Bioavailability of peptides from proteins of lupine, cowpea and amaranth, by permeability through Caco-2 cells and alterations in gene expression of the transporters associated with intestinal cholesterol absorption

• Grant number: 12/15900-4
• Support Opportunities: Regular Research Grants
• Start date: July 01, 2013
• End date: September 30, 2015
• Field of knowledge: Agronomical Sciences - Food Science and Technology - Food Science
• Principal Investigator: José Alfredo Gomes Arêas
• Grantee: José Alfredo Gomes Arêas
• Host Institution: Faculdade de Saúde Pública (FSP). Universidade de São Paulo (USP). São Paulo , SP, Brazil
• Associated researchers: Alvaro Danilo Cerda Maureira ; Gustavo Guadagnucci Fontanari ; Mario Hiroyuki Hirata ; Rosario Dominguez Crespo Hirata

Abstract

The proteins present in legumes, and other grains have been receiving great attention by presenting relevant and functional properties, mainly related to cholesterol metabolism. However, bioavailability, and particularly, its mechanisms of intestinal absorption are not well understood. The present work aims to evaluate the bioavailability of proteins from white lupin (Lupinus albus), cowpea (Vigna unguiculata L. Walp), and amaranth (Amaranthus cruentus L.), in models of human enterocytes (Caco-2 cells). Proteins will be submitted to physiological hydrolysis, which simulates protein digestion in the body, and also to industrial hydrolysis using alcalase enzyme (from microbial origin) aiming at improvement of technologies for food processing. The protein hydrolysates will be analyzed by SDS-PAGE, HPLC and LC/MS/MS. The bioavailability of the protein hydrolysates will be evaluated by experiments of in vitro permeation, using the Caco-2 cell line in transwell plates. The final products (permeates) of both experimental conditions will be identified by HPLC and LC/MS/MS. It will also be assessed the expression of genes NPC1L1, ABCG5, ABCG8, ABCA1 and ABCG1 in Caco-2 cells, in order to elucidate the possible involvement of peptide from hydrolysates on genes involved in the modulation of intracellular cholesterol homeostasis and its relationship to biological effect previously described in vivo. Through this work, we want to detect which fractions of peptides that are really bioavailable and absorbed by enterocytes and enter the bloodstream and reach target tissues, promoting its functional effect. This will enable better use of these proteins and their bioavailable fractions in formulations or as part of processed foods. (AU)

Scientific publications

(7)

(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)

MARQUES, MARCELO RODRIGUES; FONTANARI, GUSTAVO GUADAGNUCCI; PIMENTA, DANIEL CARVALHO; SOARES-FREITAS, ROSANA MANOLIO; GOMES AREAS, JOSE ALFREDO. Proteolytic hydrolysis of cowpea proteins is able to release peptides with hypocholesterolemic activity. Food Research International, v. 77, n. 1, SI, p. 43-48, NOV 2015. (12/15900-4, 11/04179-0)

KAROLINE DE MACÊDO GONÇALVES FROTA; LAYS ARNAUD ROSAL LOPES; IZABEL CRISTINA VERAS SILVA; JOSÉ ALFREDO GOMES ARÊAS. PERFIL MOLECULAR, PUREZA E ATIVIDADE DO INIBIDOR DE TRIPSINA EM ISOLADOS PROTEICOS DE FEIJÃO-CAUPI. Rev. Caatinga, v. 31, n. 1, p. 202-208, 2018-03-00. (12/15900-4, 16/08758-8)

GONCALVES FROTA, KAROLINE DE MACEDO; ROSAL LOPES, LAYS ARNAUD; VERAS SILVA, IZABEL CRISTINA; AREAS, ALFREDO GOMES. MOLECULAR PROFILE, PURITY AND PRESENCE OF TRYPSIN INHIBITORS IN COWPEA PROTEIN ISOLATES. REVISTA CAATINGA, v. 31, n. 1, p. 202-208, JAN-MAR 2018. (16/08758-8, 12/15900-4)

MARQUES, MARCELO RODRIGUES; CERDA, ALVARO; FONTANARI, GUSTAVO GUADAGNUCCI; PIMENTA, DANIEL CARVALHO; SOARES-FREITAS, ROSANA MANOLIO; HIRATA, MARIO HIROYUKI; CRESPO HIRATA, ROSARIO DOMINGUEZ; GOMES AREAS, JOSE ALFREDO. Transport of cowpea bean derived peptides and their modulator effects on mRNA expression of cholesterol-related genes in Caco-2 and HepG2 cells. Food Research International, v. 107, p. 165-171, MAY 2018. (12/15900-4, 13/09304-2)

MARQUES, MARCELO RODRIGUES; FONTANARI, GUSTAVO GUADAGNUCCI; PIMENTA, DANIEL CARVALHO; SOARES-FREITAS, ROSANA MANOLIO; GOMES AREAS, JOSE ALFREDO. Proteolytic hydrolysis of cowpea proteins is able to release peptides with hypocholesterolemic activity. Food Research International, v. 77, p. 6-pg., 2015-11-01. (11/04179-0, 12/15900-4)

GONCALVES FROTA, KAROLINE DE MACEDO; ROSAL LOPES, LAYS ARNAUD; VERAS SILVA, IZABEL CRISTINA; GOMES AREAS, JOSE ALFREDO. Nutritional quality of the protein of Vigna unguiculata L. Walp and its protein isolate. Rev. Ciênc. Agron., v. 48, n. 5, p. 792-798, 2017. (12/15900-4, 16/08758-8)

GONCALVES FROTA, KAROLINE DE MACEDO; DOS SANTOS, RAUL DIAS; RIBEIRO, VALDENIR QUEIROZ; GOMES AREAS, JOSE ALFREDO. Cowpea protein reduces LDL-cholesterol and apolipoprotein B concentrations, but does not improve biomarkers of inflammation or endothelial dysfunction in adults with moderate hypercholesterolemia. NUTRICION HOSPITALARIA, v. 31, n. 4, p. 1611-1619, APR 2015. (12/15900-4, 07/05977-1)