Peptídeo do hidrolisado protéico de RuBisCO e sua aplicação na preservação de carne bovina

The objective of this work was to purify the small (705 Da) and hydrophilic antimicrobial Arg-Asp-Arg-Phe-Leu peptide from RuBisCO protein hydrolysate and to evaluate its effect on the microbiological and oxidative stability of beef mince during refrigeration. RuBisCO was obtained from alfafa green juice. The peptide extract was fractionated using RP-HPLC, and the active fractions were analyzed by liquid chromatography, electrospray ionization, and tandem mass spectrometry (LC-ESI-MS). Beef mince was analyzed in the following treatments: negative control, meat with two different BHT concentrations of 0.1 and 0.5% (w/w), and meat with two different Arg-Asp-Arg-Phe-Leu peptide concentrations of 0.1 and 0.5% (w/w). Lipid oxidation using the thio-barbituric acid-reactive substance (TBARS) values were significantly affected by the storage period and the concentration of bioactive peptide. Arg-Asp-Arg-Phe-Leu, a small antibacterial peptide from RuBisCO, can be isolated and purified by HPLC from alfafa green juice with retention time between 10 and 50 min, which corresponds to antimicrobial peptides. RuBisCO peptide Arg-Asp-Arg-Phe-Leu 0.5% increases oxidative stability of beef mince during refrigeration. RuBisCO peptide Arg-Asp-Arg-Phe-Leu inhibit microbial growth under refrigeration for 11 days.O objetivo deste trabalho foi purificar peptídeo antimicrobiano pequeno (705 Da) e hidrofílico Arg-Asp-Arg-Phe-Leu a partir do hidrolisado proteico RuBisCO e avaliar seu efeito na estabilidade microbiológica e oxidativa de carne bovina moída durante refrigeração. RuBisCO foi obtida a partir do suco verde de alfafa. O extrato peptídico foi fracionado utilizando RP-HPLC, e as frações ativas foram analisadas por cromatografia líquida, ionização por eletrospray e espectrometria de massa em tandem (LC-ESI-MS). A carne bovina moída foi analisada nos seguintes tratamentos: controle negativo, carne com duas concentrações diferentes de BHT de 0,1 e 0,5% (p/p) e duas concentrações de peptideo Arg-Asp-Arg-Phe-Leu de 0,1 e 0,5% (p/p). Os valores de oxidação lipídica utilizando a substância reativa ao ácido tiobarbitúrico (TBARS) foram significativamente afetados pelo período de armazenamento e pela concentração do peptídeo bioativo. Arg-Asp-Arg-Phe-Leu, um pequeno peptídeo de RuBisCO, pode ser isolado e purificado por HPLC de suco verde de alfafa com tempos de retenção de 10 e 50 min, o que corresponde ao peptídeo antimicrobiano. O peptídeo Arg-Asp-Arg-Phe-Leu 0,5% aumenta a estabilidade de oxidação da carne moída durante a refrigeração. O peptídeo Arg-Asp-Arg-Phe-Leu de RuBisCO inibe o crescimento microbiano sob refrigeração durante 11 dias

Peptide from RuBisCO protein hydrolysate and its application in beef meat preservation

Abstract The objective of this work was to purify the small (705 Da) and hydrophilic antimicrobial Arg-Asp-Arg-Phe-Leu peptide from RuBisCO protein hydrolysate and to evaluate its effect on the microbiological and oxidative stability of beef mince during refrigeration. RuBisCO was obtained from alfafa green juice. The peptide extract was fractionated using RP-HPLC, and the active fractions were analyzed by liquid chromatography, electrospray ionization, and tandem mass spectrometry (LC-ESI-MS). Beef mince was analyzed in the following treatments: negative control, meat with two different BHT concentrations of 0.1 and 0.5% (w/w), and meat with two different Arg-Asp-Arg-Phe-Leu peptide concentrations of 0.1 and 0.5% (w/w). Lipid oxidation using the thio-barbituric acid-reactive substance (TBARS) values were significantly affected by the storage period and the concentration of bioactive peptide. Arg-Asp-Arg-Phe-Leu, a small antibacterial peptide from RuBisCO, can be isolated and purified by HPLC from alfafa green juice with retention time between 10 and 50 min, which corresponds to antimicrobial peptides. RuBisCO peptide Arg-Asp-Arg-Phe-Leu 0.5% increases oxidative stability of beef mince during refrigeration. RuBisCO peptide Arg-Asp-Arg-Phe-Leu inhibit microbial growth under refrigeration for 11 days

Alterations in juvenile diploid and triploid African catfish skin gelatin yield and amino acid composition: effects of chlorpyrifos and butachlor exposures

Skin is a major by-product of the fisheries and aquaculture industries and is a valuable source of gelatin. This study examined the effect of triploidization on gelatin yield and proximate composition of the skin of African catfish (Clarias gariepinus). We further investigated the effects of two commonly used pesticides , chlorpyrifos (CPF) and butachlor (BUC), on the skin gelatin yield and amino acid composition in juvenile full-sibling diploid and triploid African catfish. In two separate experiments, diploid and triploid C. gariepinus were exposed for 21 days to graded CPF [mean measured: 10, 16, or 31 mg/L] or BUC concentrations [Mean measured: 22, 44, or 60 mg/L]. No differences in skin gelatin yield, amino acid or proximate compositions were observed between diploid and triploid control groups. None of the pesticide treatments affected the measured parameters in diploid fish. In triploids, however, gelatin yield was affected by CPF treatments while amino acid composition remained unchanged. Butachlor treatments did not alter any of the measured variables in triploid fish. To our knowledge, this study is the first to investigate changes in the skin gelatin yield and amino acid composition in any animal as a response to polyploidization and/or contaminant exposure

