Antimicrobial activity of an aspartic protease from Salpichroa origanifolia fruits

Abstract: Plant proteases play a fundamental role in several processes like growth, development and in response to biotic and abiotic stress. In particular, aspartic proteases (AP) are expressed in different plant organs and have antimicrobial activity. Previously, we purified an AP from Salpichroa origanifolia fruits called salpichroin. The aim of this work was to determine the cytotoxic activity of this enzyme on selected plant and human pathogens. For this purpose, the growth of the selected pathogens was analysed after exposure to different concentrations of salpichroin. The results showed that the enzyme was capable of inhibiting Fusarium solani and Staphylococcus aureus in a dose-dependent manner. It was determined that 1·2 μmol l−1 of salpichroin was necessary to inhibit 50% of conidial germination, and the minimal bactericidal concentration was between 1·9 and 2·5 μmol l−1. Using SYTOX Green dye we were able to demonstrate that salpichroin cause membrane permeabilization. Moreover, the enzyme treated with its specific inhibitor pepstatin A did not lose its antibacterial activity. This finding demonstrates that the cytotoxic activity of salpichroin is due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of the AP could represent a potential alternative for the control of pathogens that affect humans or crops of economic interest. Significance and Impact of the Study: This study provides insights into the antimicrobial activity of an aspartic protease isolated from Salpichroa origanifolia fruits on plant and human pathogens. The proteinase inhibited Fusarium solani and Staphylococcus aureus in a dose-dependent manner due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of salpichroin suggests its potential applications as an important tool for the control of pathogenic micro-organisms affecting humans and crops of economic interest. Therefore, it would represent a new alternative to avoid the problems of environmental pollution and antimicrobial resistance.Fil: Díaz, María Eugenia. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidad Nacional de Luján; ArgentinaFil: Rocha, Gabriela Fernanda. Universidad Nacional de Luján; ArgentinaFil: Kise, Francisco. Universidad Nacional de Luján; ArgentinaFil: Rosso, A. M.. Universidad Nacional de Luján; ArgentinaFil: Guevara, Maria Gabriela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Mar del Plata. Instituto de Investigaciones Biológicas. Universidad Nacional de Mar del Plata. Facultad de Ciencias Exactas y Naturales. Instituto de Investigaciones Biológicas; ArgentinaFil: Parisi, M.G.. Universidad Nacional de Luján; Argentin

Aprovechamiento del expeller de chía (Salvia hispanica L.) como potencial fuente de péptidos bioactivos generados mediante hidrólisis enzimática con papaína comercial

En las últimas décadas los procesos industriales para la obtención de aceites a base de semillas han cobrado gran importancia en el mercado, generando como producto secundario del proceso de extrusado-prensado en frío una masa compacta denominada expeller. En el caso de la chía (Salvia hispánica L.) el expeller obtenido mediante el procesamiento de semillas es empleado como alimento balanceado para el ganado. Si bien el estudio de las ventajas nutricionales que otorga la incorporación de semillas de chía en la dieta humana se encuentra en auge, no se presentan hasta la fecha investigaciones para la reutilización del expeller producto del procesamiento industrial. El objetivo de nuestro trabajo es el estudio de los biopéptidos generados mediante hidrólisis enzimática a partir de expeller de chía, con el fin de generar un valor agregado a dicho material y brindar en un futuro la posibilidad de reutilizarlo e insertarlo en el mercado como suplemento alimenticio. Siguiendo este objetivo se realizó la hidrólisis enzimática del expeller de chía durante veinte horas con papaína comercial (10 UE/mg), alcanzando un grado de hidrólisis de 15,2% estimado por el método de pH-STAT y determinación de péptidos solubles. Se observó que la hidrólisis se producía en las primeras dos horas del proceso, resultado que además se confirmó por SDS-PAGE. Este hidrolizado fue fraccionado por ultrafiltración obteniendo tres fracciones peptídicas de tamaños: mayor a 10 kDa, entre 3 y 10 kDa y menor a 3 kDa, en las cuáles se estimó la actividad antioxidante in vitro por el método de DPPH encontrando que tanto la muestra de partida como las distintas fracciones presentaban elevada actividad antirradicalaria. Se encontró que la concentración de expeller de chía sin tratamiento que producía un 50% de actividad antirradicalaria (IC50) era de 641.61 μg/ml, mientras que para los hidrolizados de peso molecular entre 3 y 10 kDa el IC50 era de 312.22 μg/ml, observándose que esta fracción de hidrolizado de expeller de chía presentaba péptidos con mayor actividad que el expeller de partida. No se observó incremento significativo en la actividad antioxidante en la fracción de peso molecular mayor a 10 kDa ni en la inferior a 3 kDa, probablemente debido a la baja concentración obtenida en el tiempo de hidrólisis en este último caso. Finalmente se realizó un estudio proteómico mediante espectrometría de masas MALDI-TOF de los hidrolizados a distintos tiempos y se evaluó en distintos rangos de pesos moleculares la aparición y desaparición de señales correspondientes a los péptidos generados por la hidrólisis enzimática. Finalmente, en este trabajo comprobamos la generación de un elevado contenido proteico del hidrolizado de chía generado con papaína comercial y confirmamos la presencia de péptidos bioactivos con actividad antioxidante en las fracciones generadas, presentando un marco inicial para continuar el estudio de dichos péptidos y evaluar su posible incorporación como suplementos dietarios en matrices alimentarias.Centro de Investigación de Proteínas Vegetale

