Peptide Hydrolysis Research Articles

Exploring novel peptides in adzuki bean and mung bean hydrolysates with potent antibacterial activity

SummaryThis study delved into the antimicrobial potential of hydrolysates from adzuki bean and mung bean protein concentrates, created using Alcalase and Flavourzyme enzymes. Targeting common foodborne pathogens (Salmonella Typhimurium, Salmonella Enteritidis, Vibrio parahaemolyticus, Staphylococcus aureus, and Bacillus subtilis) and Escherichia coli, the research underscored the crucial role of peptidase in generating antibacterial peptides. Fractions F2 of Flavourzyme hydrolysed adzuki bean (ABF) and F4 of Flavourzyme hydrolysed mung bean (MBF) showed the highest antibacterial activity against S. Typhimurium and S. aureus, respectively, whereas F2 of Alcalase hydrolysed mung bean (MBA) exhibited very poor activity against S. aureus. TripleTOF LC–MS/MS analysis revealed small cationic antibacterial peptides, with lysine being prominent against S. aureus and proline–proline against S. Typhimurium. These findings highlighted the significant contribution of protease enzymes in fostering antibacterial peptide production, offering valuable insights for developing effective antimicrobial agents in the context of food safety.

Identification, characteristics and molecular docking studies of flavor peptides in enzymatic hydrolysates of Capparis masaikai Levl

Identification, characteristics and molecular docking studies of flavor peptides in enzymatic hydrolysates of Capparis masaikai Levl

Antihypertensive effect of chicken blood peptide hydrolysate in streptozotocin-induced diabetic rats

Chicken blood peptide hydrolysate (CBH) has been reported to exert antioxidant, angiotensin 1-converting enzyme (ACE) inhibitory, and cholesterol-lowering activities. Antihypertensive effect of the CBPH was also demonstrated in spontaneously hypertensive rats. This study aimed to assess the anti-diabetic and antihypertensive effect of the CBH in streptozotocin (STZ)-induced diabetic rats. CBH was prepared from fresh chicken blood collected from an industrial slaughterhouse in Nakhon Ratchasima, Thailand. Adult male Wistar rats were randomly divided into 2 main groups: normal and STZ-induced diabetes mellitus (DM) groups. Five subgroups of each main group were given distilled water, metformin (200 mg/kg), and CBH (300, 600, 1200 mg/kg) by oral gavage for 28 days (n=10 per each group). Fasting blood sugar level and OGTT were evaluated before and at the end of experiment. Each rat underwent monitoring of blood pressure using femoral artery. STZ-induced diabetic rats showed higher fasting blood glucose concentration and area under curve from OGTT than normal rats, which was not reduced by all treatments. STZ-induced diabetic rats showed higher mean arterial blood pressure than normal rats, which was significantly reduced by metformin and CBH (300 and 1200 mg/kg) (p<0.05). The blood pressure-lowering effect of CBH suggested the potential for the application of CBH in the management of diabetic hypertension. However, attention must be paid to concerns about bird flu in certain places and prohibition of the use of blood product. This work was supported by Suranaree University of Technology (SUT), Thailand Science Research and Innovation (TSRI), and National Science Research and Innovation Fund (NSRF) (FF1-111-65-12-30(08)). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Protein identification and potential bioactive peptides from pumpkin (Cucurbita maxima) seeds.

