Bioactive Enzymes Research Articles

Metabolic Carcinogenesis.

Several types of environmental, chemical and metabolic carcinogens exist both exogenously and endogenously. Humans are daily exposed to aforementioned carcinogens through various sources such as through water, air and food or through metabolic and inflammatory products. This chapter will summarize the links between exogenous and endogenous carcinogen exposure and their metabolism with the cancer pathogenesis and associated risks. This chapter will also cover the carcinogens acquired through lifestyle factors like tobacco use and occupational exposures to different chemicals like asbestos, arsenic, chloroform, vinyl chloride, etc. Moreover, environmental carcinogens such as radiation, sunlight, diet, smoke, etc. will also be discussed in this chapter. Furthermore, there are certain carcinogens that require bio-activation and various human enzymes that play a vital role in the metabolic carcinogenesis will also be recapitulated. Necessary preventive measures against carcinogenic exposure from the exogenous environment are significant to be taken into account to reduce the risks associated with the carcinogens.

Penicillium chrysogenum: Beyond the penicillin.

Almost one century after the Sir Alexander Fleming's fortuitous discovery of penicillin and the identification of the fungal producer as Penicillium notatum, later Penicillium chrysogenum (currently reidentified as Penicillium rubens), the molecular mechanisms behind the massive production of penicillin titers by industrial strains could be considered almost fully characterized. However, this filamentous fungus is not only circumscribed to penicillin, and instead, it seems to be full of surprises, thereby producing important metabolites and providing expanded biotechnological applications. This review, in addition to summarizing the classical role of P. chrysogenum as penicillin producer, highlights its ability to generate an array of additional bioactive secondary metabolites and enzymes, together with the use of this microorganism in relevant biotechnological processes, such as bioremediation, biocontrol, production of bioactive nanoparticles and compounds with pharmaceutical interest, revalorization of agricultural and food-derived wastes or the enhancement of food industrial processes and the agricultural production.

Cyclophosphamide Pharmacogenomic Variation in Cancer Treatment and Its Effect on Bioactivation and Pharmacokinetics.

Cyclophosphamide (Cy) is a prodrug that is mainly bioactivated by cytochrome P450 (CYP) 2B6 enzyme. Several other enzymes are also involved in its bioactivation and affect its kinetics. Previous studies have shown the effect of the enzymes' genetic polymorphisms on Cy kinetics and its clinical outcome. These results were controversial primarily because of the involvement of several interacting enzymes in the Cy metabolic pathway, which can also be affected by several clinical factors as well as other drug interactions. In this review article, we present the effect of CYP2B6 polymorphisms on Cy kinetics since it is the main bioactivating enzyme, as well as discussing all previously reported enzymes and clinical factors that can alter Cy efficacy. Additionally, we present explanations for key Cy side effects related to the nature and site of its bioactivation. Finally, we discuss the role of busulphan in conditioning regimens in the Cy metabolic pathway as a clinical example of drug-drug interactions involving several enzymes. By the end of this article, our aim is to have provided a comprehensive summary of Cy pharmacogenomics and the effect on its kinetics. The utility of these findings in the development of new strategies for Cy personalized patient dose adjustment will aid in the future optimization of patient specific Cy dosages and ultimately in improving clinical outcomes. In conclusion, CYP2B6 and several other enzyme polymorphisms can alter Cy kinetics and consequently the clinical outcomes. However, the precise quantification of Cy kinetics in any individual patient is complex as it is clearly under multifactorial genetic control. Additionally, other clinical factors such as the patient's age, diagnosis, concomitant medications, and clinical status should also be considered.

Synthesis and antibacterial activities of bromelain stabilized fluorescent copper nanoclusters against E. coli and S. aureus

Bromelain, a bioactive proteolytic enzyme is composed of a mixture of cysteine protease. It is obtained from fruits and stems of pineapple. Cu in bulk as well as in nano form and bromelain individually demonstrate antibacterial activities. With the aim to make bromelain and Cu act synergistically against bacteria, in this work, bromelain-capped copper nanoclusters (Bro-CuNCs) were synthesized for the first time by adhering to the principles of green chemistry. Synthesized nanoclusters exhibit a quantum yield (QY) of 4.47% in an aqueous medium. These nanoclusters were characterized by several spectroscopy and microscopy techniques such as UV–VIS spectroscopy, Fluorescence spectroscopy, FT-IR spectroscopy, and HR-TEM. The findings reveal that Bro-CuNCs interact more effectively with the Gram-negative strain of E. coli than the Gram-positive strain of S. aureus. Additionally, the Bro-CuNCs exhibit an enhancement of fluorescence intensity when exposed to E. coli, while demonstrating a diminution in the presence of S. aureus. Surprisingly, they also demonstrate efficacy against Gram-negative bacteria, P. aeruginosa. This strain is inherently resistant to conventional antibiotics such as penicillin.