Searches for transverse momentum dependent flow vector fluctuations in Pb-Pb and p-Pb collisions at the LHC

CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFINANCIADORA DE ESTUDOS E PROJETOS - FINEPFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPThe measurement of azimuthal correlations of charged particles is presented for Pb-Pb collisions at root S-NN 2.76 TeV and p-Pb collisions at root S-NN 5.02 TeV with the ALICE detector at the CERN Large Hadron Collider. These correlations are measured for the second, third and fourth order flow vector in the pseudorapidity region vertical bar eta vertical bar 0.8 as a function of centrality and transverse momentum pT using two observables, to search for evidence of PT-dependent flow vector fluctuations. For Ph-Ph collisions at 2.76 TeV, the measurements indicate that PT-dependent fluctuations are only present for the second order flow vector. Similar results have been found for p-Pb collisions at 5.02 TeV. These measurements are compared to hydrodynamic model calculations with event-by-event geometry fluctuations in the initial state to constrain the initial conditions and transport properties of the matter created in Ph-Ph and p-Pb collisions.9133CONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFINANCIADORA DE ESTUDOS E PROJETOS - FINEPFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPCONSELHO NACIONAL DE DESENVOLVIMENTO CIENTÍFICO E TECNOLÓGICO - CNPQFINANCIADORA DE ESTUDOS E PROJETOS - FINEPFUNDAÇÃO DE AMPARO À PESQUISA DO ESTADO DE SÃO PAULO - FAPESPSem informaçãoSem informaçãoSem informaçãoThe ALICE collaboration would like to thank all its engineers and technicians for their invaluable contributions to the construction of the experiment and the CERN accelerator teams for the outstanding performance of the LHC complex. The ALICE collaboration gratefully acknowledges the resources and support provided by all Grid centres and the Worldwide LHC Computing Grid (WLCG) collaboration. The ALICE collaboration acknowledges the following funding agencies for their support in building and running the ALICE detector: A.I. Alikhanyan National Science Laboratory (Yerevan Physics Institute) Foundation (ANSL), State Committee of Science and World Federation of Scientists (WFS), Armenia; Austrian Academy of Sciences and Nationalstiftung fur Forschung, Technologie und Entwicklung, Austria; Ministry of Communications and High Technologies, National Nuclear Research Center, Azerbaijan; Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq), Universidade Federal do Rio Grande do Sul (UFRGS), Financiadora de Estudos e Projetos (Finep) and Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP), Brazil; Ministry of Science & Technology of China (MSTC), National Natural Science Foundation of China (NSFC) and Ministry of Education of China (MOEC), China; Ministry of Science, Education and Sport and Croatian Science Foundation, Croatia; Ministry of Education, Youth and Sports of the Czech Republic, Czech Republic; The Danish Council for Independent Research Natural Sciences, the Carlsberg Foundation and Danish National Research