Study On Enzymatic Extraction Process Of Aconitum Alkaloids From Prepared Radix Aconiti

Owing to the deletion of petrochemical resources and the energy shortage, environment and society crisis were acceleratly deteriorating. As a promising strategy for developing green chemistry and green chemical engineering, biocatalysis technology will exert significant influence on medicine, agriculture, food, life science and so on. Biocatalysis is a new technology for extracting and separating active constituents from natural plants via enzyme catalysis. The plant cell walls usually hinder the extraction of effective constituents. If proper enzymes are employed, the cell walls will be decomposed, the extraction rate will be improved and the cost thus be reduced.Up to now, no studies of cellulose-assisted extraction of aconitum alkaloids have been reported both at home and broad. This preliminarily study was to explore the reaction condition and underlying mechanism of aconitum alkaloids catalyzed and transformed by cellulase.In this present study, the extraction rate of aconitine and aconitum alkaloids were regarded as indexes. The impact of aconitum alkaloids rate through enzyme-catalyzed reaction was studied by single factor experiment to determine the scope of the temperature, pH, cellulase dosage and reaction time. The optimum conditions of enzymatic extraction have been obtained by orthogonal test and compared with acid extraction, macroporous adsorbing resins extraction, semi-bionic extraction and alcohol extraction. Cellulase was recruited to pretreat the prepared Radix Aconiti, subsequently the aconitum alkaloids were extracted by alcohol. The optimum catalysis conditions of cellulase and its effect on the extraction rate of aconitum alkaloids were studied. The extraction rate and purity by cellulase catalysis synergism method was compared with others. Meanwhile, the effect of cellulase catalysis on conventional alcohol extraction and the mechanism of cellulase catalysis were also studied.The optimum conditions that cellulase catalyzed and transformed aconitum alkaloids were at 45℃, pH 4.5, with cellulase dosage of 0.8mg/g(0.01g cellulase in 1 g material) and 4 h reaction time of cellulase. The extraction rates of aconitine and aconitum alkaloids of semi-bionic extraction were 0.002 447%, 0.244 410% respectively. Compared with the acid extraction, the extraction rates of aconitine and aconitum alkaloids of cellulose enzyme extraction increased by 84.4%, 59.3% respectively. Compared with the macroporous adsorbing resins extraction, the extraction rates of aconitine and aconitum alkaloids increased by 61.1%, 41.4%, respectively. Compared with the semi-bionic extraction, the extraction rates of aconitine and aconitum alkaloids increased by 41.0%, 29.1%, respectively. Compared with the alcohol extraction, the extraction rates of aconitine and aconitum alkaloids increased by30.9%, 21.8%, respectively. Compared with acid extraction, macroporous adsorbing resins extraction, semi-bionic extraction, alcohol extraction, the cellulose enzyme extraction rates of aconitine and aconitum alkaloids were much higher. It indicated that the enzyme catalysis destroyβ-D-glucoside bond of the cell wall and decomposed cellulose and minished mass transfer resistance. It made aconitum alkaloids be taken out easier. The yield rate of aconitum alkaloids improved obviously after the prepared radix aconiti was pretreated by cellulase. Cellulase catalysis method significantly improve the extration rate of aconitum alkaloids, which has advantages of mild conditions, short time and so on. This study suggested that the cellulase catalysis technology is promising in extracting active ingredients from natural plants and also feasible in practical production.