Production Of Bioactive Peptides Research Articles

Cilia Provide a Platform for the Generation, Regulated Secretion, and Reception of Peptidergic Signals.

Cilia are microtubule-based cellular projections that act as motile, sensory, and secretory organelles. These structures receive information from the environment and transmit downstream signals to the cell body. Cilia also release vesicular ectosomes that bud from the ciliary membrane and carry an array of bioactive enzymes and peptide products. Peptidergic signals represent an ancient mode of intercellular communication, and in metazoans are involved in the maintenance of cellular homeostasis and various other physiological processes and responses. Numerous peptide receptors, subtilisin-like proteases, the peptide-amidating enzyme, and bioactive amidated peptide products have been localized to these organelles. In this review, we detail how cilia serve as specialized signaling organelles and act as a platform for the regulated processing and secretion of peptidergic signals. We especially focus on the processing and trafficking pathways by which a peptide precursor from the green alga Chlamydomonas reinhardtii is converted into an amidated bioactive product-a chemotactic modulator-and released from cilia in ectosomes. Biochemical dissection of this complex ciliary secretory pathway provides a paradigm for understanding cilia-based peptidergic signaling in mammals and other eukaryotes.

Investigations on the chemical and bioactivity profile of Bromeliaceae Juss

Bromeliaceae Juss., in several cultures for thousands of years it has been used for food, fibers, in ceremonies, medicines and ornamental plants. The phytochemical composition of the species of the family is chemodiverse, with the presence of fatty acids, phenolic compounds, vitamins, triterpenes and enzymes, exerting antibacterial, antimicrobial, hypoglycemic, antileukemic, proteolytic activities, non-polar natural products and production of bioactive peptides. As a result, the family has great chemical and pharmacological potential in order to contribute to scientific knowledge and the development of new drugs.

Extracellular production of antifungal peptides from oxidative endotoxin-free E. coli and application.

Antifungal peptides (AFPs) can be used as novel preservatives, but achieving large-scale production and application remains a long-term challenge. In this study, we developed a hybrid peptide MD (metchnikowin-drosomycin fusion) secreted into Escherichia coli supernatant, demonstrating strong inhibitory activity against Aspergillus flavus and Botrytis cinerea. The fusion tag did not impact its activity. Moreover, an endotoxin-free and oxidative leaky strain was developed by knockingout the trxB, gor, and lpp genes of endotoxin-free E. coli ClearColi-BL21(DE3). This strain facilitates the proper folding of multi-disulfide bond proteins and promotes the extracellular production of recombinant bioactive AFP MD, achieving efficient production of endotoxin-free MD. In addition, temperature control replaces chemical inducers to further reduce production costs and circumvent the toxicity of inducers. This extracellularly produced MD exhibited favorable effectiveness in inhibiting fruit mold growth, and its safety was preliminarily established by gavage testing in mice, suggesting that it can be developed into a green and sustainable fruit fungicide. In conclusion, this study provides novel approaches and systematic concepts for producing extracellularly active proteins or peptides with industrial significance. KEY POINTS: • First report of extracellular production of bioactive antifungal peptide in Escherichia coli. • The hybrid antifungal peptide MD showed strong inhibitory activity against Aspergillus flavus and Botrytis cinerea, and the activity was not affected by the fusion tag. • Endotoxin-free oxidative Escherichia coli suitable for the expression of multi-disulfide bond proteins was constructed.

