Revista Mexicana de Ingeniería Química ISSN: 1665-2738 [email protected] Universidad Autónoma Metropolitana Unidad Iztapalapa México Tovar-Jiménez, X.; Muro-Urista, C.R; Tellez-Jurado, A.; Mercado-Flores, Y.; Abreu- Corona, A.; Arana-Cuenca, A. HYDROLYSATE ANTIMICROBIAL ACTIVITY RELEASED FROM BOVINE WHEY PROTEIN CONCENTRATE BY THE ASPARTYL PROTEASE Eap1 OF Sporisorium reilianum Revista Mexicana de Ingeniería Química, vol. 16, núm. 1, 2017, pp. 11-18 Universidad Autónoma Metropolitana Unidad Iztapalapa Distrito Federal, México Available in: http://www.redalyc.org/articulo.oa?id=62049878002 How to cite Complete issue Scientific Information System More information about this article Network of Scientific Journals from Latin America, the Caribbean, Spain and Portugal Journal's homepage in redalyc.org Non-profit academic project, developed under the open access initiative Vol. 16, No. 1 (2017) 11-18 Revista Mexicana de Ingeniería Química HYDROLYSATE ANTIMICROBIAL ACTIVITYCONTENIDO RELEASED FROM BOVINE WHEY PROTEINVolumen CONCENTRATE 8, número 3, 2009 BY / THE Volume ASPARTYL 8, number 3, PROTEASE2009 Eap1 OF Sporisorium reilianum HIDROLIZADOS CON ACTIVIDAD ANTIMICROBIANA LIBERADOS POR LA ACCION´ DE213 LA Derivation ASPARTIL and application PROTEASA of the Stefan-Maxwell Eap1 equations DE Sporisorium reilianum A PARTIR DE UN CONCENTRADO PROTEICO DEL SUERO LACTEO´ BOVINO (Desarrollo y aplicación de las ecuaciones de Stefan-Maxwell) 1 2 1 1 1 X. Tovar-Jim enezStephen´ * ,Whitaker C.R Muro-Urista , A. Tellez-Jurado , Y. Mercado-Flores , A. Abreu-Corona , A. 1 Arana-Cuenca 1 Universidad Polit´ecnicade Pachuca, Carr. Pachuca-Cd. Sahag´unKm. 20, Zempoala, Hidalgo. M´exico. 2 DepartamentoBiotecnología de Ingenier´ıaQu´ımicae / Biotechnology Investigaci´on.Instituto Tecnol´ogicode Toluca. Av. Tecnol´ogicos/n Ex-Rancho la Virgen. Toluca, M´exico.C.P. 52140. 245 Modelado de laReceived biodegradación May 10, en 2016; biorreactores Accepted de November lodos de hidrocarburos 21, 2016 totales del petróleo intemperizados en suelos y sedimentos Abstract (Biodegradation modeling of sludge bioreactors of total petroleum hydrocarbons weathering in soil Whey, a by-product of cheese processing, has important nutritional qualities, as it is a rich source of proteins balanced with amino acids. As such,and sediments) it has a broad range of functional and biological properties that can be exploited biotechnologically for diverse applications.S.A. Medina-Moreno, Several studies S. have Huerta-Ochoa, demonstrated C.A. that Lucho-Constantino, the enzymatic hydrolysis L. Aguilera-Vázquez, of whey proteins A. Jiménez- releases peptides that can perform biologicalGonzález y activities, M. Gutiérrez-Rojas depending on the precise enzymatic process applied and the specific protein used. The present study259 examinesCrecimiento, the sobrevivencia antimicrobial yactivity adaptación obtained de Bifidobacterium from hydrolysates infantis of a proteincondiciones concentrates ácidas of bovine whey by means of the aspartyl protease Eap1. It demonstrates antimicrobial activity against the following human non-opportunistic (Salmonella sp.) and(Growth, opportunistic survival and pathogens adaptation (Staphylococcus of Bifidobacterium aureus infantis, Klebsiella to acidic pneumoniae conditions) , Pseudomonas aeruginosa, Escherichia coli andL. CandidaMayorga-Reyes, albicans P.), andBustamante-Camilo, then compares theA. findingsGutiérrez-Nava, obtained E. to Barranco-Florido the action of two y commercial A. Azaola- enzymes: trypsin (TP) and chymotrypsinEspinosa (CMTP). Results indicate that the antimicrobial activity of the hydrolysates produced with Eap1 was higher than that generated by TP and CMTP. Also, the study determined that the Eap1 products were the only 265 Statistical approach to optimization of ethanol fermentation by Saccharomyces cerevisiae in the ones that inhibited growth of the yeast C. albicans. Keywords: antimicrobialpresence protein of Valfor® hydrolysates, zeolite NaA protein concentrate from bovine whey, aspartyl protease of Sporisorium reilianum, trypsin, chymotrypsin. (Optimización estadística de la fermentación etanólica de Saccharomyces cerevisiae en presencia de Resumen zeolita Valfor® zeolite NaA) El suero lacteo´ es un subproducto de la elaboracion´ de queso con importantes cualidades nutrimentales, ya que representa una fuente rica en proteG. Inei-Shizukawa,´ınas y equilibrada H. A. enVelasco-Bedrán, aminoacidos´ queG. F. poseen Gutiérrez-López amplio rango and deH. propiedadesHernández-Sánchez funcionales y biologicas´ que pueden ser aprovechadas biotecnologicamente´ para diversas aplicaciones. A traves´ de diversos estudios se ha podido comprobar queIngeniería la hidrolisis´ de procesos enzimatica´ / Process de las engineering