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.

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.

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.

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.
METHODS: 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.
RESULTS: 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.
CONCLUSIONS: The article offers a comprehensive exploration of tyrosinase, encompassing its structural features, evolutionary origins, biochemical characteristics, and contemporary applications in various fields.

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% β-sheet, 1.4 ± 0.2% β-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.

This study was aimed to evaluate the potential of high-humidity hot air impingement cooking (HHAIC) on Penaeus vannamei, focusing on its drying characteristics, microstructure, water distribution, enzyme activity, astaxanthin content, antioxidant capacity, color, and Maillard reaction. Results demonstrated that a 3 min HHAIC significantly improved the shrimp\'s color and optimized astaxanthin content with a notable increase in scavenging capacity based on an in-vitro as antioxidation activity evaluation. Compared to the untreated samples, HHAIC could significantly inactivate polyphenol oxidase by 95.76%. Also, it suppressed the Maillard reaction by decreasing 5-hydroxymethylfurfural content and shortened the drying time by 40%. In addition, the low-field nuclear magnetic resonance and microstructure analysis showed alterations in the shrimp muscle fiber structure and water distribution. This study indicated that HHAIC could elevate quality, enhance appearance, and reduce the processing time of dried shrimp, presenting valuable implications for industry progress.

The present work evaluated kiwi juice addition alongside pasteurization (at 85 °C for 5 min) or microwave treatment (for 3 min) on the quality improvement of sugarcane juice. The juice was treated in the presence of kiwi juice (0-8%), and its physicochemical properties and microbial load were compared with raw juice. The study also highlighted the key enzymes causing sugarcane juice discoloration, peroxidase (POD) and polyphenol oxidase (PPO), by quantifying kiwi juice constituents using GC-MS and monitoring their effects by molecular docking. Kiwi addition considerably raised (p < 0.05) acidity, ascorbic acid (54.28%), and phenolic compounds (32%), and decreased the POD and PPO activity of raw cane juice. Pasteurization in the presence of kiwi, rather than microwave treatment, has significantly (p < 0.05) increased the phenolic compounds and reduced POD and PPO activities until barley was detected. Molecular docking revealed that heptacosane, oleic acid, and melezitose are the primary kiwi components responsible for enzyme inactivation.

The development of natural inhibitors of polyphenol oxidase (PPO) is crucial in the prevention of enzymatic browning in fresh foods. However, few studies have focused on the effect of subsequent sterilization on their inhibition efficiency. This study investigated the influence and mechanism of high hydrostatic pressure (HHP) on the inhibition of PPO by epigallocatechin gallate (EGCG), cyanidin-3-O-glucoside (C3G), and ferulic acid. Results showed that under the conditions of 550 MPa/30 min, the activity of EGCG-PPO decreased to 55.92%, C3G-PPO decreased to 81.80%, whereas the activity of FA-PPO remained stable. Spectroscopic experiments displayed that HHP intensified the secondary structure transformation and fluorescence quenching of PPO. Molecular dynamics simulations revealed that at 550 MPa, the surface interaction between PPO with EGCG or C3G increased, potentially leading to a reduction in their activity. In contrast, FA-PPO demonstrated conformational stability. This study can provide a reference for the future industrial application of natural inhibitors.

Acteoside is a bioactive phenylethanoid glycoside widely distributed throughout the plant kingdom. Because of its two catechol moieties, acteoside displays a variety of beneficial activities. The biosynthetic pathway of acteoside has been largely elucidated, but the assembly logic of two catechol moieties in acteoside remains unclear. Here, we identified a novel polyphenol oxidase OfPPO2 from Osmanthus fragrans, which could hydroxylate various monophenolic substrates, including tyrosine, tyrosol, tyramine, 4-hydroxyphenylacetaldehyde, salidroside, and osmanthuside A, leading to the formation of corresponding catechol-containing intermediates for acteoside biosynthesis. OfPPO2 could also convert osmanthuside B into acteoside, creating catechol moieties directly via post-modification of the acteoside skeleton. The reverse transcription-quantitative polymerase chain reaction (RT-qPCR) analysis and subcellular localization assay further support the involvement of OfPPO2 in acteoside biosynthesis in planta. These findings suggest that the biosynthesis of acteoside in O. fragrans may follow \"parallel routes\" rather than the conventionally considered linear route. In support of this hypothesis, the glycosyltransferase OfUGT and the acyltransferase OfAT could direct the flux of diphenolic intermediates generated by OfPPO2 into acteoside. Significantly, OfPPO2 and its orthologs constitute a functionally conserved enzyme family that evolved independently from other known biosynthetic enzymes of acteoside, implying that the substrate promiscuity of this PPO family may offer acteoside-producing plants alternative ways to synthesize acteoside. Overall, this work expands our understanding of parallel pathways plants may employ to efficiently synthesize acteoside, a strategy that may contribute to plants\' adaptation to environmental challenges.

In this study, inactivation of mushroom polyphenol oxidase (PPO) by low intensity direct current (DC) electric field and its molecular mechanism were investigated. In the experiments under 3 V/cm, 5 V/cm, 7 V/cm and 9 V/cm electric fields, PPOs were all completely inactivated after different exposure times. Under 1 V/cm, a residual activity of 11.88 % remained. The inactivation kinetics confirms to Weibull model. Under 1-7 V/cm, n value closes to a constant about 1.3. The structural analysis of PPO under 3 V/cm and 5 V/cm by fluorescence emission spectroscopy and molecular dynamics (MD) simulation showed that the tertiary structure was slightly changed with increased radius of gyration, higher potential energy and rate of C-alpha fluctuation. After exposure to the electric field, most of the hydrophobic tryptophan (TRP) residues turned to the hydrophilic surface, resulting the fluorescence red-shifted and quenched. Molecular docking indicated that the receptor binding domain of catechol in PPO was changed. PPO under electric field was MD simulated the first time, revealing the changing mechanism of the electric field itself on PPO, a binuclear copper enzyme, which has a metallic center. All these suggest that the low intensity DC electric field would be a promising option for enzymatic browning inhibition or even enzyme activity inactivation.