Pumpkin is an economically important crop all over the world. Approximately, 18%-21% of pumpkins, consisting of peels and seeds by-products, are wasted during processing. In addition, the seeds are rich in protein and have the potency of bioactive peptide production. This study aims to recognize the proteins and investigate the potential bioactive peptides from pumpkin (Cucurbita maxima) seeds. Pumpkin seeds were subjected to hot air drying (HAD) at 55°C for 12 h and freeze-drying (FD) at -80°C for 54 h before they were powdered, analyzed, and precipitated by isoelectric point to obtain pumpkin seed protein isolates (PSPI). PSPI comprised 11S globulin subunit beta, 2S seed storage albumin, and chaperonin CPN60-1. To generate hydrolysate peptides, PSPI was hydrolyzed using papain, pepsin, and bromelain. FD group pepsin hydrolysates had the highest peptide content of 420.83 mg/g. ACE inhibition and DPP-IV inhibition activity were analyzed for each enzymatic hydrolysate. The pepsin hydrolyzed sample exhibited the highest ACE inhibition of 70.26%, and the papain hydrolyzed sample exhibited the highest DPP-IV inhibition of 30.51%. The simulated gastrointestinal digestion (SGID) conducted by pepsin and pancreatin increased ACE inhibitory activity from 76.93% to 78.34%, and DPP-IV inhibited activity increased from 58.62% to 77.13%. Pepsin and papain hydrolysates were fractionated using ultrafiltration to measure ACE and DPP-IV inhibition activity. The highest free radical scavenging abilities were exhibited by the <1 kDa hydrolysate fractions with 78.34% ACE inhibitory activities and 79.55% DPP-IV inhibitory activities. This research revealed that pumpkin seeds had the potency to produce bioactive peptides.

Highly defective ultra-small tetravalent MOF nanocrystals

The size and defects in crystalline inorganic materials are of importance in many applications, particularly catalysis, as it often results in enhanced/emerging properties. So far, applying the strategy of modulation chemistry has been unable to afford high-quality functional Metal–Organic Frameworks (MOFs) nanocrystals with minimized size while exhibiting maximized defects. We report here a general sustainable strategy for the design of highly defective and ultra-small tetravalent MOFs (Zr, Hf) crystals (ca. 35% missing linker, 4–6 nm). Advanced characterizations have been performed to shed light on the main factors governing the crystallization mechanism and to identify the nature of the defects. The ultra-small nanoMOFs showed exceptional performance in peptide hydrolysis reaction, including high reactivity, selectivity, diffusion, stability, and show emerging tailorable reactivity and selectivity towards peptide bond formation simply by changing the reaction solvent. Therefore, these highly defective ultra-small M(IV)-MOFs particles open new perspectives for the development of heterogeneous MOF catalysts with dual functions.

Biological properties of LMW-peptide fractions from apricot kernel protein: Nutritional, antibacterial and ACE-inhibitory activities

In this study, angiotensin-converting enzyme (ACE) inhibitory, antioxidant and antibacterial activities, and nutritional quality of low molecular weights (LMWs) peptide fractions (PFs) from apricot-kernel (Prunus armeniaca cv. Tabarzeh) with PFs <3, 3–10, 10–30 and 30–100 kDa were investigated. The ratio of essential to total (E/T) amino acids (∼34.5 %) and PER value (1.8–1.9) indicated the high nutritional and digestibility of peptide fractions. Antioxidant indices were affected by enzymatic hydrolysis, molecular weight (Mw) and concentration of peptides. In general, the highest inhibition of DPPH− (89.2 %), ABTS+ (93.1 %), TEAC (2.52 mM), OH• (78.1 %), NO• (70.4 %), total antioxidant activity (2.01), reducing power (1.27), ACE inhibitory (75.1 %), chelation of Fe2+ (90.2 %) and Cu2+ (40.2 %) metal-ions were related to low Mw fractions (PF-3 and PF-10). The hydrolysates and primary protein showed enhanced antibacterial effects against Escherichia coli and Bacillus cereus with diameter of inhibition zones of 17 and 14 mm, respectively. Overall, apricot-kernel peptides can be considered as a nutrient source, natural antioxidant and preservative with ACE-inhibitory effect in dietary supplements and food formulations.

