Ficin: A protease extract with relevance in biotechnology and biocatalysis.

This study evaluated transepithelial transport mechanisms of Ile-Gln-Pro (IQP) and Val-Glu-Pro (VEP), two ACE-inhibitory peptides derived from Spirulina platensis, using human intestinal Caco-2 cell monolayers. IQP and VEP were absorbed intact through Caco-2 cell monolayers with Papp values of 7.48 ± 0.58 × 10-6 and 5.05 ± 0.74 × 10-6 cm/s, respectively. The transport of IQP and VEP were affected neither by Gly-Pro nor by wortmannin, indicating that they were not PepT1-mediated and did not involve endocytosis. However, transport of IQP and VEP were increased significantly by sodium deoxycholate, suggesting that the major transport mechanism was paracellular. In addition, the increased transport of VEP and IQP were followed with the addition of sodium azide, suggesting influence of energy to the process. The transport of VEP was also increased by verapamil, indicating an apical-to-basolateral flux mediated by P-gp.; Practical Application: Bioactive peptides derived from food proteins have been considered as potentially ideal products to reduce hypertension because of their safety and positive impacts on health. IQP and VEP are the 2 ACE inhibitory peptides derived from Spirulina platensis, a kind of edible cyanobacteria with rich nutrition and multiple physiological functions, and were demonstrated to inhibit ACE and lower blood pressure in spontaneously hypertensive rats. However, it is prerequisite that such bioactive peptides must be absorbed intact across the intestinal epithelium, so as to exert antihypertensive effects in vivo. This study evaluated transepithelial transport mechanisms of IQP and VEP. It contributes to the study of Spirulina in lowering blood pressure and supports the development of bioactive peptide products.; © 2018 Institute of Food Technologists®.

Transport of ACE Inhibitory Peptides Ile-Gln-Pro and Val-Glu-Pro Derived from Spirulina platensis Across Caco-2 Monolayers.

The currently available oral anti-Xa agents are claimed to produce their anticoagulant and antithrombotic effects solely by the inhibition of factor Xa. This study profiled various anti-Xa drugs in routinely used laboratory assays to demonstrate that their effects are not solely related to the anti-Xa activities. Apixaban, betrixaban, edoxaban, and rivaroxaban were obtained commercially. Native and citrated whole blood was used for the activated clotting time (ACT) and thromboelastography (TEG). Citrated plasma was used for monitoring the prothrombin time (PT), activated partial thromboplastin time (aPTT), Heptest, and prothrombinase-induced clotting time (PiCT) tests. An amidolytic method was used for the determination of anti-Xa effects. Thrombin-induced fibrinokinetics was monitored optically. Thrombin generation studies were carried out using the calibrated automated thrombogram. All of the anti-Xa agents produced concentration- and assay-dependent effects. In the ACT at 2.5 μg/mL and TEG at 1.0 μg/mL, edoxaban exhibited the strongest anticoagulation effect. In the PiCT, PT, and aPTT assay at 1 μg/mL, edoxaban showed stronger effects than other agents. The half maximal inhibitory concentration of these agents for the inhibition of factor Xa ranged from 340 to >1000 ng/mL. In the thrombin generation inhibition assay, apixaban showed the strongest activity. In the fibrinokinetics, different anti-Xa agents produced varying degrees of inhibition. These results demonstrate that the measured anti-Xa activity alone does not fully reflect the overall biologic spectrum of these agents.

https://journals.sagepub.com/doi/pdf/10.1177/1076029619847524

Factor Xa Inhibitory Profile of Apixaban, Betrixaban, Edoxaban, and Rivaroxaban Does Not Fully Reflect Their Biologic Spectrum.

The aim of this work is to isolate and study the performance of an alkaline protease from fruit by-products of Bromelia antiachanta Bertol. for its potential application as an additive of laundry detergent formulations. Its behavior is compared with Pura Felt 4000 (Genencor Argentina). Antiacanthain, a cysteine protease from fruit by-products of B. antiachanta, showed a remarkable stability (50 ± 0.82% to 177 ± 0.82% residual activity) in buffer with non-ionic and cationic surfactants (0.1, 0.4 and 1.0%), metal chelation (1, 5 and 10 mM EDTA), antiredepositant (100 mM Na2CO3) and bleaching (5% H2O2) agents. In commercial detergents, antiacanthain retained 100 ± 1.25% to 120 ± 0.82% of its maximum activity at 60 °C and removed stains from cloths in a short time period. Antiacanthain fulfilled all the requirements as detergent additive: high activity and stability in a broad temperature range (25–60 °C) and alkaline pH (5–9), and very good compatibility with commercial detergents.

