Identification of novel natural anti-browning agents based on phenotypic and metabolites differences in potato cultivars.

This study aimed to elucidate the changes of browning-related metabolite in fresh-cut potato and to identify anti-browning agents. Metabolomics and weighted correlation network analysis (WGCNA) were used to identify metabolites and correlate them with potato browning traits. A total of 79 browning trait-positive-related metabolites and 19 browning trait-negative-related metabolites were obtained from four key modules via WGCNA. The accumulation of metabolites with rich reducing groups and acidic groups were found to enhance anti-browning activity in potatoes. Among these metabolites, only L-pyroglutamic acid (L-PA) and ascorbic acid had variable importance for the projection (VIP) values greater than 1.5. In addition, it was found that L-PA inhibited polyphenol oxidase (PPO) activity by lowering pH and interacting with amino acid residues of PPO. L-PA also inhibited the growth of microorganisms in fresh-cut potato. Our results show that L-AP is an effective novel anti-browning agent with antibacterial activity.

Inhibition of polyphenol oxidase for preventing browning in edible mushrooms: A review.

Edible mushrooms are rich in nutrients and bioactive compounds, but their browning affects their quality and commercial value. This article reviews various methods to inhibit polyphenol oxidase (PPO)-induced browning in mushrooms. Physical methods such as heat treatment, low temperatures, irradiation, and ultrasound effectively reduce PPO activity but may affect mushroom texture and flavor. Chemical inhibitors, including synthetic chemicals and natural plant extracts, provide effective PPO inhibition but require careful monitoring of their content. Biological methods, including gene editing and microbial fermentation, show promise in targeting PPO genes and enhancing antioxidant production. Combining these methods offers a comprehensive strategy for preserving mushroom quality, extending shelf life, and maintaining nutritional value. PRACTICAL APPLICATION: These approaches can be applied in the food industry to improve post-harvest mushroom preservation, enhance product quality, and reduce waste, benefiting both producers and consumers. Further research and innovation are needed to optimize the practical application of these methods in large-scale processing and storage conditions.

Biodegradation of organic micropollutants by anoxic denitrification: Roles of extracellular polymeric substance adsorption, enzyme catalysis, and reactive oxygen species oxidation.

The control of organic micropollutants (OMPs) in water environments have received significant attention. Denitrification was reported to exhibit good efficiency to remove OMPs, and the mechanisms involved in are too intricate to be well illustrated. In this study, we selected nitrobenzene [NB] and bisphenol A [BPA] as model pollutants and aimed to unravel the mechanisms of Paracoccus Denitrificans in the removal of OMPs, with a specific emphasis on aerobic behavior during denitrification processes. We demonstrated the formation of extracellular superoxide radicals, i.e., extracellular •O2-, using a chemiluminescence probe and found that extracellular polymeric substance adsorption, extracellular •O2-, and microbial assimilation contributed approximately 40 %, 10 %, and 50 % to OMPs removal, respectively. Transcriptome analysis further revealed the high expression and enrichment of several pathways, such as drug metabolism-other enzymes, of which a typical aerobic enzyme of polyphenol oxidase [PPO] participates in the degradation of NB and BPA. Importantly, all the immediate products showed a significant decrease in toxicity during the aerobic activity-related OMPs degradation process based on the proposed degradation pathways. This study demonstrates the formation of extracellular •O2- and the mechanisms of extracellular •O2-- and PPO-mediated OMPs biodegradation, and offers new insights into OMPs control in widely-used denitrification treatment processes.

In Silico Analysis of the Molecular Interaction between Anthocyanase, Peroxidase and Polyphenol Oxidase with Anthocyanins Found in Cranberries.