Production of functional recombinant roseltide rT1 antimicrobial peptide in tobacco plants

Plant-derived peptides represent a promising group of natural compounds with broad industrial and pharmaceutical applications. Low-efficiency production level is the major obstacle to the commercial production of such bioactive peptides. Today, recombinant techniques have been developed for fast and cost-effective production of high-quality peptides for various applications in the chemical and food industries. The roseltide rT1 is a plant peptide with different antimicrobial properties and therapeutic applications in the prevention and treatment of inflammatory lung diseases by inhibiting human neutrophil elastases. Here, we report the expression of functional recombinant roseltide rT1 peptide in tobacco plants. Transgenic plants were generated by the Agrobacterium-mediated transformation method followed by molecular analysis of transgenic plants to demonstrate successful integration and expression of recombinant rT1 peptide. Protein extracts of transgenic plants expressing a single-copy rT1 gene showed efficient antimicrobial properties as verified by growth inhibition of different bacterial strains. Our results illustrate that plant-derived recombinant rT1 peptide is a promising alternative for rapid and cost-effective production of this important antimicrobial peptide for application in therapeutic and food industries.

Lentils protein isolate as a fermenting substrate for the production of bioactive peptides by lactic acid bacteria and neglected yeast species.

In the current trend where plant-based foods are preferred over animal-based foods, pulses represent an alternative source of protein but also of bioactive peptides (BPs). We investigated the pattern of protein hydrolysis during fermentation of red lentils protein isolate (RLPI) with various lactic acid bacteria and yeast strains. Hanseniaspora uvarum SY1 and Fructilactobacillus sanfranciscensis E10 were the most proteolytic microorganisms. H. uvarum SY1 led to the highest antiradical, angiotensin-converting enzyme-inhibitory and antifungal activities, as found in low molecular weight water soluble extracts (LMW-WSE). The 2039 peptide sequences identified by LMW-WSE were screened using BIOPEP UWM database, and 36 sequences matched with known BPs. Fermentation of RLPI by lactic acid bacteria and yeasts generated 12 peptides undetected in raw RLPI. Besides, H. uvarum SY1 led to the highest abundance (peak areas) of BPs, in particular with antioxidant and ACE-inhibitory activities. The amino acid sequences LVR and LVL, identified in the fermented RLPI, represent novel findings, as they were detected for the first time in substrates subjected to microbial fermentation. KVI, another BP highly characteristic of RLPI-SY1, was previously observed only in dried bonito. 44 novel potential BPs, worthy of further characterization, were correlated with antifungal activity.

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.

Production of bioactive peptides with antioxidant and antihypertensive activities from wheat gluten using Withania coagulans

Production of bioactive peptides with antioxidant and antihypertensive activities from wheat gluten using Withania coagulans

Bioactive Peptides from Velvet Bean Tempe: Neutrase-Catalyzed Production in Membrane Reactor

Velvet beans are potential sources of parent proteins for bioactive peptide production. In this study, a combination of fermentation and neutrase-catalyzed continuous hydrolysis in an enzymatic membrane reactor was performed to produce antioxidative and angiotensin I-converting enzyme inhibitory (ACEi) peptides. The optimum operating conditions were τ = 6 h and [E]/[S] = 7.5%. The resulting permeate, which was a<10-kDa fraction, exhibited antioxidant activity at 0.38 mg ascorbic acid equivalent antioxidant capacity (AEAC)/mL (2,2-diphenyl-1-picrylhydrazyl, DPPH inhibition) and 0.26 mg AEAC/mL (ferric reducing antioxidant power, FRAP), and ACEi activity of 81.02%. Further fractionation of the permeate increased the ACEi activity in which 2-kDa fraction showed the most potent activity (IC50 = 0.23 µg protein/mL). The IC50 value of the outcome was comparable to those reported in the literature for velvet bean-based peptides. Furthermore, this study suggests that neutrase is a good catalyst candidate for the synthesis of bioactive peptides from velvet beans.

