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There is currently no appropriate cell model suitable for evaluating the insulinotropic effects of DPP-4 inhibitory peptides (DPP-4IPs) mediated by active glucagon-like peptide-1 7-36 (active GLP-1). The study aims to evaluate the transepithelial transport of IPYWTY on its in situ insulinotropic effects by using a 2D and dual-layered coculture cell model that consists of Caco-2 and NCI-H716 cells on the apical (AP) side and β-TC-6 cells on the basolateral (BL) side. During transportation, IPYWTY was absorbed in its intact form through PepT1 and paracellular transport. Meanwhile, it was degraded to several peptide fragments, including PYWTY, YWTY, WTY, and IPY, which decreased its in situ DPP-4 inhibitory activity. IPYWTY does not directly stimulate insulin release in β-TC-6 cells, while it increased the active GLP-1 level from 76.57 ± 15.16 to 95.63 ± 1.99 pM (1.25 times) in NCI-H716 cells. Interestingly, IPYWTY indirectly increased insulin levels from 426.91 ± 6.07 to 573.94 ± 2.97 μIU/mL (1.34 times) in the 2D and dual-layered coculture cell model for its dual function of stimulating active GLP-1 secretion and DPP-4 inhibition. These results suggested that the 2D and dual-layered coculture cell model is an alternative strategy for effectively evaluating the insulinotropic effects of DPP-4IPs mediated by active GLP-1.

Anchovy (Engraulis japonicus) cooking wastewater (ACWW) is a by-product resulted from the production of boiled-dried anchovies in the seafood processing industry. In this study, the protein hydrolysate of ACWW (ACWWPH) was found to have antimicrobial activity after enzymatic hydrolysis with Protamex. For the targeted screening of antibacterial peptides, liposomes constructed from Staphylococcus aureus membrane lipids were used in an equilibrium dialysis system. The hydrolysate was further purified by liposome equilibrium dialysis combined with high performance liquid chromatography. The purified antimicrobial peptide (ACWWP1) was determined to be GLSRLFTALK, with a molecular weight of 1104.6622Da. The peptide exhibited no haemolytic activity up to a concentration of 512μg/ml. It displayed a dose-dependent bactericidal effect in reconstituted milk. The change in cell surface hydrophobicity and membrane-permeable action of the purified ACWWP1 may have contributed to the antibacterial effect. This study suggests that liposome equilibrium dialysis can be used for the targeted screening of antimicrobial peptides., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Dipeptidyl Peptidase IV (DPP-IV) inhibitory peptides are attracting increasing attention, owing to their potential role in glycemic regulation by preventing the inactivation of incretins. However, few reviews have summarized the current understanding of DPP-IV inhibitory peptides and their knowledge gaps. This paper reviews the production, identification and structure-activity relationships (SAR) of DPP-IV inhibitory peptides. Importantly, their bioavailability and hypoglycemic effects are critically discussed. Unlike the traditional method to identifying peptides after separation step by step, the bioinformatics approach identifies peptides via virtual screening that is more convenient and efficient. In addition, the bioinformatics approach was also used to investigate the SAR of peptides. Peptides with proline (Pro) or alanine (Ala) residue at the second position of N-terminal are exhibit strong DPP-IV inhibitory activity. Besides, the bioavailability of DPP-IV inhibitory peptides is related to their gastrointestinal stability and cellular permeability, and in vivo studies showed that the glucose homeostasis has been improved by these peptides. Especially, the intestinal transport of DPP-IV inhibitory peptides and cell biological assays used to evaluate their potential role in glycemic regulation are innovatively summarized. For further successful development of DPP-IV inhibitory peptides in glycemic regulation, future study should elucidate their SAR and in vivo hypoglycemic effects. [ABSTRACT FROM AUTHOR]