Anthocyanins are bioactive compounds responsible for various physiological processes in plants and provide characteristic colors to fruits and flowers. Their biosynthetic pathway is well understood; however, the enzymatic degradation mechanism is less explored. Anthocyanase (ß-glucosidase (BGL)), peroxidase (POD), and polyphenol oxidase (PPO) are enzymes involved in degrading anthocyanins in plants such as petunias, eggplants, and Sicilian oranges. The aim of this work was to investigate the physicochemical interactions between these enzymes and the identified anthocyanins (via UPLC-MS/MS) in cranberry (Vaccinium macrocarpon) through molecular docking to identify the residues likely involved in anthocyanin degradation. Three-dimensional models were constructed using the AlphaFold2 server based on consensus sequences specific to each enzyme. The models with the highest confidence scores (pLDDT) were selected, with BGL, POD, and PPO achieving scores of 87.6, 94.8, and 84.1, respectively. These models were then refined using molecular dynamics for 100 ns. Additionally, UPLC-MS/MS analysis identified various flavonoids in cranberries, including cyanidin, delphinidin, procyanidin B2 and B4, petunidin, pelargonidin, peonidin, and malvidin, providing important experimental data to support the study. Molecular docking simulations revealed the most stable interactions between anthocyanase and the anthocyanins cyanidin 3-arabinoside and cyanidin 3-glucoside, with a favorable ΔG of interaction between -9.3 and -9.2 kcal/mol. This study contributes to proposing a degradation mechanism and seeking inhibitors to prevent fruit discoloration.

Lily bulb polyphenol oxidase obtained via an optimized multi-stage separation strategy for structural analysis and browning mechanism elucidation.

An optimized multi-stage separation strategy was developed to purify lily bulb polyphenol oxidase (PPO) for revealing its molecular structure. The PPO was purified 14.64-fold with high specific activity of 153,900 U/mg via optimized conditions of phosphate buffer pH (6.5), solid-liquid ratio (1:3), PVPP content (2 %), extraction time (4 h), followed by 30 %-50 % ammonium sulfate, diethylaminoethyl ion-exchange chromatography (0.1 M NaCl), and size exclusion chromatography. The PPO was identified as a dimeric protein with molecular weight of 135 kDa, containing 58.79 % random coil, 20.78 % α-helix, 17.41 % ß-folding, and 3.02 % ß-corner. The three-dimensional structure via homology modeling suggested that active center CuA bound to His151, His172, and His181, CuB bound to His307, His311, and His341. Furthermore, molecular docking indicated that its Phe337 and Tyr312 residues were catalytic cavity gates of catechol and 4-methylcatechol, respectively. Therefore, this study successfully analyzed purified PPO structure and further provided a theoretical foundation for its browning mechanism.

Specific Substrate Activity of Lotus Root Polyphenol Oxidase: Insights from Gaussian-Accelerated Molecular Dynamics and Markov State Models.

Polyphenol oxidase (PPO) plays a key role in the enzymatic browning process, and this study employed Gaussian-accelerated molecular dynamics (GaMD) simulations to investigate the catalytic efficiency mechanisms of lotus root PPO with different substrates, including catechin, epicatechin, and chlorogenic acid, as well as the inhibitor oxalic acid. Key findings reveal significant conformational changes in PPO that correlate with its enzymatic activity. Upon substrate binding, the alpha-helix in the Q53-D63 region near the copper ion extends, likely stabilizing the active site and enhancing catalysis. In contrast, this helix is disrupted in the presence of the inhibitor, resulting in a decrease in enzymatic efficiency. Additionally, the F350-V378 region, which covers the substrate-binding site, forms an alpha-helix upon substrate binding, further stabilizing the substrate and promoting catalytic function. However, this alpha-helix does not form when the inhibitor is bound, destabilizing the binding site and contributing to inhibition. These findings offer new insights into the substrate-specific and inhibitor-induced structural dynamics of lotus root PPO, providing valuable information for enhancing food processing and preservation techniques.

Polyphenol Oxidase Activity on Guaiacyl and Syringyl Lignin Units.

The natural heterogeneity of guaiacyl (G) and syringyl (S) compounds resulting from lignin processing hampers their direct use as plant-based chemicals and materials. Herein, we explore six short polyphenol oxidases (PPOs) from lignocellulose-degrading ascomycetes for their capacity to react with G-type and S-type phenolic compounds. All six PPOs catalyze the ortho-hydroxylation of G-type compounds (guaiacol, vanillic acid, and ferulic acid), forming the corresponding methoxy-ortho-diphenols. Remarkably, a subset of these PPOs is also active towards S-compounds (syringol, syringic acid, and sinapic acid) resulting in identical methoxy-ortho-diphenols. Assays with 18O2 demonstrate that these PPOs in particular catalyze ortho-hydroxylation and ortho-demethoxylation of S-compounds and generate methanol as a co-product. Oxidative (ortho-) demethoxylation of S-compounds is a novel reaction for PPOs, which we propose occurs by a distinct reaction mechanism as compared to aryl-O-demethylases. We further show that addition of a reducing agent can steer the PPO reaction to form methoxy-ortho-diphenols from both G- and S-type substrates rather than reactive quinones that lead to unfavorable polymerization. Application of PPOs opens for new routes to reduce the heterogeneity and methoxylation degree of mixtures of G and S lignin-derived compounds.