Differential Correlation of Transcriptome Data Reveals Gene Pairs and Pathways Involved in Treatment of Citrobacter rodentium Infection with Bioactive Punicalagin

This study is part of the work investigating bioactive fruit enzymes as sustainable alternatives to parasite anthelmintics that can help reverse the trend of lost efficacy. The study looked to define biological and molecular interactions that demonstrate the ability of the pomegranate extract punicalagin against intracellular parasites. The study compared transcriptomic reads of two distinct conditions. Condition A was treated with punicalagin (PA) and challenged with Citrobacter rodentium, while condition B (CM) consisted of a group that was challenged and given mock treatment of PBS. To understand the effect of punicalagin on transcriptomic changes between conditions, a differential correlation analysis was conducted. The analysis examined the regulatory connections of genes expressed between different treatment conditions by statistically querying the relationship between correlated gene pairs and modules in differing conditions. The results indicated that punicalagin treatment had strong positive correlations with the over-enriched gene ontology (GO) terms related to oxidoreductase activity and lipid metabolism. However, the GO terms for immune and cytokine responses were strongly correlated with no punicalagin treatment. The results matched previous studies that showed punicalagin to have potent antioxidant and antiparasitic effects when used to treat parasitic infections in mice and livestock. Overall, the results indicated that punicalagin enhanced the effect of tissue-resident genes.

Induction of bioactive constituents and antioxidant enzyme activities in Achillea fragrantissima (Forskal) callus cultures using ZnO nanoparticles

The medicinally effective plant Achillea fragrantissima exhibits a magnitude of pharmacological activities. In this study, the effects of different ZnONP concentrations on antioxidant enzymes, bioactive secondary metabolites, redox potential, and molecular changes in A. fragrantissima callus cultures were investigated. First, the concentrations of the growth regulators 2,4-D and BA were optimized using Murashige and Skoog (MS) medium. The MS medium was then administered with 2,4-D and BA at its optimal dosage (1.0 mg.L−1); afterward, different ZnONP supplements (0.0, 5.0, 10.0, 15.0, and 20.0 mg.L−1) were added. ZnONPs resulted in many physiological and molecular responses. ZnONPs significantly increased POD, APX, and SOD activities. While 10.0 mg.L−1 ZnONPs significantly increased POD and APX activities, 15.0 mg.L−1 ZnONPs significantly increased SOD. However, CAT activity gradually decreased with ZnONPs. Metabolically, ZnONPs increased phenolics, flavonoids, alkaloids, and saponin levels. Phenolic levels peaked at 20.0 mg.L−1, flavonoids at 15.0 mg.L−1, and alkaloids and saponins at 10.0 mg.L−1. Terpenoids were more prevalent at lower levels of ZnONPs. With 15.0 and 10.0 mg.L−1 giving the maximum activity, ZnONPs enhanced the DPPH activity and TAC of the callus culture extracts, respectively. RAPD and ISSR fingerprinting were applied using 12 random and ISSR primers to evaluate the genetic stability of ZnONP-induced callus cultures. Six RAPD primers showed 83% polymorphism while the seven ISSR primers achieved 30% polymorphism. Consequently, DNA mutations may have been induced by ZnONPs and caused DNA fragments to either appear or disappear in RAPD and ISSR callus profiles. The dendrogram based on RAPD and ISSR combined data showed that by increasing ZnONP concentration the genetic differentiation among callus cultures was elevated. In conclusion, higher accumulation of secondary metabolites and redox activity were increased in A. fragrantissima callus cultures using low ZnONPs (10.0 mg.L−1) concentration.