Enzymatic Method of Bioactive Peptides Production from Milk Protein Raw Materials: A Scoping Review

Introduction: For 14 years, interest in the use of dairy raw materials for the production of bioactive peptides (BPs) with antihypertensive, antioxidant and antidiabetic effects has been in a phase of rapid growth because of the need to prevent different diseases. Particular attention is being paid to enzymatic production method, as it allows for a targeted process of BP extraction with specified properties.The purpose of this scoping review was to summarize and systematize the experience of international research over the last 14 years in the field of FG as a method to extract BPs from milk protein substrates.Materials and Methods: This scoping review was performed in accordance with the PRISMA-ScR guidelines. Scientific papers were searched in the Google Scholar database, because allows searching for scientific sources without restrictions on their format and type of scientific data presentation (book, dissertation, article), as well as includes scientific sources from other databases and performs relevance sorting.Results: The search identified 72 sources published between 2007 and 2021 in English. Included sources are devoted to hydrolysis by starter bacteria enzymes (SBE) - 50% and by commercial enzyme preparations (CEP) - 50%.Conclusion: There are a number of challenges that need to be addressed in order to increase the level of commercialization of BP production through directed hydrolysis. Creation of databases; building up the research base on co-fermentation by microorganisms and commercial enzymes; conducting PCR studies of microbial protease genes will increase the practical applicability of research and the validity of conducting more evidence-based in vivo and clinical studies.

The Production of Bioactive Peptides by Optimization of Enzymatic Hydrolysis Process of Protein from Tilapia Fish Skin Waste (Oreochromis niloticus, Linnaeus 1758) using Alcalase 2.4.L

Aims: This study aimed at developing bioactive peptides by optimization of the enzymatic hydrolysis process of protein from tilapia fish skin waste (Oreochromis niloticus, Linnaeus 1758) using alcalase 2.4.L. Background: Natural bioactive peptides are considered to have low toxicity and therapeutic properties as antioxidants. Objective: The conditions of protein hydrolysis obtained from tilapia fish skin waste (Oreochromis niloticus, Linnaeus 1758) were optimized using alcalase 2.4.l Methods: In this study, the hydrolysis of protein obtained from tilapia fish skin waste (TFSW) was optimized using alcalase 2.4.L by central composite design (CCD). Degree of hydrolysis (DH), radical scavenging activities (DPPH), and ferric-reducing antioxidant power (FRAP) were used as dependent variables, whereas temperature, pH, and proportion of enzyme to the substrate (PE%) as independent variables. Results: The optimum degree of hydrolysis DH%, DPPH, and FRAP were achieved at a temperature of 58.4 ℃, a pH of 8.7, except for DPPH, which was achieved at a pH of 7.0. Conclusion: The present work demonstrated that TFSW could be used as a source to produce bioactive peptides with significant antioxidant activities under specific conditions of enzymatic hydrolysis.

Bifidobacteria in Fermented Dairy Foods: A Health Beneficial Outlook.

Bifidobacteria, frequently present in the human gastrointestinal tract, play a crucial role in preserving gut health and are mostly recognized as beneficial probiotic microorganisms. They are associated with fermenting complex carbohydrates, resulting in the production of short-chain fatty acids, bioactive peptides, exopolysaccharides, and vitamins, which provide energy and contribute to gut homeostasis. In light of these findings, research in food processing technologies has harnessed probiotic bacteria such as lactobacilli and bifidobacteria for the formulation of a wide range of fermented dairy products, ensuring their maximum survival and contributing to the development of distinctive quality characteristics and therapeutic benefits. Despite the increased interest in probiotic dairy products, introducing bifidobacteria into the dairy food chain has proved to be complicated. However, survival of Bifidobacterium species is conditioned by strain of bacteria used, metabolic interactions with lactic acid bacteria (LAB), fermentation parameters, and the temperature of storage and preservation of the dairy products. Furthermore, fortification of dairy foods and whey beverages with bifidobacteria have ability to change physicochemical and rheological properties beyond economic value of dairy products. In summary, this review underscores the significance of bifidobacteria as probiotics in diverse fermented dairy foods and accentuates their positive impact on human health. By enhancing our comprehension of the beneficial repercussions associated with the consumption of bifidobacteria-rich products, we aim to encourage individuals to embrace these probiotics as a means of promoting holistic health.