Phenolic profile, antioxidant capacity and oxidoreductase enzyme activity in autochthonous grape varieties from Bosnia and Herzegovina.

The variability of phenolic compounds among grape varieties has an important role in selecting winemaking techniques, but the use of phenolic profiles for quality control is still fragmented and incomplete. Given the recent climate change and global warming, biochemical characterisation of secondary metabolites in autochthonous grape varieties is a very important factor for their preservation and sustainable agriculture. Two autochthonous grape varieties from the western Herzegovina region in Bosnia and Herzegovina have been selected for the research targeting at the evaluation of their phenolic profiles, antioxidant activities, and the correlation with oxidoreductase enzymes polyphenol oxidase and Class III peroxidase, in different berry tissues. The obtained results indicate a similar qualitative profile of phenolic compounds in exocarp and mesocarp in both varieties, but their concentrations and antioxidant activity vary significantly. The correlation between phenolic compounds and oxidoreductase enzyme activities in different grape berry tissues is discussed in this article.

Purification of polyphenol oxidase from tea (Camellia sinensis) using three-phase partitioning with a green deep eutectic solvent.

In this study, tea polyphenol oxidase (PPO) was purified via three-phase partitioning (TPP) using a deep eutectic solvent (DES) instead of t-butanol. First, the properties of 13 types of synthesized DESs were characterized, and DES-7 (thymol/dodecanoic acid) was selected as the best alternative solvent. The process parameters were optimized using response surface methodology. The experimental results revealed that when the (NH4)2SO4 concentration, DES to crude extract ratio, extraction time, and pH were 41%, 0.5:1, 75 min, and 5.6, respectively, the recovery and purification fold of tea PPO were 78.44% and 8.26, respectively. SDS-PAGE and native-PAGE were used to analyze the PPO before and after purification of the TTP system, and the molecular weight and purification effect of PPO were detected. Moreover, the DES could be recovered and recycled. The results indicate an environmentally friendly and stable DES, and provide a reference for the large-scale application of TPP to extract PPO.

Biosynthesis of Piceatannol from Resveratrol in Grapevine Can Be Mediated by Cresolase-Dependent Ortho-Hydroxylation Activity of Polyphenol Oxidase.

Piceatannol is a naturally occurring hydroxylated analogue of the stilbene phytoalexin resveratrol that can be found in grape fruit and derived products. Piceatannol has aroused great interest as it has been shown to surpass some human health-beneficial properties of resveratrol including antioxidant activity, several pharmacological activities and also bioavailability. The plant biosynthetic pathway of piceatannol is still poorly understood, which is a bottleneck for the development of both plant defence and bioproduction strategies. Cell cultures of Vitis vinifera cv. Gamay, when elicited with dimethyl-ß-cyclodextrin (MBCD) and methyl jasmonate (MeJA), lead to large increases in the accumulation of resveratrol, and after 120 h of elicitation, piceatannol is also detected due to the regiospecific hydroxylation of resveratrol. Therefore, an ortho-hydroxylase must participate in the biosynthesis of piceatannol. Herein, three possible types of resveratrol hydroxylation enzymatic reactions have been tested, specifically, a reaction catalyzed by an NADPH-dependent cytochrome, P450 hydroxylase, a 2-oxoglutarate-dependent dioxygenase and ortho-hydroxylation, similar to polyphenol oxidase (PPO) cresolase activity. Compared with P450 hydoxylase and the dioxygenase activities, PPO displayed the highest specific activity detected either in the crude extract, the particulate or the soluble fraction obtained from cell cultures elicited with MBCD and MeJA for 120 h. The overall yield of PPO activity present in the crude extract (107.42 EU) was distributed mostly in the soluble fraction (66.15 EU) rather than in the particulate fraction (3.71 EU). Thus, partial purification of the soluble fraction by precipitation with ammonium sulphate, dialysis and ion exchange chromatography was carried out. The soluble fraction precipitated with 80% ammonium sulphate and the chromatographic fractions also showed high levels of PPO activity, and the presence of the PPO protein was confirmed by Western blot and LC-MS/MS. In addition, a kinetic characterization of the cresolase activity of partially purified PPO was carried out for the resveratrol substrate, including Vmax and Km parameters. The Km value was 118.35 ± 49.84 µM, and the Vmax value was 2.18 ± 0.46 µmol min-1 mg-1.