Study of antitumor effects of human placenta hydrolysate on PC-3, OAW-42, BT-474 cell cultures

To investigate the antitumor effects of human placenta hydrolysate (HPH) peptides on three hormone-dependent human cell lines: prostate adenocarcinoma, breast carcinoma, and ovarian cancer by metabolic analysis of cell cultures. The effect of HPH on tumor and control tumor cell lines was evaluated. Study stages: (A) de novo peptide sequencing by collision-induced dissociation mass spectrometry; (B) detection of peptides with anti-tumor properties; (C) expert analysis of the obtained lists of peptides. Dose-dependent cytotoxic effects of HPH on three tumor cell lines are shown: PC-3 (human prostate adenocarcinomas), OAW-42 (human ovarian cancer), BT-474 (human breast carcinomas), and IC50 constants (1.3-2.8 mg/ml) were obtained. The analysis of the HPH peptide fraction showed more than 70 peptides with antitumor properties in the composition of this HPH, including kinase inhibitors: mitogen-activated protein kinases, kappa-bi nuclear factor inhibitor kinase, AKT serine/threonine kinase 1, protein kinase C zeta, interleukin-1 receptor-associated kinase 4 and cyclin-dependent kinase 1. The results of the study indicate not only the oncological safety of the HPH used in therapy but also the mild antitumor effects of this HPH at high concentrations.

BDNF- and VEGF-Responsive Stimulus to an NGF Mimic Cyclic Peptide with Copper Ionophore Capability and Ctr1/CCS-Driven Signaling.

Neurotrophins are a family of growth factors that play a key role in the development and regulation of the functioning of the central nervous system. Their use as drugs is made difficult by their poor stability, cellular permeability, and side effects. Continuing our effort to use peptides that mimic the neurotrophic growth factor (NGF), the family model protein, and specifically the N-terminus of the protein, here we report on the spectroscopic characterization and resistance to hydrolysis of the 14-membered cyclic peptide reproducing the N-terminus sequence (SSSHPIFHRGEFSV (c-NGF(1-14)). Far-UV CD spectra and a computational study show that this peptide has a rigid conformation and left-handed chirality typical of polyproline II that favors its interaction with the D5 domain of the NGF receptor TrkA. c-NGF(1-14) is able to bind Cu2+ with good affinity; the resulting complexes have been characterized by potentiometric and spectroscopic measurements. Experiments on PC12 cells show that c-NGF(1-14) acts as an ionophore, influencing the degree and the localization of both the membrane transporter (Ctr1) and the copper intracellular transporter (CCS). c-NGF(1-14) induces PC12 differentiation, mimics the protein in TrkA phosphorylation, and activates the kinase cascade, inducing Erk1/2 phosphorylation. c-NGF(1-14) biological activities are enhanced when the peptide interacts with Cu2+ even with the submicromolar quantities present in the culture media as demonstrated by ICP-OES measurements. Finally, c-NGF(1-14) and Cu2+ concur to activate the cAMP response element-binding protein CREB that, in turn, induces the brain-derived neurotrophic factor (BDNF) and the vascular endothelial growth factor (VEGF) release.

Utilization of peanut protein isolate amyloid-like fibers as stabilizers for high internal phase Pickering emulsions

This paper focuses on modifying peanut protein isolate (PPI) through fibrillation to enhance its emulsification properties and investigates the potential application of the fibers in stabilizing high internal phase Pickering emulsions (HIPPEs). Th T result indicates that the optimal conditions for peanut protein isolate amyloid-like fibers (PAFs) preparation are pH=2, 90 °C, and 20 h. Electrophoresis and Transmission electron microscope demonstrate that PPI undergoes peptide hydrolysis and self-assembly during the reaction process, leading to the formation of an elongated linear structure with a diameter approximately 14 nm. Furthermore, infrared spectroscopy, X-ray diffraction, and contact angle results further demonstrate an increase in β-sheet content, crystallinity, and surface hydrophilicity compared to PPI. Notably, the emulsification activity index (EAI) of PAFs (78.62 ± 1.68 m2/g) was found to be increased up to threefold compared to that of PPI (27.89 ± 1.98 m2/g). According to visual observation and microstructural analysis, the addition of 1% PAFs has been found to effectively stabilize HIPPEs (φdisp = 0.75) due to the presence of dense protective layers formed by PAFs. Additionally, the particle size results (D[3,2] = 17.02 ± 2.02 μm) also indicate that the oil droplets are tightly enveloped by the dense protective layers formed by PAFs. Moreover, rheological data indicates that HIPPEs stabilized using PAFs exhibit excellent solid-like viscoelastic behavior. Overall, these findings suggest that the utilization of PAFs has significant potential in enhancing the stability of HIPPEs.