Exploring the Release of Elastin Peptides Generated from Enzymatic Hydrolysis of Bovine Elastin via Peptide Mapping.

To enhance the understanding of enzymatic hydrolysis and to accelerate the discovery of key bioactive peptides within enzymatic products, this research focused on elastin as the substrate and investigated the variations in peptide profiles and the production of key bioactive peptides (those exceeding 5% of the total) and their impacts on the biological activity of the hydrolysates. Through the application of advanced analytical techniques, such as stop-flow two-dimensional liquid chromatography and ultra-high-performance liquid chromatography-tandem mass spectrometry, the research tracks the release and profiles of peptides within elastin hydrolysates (EHs). Despite uniform peptide compositions, significant disparities in peptide concentrations were detected across the hydrolysates, hinting at varying levels of bioactive efficacy. A comprehensive identification process pinpointed 403 peptides within the EHs, with 18 peptides surpassing 5% in theoretical maximum content, signaling their crucial role in the hydrolysate's bioactivity. Of particular interest, certain peptides containing sequences of alanine, valine, and glycine were released in higher quantities, suggesting Alcalase® 2.4L's preference for these residues. The analysis not only confirms the peptides' dose-responsive elastase inhibitory potential but also underscores the nuanced interplay between peptide content, biological function, and their collective synergy. The study sets the stage for future research aimed at refining enzymatic treatments to fully exploit the bioactive properties of elastin.

Cell-free biosynthesis combined with deep learning accelerates de novo-development of antimicrobial peptides

Bioactive peptides are key molecules in health and medicine. Deep learning holds a big promise for the discovery and design of bioactive peptides. Yet, suitable experimental approaches are required to validate candidates in high throughput and at low cost. Here, we established a cell-free protein synthesis (CFPS) pipeline for the rapid and inexpensive production of antimicrobial peptides (AMPs) directly from DNA templates. To validate our platform, we used deep learning to design thousands of AMPs de novo. Using computational methods, we prioritized 500 candidates that we produced and screened with our CFPS pipeline. We identified 30 functional AMPs, which we characterized further through molecular dynamics simulations, antimicrobial activity and toxicity. Notably, six de novo-AMPs feature broad-spectrum activity against multidrug-resistant pathogens and do not develop bacterial resistance. Our work demonstrates the potential of CFPS for high throughput and low-cost production and testing of bioactive peptides within less than 24 h.

Production of Bioactive Peptides from Hake By-Catches: Optimization and Scale-Up of Enzymatic Hydrolysis Process.

Fish by-catches, along with other fish side-streams, were previously used as raw material for the production of fishmeal and fish oil but appropriate handling allows their use in more valuable options. The aim of this research was to valorize undersized hake (Merluccius merluccius) as a model of using fish by-catch from the Bay of Biscay to produce protein hydrolysates with bioactivities. Six enzymes, with different proteolytic activities (endo- or exoproteases) and specificities, were tested to produce protein hydrolysates. Products obtained with an endoprotease of serine resulted in the most promising results in terms of protein extraction yield (68%), with an average molecular weight of 2.5 kDa, and bioactivity yield (antioxidant activity = 88.5 mg TE antioxidant capacity/g fish protein; antihypertensive activity = 47% inhibition at 1 mg/mL). Then, process conditions for the use of this enzyme to produce bioactive products were optimized using Box-Behnken design. The most favorable process conditions (time = 2 h, solids = 50% and enzyme/substrate = 2% with respect to protein) were scaled up (from 0.5 L to 150 L reactor) to confirm laboratory scale and model forecasts. The results obtained in the pilot-scale testing matched the outcomes predicted by the model, confirming the technical viability of the proposed process.

Recent Trends in the Application of Oilseed-Derived Protein Hydrolysates as Functional Foods.