prote´ınas sericas´ conduce la liberacion´ de peptidos´ que pueden presentar actividad biologica,´271 Localización dicha actividad de una dependeplanta industrial: del proceso Revisión enzim críticaatico´ y adecuación utilizado y de la los prote criterios´ına de empleados origen. en El presente trabajo muestra la actividad antimicrobiana obtenida de hidrolizados de concentrados proteicos de suero lacteo´ bovino, esta decisión mediante una aspartil proteasa (Eap1). La actividad antimicrobiana fue probada contra patogenos´ humanos no oportunistas (Plant site selection: Critical review and adequation criteria used in this decision) (Salmonella sp.) y oportunistas (Staphylococcus aureus, Klebsiella pneumoniae, Pseudomonas aeruginosa, Escherichia coli y Candida albicansJ.R. Medina,) y comparada R.L. Romero con y laG.A. obtenida Pérez por accion´ de enzimas comerciales, tripsina (TP) y quimotripsina (QMTP). Los resultados indicaron que Eap1 produjo hidrolizados con mayor actividad antimicrobiana comparada con los generados por TP y QMTP. Ademas,´ se encontro´ que los productos provenientes de Eap1 fueron los unicos´ en inhibir el crecimiento de la levadura C. albicans. Palabras clave: hidrolizados proteicos antimicrobianos, concentrado proteico del suero lacteo´ bovino, aspartil proteasa de Sporisorium reilianum, tripsina, quimotripsina. * Corresponding author. E-mail: [email protected] Publicado por la Academia Mexicana de Investigacion´ y Docencia en Ingenier´ıa Qu´ımica A.C. 11 Tovar-Jim´enezet al./ Revista Mexicana deIngenier ´ıaQu ´ımica Vol. 16, No. 1 (2017) 11-18 1 Introduction porcine gastric mucosa, alcalase from Bacillus licheniformis, chymotrypsin (CMTP), chymosin Today, the food sciences are promoting the use of or trypsin (TP) of bovine origin, all inhibit both functional foods that have the potential to improve gram-positive (Staphylococcus, Bacillus, Listeria, health by decreasing the risks of illness in humans. For Streptococcus) and gram-negative (Escherichia, this reason, some food industries have incorporated Klebsiella, Pseudomonas, Salmonella, etc.) bacteria, functional ingredients like peptides extracted from as well as pathogenic yeasts such as Candida albicans animal and vegetable sources into their products (McCann et al., 2006; Szwajkowska et al., 2011; (Chatterjee et al., 2015). The benefits of these Elbarbary et al., 2012). However, few studies have products reside, primarily, in their capacity to reduce used aspartyl proteases, which acquire affinity by and/or prevent gastrointestinal illnesses. hydrolyzing in the adjacent carbon where the amino One potential source for obtaining functional acids phenylalanine, proline, tyrosine and leucine are ingredients are whey proteins, thanks not only to found (Narayanan et al., 2007). These amino acids are their nutritional effects due to their amino acid proximal to positively-charged amino acids in proteins content, but also because, in many cases, they present in whey, like LF and lactalbumin (Muro-Urista show biological and physiological effects in vivo et al., 2011). (Baro´ et al., 2001). Specifically, the presence The present study set out to determine the of leucine, isoleucine, tyrosine, methionine, proline antimicrobial activity of peptides contained in the and valine in the structure of α-lactalbumin (α- hydrolysates of protein concentrates from bovine La), β-lactoglobulin (β-Lg), immunoglobulins (Ig), whey against important pathogens that affect humans: albumin (BSA), lactoferrin (LF) and lactoperoxidase Salmonella sp., Staphylococcus aureus, Klebsiella (LP), promises possible applications after performing pneumoniae, Pseudomonas aeruginosa, Escherichia hydrolysis to obtain bioactive peptides (Muro-Urista et coli and Candida albicans. The enzymatic al., 2011; Sinha et al., 2007) that can be incorporated hydrolysates were obtained by the action of the into foods as functional ingredients (Torruco-Uco et purified enzyme Eap1 from S. reilianum, which has al., 2008). Bioactive peptides are defined as specific recently shown high proteolytic capacity (Mandujano- fragments of proteins of animal or vegetable origin Gonzalez´ et al., 2013). The peptides obtained that have a positive impact on bodily functions or were then compared to hydrolysates obtained from conditions that can affect human health beyond areas the commercial proteases most often used for this of basic nutrition (Alvarado-Carrasco and Guerra purpose: TP and CMTP from bovine pancreases. 2010; Gonzalez-Olivares´ et al., 2011). Among the biological activities that have been studied in relation to these compounds are their antimicrobial, 2 Materials and methods antioxidant and antihypertensive properties. These peptides are usually obtained from proteins using 2.1 Obtaining protein concentrates