Microbial Diversity in Bee Species and Bee Products: Pseudomonads Contribution to Bee Well-Being and the Biological Activity Exerted by Honey Bee Products: A Narrative Review

Bees are important pollinators worldwide, promoting sustainability in agriculture and natural ecosystems. Moreover, honey bees produce a variety of honey bee products (beehive products). Honey is the main edible bee product. The consumption of pollen, bee bread, royal jelly, and propolis is becoming more popular nowadays. All these products are characterized by high nutritional value and/or bioactivity. A high microbial diversity has been reported in bees and beehive products, forming distinct microbial communities. The honey bee gut microbiome actively promotes good health and nutrient availability for the host. Furthermore, it prevents food spoilage and contributes to the maintenance of good hygiene conditions in the hive. Pseudomonads are often reported in investigations on bee and bee product microbiomes. Diverse Pseudomonas species demonstrate high metabolic adaptability, producing a wide range of bioactive enzymes and secondary metabolites. Several studies have provided evidence that Pseudomonads might play a role in bee well-being and the bioactivity exerted by honey bee products, though further research is warranted to fully understand the effects and mechanisms. The aim of this narrative review is to highlight the importance of Pseudomonads in the context of up-to-date knowledge regarding the bee and bee product microbiomes.

Bio-Fabricated Silver Nanoparticles from the Leaf Extract of the Poisonous Plant, Holigarna arnottiana: Assessment of Antimicrobial, Antimitotic, Anticancer, and Radical-Scavenging Properties.

This study presents a novel approach to fabricate silver nanoparticles (AgNPs) using the poisonous plant, Holigarna arnottiana leaf extract. The formation of AgNPs was confirmed by a color change from green to dark brown and validated by UV analysis. FTIR analysis identified functional groups on the AgNPs, while Zeta potential analysis assessed their stability. TEM analysis established an average diameter of 18 nm and a spherical morphology for the nanoparticles. LC MS analysis coupled with database searches revealed the presence of diverse bioactive compounds, including flavonoids, nucleotides, dipeptides, enzymes, and glycosides. These compounds are postulated to act as reducing agents in the leaf extract-mediated synthesis process. Moreover, the bio-fabricated AgNPs exhibited noteworthy anticancer properties against DLA cells. In addition, AgNPs displayed substantial antimitotic effects in an assay involving Allium cepa root cells. These findings underscore the potential of the AgNPs as cytotoxic agents. The biosynthesized AgNPs showed antimicrobial activity against various bacterial pathogens, including Escherichia coli, Klebsiella pneumoniae, and Staphylococcus aureus. Furthermore, the AgNPs exhibited outstanding radical-scavenging properties in the DPPH assay, suggesting their potential application in antioxidant therapies. The study collectively highlights the successful synthesis of AgNPs through a green, biocompatible approach, and demonstrates their promising potential for anticancer, antimitotic, and radical-scavenging applications.

Mn3O4 nanozymes prevent acetaminophen-induced acute liver injury by attenuating oxidative stress and countering inflammation

Acetaminophen (APAP) overdose is steadily becoming the chief reason for drug-induced acute liver failure, yet limited treatment is currently clinically available. Considering that the mechanism of APAP-induced hepatotoxicity is inseparable from oxidative stress and inflammation, a biocompatible Mn3O4 nanozyme mimicking superoxide dismutase (SOD) and catalase (CAT) activities and possessing reactive oxygen species (ROS)–scavenging capacity and antiapoptotic properties, is reported herein as a promising nanodrug to treat APAP-induced liver injury (AILI). Possessing bioactive enzyme–like functions, Mn3O4 nanoparticles (NPs) can not only reduce the oxidative stress on the liver by decreasing ROS accumulation but also downregulate the infiltration of inflammatory macrophages that secrete proinflammatory cytokines (tumor necrosis factor-α, interleukin-1β, and interleukin-6). Notably, the bifunctional Mn3O4 NPs mediate nuclear factor–erythroid 2 p45-related factor 2 signaling pathway activation and nuclear factor kappa B signaling pathway inhibition to effectively prevent the already fragile APAP-overdosed murine hepatocytes from being attacked again, thus mitigating hepatocyte apoptosis and alleviating APAP-induced liver damage. Thus, the Mn3O4 nanozyme (Mn3O4 NPs) evaluated in this study has potential preventive and therapeutic effects on AILI.