Oilseed-derived proteins have emerged as an excellent alternative to animal sources for the production of bioactive peptides. The bioactivities exhibited by peptides derived from plant proteins encompass a wide range of health-promoting and disease-preventing effects. Peptides demonstrate potential capabilities in managing diseases associated with free radicals and regulating blood pressure. They can also exhibit properties that lower blood sugar levels and modify immune responses. In addition to their bioactivities, plant-derived bioactive peptides also possess various functional properties that contribute to their versatility. An illustration of this potential can be the ability of peptides to significantly improve food preservation and reduce lipid content. Consequently, plant-derived bioactive peptides hold great promise as ingredients to develop functional products. This comprehensive review aims to provide an overview of the research progress made in the elucidation of the biological activities and functional properties of oilseed-derived proteins. The ultimate objective is to enhance the understanding of plant-derived bioactive peptides and provide valuable insights for further research and use in the food and medicine industries.

Obtaining New Candidate Peptides for Biological Anticancer Drugs from Enzymatic Hydrolysis of Human and Bovine Hemoglobin.

Enzymatic hydrolysis of bovine and human hemoglobin generates a diversity of bioactive peptides, mainly recognized for their antimicrobial properties. However, antimicrobial peptides stand out for their ability to specifically target cancer cells while preserving rapidly proliferating healthy cells. This study focuses on the production of bioactive peptides from hemoglobin and evaluates their anticancer potential using two distinct approaches. The first approach is based on the use of a rapid screening method aimed at blocking host cell protein synthesis to evaluate candidate anticancer peptides, using Lepidium sativum seed germination as an indicator. The results show that: (1) The degree of hydrolysis (DH) significantly influences the production of bioactive peptides. DH levels of 3 to 10% produce a considerably stronger inhibition of radicle growth than DH 0 (the native form of hemoglobin), with an intensity three to four times greater. (2) Certain peptide fractions of bovine hemoglobin have a higher activity than those of human hemoglobin. (3) The structural characteristics of peptides (random coil or alpha helix) play a crucial role in the biological effects observed. (4) The α137-141 peptide, the target of the study, was the most active of the fractions obtained from bovine hemoglobin (IC50 = 29 ± 1 µg/mL) and human hemoglobin (IC50 = 48 ± 2 µg/mL), proving to be 10 to 15 times more potent than the other hemoglobin fractions, attributed to its strong antimicrobial potential. The second approach to assessing anticancer activity is based on the preliminary in vitro analysis of hydrolysates and their peptide fractions, with a focus on the eL42 protein. This protein is of major interest due to its overexpression in all cancer cells, making it an attractive potential target for the development of anticancer molecules. With this in mind, astudy was undertaken using a method for labeling formylase (formyl-methionyl-tRNA transformylase (FMTS)) with oxidized tRNA. This approach was chosen because of the similarities in the interaction between formylase and the eL42 protein with oxidized tRNA. The results obtained not only confirmed the previous conclusions but also reinforced the hypothesis that the inhibition of protein synthesis plays a key role in the anticancer mechanism of these peptides. Indeed, the data suggest that samples containing α137-141 peptide (NKT) and total hydrolysates may have modulatory effects on the interaction between FMTS and oxidized tRNA. This observation highlights the possibility that the latter could influence molecular binding mechanisms, potentially resulting in a competitive situation where the ability of substrate tRNA to bind efficiently to ribosomal protein is compromised in their presence. Ultimately, these results suggest the feasibility of obtaining candidate peptides for biological anticancer drugs from both human and bovine hemoglobin sources. These scientific advances show new hope in the fight against cancer, which affects a large number of people around the world.

Nutritional composition, health benefits and bio-active compounds of chickpea (Cicer arietinum L.).

