Properties of lectin from tissue culture of Ungernia victoris and its ultrasonic extraction

Abstract

The aim was to investigate biomass extracts of U. victoris strain UV-2 tissue culture for the content of lectins, to study their general characteristics and to optimize the method of ultrasonic extraction to obtain a soluble form of the lectin. Methods. Tissue culture method, direct hemagglutination test for detection of lectin activity (LA), determination of lectin carbohydrate specificity, ultrasonic extraction method. The results. Lectin activity was detected in the soluble (supernatant) and insoluble (sediment) fractions of the extract of biomass U. victoris tissue culture. In the soluble fraction LA was significantly less compared to that in the sediment, which indicated the presence of the membrane-bound form of the lectin. LA was characterized by pronounced species specificity: the reaction with mouse erythrocytes was the most intense. The study of carbohydrate specificity revealed a weak affinity for monosugars (galactose and galactosamine) and pronounced suppression of LA in the case of polysaccharides - hyaluronic acid, heparin and mucin. The possibilities of the ultrasound extraction method for separation the detected lectin from the the cell surface in the sediment fraction were investigated and the extraction procedure was optimized. The transition of LA into the soluble phase under the influence of ultrasound depended on the following parameters: the concentration and volume of the extract, as well as the time of exposure to ultrasound. It was established that the LA of the soluble phase doubled compared to the initial one with an extract concentration of 50 mg/ml and a volume of 20 ml under the action of ultrasound 40 kHz/70 W in the interval of 15-45 minutes. Longer ultrasound treatment had a negative effect on LA. Conclusions. Lectin activity of U. victoris tissue culture biomass extracts was discovered for the first time, which can become a promising source of lectin with a wide spectrum of pharmacological properties. The general characteristics of the lectin were given, and its species and carbohydrate specificity was revealed. The lectin was generally characterized and its species and carbohydrate specificity were established. Possibilities of ultrasound extraction were used and the procedure was optimized to obtain a soluble form of the lectin, necessary for its further fundamental and practical research.