Aplicaciones de las herramientas proteómicas en tecnología de la alimentación

Las técnicas proteómicas se usan cada vez más en la valoración de alimentos tanto procesados como en crudo, así como en el control, la optimización y el desarrollo de nuevos productos en tecnología y biotecnología de alimentos.Peer Reviewe

Lactobacillus sakei CRL1862 improves safety and protein hydrolysis in meat systems

Aims The capacity of Lactobacillus sakei CRL1862 to prevent the growth of pathogens and its ability to degrade sarcoplasmic and myofibrillar proteins in pork meat systems was evaluated. In addition, basic safety aspects of Lact. sakei CRL1862 such as production of biogenic amines and antibiotic susceptibility were addressed. Methods and Results The bacteriocin-producing Lact. sakei CRL1862 showed respectively bactericide and bacteriostatic effect against Listeria monocytogenes and Staphylococcus aureus in beaker sausage assay during 9 days of storage at 22°C. The hydrolytic effect of Lact. sakei CRL1862 on protein extracts was evaluated by SDS-PAGE and reverse phase HPLC. A more pronounced proteolysis was evidenced in inoculated sarcoplasmic proteins compared with myofibrillar extracts with the generation of predominantly hydrophilic peptides and increase of total free amino acids concentration. Lactobacillus sakei CRL1862 produced neither histamine nor tyrosine and exhibited no resistance to the antibiotics assayed. Conclusions Lactobacillus sakei CRL1862 effectively controlled the growth of L. monocytogenes and Staph. aureus; moreover, it was able to hydrolyse pork meat extracts generating peptides and amino acids, which may improve hygienic and sensorial attributes of fermented meat products. Significance and Impact of the Study The use of an integrated approach to evaluate the major traits of Lact. sakei CRL1862 showed it can be applied as an autochthonous functional starter in meat fermentation.Fil: Castellano, Patricia Haydee. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Aristoy, Maria Concepción. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; EspañaFil: Sentandreu, Miguel Angel. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; EspañaFil: Vignolo, Graciela Margarita. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Toldrá, Fidel. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; Españ

Dr. Ahmed Ouali, 1948–2020

International audienceAhmed Ouali was born on October 4, 1948 in Tigzirt, Tizi-Ouzou, Algeria. In 1952, he moved with his parents to Montluçon, France. In 1974, he was trained and graduated with a bachelor's degree in Biochemistry at the University of Lyon. He then, in 1976, earned a joint Ph.D. in Animal Science at the University of Blaise Pascal (Clermont-Ferrand) where he studied at the National Institute of Agricultural Research (INRA, Theix). The title of his doctorate thesis was “The role of muscle proteases on meat tenderization”. Subsequently, he was employed in a private laboratory for medical analysis from 1976 to 1978 and thereafter at the Meat Research Laboratory group at INRA, Theix as a permanent researcher. In 1990, he was appointed as a research director and led the “Biochemistry and Functions of Muscle Proteins” unit for 8 years. The Meat Research Station focused their research on many topics including colour and protein oxidation; enzymology and tenderness; and muscle protein functionalities. During his entire scientific career at INRA, but before his retirement on October 2013, Ahmed was living in Clermont-Ferrand, the city of the famous volcanic chain of the Puy-de-Dôme, with his wife Anne-Marie with whom he had two lovely children: Armelle (41 years) and Gäel (38 years). In 2019, they moved to their new house in Montpellier in the South of France

Generation of ACE inhibitory dipeptides by pork muscle dipeptidyl peptidases I and III

The proteolytic action of dipeptidyl peptidases I and III along the processing of dry-cured ham can generate dipeptides which, some of them, are able to considerably inhibit the activity of angiotensin-I converting enzyme. This could help to maintain an adequate systolic blood pressure even if dry-cured ham is present in the diet. This would be specially positive for hypertensive population from countries like Spain, having great tradition in consumption of dry-cured meat productsI3P contract from the European Social Fund and by a Marie Curie grant (MERG-CT-2004-510652) from European Commisio

Peptidomics on farm animal research

Although Peptidomics is a discipline complementary to proteomics, since nowadays both mainly rely on analytical strategies based on mass spectrometry, there are fundamental differences. In this chapter, we discuss these differences along with the application of these technologies for the study of the different stages of meat production, from storage to processing to unravel mechanisms that will allow reaching high quality and safer meat products. The use of Peptidomics and the related high throughput technologies, now relying on mass spectrometry but once also on N-terminal sequencing, are discussed. Clear examples are provided dealing with relevant studies on meat proteolysis and peptide generation occurring during ageing, as well as those produced during ripening of meat products by endogenous and microbial enzymes. Also the involvement of this phenomenon in the development of taste active compounds is addressed. Finally, the application of novel Omics technologies on bacterial identification in food for diagnosis and safety purposes is presented, putting emphasis on their potential advantages and future perspectives.Fil: Fadda, Silvina G.. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Tucumán. Centro de Referencia para Lactobacilos; ArgentinaFil: Sentandreu, Enrique. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; EspañaFil: Sentandreu Vicente, Miguel Angel. Consejo Superior de Investigaciones Científicas. Instituto de Agroquímica y Tecnología de Alimentos; Españ