Antifungal screening of selenium nanoparticles biosynthesized by microcystin-producing Desmonostoc alborizicum

Metal nanoparticles exhibit excellent antifungal abilities and are seen as a good substitute for controlling different kinds of fungi. Of all known taxa, cyanobacteria have received significant consideration as nanobiofactories, as a result of the cellular assimilation of heavy metals from the environment. The cellular bioactive enzymes, polysaccharides and pigments can be used as reducers and coatings during biosynthesis. The probability of the antifungal activity of selenium nanoparticles (SeNPs) to prevent plant fungi that can affect humans was evaluated and a toxic Iranian cyanobacterial strain of Desmonostoc alborizicum was used to study the biotechnology of SeNP synthesis for the first time. Characterization of nanoparticles with a UV-Vis spectrophotometer showed the formation of SeNPs in the range of 271–275 nm with the appearance of an orange color. Morphological examination of nanoparticles with Transmission Electron Microscopy (TEM), revealed the spherical shape of nanoparticles. The results of X-Ray Diffraction (XRD) showed 7 peaks and a hexagonal structure of average crystal size equal to 58.8 nm. The dispersion index of SeNPs was reported as 0.635, which indicated the homogeneity of the nanoparticle droplet size. The zeta potential of the nanoparticles was + 22.7. Fourier-transform infrared spectroscopy (FTIR) analysis exhibited a sharp and intense peak located at the wave number of 404 cm− 1, related to the SeNPs synthesized in this research. The results of the antifungal activity of SeNPs showed among the investigated fungi, Pythium ultimum had the highest resistance to SeNPs (14.66 ± 0.52 µg/ml), while Alternaria alternata showed the highest sensitivity (9.66 ± 0.51 µg/ml) (p < 0.05). To the best of our knowledge this is the first report concerning the characterization and antifungal screening of SeNPs biosynthesized by Iranian cyanobacteria, which could be used as effective candidates in medical applications.

A Novel Dual-Reporter System Reveals Distinct Characteristics of Exosome-Mediated Protein Secretion in Human Cells

BackgroundExosomes, a special subtype of extracellular vesicles derived from human cells, serve as vital mediators of intercellular communication by transporting diverse bioactive cargos, including proteins and enzymes. However, the underlying mechanisms governing exosome secretion and regulation remain poorly understood. In this study, we employed a dual-reporter system consisting of bioluminescent Gaussia luciferase and fluorescent proteins to investigate the dynamics and regulation of exosome secretion in cultured human cells.ResultsOur results demonstrated that the engineered dual-reporters effectively monitored both exosome-mediated and ER-Golgi-mediated secretory pathways in a specific and quantitative manner. Notably, we observed distinct characteristics of exosome-mediated protein secretion, including significantly lower capacity and different dynamics compared to the ER-Golgi pathway. This phenomenon was observed in human kidney 293T cells and liver HepG2 cells, emphasizing the conserved nature of exosome-mediated secretion across cell types. Furthermore, we investigated the impact of brefeldin A (BFA), an inhibitor of ER-to-Golgi membrane trafficking, on protein secretion. Interestingly, BFA inhibited protein secretion via the ER-Golgi pathway while stimulating exosome-mediated protein secretion under same experimental conditions.ConclusionsCollectively, our study highlights the utility of the dual-reporter system for real-time monitoring and quantitative analysis of protein secretion through conventional ER-Golgi and unconventional exosome pathways. Moreover, our findings unveil distinct features of exosome-mediated protein secretion, shedding light on its differential capacity, dynamics, and regulatory mechanisms compared to ER-Golgi-mediated proteins in human cells.

Bioactive angiotensin converting enzyme inhibitory activity and antihypertensive activity derived from fish protein hydrolysate: a systematic review

In the fish processing sector, fish have been processed in large quantities, are generated and discarded into the sea or dumped into the oceans. However, by employing effective strategies, this fishery waste can be utilized and converted into fish protein hydrolysate (FPH). FPH is a rich source of amino acids and peptides that have biological properties such as angiotensin-converting enzyme (ACE) inhibition, antimicrobial, antioxidant, and anticancer activity. The FPH has been used as a functional food perspective related to improving human health, primarily by maintaining blood pressure and normal heart function as measured by antihypertensive activity. FPH can be produced by various fish species, parts, and hydrolysis methods. Several studies have been published on the acceptability of FPH in obtaining bioactive properties from various fish, each using a different method to obtain bioactive properties. FPH is commonly produced by fish species such as Atlantic Cod, Lizard Fish, Atlantic Chub Mackerel, Atlantic Horse Mackerel, and Kawakawa (Mackerel Tuna). Furthermore, we summarized the various methodologies used by various researchers based on raw material data collection, method of production, ACE-inhibitory assay, ACE-inhibitory activity, degree of hydrolysis, molecular weight, purification method, and ACE-I peptide structure in this review.