Implementation of the One Health approach to fight arbovirus infections in the Mediterranean and Black Sea Region: Assessing integrated surveillance in Serbia, Tunisia and Georgia

Background In the Mediterranean and Black Sea Region, arbovirus infections are emerging infectious diseases. Their surveillance can benefit from one health inter-sectoral collaboration; however, no standardized methodology exists to study One Health surveillance. Methods We designed a situation analysis study to document how integration of laboratory/clinical human, animal and entomological surveillance of arboviruses was being implemented in the Region. We applied a framework designed to assess three levels of integration: policy/institutional, data collection/data analysis and dissemination. We tested the use of Business Process Modelling Notation (BPMN) to graphically present evidence of inter-sectoral integration. Results Serbia, Tunisia and Georgia participated in the study. West Nile Virus surveillance was analysed in Serbia and Tunisia, Crimea-Congo Haemorrhagic Fever surveillance in Georgia. Our framework enabled a standardized analysis of One Health surveillance integration, and BPMN was easily understandable and conducive to detailed discussions among different actors/institutions. In all countries, we observed integration across sectors and levels except in data collection and data analysis. Data collection was interoperable only in Georgia without integrated analysis. In all countries, surveillance was mainly oriented towards outbreak response, triggered by an index human case. Discussion The three surveillance systems we observed prove that integrated surveillance can be operationalized with a diverse spectrum of options. However, in all countries, the integrated use of data for early warning and inter-sectoral priority setting is pioneeristic. We also noted that early warning before human case occurrence is recurrently not operationally prioritized

Development of Bioprocesses for Potential Functional Ingredients from Marine products

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Angiotensin-I-converting enzyme inhibitory and antioxidant activities of protein hydrolysate from muscle of barbel (barbus callensis)

The present study investigated angiotensin-I-converting enzyme (ACE) inhibitory and antioxidant activities of barbel muscle protein hydrolysate prepared with Alcalase. The barbel muscle protein hydrolysate displayed a high ACE inhibitory activity (C I 50 = 0.92 mg/mL). The antioxidant activities of protein hydrolysate at different concentrations were evaluated using various in vitro antioxidant assays, including 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical method and reducing power assay. The barbel muscle protein hydrolysate exhibited an important radical scavenging effect and reducing power. These results obtained by in vitro systems obviously established the antioxidant potency of barbel hydrolysate to donate electron or hydrogen atom to reduce the free radical. Furthermore, these bioactive substances can be exploited into functional foods or used as source of nutraceuticals. © 2013 Assaad Sila et al.Peer Reviewe

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Identification of ACE I-Inhibitory Peptides Released by the Hydrolysis of Tub Gurnard (<i>Chelidonichthys lucerna</i>) Skin Proteins and the Impact of Their In Silico Gastrointestinal Digestion

Tub gurnard is a highly abundant fishery species caught as a discard in the Mediterranean Sea. This work proposes its valorisation through the release of potential antihypertensive peptides and glycosaminoglycans (GAGs) through the controlled hydrolysis of tub gurnard skin proteins. Four proteases (Esperase, Alcalase, Trypsin and Pronase E) were used to obtain potent angiotensin converting enzyme I (ACE)-inhibitory hydrolysates. Peptides and GAGs were separated and evaluated for their antihypertensive potential by fluorometry. The peptide-rich fractions derived from the Esperase and Alcalase hydrolysates showed very low IC50 values (47 and 68 μg/mL, respectively). Only the GAGs from the Trypsin and Esperase hydrolysates were relevant ACE inhibitors (63 and 52% at 1 mg/mL, respectively). The peptide composition of the most potent ACE-inhibitory fractions derived from the Esperase and Alcalase hydrolysates (IC50 values of 33 and 29 μg/mL, respectively) was analysed by RP-LC-ESI-MS/MS. The analysis suggests that the ACE-inhibitory activity is related to the peptide hydrophobicity, as well as to the presence of specific residues at any of the last four C-terminal positions. The in silico gastrointestinal digestion of these fractions yielded small peptides with antihypertensive potential

Physicochemical and biological properties of eel by-products protein hydrolysates: Potential application to meat product preservation

European eel by-products have been hydrolysed in order to produce novel bioactive peptides with antibacterial and antioxidant activity. Protein hydrolysates from Anguilla by-products obtained by treatment with Savinase® (PHAB-s) have high protein content than that produced with Protamex® (PHAB-p). Furthemore, both protein hydrolysates had a high percentage of essential amino acids. In addition, results indicated that PHAB-s and PHAB-p exhibited different degrees of antioxidant activities, evaluated by several antioxidant assays. Thus, PHAB-s displayed high antioxidant activity, in terms of DPPH-radical scavenging effect (IC50 = 5.2 mg/ml), β-carotene bleaching inhibitory effect (CI50 = 4.4 mg/ml) and metal chelating activity. At 10 mg/ml, PHAB-p showed the highest reducing power. Antibacterial activities of PHAB-s and PHAB-p were also evaluated. PHAB-s showed the most important inhibitory effects against seven bacterial species. The application of PHAB-s and PHAB-p on minced meat as preservative reduced the lipid oxidation and inhibited the microbial growth during chilled storage for 11 days.This work was funded by the Ministry of Higher Education and Scientific Research, Tunisia and the Spanish Ministry of Economy, Industry and Competitiveness under project AGL2014-52825-R.Peer reviewe