ABSTRACT
Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.
Subject(s)
Animals , Allergens/drug effects , Aspergillus niger/growth & development , Calcium Compounds/pharmacology , Ricinus communis/drug effects , Inactivation, Metabolic , Allergens/toxicity , Aspergillus niger/drug effects , Chlorocebus aethiops , Ricinus communis/toxicity , Cell Death/drug effects , Cell Degranulation/drug effects , Enzyme Activation , Fermentation , L-Lactate Dehydrogenase/metabolism , Mast Cells/drug effects , Ricin/isolation & purification , Ricin/toxicity , Time Factors , Toxicity Tests , /isolation & purification , /toxicity , Vero CellsABSTRACT
Ricinus communis L. is of great economic importance due to the oil extracted from its seeds. Castor oil has been used for pharmaceutical and industrial applications, as a lubricant or coating agent, as a component of plastic products, as a fungicide or in the synthesis of biodiesel fuels. After oil extraction, a castor cake with a large amount of protein is obtained. However, this by-product cannot be used as animal feed due to the presence of toxic (ricin) and allergenic (2S albumin) proteins. Here, we propose two processes for detoxification and allergen inactivation of the castor cake. In addition, we establish a biological test to detect ricin and validate these detoxification processes. In this test, Vero cells were treated with ricin, and cell death was assessed by cell counting and measurement of lactate dehydrogenase activity. The limit of detection of the Vero cell assay was 10 ng/mL using a concentration of 1.6 x 10(5) cells/well. Solid-state fermentation (SSF) and treatment with calcium compounds were used as cake detoxification processes. For SSF, Aspergillus niger was grown using a castor cake as a substrate, and this cake was analyzed after 24, 48, 72, and 96 h of SSF. Ricin was eliminated after 24 h of SSF treatment. The cake was treated with 4 or 8% Ca(OH)2 or CaO, and both the toxicity and the allergenic properties were entirely abolished. A by-product free of toxicity and allergens was obtained.
Subject(s)
Allergens/drug effects , Aspergillus niger/growth & development , Calcium Compounds/pharmacology , Inactivation, Metabolic , Ricinus communis/drug effects , 2S Albumins, Plant/isolation & purification , 2S Albumins, Plant/toxicity , Allergens/toxicity , Animals , Aspergillus niger/drug effects , Ricinus communis/toxicity , Cell Death/drug effects , Cell Degranulation/drug effects , Chlorocebus aethiops , Enzyme Activation , Fermentation , L-Lactate Dehydrogenase/metabolism , Mast Cells/drug effects , Ricin/isolation & purification , Ricin/toxicity , Time Factors , Toxicity Tests , Vero CellsABSTRACT
Ric c 1 and Ric c 3 are the major castor bean allergens. In order to identify continuous IgE-epitopes in Ric c 1 and Ric c 3, pools of sera from rats immunized with a pool of 2S albumin from these seeds, Ric c 1 and Ric c 3 overlapping synthetic peptides, were used to screen for IgE-binding epitopes. The allergenic properties were monitored by mast cell degranulation assays, histamine quantification and human-IgE binding. Large and small chains isolated from these proteins present allergenic properties. Four continuous epitopes were identified in Ric c 3 and two in Ric c 1. This knowledge may allow the induction of protective antibody responses to antagonize the IgE recognition.