Efficacy of Aspergillus tubingensis GX3' Fermentation against Meloidogyne enterolobii in Tomato (Solanum lycopersicum L.).

Meloidogyne enterolobii is one of the most virulent root-knot nematodes (RKNs). Aspergillus tubingensis Raoul Mosseray, 1934, is used to produce bioactive substances, enzymes, and secondary metabolites. However, no research has been conducted yet on the efficacy of A. tubingensis against plant-parasitic nematodes. Thus, the novel research was planned to evaluate the biocontrol efficacy of A. tubingensis fermentation against M. enterolobii. The findings showed that egg hatching inhibition and mortality of M. enterolobii increased with increasing concentration of fermentation and exposure time. The maximum second-stage juveniles (J2s) mortality was achieved via 100% fermentation at 72 h. Similarly, 100% fermentation inhibited 99.9% of egg hatching at 8 d. A. tubingensis fermentation increased plant biomass, decreased second-stage juvenile invasion, and inhibited nematode development and reproduction in greenhouse conditions. A. tubingensis reduced J2 invasion into tomato roots by 42.84% with CS+ (coated seeds plants with nematodes inoculum) and 27.04% with T+ (100% fermentation broth and nematodes inoculum both) treatments. Moreover, CS+ and T+ treatments decreased nematode development by 54.31% and 21.48%, respectively. It is concluded that the A. tubingensis GX3 strain can be used as a novel microbial biocontrol agent against M. enterolobii.

An integrated microfluidic-based biosensor using a magnetically controlled MNPs-enzyme microreactor to determine cholesterol in serum with fluorometric detection

This work provides a microfluidic-based biosensor to determine total cholesterol in serum based on integrating the reaction/detection zone of a microfluidic chip of a magnetically retained enzyme microreactor (MREµR) coupled with the remote fluorometric detection through a bifurcated fiber-optic bundle (BFOB) connected with a conventional spectrofluorometer. The method is based on developing the enzymatic hydrolysis and oxidation of cholesterol at microscale size using both enzymes (cholesterol esterase (ChE) and cholesterol oxidase (ChOx)) immobilized on magnetic nanoparticles (MNPs). The biocatalyst reactions were followed by monitoring the fluorescence decreasing by the naphtofluorescein (NF) oxidation in the presence of the previous H2O2 formed. This microfluidic biosensor supposes the physical integration of a minimal MREµR as a bioactive enzyme area and the focused BFOB connected with the spectrofluorometer detector. The MREµR was formed by a 1 mm length of magnetic retained 2:1 ChE-MNP/ChOx-MNP mixture. The dynamic range of the calibration graph was 0.005–10 mmol L−1, expressed as total cholesterol concentration with a detection limit of 1.1 µmol L−1 (r2 = 0.9999, sy/x = 0.03, n = 10, r = 3). The precision expressed as the relative standard deviation (RSD%) was between 1.3 and 2.1%. The microfluidic-based biosensors showed a sampling frequency estimated at 30 h−1. The method was applied to determine cholesterol in serum samples with recovery values between 94.8 and 102%. The results of the cholesterol determination in serum were also tested by correlation with those obtained using the other two previous methods.Graphical abstract

Diving into Fish Valorisation: Review Opportunities and Analyzing Azorean Fish Data

In response to the exponential growth in world population, there has been a striking surge in the volume of discarded fish worldwide. This surge is particularly evident in the fish processing industry, where a substantial amount of waste is generated, posing significant environmental concerns. Consequently, the repurposing and utilisation of these waste materials have emerged as pivotal processes for the preservation of marine resources. By employing innovative strategies, valuable products can be extracted from these fish by-products, offering not only economic advantages but also contributing to mitigating environmental impacts. This comprehensive literature review focuses on exploring diverse avenues for using fish waste and extracting high-value materials such as bioactive peptides, collagen, and enzymes, elucidating their potential applications across various industries. The literature review also demonstrates the possibility of extracting various bio-compounds from highly diverse fish waste. It has been observed that there is a need for optimisation of extraction protocols, as the variation in extraction methods and respective conditions significantly affects the extraction yields of the products. Moreover, considering our specific interest in the fish species endemic to The Azores, a meticulous characterisation will be conducted, as there is limited knowledge about waste utilisation processes specific to this archipelago.