Chickpea (Cicer arietinum L.), an annual plant of the family Fabaceae is mainly grown in semiarid and temperate regions. Among pulses, cultivated worldwide chickpeas are considered an inexpensive and rich source of protein. Chickpea is a good source of protein and carbohydrate, fiber, and important source of essential minerals and vitamins. The quality of protein is better among other pulses. Consumption of chickpeas is related to beneficial health outcomes. Dietary peptides from the protein of chickpeas gaining more attention. Peptides can be obtained through acid, alkali, and enzymatic hydrolysis. Among all these, enzymatic hydrolysis is considered safe. Various enzymes are used for the production of peptides, i.e., flavorzyme, chymotrypsin, pepsin, alcalase, papain, and trypsin either alone or in combinations. Chickpea hydrolysate and peptides have various bioactivity including angiotensin 1-converting enzyme inhibition, digestive diseases, hypocholesterolemic, CVD, antioxidant activity, type 2 diabetes, anti-inflammatory, antimicrobial, and anticarcinogenic activity. This review summarizes the nutritional composition and bioactivity of hydrolysate and peptides obtained from chickpea protein. The literature shows that chickpea peptides and hydrolysate have various functional activities. But due to the limited research and technology, the sequences of peptides are unknown, due to which it is difficult to conduct the mechanism studies that how these peptides interact. Therefore, emphasis must be given to the optimization of the production of chickpea bioactive peptides, in vivo studies of chickpea bioactivity, and conducting human study trials to check the bioactivity of these peptides and hydrolysate.

A Versatile Elastin-Like Carrier for Bioactive Antimicrobial Peptide Production and Delivery.

Elastin-like polypeptidesare biotechnological protein and peptide carriers that offer a vast scope of applicability. This work aims to build a model for the expression of antimicrobial peptides (AMPs) by genetically engineering the Human Elastin-like Polypeptideplatform developed in the lab. The well-characterized AMP indolicidin is selected as an example of an antimicrobial domain for the recombinant fusion at the C-terminus of the carrier. The fusion construct has been designed to allow the release of the antimicrobial domain. The expression product has been purified and its physicochemical and antimicrobial properties has been characterized. Taking advantage of the self-assembling and matrix-forming properties of the recombinant biopolymer, the materials that are obtained have been evaluated for antimicrobial activity toward bacterial-strain models. This approach represents a cost-effective strategy for the production of smart components and materials endowed with antimicrobial capacity triggered by external stimuli.

Peptidomics approaches to the discovery and ACE inhibitory effect of casein peptides derived from fermented bovine milk by kefir grains

IntroductionKefir grains with efficient proteolytic system is an excellent starter culture for the production of bioactive peptides and milk products. This study explores the casein peptides derived from fermented bovine milk by kefir grains using the peptidomics approaches. The angiotensin converting enzyme (ACE) inhibitory activity of these peptides were also investigated.MethodsAfter fermentation, peptidomics based on the LC-MS/MS was used to investigate the dynamic profile and the structure specificity of generated peptides. The ACE inhibitory activity of peptides was determined by measuring the amount of hippuric acid (HA) by a spectrophotometer at 228 nm.ResultsThe results indicated that the cell envelope proteinases (CEPs) were the PI-/PIII-type. A total of 122 peptides were identified. The β-casein was preferentially hydrolyzed by kefir grains, and the main hydrolysis regions were f57-93, f132-160 and f192-209. The αs1-, and κ-casein were also hydrolyzed by a weaker degree. In the process of fermentation, the accumulated peptides increased with the fermentation time. The fermentation products exhibited ACE inhibitory activity, and this bioactivity remained 63% after simulated gastrointestinal (GI) digestion in vitro. Additionally, 14 Pro-containing peptides with ACE inhibitory activity were also identified.ConclusionThese results provide new insights and evidence to investigate the bioactive milk peptides generated by kefir grains fermentation, as well as a reference for the development of functional foods.

Production of Coffee oil and Bioactive Peptides from Spent Coffee Grounds via Supercritical Carbon Dioxide Extraction and Enzymatic Hydrolysis

Production of Coffee oil and Bioactive Peptides from Spent Coffee Grounds via Supercritical Carbon Dioxide Extraction and Enzymatic Hydrolysis