Dipeptidyl Peptidase 4 inhibitory peptides (DPP-4IPs) could exhibit their hypoglycemic effects by preventing Glucagon-like peptide 1 (GLP-1) degradation. However, identifying DPP-4IPs by traditional approach is laborious. Therefore, this study aims to rapidly identify DPP-4IPs by an in silico method. After in silico digestion, 509 peptide fragments were obtained from pea proteins. Subsequently, two novel DPP-4IPs SPGDVF and EPF with the in vitro half-maximal inhibitory concentrations (IC 50) values of 277.61 and 406.47 μM were obtained by virtual screening and molecular docking. Interestingly, their in situ DPP-4 IC 50 values in Caco-2 cells were increased to 918.82 and 1868.27 μM, respectively. Lineweaver−Burk double-reciprocal plots revealed that SPGDVF and EPF were competitive and mixed-type DPP-4IPs, respectively. Significantly, molecular docking suggested that SPGDVF could bond with DPP-4 active pockets. While EPF only bound outside of these active pockets, which may be the reason that EPF was weaker than SPGDVF in DPP-4 inhibition. Overall, this work provides a convenient strategy for identifying DPP-4IPs from food proteins. [Display omitted] • A total of 509 peptide fragments were obtained from pea proteins by in silico digestion. • Two novel DPP-4 inhibitory peptides SPGDVF and EPF were virtual screened. • SPGDVF that bound to the active pockets exhibit stronger DPP-4 inhibitory activity than EPF that bound outside the active pockets. • SPGDVF and EPF exhibited the competitive and mixed-type inhibition mode to DPP-4. [ABSTRACT FROM AUTHOR]

In order to recover and utilize peanut protein resource, peanut meal was fermented usingBacillus subtilisto prepare antioxidant peptides in this study. The antioxidant activities of the peanut peptides were evaluated via sixin vitrotests, namely, 1,1-diphenyl-2-picrylhydrazyl radical scavenging activity, 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) scavenging activity, inhibition of linoleic acid autoxidation, reducing power, and metal chelating capacity. The peanut peptides showed 63.28% of scavenging effect on 1,1-diphenyl-2-picrylhydrazyl radical, 70.67% of scavenging effect on 2,2′-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) and 39.32% of superoxide anion radical scavenging activity at 1.0 mg/ml, and 62.69% of inhibition on linoleic acid autooxidation at 0.8 mg/ml. Besides, the peanut peptides exhibited 32.8% of reducing power and 60.44% of Fe2+-chelating capacity at 2 mg/ml. Then the peanut peptides were desalted with macroporous resin; the fraction eluted with 75% ethanol showed the highest scavenging activity on 1,1-diphenyl-2-picrylhydrazyl radical. This fraction was subject to further purification using gel filtration chromatography and reverse phase-high performance liquid chromatography. At last, a peptide was gained, and it was identified to be Tyr-Pro with matrix-assisted laser desorption/ionization time-of-flight/time-of-flight mass spectrometry. The results indicated that it is feasible to prepare natural antioxidants from peanut meal usingB. subtilisfermentation. [ABSTRACT FROM AUTHOR]

To study the influence of degree of hydrolysis (DH) on antioxidant properties of peanut peptides, the peanut meal was fermented by Bacillus subtilis. The fermentation time was 48, 72, and 96 h, respectively, to prepare peanut peptides at different degree of hydrolysis. The peanut peptides (11.18, 16.20, and 21.41% of DH, respectively) were extracted from fermentation liquid. The antioxidant properties of these peanut peptides were evaluated based on DPPH radical (1,1-Diphenyl-2-picrylhydrazyl radical) scavenging activity, superoxide anion-scavenging activity, reducing power, metal-chelating activity, and inhibition of linoleic acid autooxidation. Peanut peptides (21.41% of DH) at 1 mg/mL exhibited 80.86 and 29.35% of scavenging concentration percent on DPPH and superoxide anion radical, respectively. In addition, the reducing power of peanut peptides was 0.368 at 2 mg/mL, and they possessed 76.32% of Fe-chelation ability at 2 mg/mL and 63.75% of inhibition of linoleic acid autooxidation at 0.8 mg/mL. The antioxidant activities of the peanut peptides (21.41% of DH) were stronger compared with others (11.18 and 16.20% of DH), and this indicated the antioxidant activities of peanut peptides increased with increasing DH ( p < 0.05). To know much about the peanut peptides, they were subjected to amino acid analysis and determination of molecular weight distribution. Some acidic amino acids and essential amino acids were found, and the average molecular weight distributions were concentrated in <1,400 Da (86.96%). Combined with the results of the amino acid profiles, the peanut peptides were believed to have high nutritive value besides antioxidant activities. [ABSTRACT FROM AUTHOR]