Valorization of fruit by-products of Bromelia antiacantha Bertol.: Protease obtaining and its potential as additive for laundry detergents

The Functional Food Center (FFC) has previously defined functional food as foods which have scientifically-proven benefits toward improving general health and ameliorating the effects of chronic diseases. Given this useful working definition of functional food, it’s now imperative to delineate and establish a field of functional food science. Currently, there are no published articles on what functional food science is. Defining functional food science is necessary so that a legitimate field can be established and recognized by the government as well as a global network of researchers and scientists. In this way, appropriate funding and advancements can be made, and functional foods can make their way towards significantly improving the lives of people in the United States and also around the world, as is the goal. As a result of this need, functional food science will be defined as the process of preparing functional foods as well as a field intersecting the realms of health and medical treatment. In this sense, functional food science not only deals with preparing functional foods but also with the efficacy of their health benefits. More specifically, as sciences are often defined by their processes, functional food science can be defined as the comprehensive, aggregate, interdisciplinary, and collaborative processes of: establishing a target and finding a bioactive compound that enables that effect, finding the correlated measurable biomarker, running testing to find proper dosage and effectiveness, performing clinical trials to ensure efficacy, creating the functional food with an informative label, releasing to market and running epidemiological studies to verify it. Although this understanding and definition is nascent, this can be a starting ground for the FDA and other governmental bodies, as well as the scientific and functional food community, to develop a robust, extrapolatable, and useful outline of functional food science. With an established field of functional food science, functional foods can be researched, produced, and made official and trustworthy, so that there can be large positive impacts on public health, both in disease prevention and in maintenance of good health.

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FFC’s Advancement of the Establishment of Functional Food Science

Bioactive peptides are protein-specific portions from 2 up to some dozens of amino acid residues with desirable biological activities, based on their composition and sequence. Databases report about 3000 peptide sequences with potential activities (antioxidant, antihypertensive, antithrombotic, antiadipogenic, antimicrobial, antiinflammatory, and antitumoral). It is assumed that peptides have high specificity in target tissues; no or little toxicity, do not accumulation in the organism, and easily degrade in the environment. However, their usefulness as nutraceuticals is limited by the low bioavailability due to the gastrointestinal digestion, poor transportability through the membranes, and rapid plasma clearance. This chapter reviews the different technologies for the production of bioactive peptides. Although the size of the molecule will determine the technology most suitable for its production, this chapter emphasizes the chemical and enzymatic synthesis. Finally, some strategies to enhance absorption, effectiveness, and bioavailability, and to study molecular mechanisms of activity and safety of these functional ingredients are discussed.

Bioactive peptides as functional food ingredients

Proteolytic enzymes (proteases) comprise a group of hydrolases (EC 3.4, NC-IUBMB) which share the common feature of acting on peptide bonds. Proteases are among the best-studied enzymes in terms of structure–function relationship (Krowarsch et al. 2005). Proteases, by catalyzing the cleavage of other proteins and even themselves, have an enormous physiological significance, their coding genes representing as much as 2% of the total human genome (Schilling and Overall 2008).

Peptide Synthesis Using Proteases as Catalyst

In this study we report the enzymatic synthesis of N-α-[Carbobenzyloxy]-Tyr-Gln-Gln (Z-YQQ), a new anticoagulant tripeptide. It was obtained using phytoproteases from the stems and petioles of Asclepias curassavica L. as catalyst in an aqueous-organic biphasic system formed by 50% (v/v) ethyl acetate and 0.1 M Tris-HCl buffer pH 8. The resulting peptide was compared with the analogous peptide Tyr-Gln-Gln (YQQ) produced by solid-phase chemical synthesis. The in vitro anticoagulant activity of the aforementioned peptides was determined using Wiener Lab Test (Wiener, Argentina). The toxicological activity of the peptides was also determined. The enzymatically synthesized Z-YQQ peptide acted on the extrinsic pathway of the coagulation cascade, delaying the conversion time of prothrombin to thrombin and fibrinogen to fibrin by 136 and 50%, respectively, with respect to the controls. The chemically synthesized YQQ peptide acted specifically on the intrinsic pathway of the coagulation cascade, affecting factors VIII, IX, XI, and XII from such cascade, and increasing the coagulation time by 105% with respect to the control. The results suggest that two new anticoagulant peptides (Z-YQQ and YQQ) can be useful for safe pharmaceutical applications. Nevertheless, some aspects related to peptide production should be optimized. © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 2018 © 2018 American Institute of Chemical Engineers Biotechnol. Prog., 34:1093-1101, 2018.

Anabella Lucía Origone

Papers

Bioactive peptides as functional food ingredients

Peptide Synthesis Using Proteases as Catalyst

Enzymatic and chemical synthesis of new anticoagulant peptides.