ABSTRACT
Objectives: To investigate the effect of different mixtures from nano casein phosphopeptides (CPP), amorphous calcium phosphate (ACP), probiotic Lactobacillus rhamnosus B-445 (L. rhamnosus) and casein glycomacropeptide (GMP) against Streptococcus mutans (S. mutans) growth and its adhesion. Methods: CPP was prepared from the tryptic digest of bovine casein and GMP by the action of chymosin on casein solution. Four mixtures namely: Group 1) Nano CPP; Group 2) Nano CPP+ ACP; Group 3)Nano CPP+ ACP + L. rhamnosus; and Group 4)Nano CPP+ ACP+ GMP were prepared and tested for its inhibitory activity against S. mutans growth and its adhesion to saliva-treated glass surfaces in comparison with a commercial product (GC MI paste plus) and chlorhexidine (0.2%) as a positive control. The particle size and zeta potential of nano CPP and its complex with ACP were evaluated. Furthermore, the viability of L. rhamnosusin its mixture was determined during two weeks of storage at pH 6.8 and 8 respectively. Results: Revealed that Nano CPP had an average particle size (7.75 nm) and zeta potential (-8.43 mV) lower than that of CPP+ACP mixture. Probiotic containing mixture exhibited inhibitory activity slightly less than the positive control at pH 6.8. All tested mixtures reduced the adhesion of S. mutans to saliva-treated glass surfaces and the highest was that containing probiotic and GMP. L. rhamnosus showed acceptable stability in CPP+ACP mixture during storage period. Conclusions: All these findings suggest the use of probiotic, CPP+ACP mixture as a dental anticariogenic and remineralizing agents.
ABSTRACT
Muitos métodos têm sido empregados na produção de peptídeos bioativos para a promoção da saúde. O objetivo deste estudo foi produzir caseinofosfopeptídeos por hidrólise tríptica do caseinato de sódio usando diferentes temperaturas (37 e 50 °C) e tempos de reação (2 e 4 h), caracterizá-los e analisar sua influência nas atividades citotóxica, antimicrobiana e antioxidante. Caseinofosfopeptídeos foram caracterizados através da composição centesimal, eletroforese em gel de poliacrilamida, Espectrometria de Massas e Cromatografia Líquida de Alta Eficiência. Toxicidade para leucócitos humanos, atividade antimicrobiana utilizando o teste de microdiluição em caldo e determinação da capacidade antioxidante pelo método de espécies reativas ao ácido tiobarbitúrico foram os ensaios biológicos realizados. Os resultados mostraram que as quatro frações peptídicas obtidas apresentaram-se com baixo peso molecular e elevados teores proteico e mineral; quanto ao perfil aminoacídico, apresentaram elevadas e diferenciadas quantidades de ácido glutâmico e serina, que pouco variaram de acordo com o processo de obtenção; não se mostraram tóxicos para leucócitos humanos; demonstraram atividade antimicrobiana para Escherichia coli e Salmonella Enteritidis e elevada capacidade antioxidante. Os resultados físico-químicos das frações de caseinofosfopeptídeos demonstraram elevada composição nutricional em termos de proteína e, principalmente, cálcio. O conjunto de dados indicou que alterações no tempo e na temperatura de reação para a obtenção dos hidrolisados não interferem nas suas qualidades biológicas, mostrando serem seguros para a promoção da saúde e para a aplicação em situações especiais, que envolvem pacientes desnutridos, imunossuprimidos, com comprometimento ósseo ou gastrintestinal decorrentes de inflamações e infecções.
Several methods have been employed for the production of bioactive peptides for health promotion. The aim of this study was to produce and characterize Casein phosphopeptides obtained by sodium caseinate tryptic hydrolysis under different temperatures (37 and 50 °C) and reaction times (2 and 4 h), and evaluate their biological capabilities. They have been characterized by assessing their centesimal composition, by polyacrylamide gel electrophoresis, Mass Spectrometry, and High-Performance Liquid Chromatography. The biological activities tested included toxicity for human leukocytes, antimicrobial assay using the microdilution test, and determination of the antioxidant capacity by the thiobarbituric acid reactive species method. The results showed that the four fractions obtained were of low molecular weight with high protein and mineral contents; their amino acid profile showed high and differentiated amounts of glutamic acid and serine independent of the methodological procedures. The results also showed no toxicity for human peripheral leukocytes, demonstrated antimicrobial activity against Escherichia coli and Salmonella Enteritidis as well as high antioxidant capacity. The results of the physicochemical Casein phosphopeptides' fractions showed high nutritional composition in terms of protein and, particularly, calcium. The biological assays indicated that time and temperature changes in the process for obtaining casein hydrolysates have not interfered with their biological qualities. In addition, they have proven safe in promoting health in special conditions involving malnourished and/or, immunocompromised patients or those with bone and/or gastrointestinal impairment due to inflammations and infections.