Foundation (DNRF), Denmark; Helsinki Institute of Physics (HIP), Finland; Commissariat a l'Energie Atomique (CEA) and Institut National de Physique Nucleaire et de Physique des Particules (IN2P3) and Centre National de la Recherche Scientifique (CNRS), France; Bundesministerium fur Bildung, Wissenschaft, Forschung und Technologie (BMBF) and GSI Helmholtzzentrum fur Schwerionenforschung GmbH, Germany; General Secretariat for Research and Technology, Ministry of Education, Research and Religions, Greece; National Research, Development and Innovation Office, Hungary; Department of Atomic Energy Government of India (DAE) and Council of Scientific and Industrial Research (CSIR), New Delhi, India; Indonesian Institute of Science, Indonesia; Centro Fermi Museo Storico della Fisica e Centro Studi e Ricerche Enrico Fermi and Istituto Nazionale di Fisica Nucleare (INFN), Italy; Institute for Innovative Science and Technology, Nagasaki Institute of Applied Science (IIST), Japan Society for the Promotion of Science (JSPS) KAKENHI and Japanese Ministry of Education, Culture, Sports, Science and Technology (MEXT), Japan; Consejo Nacional de Ciencia (CONACYT) y Tecnologia, through Fondo de Cooperacion Internacional en Ciencia y Tecnologia (FONCICYT) and Direccion General de Asuntos del Personal Academico (DGAPA), Mexico; Nederlandse Organisatie voor Wetenschappelijk Onderzoek (NWO), Netherlands; The Research Council of Norway, Norway; Commission on Science and Technology for Sustainable Development in the South (COMSATS), Pakistan; Pontificia Universidad Catolica del Peril, Peru; Ministry of Science and Higher Education and National Science Centre, Poland; Korea Institute of Science and Technology Information and National Research Foundation of Korea (NRF), Republic of Korea; Ministry of Education and Scientific Research, Institute of Atomic Physics and Romanian National Agency for Science, Technology and Innovation, Romania; Joint Institute for Nuclear Research (JINR), Ministry of Education and Science of the Russian Federation and National Research Centre Kurchatov Institute, Russia; Ministry of Education, Science, Research and Sport of the Slovak Republic, Slovakia; National Research Foundation of South Africa, South Africa; Centro de Aplicaciones Tecnologicas y Desarrollo Nuclear (CEADEN), Cubaenergia, Cuba, Ministerio de Ciencia e Innovacion and Centro de Investigaciones Energeticas, Medioambientales y Tecnologicas (CIEMAT), Spain; Swedish Research Council (VR) and Knut & Alice Wallenberg Foundation (KAW), Sweden; European Organization for Nuclear Research, Switzerland; National Science and Technology Development Agency (NSDTA), Suranaree University of Technology (SUT) and Office of the Higher Education Commission under NRU project of Thailand, Thailand; Turkish Atomic Energy Agency (TAEK), Turkey; National Academy of Sciences of Ukraine, Ukraine; Science and Technology Facilities Council (STFC), United Kingdom; National Science Foundation of the United States of America (NSF) and United States Department of Energy, Office of Nuclear Physics (DOE NP), United States of America