Investigation of the peptides with calcium chelating capacity in hydrolysate derived from spent hen meat.

Calcium peptide chelates are developed as efficient supplements for preventing calcium deficiency. Spent hen meat (SHM) contains a high percentage of proteins but is generally wasted due to the disadvantages such as hard texture. We chose the underutilized SHM to produce peptides to bind calcium by proteolysis and aimed to investigate chelation between calcium and peptides in hydrolysate for a sustainable purpose. The optimized proteolysis conditions calculated from the result of response surface methodology for two-step hydrolysis were 0.30% (wenzyme/wmeat) for papain with a hydrolysis time of 3.5h and 0.18% (wenzyme/wmeat) for flavourzyme with a hydrolysis time of 2.8h. The enzymatic hydrolysate (EH) showed a binding capacity of 63.8±1.8mg calcium/g protein. Ethanol separation for EH improved the capacity up to a higher value of 68.6±0.6mg calcium/g protein with a high association constant of 420M-1 (25°C) indicating high stability. The separated fraction with a higher amount of Glu, Asp, Lys, and Arg had higher calcium-binding capacity, which was related to the number of ─COOH and ─NH2 groups in peptide side chains according to the result from amino acid analysis and Fourier transform infrared spectroscopy. Two-step enzymatic hydrolysis and ethanol separation were an efficient combination to produce peptide mixtures derived from SHM with high calcium-binding capacity. The high percentage of hydrophilic amino acids in the separated fraction was concluded to increase calcium-binding capacity. This work provides foundations for increasing spent hen utilization and developing calcium peptide chelates based on underutilized meat.

Isolation and antioxidant activity of peptides from Chinese hairy tofu.

Hairy tofu is a famous Chinese snack that is made from soybeans and rich in various nutrients. In order to further explore the antioxidant peptides of hairy tofu hydrolysates, seven proteases were used to hydrolyze hairy tofu. The results of in vitro radical scavenging activity showed that hairy tofu hydrolysates obtained by pancreatin exhibited the highest antioxidant activity. After Sephadex G-25 gel filtration and reversed-phase high-performance liquid chromatography (RP-HPLC), 97 peptides were identified in the most antioxidant fraction using liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). Among them, nine peptides were synthesized and their antioxidant activities were assessed using a H2O2-induced oxidative 293T cell model. Finally, four peptides (QCESHK, LAWNEGR, NLQGENEWDQK, and FTEMWR) at concentrations of < 50 μg/ml significantly decreased the malondialdehyde content compared with the model group, displaying in vivo antioxidant activity and low cytotoxicity. Overall, this research provided the choice of using hairy tofu peptides as antioxidant products in the pharmaceutical and food industries.

Enhanced methane production with co-feeding spent coffee grounds using spare capacity of existing anaerobic food waste digesters

With increasing coffee consumption worldwide, the efficient and sustainable management of spent coffee grounds (SCG) has become increasingly challenging. This study investigated the anaerobic co-digestion of small amounts of SCG with food waste (FW) at increasing co-feeding ratios of 1:100–1:10 (volatile solids basis) to assess the possibility of SCG treatment using the spare capacity of existing anaerobic digesters. Co-feeding SCG increased methane production compared to FW mono-digestion in the tested range of co-feeding ratios without compromising process stability. Methane yield did not further increase when the SCG/FW ratio increased above 4%, and process failure occurred at a 1:10 co-feeding ratio without trace element supplementation. The enhanced methanogenic performance was attributed to increased protein removal efficiency, which was potentially related to the promotion of peptide hydrolysis. The overall results suggest that co-feeding appropriate small amounts of SCG to FW digesters can be a realistic sustainable option for SCG management.