The role of processing on phenolic bioaccessibility and antioxidant capacity of apple derivatives.

Fruit derivatives are commonly obtained by applying processing operations deemed responsible for the loss of phenol compounds, but very little information is available on the fate of phenols upon digestion of these products. The present study evaluated the effect of thermal and mechanical treatments, commonly applied to turn apple pulp into puree and homogenate, on phenolic bioaccessibility and antioxidant activity. Despite a 20 % decrease in polyphenols due to processing, their bioaccessibility was higher in apple derivatives (>20 %) compared to pulp (∼2 %). Polyphenol oxidase (PPO), inactivated by thermal treatments in apple derivatives but not in the pulp, was hypothesized to be responsible for this difference. Results acquired on an unprocessed PPO-free apple model, only featuring quercetin-3-glucoside and pectin, actually exhibited similar bioaccessibility as processed derivatives. The radical scavenging capacity was unaffected by the structural integrity of apples, indicating independence from the plant tissue's hierarchical arrangement. After digestion, radical scavenging capacity decreased in the real apple matrices, correlating with phenolic content, while it was retained in the apple model, further suggesting the pivotal food matrix role in modulating polyphenols bioaccessibility and subsequent biological activity. Translating these results to an industrial scale, processing conditions can be optimized not only to guarantee that the quality requirements are met, but also to achieve desired nutritional benefits.

Biochemical and toxicological characteristics of polyphenol oxidase from red palm weevil Rhynchophorus ferrugineus (Olivier) (Coleoptera: Curculionidae).

Red palm weevil (RPW) Rhynchophorus ferrugineus is the most destructive insect pests of numerous palm species in the world. The introduction of botanical extract(s) as integral part of an integrated pest management (IPM) programs against RPW will reduce the use of chemical insecticides. Polyphenol oxidase (PPO) is one of the RPW innate immune mechanisms and inhibition of such enzyme could result in a disorder of the insect's immune system. A one single PO isoenzyme has been purified from the hemolymph of the 12th instar larvae of RPW. Using L-DOPA as substrate, R. ferrugineus PPO exhibited specific activity 428 Units/mg proteins with 8.3-fold purification, optimum pH and temperature for activity at 7.5 and 40 °C, respectively and is enhanced by Cu2+ with 1.76-fold. The rank order for oxidizing R. ferrugineus PPO different substrates is catechol > pyrogallol > L-DOPA > pyrocatechuic acid and not tyrosine. The kinetic parameters Km, Vmax and Vmax/Km for L-DOPA are 3.3 mM, 1.3 µmol/ml/min, and 0.39, respectively. The catalytic efficiency of the enzyme towards catechol is 5.3-fold higher than that for L-DOPA. The enzyme completely inhibited by thiourea, ascorbic acid, dithiothreitol, and SDS. R. ferrugineus PPO is a catechol oxidase di-phenol: O2 oxidoreductase. Based on the toxicological studies of various botanical extracts, the IC50 ranged from 20 to 90 mg/ml. The enzyme completely inhibited by 50 mg/ml Cinnamomum camphora. Gallic acid, the major phenolic compound, has IC50 0.8 mM and competitively inhibited the enzyme with Ki 0.54 mM. C. camphora could be a useful natural RPW-controlling agent and used as integral part in IPM programs. This interpretation can be validated in future through an in vivo investigation.

Evaluating the efficacy of microbial antagonists in inducing resistance, promoting growth, and providing biological control against powdery mildew in wheat.