Integrative roles of sphingosine kinase in liver pathophysiology.

Bioactive sphingolipids and enzymes that metabolize sphingolipid-related substances have been considered as critical messengers in various signaling pathways. One such enzyme is the crucial lipid kinase, sphingosine kinase (SphK), which mediates the conversion of sphingosine to the potent signaling substance, sphingosine-1-phosphate. Several studies have demonstrated that SphK metabolism is strictly regulated to maintain the homeostatic balance of cells. Here, we summarize the role of SphK in the course of liver disease and illustrate its effects on both physiological and pathological conditions of the liver. SphK has been implicated in a variety of liver diseases, such assteatosis, liver fibrosis, hepatocellular carcinoma, and hepatic failure. This study may advance the understanding of the cellular and molecular foundations of liver disease and establish therapeutic approaches via SphK modulation.

MiR-766-3p and miR-671-5p attenuate aristolochic acid-induced hepatotoxicity by directly targeting the key bioactivating enzyme NQO1

Aristolochic acid (AA) as an emerging contaminant in herbal medicines or crops has been well-recognized for causing nephropathy since 1990s. Over the last decade, mounting evidence has linked AA to liver injury; however, the underlying mechanism is poorly elucidated. MicroRNAs respond to environmental stress and mediate multiple biological processes, thus showing biomarker potentials prognostically or diagnostically. In the present study, we investigated the role of miRNAs in AA-induced hepatotoxicity, specifically in regulating NQO1, the key enzyme responsible for AA bioactivation. In silico analysis showed that hsa-miR-766-3p and hsa-miR-671-5p were significantly associated with AAI exposure as well as NQO1 induction. A 28-day rat experiment of 20 mg/kg AA exposure demonstrated a 3-fold increase of NQO1 and an almost 50 % decrease of the homologous miR-671 that were accompanied with liver injury, which was consistent with in silico prediction. Further mechanistic investigation using Huh7 cells with IC50 of AAI at 146.5 µM showed both hsa-miR-766-3p and hsa-miR-671-5p were able to directly bind to and down-regulate NQO1 basal expression. In addition, both miRNAs were shown to suppress AAI-induced NQO1 upregulation in Huh7 cells at a cytotoxic concentration of 70 μM, and consequently alleviate AAI-induced cellular effects, including cytotoxicity and oxidative stress. Together, these data illustrate that miR-766-3p and miR-671-5p attenuate AAI-induced hepatotoxicity, and thus have monitoring and diagnostic potentials.

Beneficial Bio-Extract of Camellia sinensis var. assamica Fermented with a Combination of Probiotics as a Potential Ingredient for Skin Care

Biotechnology, cosmetics, and aesthetic remedies are now inextricably intertwined due to the production of alternative, more effective, and safer active ingredients. Additionally, there has been an increase in demand for natural cosmetic ingredients across the globe. Camellia sinensis var. assamica (Miang tea) is a good alternative because of several biological activities, and is commercially cultivated as a resource in northern Thailand. The process of fermentation mediated by probiotics can enhance the bioavailability of compounds, transform bioactive compounds, and decrease chemical solvent use for sustainability. This study aims to apply the functional evaluation of Miang tea bio-extracts to promote skin health. On the basis of their bioactive enzymes, β-glucosidase, and antioxidant properties, the strains Lacticaseibacillus rhamnosus (previously Lactobacillus rhamnosus), Lactiplantibacillus plantarum (previously Lactobacillus plantarum), and Saccharomyces cerevisiae were used as mixed probiotic starter cultures. The activities of white, green, and black Miang tea bio-extracts, including ferric reducing antioxidant power, lipid peroxidation, nitric oxide inhibition, tyrosinase inhibition, collagenase inhibition (MMP-1 and MMP-2), and antimicrobial activity, were all considerable after 7 days of fermentation time. Additionally, phenolic antioxidant compounds (gallic acid, epigallocatechin gallate, caffeic acid, caffeine, and p-coumaric acid) were identified. The current study’s findings can determine the most effective fermentation time and dose of bio-extract, as well as suggest improvements in bioactive compounds for use in skin care formulations. These results will be used for testing on human participants in further work.