Evaluation of different isotope dilution mass spectrometry strategies for the characterization of naturally abundant and isotopically labelled peptide standards

Natural abundance and isotopically labelled tryptic peptides are routinely employed as standards in quantitative proteomics. The certification of the peptide content is usually carried out by amino acid analysis using isotope dilution mass spectrometry (IDMS) after the acid hydrolysis of the peptide. For the validation and traceability of the amino acid analysis procedure, expensive certified peptides must be employed. In this work we evaluate different IDMS alternatives which will reduce the amount of certified peptide required for validation of the amino acid analysis procedure. In this context, the characterization of both natural and isotopically labelled synthetic angiotensin I peptides was carried out. First, we applied a fast procedure for peptide hydrolysis based on microwave-assisted digestion and employed two certified peptide reference materials SRM 998 angiotensin I and CRM 6901-b C-peptide for validation of the hydrolysis procedure. The amino acids proline, leucine, isoleucine, valine, tyrosine, arginine and phenylalanine were evaluated for their suitability for peptide certification by IDMS by both liquid chromatography with tandem mass spectrometry (LC–MS/MS) and gas chromatography with mass spectrometry (GC)–MS/MS. Then, natural angiotensin I and 13C1-labelled angiotensin I were synthesized in-house and purified by preparative liquid chromatography. The concentration of the 13C1-labelled angiotensin I peptide was established by reverse IDMS in its native form using SRM 998 angiotensin I as reference. The concentration of the natural synthesized peptide was determined by IDMS both using the 13C1-labelled peptide in its native form and by amino acid analysis showing comparable results. Finally, the synthetic naturally abundant angiotensin I peptide was employed as “in-house” standard for the validation of subsequent peptide characterization procedures. Therefore, the novelty of this work relies on, first, the development of a faster hydrolysis procedure assisted by focused microwaves, providing complete hydrolysis in 150 min, and secondly, a validation strategy combining GC–MS and LC–MS/MS that allowed us to certify the purity of an in-house-synthesized peptide standard that can be employed as quality control in further experiments.Graphical abstract

Identification, structural characterization, and molecular dynamic simulation of ACE inhibitory peptides in whey hydrolysates from Chinese Rushan cheese by-product

To realize the high-value utilization of Rushan cheese by-product, Rushan cheese whey was used as a raw material to prepare angiotensin-Ⅰ-converting enzyme inhibitory peptides (ACEIPs). After enzymatic hydrolysisn and ultrafiltration, the sequences of peptides were identified by liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS). Two novel ACE inhibitory peptides Phe-Asp-Arg-Pro-Phe-Leu (FDRPFL) and Lys-Trp-Glu-Lys-Pro-Phe (KWEKPF) were identified. Additionally, both of the peptides exhibited good water-solubility and no toxicity according to in-silico prediction. Fourier transform infrared spectroscopy results show that both FDRPFL and KWEKPF were enriched in β-turn and β-sheet structures. Lineweaver-Burk plots revealed that FDRPFL and KWEKPF exhibited non-competitive and mixed inhibition patterns, respectively. Molecular docking and MD simulation showed that hydrogen bonds and ionic bonds forces allowed FDRPFL and KWEKPF to form stable and compact complexes with ACE. In conclusion, enzymatic hydrolysis of Rushan cheese by-products yields bioactive peptides, increases the added value of whey and reduces environmental pollution.