This study evaluates the biocontrol efficacy of three bacterial strains (Pseudomonas fluorescens DTPF-3, Bacillus amyloliquefaciens DTBA-11, and Bacillus subtilis DTBS-5) and two fungal strains (Trichoderma harzianum Pusa-5SD and Aspergillus niger An-27) antagonists, along with their combinations at varying doses (5.0, 7.5, and 10.0 g/kg of seeds), against wheat powdery mildew. The most effective dose (10 g/kg seeds) was further analyzed for its impact on induced resistance and plant growth promotion under greenhouse conditions. The study measured defense enzyme activities, biochemical changes, and post-infection plant growth metrics. All tested microbial antagonists at 10 g/kg significantly reduced PM severity, with B. subtilis strain DTBS-5 outperforming others in reducing PM severity and achieving the highest biocontrol efficacy. It was followed by B. amyloliquefaciens strain DTBA-11 and P. fluorescens strain DTPF-3, with the fungal antagonists showing no significant effect. Wheat crops treated with B. subtilis strain DTBS-5 exhibited substantial increases in defense-related enzyme activities and biochemicals, suggesting an induced resistance mechanism. The study found a 45% increase in peroxidase (POD) activity, a 50% increase in catalase (CAT) activity, a 30% increase in phenolic content, and a 25% increase in soluble protein content in the wheat plants treated with microbial antagonists. The study highlights the effectiveness of microbial antagonists, particularly B. subtilis strain DTBS-5, in managing wheat PM through biocontrol, induced resistance, and enhanced plant growth, offering a sustainable alternative to chemical treatments.

Identification and Analysis of PPO Gene Family Members in Paulownia fortunei.

Polyphenol oxidase (PPO) is a common metalloproteinase in plants with important roles in plant responses to abiotic and biotic stresses. There is evidence that PPOs contribute to stress responses in Paulownia fortunei. In this study, PPO gene family members in P. fortunei were comprehensively identified and characterized using bioinformatics methods as well as analyses of phylogenetic relationships, gene and protein structure, codon usage bias, and gene expression in response to stress. The genome contained 10 PPO gene family members encoding 406-593 amino acids, with a G/C bias. Most were localized in chloroplasts. The motif structure was conserved among family members, and α-helices and random coils were the dominant elements in the secondary structure. The promoters contained many cis-acting elements, such as auxin, gibberellin, salicylic acid, abscisic acid, and photoresponsive elements. The formation of genes in this family was linked to evolutionary events, such as fragment replication. Real-time quantitative PCR results showed that PfPPO7, PfPPO10, PfPPO1, PfPPO2, PfPPO3, PfPPO4, PfPPO5, and PfPPO8 may be key genes in drought stress resistance. PfPPO1, PfPPO3, PfPPO4, and PfPPO10 were resistant stress-sensitive genes. These results provide a reliable basis for fully understanding the potential functions of these genes and the selection of resistance breeding.

Impact of the tripartite interaction between rice, sheath blight and diverse crop-associated endophytes on phenotypic and biochemical responses in rice.

Endophytes stimulate plant growth and inhibit phytopathogens. Most of the known endophytes are host-specific and only a few strains are effective for practical field use. Thus, this study focuses on the evaluation of endophytes viz., Bacillus pseudomycoides strain HP3d, Paenibacillus polymyxa strain PGSS1, B. velezensis strain A6 and P42 isolated from diverse crop ecosystems for their potential to promote plant growth and induce systemic resistance against sheath blight disease in rice. The endophytes were studied for plant growth promoting traits in vivo conditions and were found to exhibit ammonia (light to strong), siderophore (yellow zone on the CAS agar plate), indole-3-acetic acid (15.20-22.19 µg mL-1) production and phosphorus solubilization (1.2-1.5 cm). In the glasshouse, when applied individually and in combinations through various methods like seed treatment, seedling dip, and foliar spray these endophytes significantly reduced lesion size (2.06-2.37 fold) and ShB severity (2.60-2.58 fold), enhancing growth parameters viz., shoot (1.09-1.11 fold), root (1.02-1.20 fold), number of tillers (1.2-1.6 fold), shoot (80.58-82.64 %) and root (62.01-66.66 %) dry matter over untreated control. Consequently, enzyme activity viz., polyphenol oxidase (2.20-3.00 U-1min-1g-1), peroxidase (0.31-0.35 min-1g-1), superoxide dismutase (118.50-123.00 Ug-1 FW), and phenylalanine ammonia lyase (0.84-0.90 min⁻1g⁻1FW) was found to increase up to the fourth day after the pathogen challenge and subsequently decrease thereafter. Chlorophyll content post inoculation of ShB declined over time but endophyte treated plants exhibited lesser reductions over uninoculated control. Field trials corroborated the in vitro findings, demonstrating reduced ShB (1.71-1.88 fold decrease in PDI) and enhanced growth (1.1-1.2 fold increase in shoot length) over untreated controls. The combined application of seedling dip, seed treatment, and foliar spray proved to be the most optimum treatment. The findings highlight the potential of diverse crop-derived endophytes, emphasizing their non-host specificity and effectiveness as broad-spectrum bioagents in actual field conditions.

