ABSTRACT
The present report was designed to determine the antioxidant and antigenotoxic effects of phaseolin (isolated from Phaseolus vulgaris) against mouse colon and liver damage induced by azoxymethane (AOM) and its colon chemopreventive effect. Eight groups with 12 mice each were utilized for an eight-week experiment: the control group was intragastrically (ig) administered 0.9% saline solution; the positive control group was intraperitoneally (ip) injected with 7.5 mg/kg AOM twice a week (weeks three and four of the experiment); three groups were ig administered each day with phaseolin (40, 200, and 400 mg/kg); and three groups were ig administered phaseolin daily (40, 200, and 400 mg/kg) plus 7.5 mg/kg AOM twice a week in weeks three and four of the experiment. The results showed that phaseolin did not produce oxidative stress, DNA damage, or aberrant crypts; in contrast, 100% inhibition of lipoperoxidation, protein oxidation, and nitrites induction generated by AOM was found in both organs, and DPPH radical capture occurred. The two highest phaseolin doses reduced DNA damage induced by AOM in both organs by more than 90% and reduced the AOM-induced aberrant crypts by 84%. Therefore, our study demonstrated the strong in vivo antioxidant, antigenotoxic, and chemopreventive potential of phaseolin.
Subject(s)
Chemical and Drug Induced Liver Injury/drug therapy , Colitis/prevention & control , Phaseolus/chemistry , Plant Extracts/pharmacology , Protective Agents/pharmacology , Aberrant Crypt Foci/prevention & control , Animals , Antioxidants , Azoxymethane , Chemoprevention , Colitis/chemically induced , Colon , DNA Damage/drug effects , Disease Models, Animal , Male , Mice , Oxidation-Reduction/drug effects , Oxidative Stress/drug effects , Seeds/chemistryABSTRACT
Background: Although legume protein extracts are useful in food preparation and processing as foam stabilizers and as viscosity, palatability and nutrition enhancers, many legume proteins from South America have not been characterized extensively. One such legume is the ñuña bean (Phaseolus vulgaris L.), which is cooked using dry heat until the cotyledons rapidly expand with a pop. The bean is widely cultivated in the Andes, but almost unknown elsewhere. Objective & Methods: In this study, we characterized ten functional properties of a ñuña protein extract using standard food analysis methods. Results: The extract was similar to other legume protein extracts for many properties (amino acid profile, proximate analysis, yield, water absorption, color, isoelectric point, and thermogravimetric analysis). The electrophoretic analysis revealed that the sample was nearly pure phaseolin. Additionally, the ability to form foam and increase solution viscosity were comparatively low when contrasted to other extracts. Conclusion: These properties make ñuña protein extract useful as a nearly pure phaseolin nutrition enhancer in beverages where foaming and high viscosity are undesirable, such as in fortified beverages, drinkable yogurts, or protein supplements. The extract may also have relevance as a weight-loss supplement. Therefore, we expect that incorporating ñuña protein in processed foods would be a straightforward process.
Antecedentes: Los extractos proteicos de leguminosas son muy utilizados en la preparación y procesamiento de alimentos como agentes estabilizadores de espuma y viscosidad, así como potenciadores de palatabilidad y nutrición. Sin embargo, muchas proteínas de leguminosas procedentes de Sudamérica no han sido caracterizadas extensamente. Una de ellas es el frijol ñuña (Phaseolus vulgaris L.), el cual se cocina utilizando calor seco hasta que los cotiledones se expanden rápidamente y explotan. La ñuña se cultiva ampliamente en los Andes, pero es mayormente desconocida en otras partes del mundo. Objetivo y Métodos: En el presente estudio, caracterizamos diez propiedades funcionales de un extracto proteico de ñuña, utilizando métodos estándares para análisis de alimentos. Resultados: Varias propiedades del extracto analizado fueron similares a las de los extractos proteicos de otras leguminosas (perfil de aminoácidos, análisis proximal, rendimiento, absorción de agua, color, punto isoeléctrico y análisis termogravimétrico). El análisis electroforético reveló que la muestra es mayormente faseolina. Además, el extracto analizado presentó baja capacidad para formar espuma e incrementar viscosidad de una solución a comparación de los otros extractos. Conclusión: Los resultados obtenidos indican que el extracto proteico de ñuña, que es casi faseolina pura, puede ser muy útil como potenciador nutricional de bebidas en las que la espuma y alta viscosidad son indeseadas, como es el caso de bebidas fortificadas, yogures bebibles o suplementos proteicos. El extracto podría tener relevancia como suplemento para pérdida de peso. Por lo tanto, esperamos que el uso de proteína de ñuña sea un proceso sencillo en la industria de alimentos procesados.
Subject(s)
Humans , Phaseolus , Food Additives , Amino Acids, EssentialABSTRACT
Tobacco seeds can be used as a cost effective system for production of recombinant vaccines. Avian influenza is an important respiratory pathogen that causes a high degree of mortality and becomes a serious threat for the poultry industry. A safe vaccine against avian flu produced at low cost could help to prevent future outbreaks. We have genetically engineered tobacco plants to express extracellular domain of hemagglutinin protein from H5N1 avian influenza virus as an inexpensive alternative for production purposes. Two regulatory sequences of seed storage protein genes from Phaseolus vulgaris L. were used to direct the expression, yielding 3.0 mg of the viral antigen per g of seeds. The production and stability of seed-produced recombinant HA protein was characterized by different molecular techniques. The aqueous extract of tobacco seed proteins was used for subcutaneous immunization of chickens, which developed antibodies that inhibited the agglutination of erythrocytes after the second application of the antigen. The feasibility of using tobacco seeds as a vaccine carrier is discussed.
Subject(s)
Hemagglutinin Glycoproteins, Influenza Virus/genetics , Influenza Vaccines/pharmacology , Nicotiana/metabolism , Seeds/genetics , Agglutination Tests , Animals , Chickens/virology , Chromatography, High Pressure Liquid , Gene Expression Regulation, Plant , Hemagglutinin Glycoproteins, Influenza Virus/metabolism , Influenza Vaccines/genetics , Influenza Vaccines/immunology , Influenza in Birds/prevention & control , Phaseolus/genetics , Plants, Genetically Modified/genetics , Plants, Genetically Modified/metabolism , Polysaccharides/analysis , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Seeds/metabolism , Nicotiana/geneticsABSTRACT
The use of plants as heterologous hosts is one of the most promising technologies for manufacturing valuable recombinant proteins. Plant seeds, in particular, constitute ideal production platforms for long-term applications requiring a steady supply of starting material, as they combine the general advantages of plants as bioreactors with the possibility of biomass storage for long periods in a relatively small volume, thus allowing manufacturers to decouple upstream and downstream processing. In the present work we have used transgenic tobacco seeds to produce large amounts of a functionally active mouse monoclonal antibody against the Hepatitis B Virus surface antigen, fused to a KDEL endoplasmic reticulum retrieval motif, under control of regulatory sequences from common bean (Phaseolus vulgaris) seed storage proteins. The antibody accumulated to levels of 6.5 mg/g of seed in the T3 generation, and was purified by Protein A affinity chromatography combined with SEC-HPLC. N-glycan analysis indicated that, despite the KDEL signal, the seed-derived plantibody bore both high-mannose and complex-type sugars that indicate partial passage through the Golgi compartment, although its performance in the immunoaffinity purification of HBsAg was unaffected. An analysis discussing the industrial feasibility of replacing the currently used tobacco leaf-derived plantibody with this seed-derived variant is also presented.