Protein degradation by human 20S proteasomes elucidates the interplay between peptide hydrolysis and splicing

If and how proteasomes catalyze not only peptide hydrolysis but also peptide splicing is an open question that has divided the scientific community. The debate has so far been based on immunopeptidomics, in vitro digestions of synthetic polypeptides as well as ex vivo and in vivo experiments, which could only indirectly describe proteasome-catalyzed peptide splicing of full-length proteins. Here we develop a workflow—and cognate software - to analyze proteasome-generated non-spliced and spliced peptides produced from entire proteins and apply it to in vitro digestions of 15 proteins, including well-known intrinsically disordered proteins such as human tau and α-Synuclein. The results confirm that 20S proteasomes produce a sizeable variety of cis-spliced peptides, whereas trans-spliced peptides are a minority. Both peptide hydrolysis and splicing produce peptides with well-defined characteristics, which hint toward an intricate regulation of both catalytic activities. At protein level, both non-spliced and spliced peptides are not randomly localized within protein sequences, but rather concentrated in hotspots of peptide products, in part driven by protein sequence motifs and proteasomal preferences. At sequence level, the different peptide sequence preference of peptide hydrolysis and peptide splicing suggests a competition between the two catalytic activities of 20S proteasomes during protein degradation.

Nutritional and functional properties of protein hydrolysate from Tra fish by-products

The nutritional and functional properties, molecular weight distribution of protein hydrolysate from Tra fish by-products were studied. The protein hydrolysate was produced by enzymatic hydrolysis of Tra fish by-products with Protamex 0,5% at the temperature of 50 oC in 6 h. The study results showed that the Tra fish by-products protein hydrolysate had a moisture content of 6,7%, protein content of 65.8%, lipid content of 1,6% and ash content of 9,5 %. The protein hydrolysate had a total amino acid content of 52,81 g/100g of protein and essential amino acid content of 16,25 g/100g of protein. The ratio of essential amino acids to total amino acids was 30,76%. The protein hydrolysate was rich in glycine, glutamic, proline, alanine, aspartic and leucine. The most of peptides (68,25%) in Tra fish by-products protein hydrolysate had a molecular weight between 360 Da and 2.500 Da. The protein hydrolysate had solubility in the range of 90,3% - 98,2%. The solubility of protein hydrolysate was the lowest value at pH 4 and the highest value at pH 10. The foaming capacity of protein hydrolysate ranged from 19,5% to 27,8%. The emulsifying capacity of protein hydrolysate was 12,8 – 17,3 mL/g. The protein hydrolysate of Tra fish by-products possesses high nutritional value and good functional properties. The study results suggested that the protein hydrolysate from Tra fish by-products could be used as a promising food ingredient and protein source in food systems. ® 2023 Journal of Science and Technology - NTTU

Leveraging SARS-CoV-2 Main Protease (Mpro) for COVID-19 Mitigation with Selenium-Based Inhibitors.

The implementation of innovative approaches is crucial in an ongoing endeavor to mitigate the impact of COVID-19 pandemic. The present study examines the strategic application of the SARS-CoV-2 Main Protease (Mpro) as a prospective instrument in the repertoire to combat the virus. The cloning, expression, and purification of Mpro, which plays a critical role in the viral life cycle, through heterologous expression in Escherichia coli in a completely soluble form produced an active enzyme. The hydrolysis of a specific substrate peptide comprising a six-amino-acid sequence (TSAVLQ) linked to a p-nitroaniline (pNA) fragment together with the use of a fluorogenic substrate allowed us to determine effective inhibitors incorporating selenium moieties, such as benzoselenoates and carbamoselenoates. The new inhibitors revealed their potential to proficiently inhibit Mpro with IC50-s in the low micromolar range. Our study contributes to the development of a new class of protease inhibitors targeting Mpro, ultimately strengthening the antiviral arsenal against COVID-19 and possibly, related coronaviruses.