Rational Design, Synthesis, and Computational Investigation of Dihydropyridine [2,3-d] Pyrimidines as Polyphenol Oxidase Inhibitors with Improved Potency.

Polyphenol oxidase (PPO) is an industrially important enzyme associated with browning reactions. In the present study, a set of ten new dihydropyridine [2,3-d] pyrimidines (TD-Hid-1-10) were synthesized and was found to be proven characteristically by 1H NMR, 13C NMR, IR, elemental analysis, and assessed as possible PPO inhibitors. PPO was purified from banana using three-phase partitioning, achieving an 18.65-fold purification and 136.47% activity recovery. Enzyme kinetics revealed that the compounds TD-Hid-6 and TD-Hid-7 are to be the most potent inhibitors, exhibiting mixed-type inhibition profile with IC50 values of 1.14 µM, 5.29 µM respectively against purified PPO enzyme. Electronic structure calculations at the B3LYP/PBE0 level of theories using def-2 SVP, def2-TZVP basis sets with various molecular descriptors characterized the electronic behavior of studied derivatives TD-Hid-1-10. Molecular electrostatic potential (MEP) and reduced density gradient analyses of RDG-NCI provided insights into charge distributions and weak intermolecular interactions. Docking study simulations predicted binding poses within crucial amino acid sequence in the 2y9x enzyme's active site, which is typically similar in sequence to the PPO form is not allowed. Ligands were analysed in terms of binding energies, inhibitor concentrations (mM) and various molecular interactions such as H-bonds, H-carbon, π-carbon, π-sigma, π-sigma, π-π T-shaped, π-π stacked, π-alkyl, Van der Waals and Cu interactions. The lowest binding energy (-7.83 kcal/mol) and the highest inhibitory effect (1.83 mM) were shown by the ligand Td-Hid-6, which forms H-bonds with Met280 and Asn260, exhibits π-sigma interactions with His61 and π-alkyl interactions with Val283. Other ligands also showed different interactions with various amino acids; for example, the Td-Hid-1 ligand formed H-bonds with His244 and showed π-sigma interactions with His244 and Val283.

Molecular insights into the anti-spoilage effect of salicylic acid in Favorita potato processing.

Salicylic acid is a commonly used anti-spoilage agent to prevent browning and quality degradation during potato processing, yet its precise mechanism remains unclear. This study elucidates the role of StuPPO2, a functional protein in Favorita potato shreds, in relation to the anti-browning and starch degradation effects of 52 SA analogues. By employing molecular docking and Gaussian computing, SA localizes within the hydrophobic cavity of StuPPO2, facilitated by hydroxyl and carboxyl groups. The inhibitory effect depends on the distribution pattern of the maximal electrostatic surface potential, requiring hydroxyl ion potentials of >56 kcal/mol and carboxyl ion potentials of >42 kcal/mol, respectively. Multiomics analysis, corroborated by validation tests, indicates that SA synthetically suppresses activities linked to defense response, root regeneration, starch degradation, glycoalkaloids metabolism, and potato shred discoloration, thereby preserving quality. Furthermore, SA enhances antimicrobial and insect-repellent aromas, thereby countering biotic threats in potato shreds. These collective mechanisms underscore SA's anti-spoilage properties, offering theoretical foundations and potential new anti-browning agents for agricultural preservatives.

Modified Metal-Organic Framework (MOF-818) inspired by natural enzymes for intelligent detection of total antioxidants and bisphenol A in infant beverages.