Characterization of a New Hypotensive Peptide from the Venom of Snake Bothrops jararaca (Bj)

Background: Snake venom has become a key source of many bioactive peptides, enzymes, and toxins associated with blood coagulation and neuronal toxicity. In the past, a number of bradykinin potentiating peptides have been isolated from snake venom that display hypotensive activity due to their inhibitory action towards Angiotensin-Converting Enzyme (ACE). Significant interest has developed to isolate, characterize, and subsequently design peptide analogs as potent ACE-inhibitors which may find therapeutic applications for the treatment of hypertension and associated diseases. Aim: The aim of this study is to search for new bioactive peptide/s in the venom of the snake Bothrops Jararaca (Bj). Objective: The objective is to isolate and characterize new hypotensive peptides from BJ venom. Methods: We examined the venom of Bj which is known to host a range of bioactive peptides. We have isolated a new peptide (BJ-1) which displayed in vitro potent hypotensive activity. The peptide was purified via Sephadex G25 column chromatography and RP-HPLC. It was characterized by mass spectrometry, amino acid analysis, N-terminal sequencing, and chemical synthesis. Result: The peptide was identified as an octa-decapeptide with amino acid sequence as DCPSDWSSYEGHCYKPFS where the two Cys residues are likely present in free state, although they can form an internal S-S bond upon oxidation. It was fully confirmed by comparing with synthetic peptide prepared by solid phase chemistry. Both have the same molecular mass (2,108 Da) and identical bioactivity. Furthermore, we rationalize that BJ-1 may be derived from precursor protein “Coagulation factor IX/factor X binding protein (CF-IX/X-BP)” by proteolytic cleavage at the Nterminus of its B-chain within the sequence KPFS18↓E19PKN. This cleavage site contains the recognition motif of enzyme PCSK8 (Proprotein Convertase Subtilisin Kexin8) also known as Subtilisin Kexin Isozyme 1 (SKI-1) or Site 1 Protease (S1P). Despite this observation, using a synthetic peptide encompassing the proposed cleavage site and recombinant PCSK8 enzyme, we found that the enzyme responsible for generation of BJ-1 is not PCSK8. Further studies will be needed to identify the associated enzyme and fully characterize the pharmacological and biological properties of the peptide. Conclusion: Our study revealed the presence of a novel hypotensive octa-decapeptide in the venom of the snake Bothrops jararaca. It is likely derived from the A-chain of protein CF-IX/X-BP via proteolytic cleavage at the N-terminus by a protease yet to be characterized.

Impact of Novel Active Layer-by-Layer Edible Coating on the Qualitative and Biochemical Traits of Minimally Processed 'Annurca Rossa del Sud' Apple Fruit.

The color changes brought on by the enzymatic interactions of phenolic compounds with released endogenous polyphenol oxidase and the penetration of oxygen into the tissue has a significant impact on the commercialization of fresh-cut fruit, such as apples. This process causes a loss of quality in fresh-cut apples, resulting in browning of the fruit surface. By acting as a semipermeable barrier to gases and water vapor and thus lowering respiration, enzymatic browning, and water loss, edible coatings can provide a chance to increase the shelf life of fresh-cut produce. In this study, the effect of edible coatings composed of carboxymethylcellulose (CMC, 1%), sodium alginate (SA, 1%), citric acid (CA, 1%), and oxalic acid (OA, 0.5%) on fresh-cut 'Annurca Rossa del Sud' apple was studied. Four formulations of edible coatings, A. SA+CMC, B. SA+CMC+CA, C. SA+CMC+OA, and D. SA+CMC+CA+OA, were tested. Fresh-cut apples were dipped into different solutions and then stored at 4 °C, and physicochemical and biochemical analyses were performed at 0, 4, 8, and 12 days of storage. Results demonstrated that all four combinations improved the shelf-life of fresh-cut apple by slowing down the qualitative postharvest decay, total soluble solid, and titratable acidity. The browning index was highest in the control samples (82%), followed by CMC+SA (53%), CMC+SA+CA (32%), CMC+SA+OA (22%), and finally CMC+SA+CA+OA (7%) after 12 days of cold storage. Furthermore, coating application increased the bioactive compound content and antioxidant enzyme activities. Furthermore, the synergistic activity of SA+CMC+CA+OA reduces enzymatic browning, prolonging the postharvest life of minimally processed 'Annurca Rossa del Sud' apples.