Aqueous two Phase Extraction Of Lectin From Pisum Sativum Seeds

Abstract

The diversified properties of the lectins from different sources and their applications in newlinebiotechnological and pharmaceutical industries demand the economically feasible and newlinescalable purification strategy. Pisum sativum seed lectin (Psl) has specific properties like newlineanti-HIV reverse transcriptase, anti-HCV, anti-cancer, and mitogenic activities. The newlinechromatographic process, which was commercially applied for the purification of this newlinelectin, has number of limitations to implement in industrial scale. The Aqueous two phase newlineextraction (ATPE) technique, a modified liquid-liquid extraction, was adopted for the newlinerecovery and partial purification of Psl from its seed crude extract. The PEG 6000- newlinetrisodium citrate system (ATPS) was identified as a suitable system by analyzing the newlineinfluence of type of citrate salt, PEG molecular weight, phase components concentration, newlinepH, additive (NaCl) and phase volume ratio on the partitioning behavior of Psl from the newlineaqueous solution of pure Psl. The maximum partitioning coefficient of 14.5 was obtained newlinefor 18% PEG 6000-16% trisodium citrate system with 1% NaCl at pH of 8 and volume newlineratio of 1.32. Further, the optimized condition for the selective partitioning of Psl to the newlinetop phase from the crude extract of Pisum sativum seed was achieved in the ATPS newlineformed with PEG 6000-sodium citrate at 41.01% TLL, 2% NaCl at pH of 7.5 with crude newlineload of 20% and volume ratio of 0.76. The optimum value of crucial variables (PEG and newlinesalt concentration, NaCl, and pH) and their individual and interactive impact on the newlinerecovery and purity was identified using Response surface methodology. The optimum newlineprocess conditions for different goals of extraction and purification were obtained by newlineemploying the desirability-based optimization tool and were validated experimentally. newlineThe feasibility of continuous operation of the extraction process has also been newlinedemonstrated in the Rotating disc contactor. Modeling of the equilibrium characteristics newlineand the partitioning of the Psl was attempted