BACKGROUND: Accurate and quick judgement of the food quality can protect the legitimate rights of consumers. Currently, nanozymes are widely employed in the rapid detection of food due to their stability and economy. The contents of bisphenol A and antioxidant can be used to measure the quality of beverages. However, due to the complexity of the actual samples, it is still challenging to achieve the sensitive detection of both at the same time. The development of nanozyme with high enzyme activity is essential for sensitive detection of targets in complex foods. RESULTS: In this work, a novel nanomaterial (ZrTGA) was synthesized based on thioglycolic acid-modified Metal-Organic Framework (MOF-818). The interaction between Cu-S bonds and increase in the proportion of Cu1+ resulted in ZrTGA exhibiting higher peroxidase-like and polyphenol oxidase-like activities. These enzyme activities were 317 % and 200 % of the original values, respectively. With high enzyme activity can sensitively detect two important indicators of bisphenol A and antioxidants in beverages. The increased enzyme activity of ZrTGA enabled the content of both substances to be detected by smartphone extraction of RGB. Finally, through the output of the ''0″ and ''1″ signals of the logic gates, it is possible to quickly determine the level of the two substances and thus directly assess the quality of the beverages. SIGNIFICANCE: The modification of nanozyme enables the detection of substances at low concentrations based on enhancing dual-enzyme activity. The combination of mobile phone photography and logic gate technology enables the continuous detection of two important indicators in beverages, overcoming the limitations of traditional large-scale instruments. It also provides an alternative strategy for food quality detection.

Sources and Applications of Tyrosinase in Life Sciences.

BACKGROUND: Tyrosinase, often recognized as polyphenol oxidase, plays a pivotal role as an enzyme in catalyzing the formation of melanin-a complex process involving the oxidation of monophenols and o-diphenols. OBJECTIVE: Tyrosinase functions as a monooxygenase, facilitating the o-hydroxylation of monophenols to generate the corresponding catechols, as well as catalyzing the oxidation of monophenols to form the corresponding o-quinones, exhibiting diphenolase or catecholase activity. This versatile enzymatic capability is not limited to specific organisms but is found across various sources, including bacteria, fungi, plants, and mammals. METHOD: Pertinent research articles, reviews, and patents on tyrosinase were gathered through a comprehensive literature search. These materials were analyzed to gain insights into the diverse applications of tyrosinase. The review was structured by categorizing these applications and offering a thorough summary of the current state of knowledge in the field. RESULT: Based on the literature survey, tyrosinase exhibits promising potential across a spectrum of biotechnological applications. These include but are not limited to: synthesizing L-DOPA, creating innovative mixed melanins, manufacturing phenolic biosensors, deploying in food and feed industries, facilitating protein cross-linking, eliminating phenols and dyes, and serving as a biocatalyst. Moreover, immobilized tyrosinase demonstrates multiple utility avenues within the pharmaceutical sector. CONCLUSION: The article offers a comprehensive exploration of tyrosinase, encompassing its structural features, evolutionary origins, biochemical characteristics, and contemporary applications in various fields.

Sequential Pulsed Light and Ultrasound Treatments for the Inactivation of Saccharomyces cerevisiae and PPO and the Retention of Bioactive Compounds in Sweet Lime Juice.

Designing a pasteurization con dition for sweet lime juice while ensuring microbial safety, enzymatic stability, and high nutritional quality is crucial for satisfying stakeholder demands. The present research investigates the effects of matrix pH, ultrasound treatments, and sequential pulsed light on the microbial population, enzyme activity, and bioactive chemicals in sweet lime juice. The sequential pulsed light (PL: 0.6-0.84 J/cm2) and ultrasound (US: 0.2-0.4 W/cm3) treatments for sweet lime juice were optimized using response surface methodology (RSM). A three-factor full factorial design was used for this purpose. The independent variables encompassed pH (X1), PL effective fluence (X2, J/cm2), and US intensity (X3, W/cm3). The responses assessed included the inactivation of Saccharomyces cerevisiae (Y1, log cfu/mL) and polyphenol oxidase (PPO: Y2 in %) and the retention of vitamin C (Y3, %). The polynomial models were optimized using numerical optimization to attain the maximum desirability value (0.89). The optimized PL + US sample (0.8 J/cm2 + 0.4 W/cm3, respectively) at pH 3.5 resulted in a 5-log cycle reduction in S. cerevisiae count and a 90% inactivation in PPO activity and retained 95% of its vitamin C content. This optimized sample underwent further analysis, including phenolic profiling, assessment of microbial cell morphology, and examination of enzyme conformational changes. After sequential pulsed-light (0.8 J/cm2) and ultrasound (0.4 W/cm3) treatments, yeast cells showed unusual structural changes, indicating additional targets besides membranes. Following PL + US treatment, the PPO composition changed to 2.7 ± 0.1% α-helix, 33.9 ± 0.3% ß-sheet, 1.4 ± 0.2% ß-turn, and 62 ± 0.7% random coil. Impressively, the optimized PL + US sample maintained a sensory acceptance level similar to that of the untreated sample.