Searches for transverse momentum dependent flow vector fluctuations in Pb-Pb and p-Pb collisions at the LHC

The measurement of azimuthal correlations of charged particles is presented for Pb-Pb collisions at sNN=2.76 TeV and p-Pb collisions at sNN=5.02 TeV with the ALICE detector at the CERN Large Hadron Collider. These correlations are measured for the second, third and fourth order flow vector in the pseudorapidity region |η| < 0.8 as a function of centrality and transverse momentum pT using two observables, to search for evidence of pT-dependent flow vector fluctuations. For Pb-Pb collisions at 2.76 TeV, the measurements indicate that pT-dependent fluctuations are only present for the second order flow vector. Similar results have been found for p-Pb collisions at 5.02 TeV. These measurements are compared to hydrodynamic model calculations with event-by-event geometry fluctuations in the initial state to constrain the initial conditions and transport properties of the matter created in Pb–Pb and p–Pb collisions

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Potential antioxidant peptides produced from whey hydrolysis with an immobilized aspartic protease from Salpichroa origanifolia fruits

An aspartic protease from Salpichroa origanifolia fruits was successfully immobilized onto an activated support of glutaraldehyde agarose. The immobilized enzyme presented higher thermal stability than the free enzyme from 40 °C to 50 °C and high reusability, retaining 54% of the initial activity after ten cycles of the process. Whey protein concentrates (WPC) were hydrolyzed with both free and immobilized enzyme, reaching a similar degree of hydrolysis of approximately 6?8% after 20 h. In addition, the immobilized derivate hydrolyzed α-lactalbumin protein with a higher affinity than β-lactoglobulin. The hydrolysate was ultra-filtrated, and the fractions were evaluated for antioxidant activities with the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity method. The fraction containing peptides with a molecular mass below 3 kDa demonstrated a strong radical quenching effect (IC50: 0.48 mg/ml). These results suggest that hydrolyzed WPC could be considered as a promising source of natural food antioxidants for the development of functional food.Fil: Rocha, Gabriela Fernanda. Universidad Nacional de Luján. Departamento de Ciencias Básicas; ArgentinaFil: Kise, Francisco. Universidad Nacional de Luján. Departamento de Ciencias Básicas; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Rosso, Adriana Mabel. Universidad Nacional de Luján. Departamento de Ciencias Básicas; ArgentinaFil: Parisi, Monica Graciela. Universidad Nacional de Luján. Departamento de Ciencias Básicas; Argentin

Diversity of Saint Helena Island and zoogeography of zoantharians in the Atlantic Ocean: jigsaw falling into place

Diversity surveys in isolated sites, such as oceanic islands, provide biogeographic data that can improve our analyses and knowledge of evolutionary processes in the oceans. Zoantharians (Cnidaria: Anthozoa) are common and widespread components of shallow-water reefs, but distributional analyses are scarce for this group. In this study, we collected Zoantharia specimens from around Saint Helena Island (STH) in the mid-Atlantic and identified species using external morphology and molecular data. Moreover, we compiled and analysed the most comprehensive distributional data for zoantharian species in the subtropical and tropical Atlantic Ocean to date. Our results show eight zoantharian species in STH, which includes seven new records for STH waters. Furthermore, all families and genera of the suborder Brachycnemina recorded are widespread in the Atlantic Ocean, including at least four amphi-Atlantic species. The Caribbean is the richness centre in the Atlantic Ocean for zoantharian species, a pattern similar to that observed for many other subtropical/tropical marine taxa. However, Zoantharia may have a lower endemism rate in some areas than other common reef animals, for example zooxanthellate scleractinian corals and reef fishes. Moreover, zoantharian species have a more extensive distribution than close-related taxa such as zooxanthellate scleractinian corals and hydrocorals in the Atlantic Ocean.Japanese Government (MEXT)Ministry of Education, Culture, Sports, Science and Technology, Japan (MEXT)JSPSMinistry of Education, Culture, Sports, Science and Technology, Japan (MEXT)Japan Society for the Promotion of Scienc

Searches for transverse momentum dependent flow vector fluctuations in Pb-Pb and p-Pb collisions at the LHC

The measurement of azimuthal correlations of charged particles is presented for Pb-Pb collisions at root S-NN 2.76 TeV and p-Pb collisions at root S-NN 5.02 TeV with the ALICE detector at the CERN Large Hadron Collider. These correlations are measured for the second, third and fourth order flow vector in the pseudorapidity region vertical bar eta vertical bar 0.8 as a function of centrality and transverse momentum pT using two observables, to search for evidence of PT-dependent flow vector fluctuations. For Ph-Ph collisions at 2.76 TeV, the measurements indicate that PT-dependent fluctuations are only present for the second order flow vector. Similar results have been found for p-Pb collisions at 5.02 TeV. These measurements are compared to hydrodynamic model calculations with event-by-event geometry fluctuations in the initial state to constrain the initial conditions and transport properties of the matter created in Ph-Ph and p-Pb collisions

Searches for transverse momentum dependent flow vector fluctuations in Pb-Pb and p-Pb collisions at the LHC

The measurement of azimuthal correlations of charged particles is presented for Pb-Pb collisions at root S-NN 2.76 TeV and p-Pb collisions at root S-NN 5.02 TeV with the ALICE detector at the CERN Large Hadron Collider. These correlations are measured for the second, third and fourth order flow vector in the pseudorapidity region vertical bar eta vertical bar 0.8 as a function of centrality and transverse momentum pT using two observables, to search for evidence of PT-dependent flow vector fluctuations. For Ph-Ph collisions at 2.76 TeV, the measurements indicate that PT-dependent fluctuations are only present for the second order flow vector. Similar results have been found for p-Pb collisions at 5.02 TeV. These measurements are compared to hydrodynamic model calculations with event-by-event geometry fluctuations in the initial state to constrain the initial conditions and transport properties of the matter created in Ph-Ph and p-Pb collisions