Blocking agents for ELISA quantification of cathepsin L in bovine and porcine muscle crude extracts

Skimmed milk revealed to be the most efficient of the assayed blockers, as also observed by other authors. In the case of pork, this would be the only suitable blocking agent but avoiding the highest assayed muscle crude extract concentration. All the same attention must be paid, since some authors pointed out the possibility that skimmed milk may interfere the antigen-antibody reaction under certain conditions Vogt et al., 1987. For ELISA quantification of cathepsin L from bovine species, goat serum and BSA, together with skimmed milk, could be used as effective blockers always that a surcoating steps be included in addition to be present in the assay buffer.Marie Curie Individual Fellowship (Q1-CT-2002-51527

Peptidomics as a robust and reliable approach to discriminate between closely-related meat animal species

Resumen del póster presentado al Meeting on Farm Animal Proteomics celebrado en Milán (Italia) del 17 al 18 de noviembre de 2014.There is an increasing demand by consumers for clear, reliable and detailed information about the foods they consume. This is especially relevant in processed foods, where ingredients cannot be distinguished by simple visual inspection. In this context, legislation must protect consumers against misdescription and fraud, practices that can be done by food producers or traders with the objective to increase the economic gain. The scandal occurring recently in Europe about the presence of undeclared horse meat in beef products illustrates this situation and highlights the importance to dispose of robust and reliable methods capable to unambiguously identify those species that are susceptible to be employed in fraudulent practices or as proof to certify the authenticity of the higher quality meats (Sentandreu & Sentandreu 2014). In the case of meat products, there is a requirement to separately indicate and quantify the different meat species that are present in the food, what it is known as Quantitative Ingredient Declaration (QUID). In addition to this, other parts of carcass such as the liver, lung, heart or tongue, for example, cannot be considered as meat and need also to be separately indicated (Zukal & Kormendy 2007). Methods that have been traditionally employed in control laboratories to assess meat composition have mainly relied on immunoassays and DNA analysis. Even if they have remarkable advantages and performance, it is also true that they are not exempted from some important limitations, especially in the analysis of complex and/or highly processed foods. In the case of immunoassays, the lack of highly specific antibodies can promote the apparition of cross-reactions, something that becomes more probable when trying to differentiate between closely-related species such as the case of chicken (Gallus gallus) and turkey (Meleagris gallopavo) meat. In addition to this, food processing can greatly alter protein structure and consequently reduce its recognition by the antibody. Food processing can also negatively affect genetic analyses because DNA can undergo a remarkable degradation due to the use of aggressive conditions such as pH changes or thermal treatments, for example. This would increase the generation of shorter non-specific DNA fragments (Primrose et al. 2010). Current advances in mass spectrometry applied to the analysis of proteins and peptides constitute an interesting and promising alternative to the aforementioned methods for the unambiguous identification of the different types of meats that can be present in meat products. The objective of the present work was to develop a peptidomic approach capable to discriminate between chicken and turkey, two closely-related meat species, through the identification and characterization of peptide biomarkers specific of each one of these farm animals.Peer Reviewe

Proteomic pipeline for biomarker hunting of defective bovine meat assisted by liquid chromatography-mass spectrometry analysis and chemometrics

A wide variety of factors prior to slaughter may affect the stress status of beef cattle, giving rise to well-known ‘dark-cutting’ defective meats characterised by a high ultimate pH (pHu). To understand the underlying mechanisms of pHu fluctuations in beef cattle there was studied the proteome changes caused by pre-slaughter stress through a gel-free proteomic approach. Comparative peptidomic analysis was carried out on 12 loin samples at 24 h post-mortem from Longissimus thoracis et lumborum bovine muscle of crossbred animals, previously sorted into two different groups according to their pHu values: normal (pHu < 6.0) and high (pHu ≥ 6.0). Tryptic peptides from direct protein extracts were approached by combining untargeted (intact mass, MS1) and targeted (Selected Reaction Monitoring, SRM) quantitative LC-MS assays followed by chemometric analysis. Seventeen peptide biomarkers belonging to 10 different proteins appropriately discriminated sample groups assayed. Results may promote the use of this simple and effective methodology towards the creation of new insights in meat quality research.Department of Economic Development and Infrastructures of the Basque Government is acknowledged for the doctoral fellowship of C.F.G. Project RTA2014-00034-C04 (Spanish Instituto Nacional de Investigación y Tecnología Agraria, INIA), RTI2018-096162-R-C22 (Spanish Agencia Estatal de Investigación) and FEDER founds partially supported this research and to uphold contract of E. Sentandreu.Peer reviewe