrylglycerol-β- guaiacyl ether, VGE) lignin model compounds, with LiP showing the highest catalytic efficiencies (around 15 and 200 s·mM for VGE and VA, respectively), while the efficiency of the DyP was 1-3 orders of magnitude lower, and no activity was detected with the bacterial DyPs. VP and LiP also showed the highest reduction potential (1.28-1.33 V) in the rate-limiting step of the catalytic cycle (i.e., compound-II reduction to resting enzyme), estimated by stopped-flow measurements at the equilibrium, while the DyP showed the lowest value (1.23 V). We conclude that, when using realistic enzyme doses, only fungal LiP and VP, and in much lower extent fungal DyP, oxidize nonphenolic aromatics and, therefore, have the capability to act on the main moiety of the native lignin macromolecule.
Topics: Catalase; Coloring Agents; Fungal Proteins; Fungi; Lignin; Peroxidase
PubMed: 33807844
DOI: 10.3390/ijms22052629 -
Analytical and Bioanalytical Chemistry Apr 2023Lysophosphatidylcholine (LPC) can be used as a biomarker for diseases such as cancer, diabetes, atherosclerosis, and sepsis. In this study, we demonstrated the ability...
Lysophosphatidylcholine (LPC) can be used as a biomarker for diseases such as cancer, diabetes, atherosclerosis, and sepsis. In this study, we demonstrated the ability of nanozymes to displace the natural derived enzyme in enzyme-based assays for the measurement of LPC. Synthesized polyvinylpyrrolidone-stabilized platinum-ruthenium nanozymes (PVP/PtRu NZs) had a uniform size of 2.48 ± 0.24 nm and superb peroxidase-mimicking activity. We demonstrated that the nanozymes had high activity over a wide pH and temperature range and high stability after long-term storage. The LPC concentration could be accurately analyzed through the absorbance and fluorescence signals generated by the peroxidation reaction using the synthesized nanozyme with substrates such as 3,3',5,5'-tetramethylbenzidine (TMB) and 10-acetyl-3,7-dihydroxyphenoxazine (Ampliflu™ Red). LPC at a concentration of 0-400 µM was used for the analysis, and the coefficient of determination (R) was 0.977, and the limit of detection (LOD) was 23.1 µM by colorimetric assay. In the fluorometric assay, the R was 0.999, and the LOD was 8.97 µM. The spiked recovery values for the determination of LPC concentration in human serum samples were 102-115%. Based on these results, we declared that PVP/PtRu NZs had an ability comparable to that of the native enzyme horseradish peroxidase (HRP) in the enzyme-based LPC detection method.
Topics: Humans; Peroxidase; Lysophosphatidylcholines; Peroxidases; Horseradish Peroxidase; Colorimetry; Hydrogen Peroxide
PubMed: 36792781
DOI: 10.1007/s00216-023-04590-1