Evaluation of antioxidant and ACE inhibitory peptides in rice bran protein hydrolysate using non‐targeted UHPLC‐QTOF‐IMS profiling, MALDI‐TOF‐TOF‐MS/MS, and molecular docking study

SummaryIn the present study, rice bran protein (RBP) from the PB1121 variety was hydrolysed using trypsin and subjected to non‐targeted IMS profiling to identify antioxidant and antihypertensive amino acids in the resulting RBP hydrolysates. The hydrolysate was then separated into three fractions using ultrafiltration, and each fraction was tested for antioxidant and ACE inhibitory activities. The ACE activity was assessed using the tripeptide hippuryl‐histidyl‐leucine (HHL) as a model peptide via HPLC‐DAD. The fraction with the molecular weight (&gt;3000 Da) displayed the strongest antioxidant and ACE inhibitory activity. This fraction was further divided into five fractions using gel Sephadex G‐25, and the greatest levels of antioxidant and ACE inhibitory activity were found in fraction F3B. Fraction F3B was then fractionated using HPLC, and the fraction with ACE inhibitory activity (IC50 of 43 mg mL−1) was analysed using MALDI‐TOF‐TOF mass spectrometry to determine its exact molecular mass and amino acid sequence. The amino acid sequence FMKSK (phe‐met‐lys‐ser‐lys) with a Mw of 655.376 Da was detected, and the molecular docking investigation revealed that FMKSK suppresses ACE by forming strong hydrogen bonds with the active pockets of human ACE. These findings suggest that rice bran contains bioactive peptides with antioxidant and ACE inhibitory effects, making it a promising raw material for the production of beneficial products.

Toxicological evaluation of peptide hydrolysates from bovine blood meal with antioxidant and antifungal activities

This study addresses the underutilized potential of bovine blood-derived bioactive peptides. With approximately 22 million metric tons of bovine blood waste generated annually, sustainable utilization is essential. Our research focuses on exploring antifungal and antioxidant properties and includes proximate analysis of blood meal, enzymatic activity determination, and optimization of bovine blood protein hydrolysis. Optimal conditions were identified at 70 °C and pH 9, achieving a remarkable degree of hydrolysis (25.75 %). Key results demonstrate that lower molecular weight peptides exhibit enhanced antioxidant capacity, making them suitable for functional foods and nutraceuticals. While antimicrobial activity against specific strains had limitations, significant antifungal efficacy against Fusarium keratoplasticum ATCC 36031 was observed. Electrophoresis analysis confirmed the presence of peptides with a molecular weight below 14 kDa in optimally hydrolyzed samples, aligning with their bioactivity. A maximum tolerated concentration of 100 µg/mL was established through toxicity assessment using a zebrafish model, ensuring safety. This research reorganizes the abstract to address underutilized bovine blood, explores bioactive potential, and presents significant results concerning antioxidant and antifungal activity. Significantly, this study has implications for food safety, environmental sustainability, and health-related product development. The enzyme used was obtained from fermenting soybean cake with B. subtilis, and treatment selection was based on optimization studies. This research offers insights into industrial applications, environmental impact reduction, and health-related advancements.

Optimization of Hydrolysis and Identification of Bioactive Peptides in Brewery Yeast Residuals

Industrial waste products are of biotechnological interest due to their abundance and can be utilized as protein sources for the production of bioactive peptides through hydrolysis. The utilization of yeast residues from the brewing industry has been demonstrated to be a viable method for obtaining protein hydrolysates. The degree of hydrolysis and soluble total protein of the hydrolysates were evaluated for optimization purposes to achieve a product with a high proportion of soluble organic nitrogen, increased levels of bioactive peptides and amino acids. Both alkaline and enzymatic hydrolysis processes were tested. The hydrolyses were carried out with the aim of large-scale implementation. The treatments were effective in liberating intracellular content and producing biologically functional hydrolysates, with degree of hydrolysis values ranging from 28 to 63% and total protein from 2.95 to 8.28% at the optimum points confirmed by statistical analysis (analysis of variance (ANOVA)). The hydrolysate produced with Alcalase® exhibited the highest peptide intensity, while the hydrolysate produced with Flavourzyme® showed the highest concentration of free amino acids. The peptides were identified by mass spectrometry and were found to have biological activities according to the Biopep database. This study presents a quick and economically feasible approach for the implementation of a pilot project for the reuse of this waste.