Aqueous two phase based selective extraction of mannose/glucose specific lectin from Indian cultivar of Pisum sativum seed
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Date
2019
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Elsevier B.V.
Abstract
Pisum sativum lectin (Psl) being a high-value protein has marked its application in the biomedical and therapeutic field. Aqueous two phase extraction (ATPE) was implemented as a selective partitioning technique for the partial purification of Psl from its seeds. PEG/citrate based biodegradable aqueous two phase system (ATPS) was screened and the factors such as the type and concentration of citrate salts, molar mass and concentration of polyethylene glycol (PEG), tie line length (TLL) and additive (NaCl) concentration, pH, crude load and volume ratio were studied for the selective partition of Psl. The Psl was successfully extracted to the top phase in the ATPS formed with 18% PEG 6000/16% sodium citrate at 41.01% TLL, 2% NaCl and pH of 7.5. A volume ratio of 0.76 and a crude load of 20% showed maximum activity yield of 122.12% with the purification factor of 16.26. The subunits of Psl namely ? and ? were identified with a molecular weight of 6 and 18 kDa respectively during the purity analysis using SDS PAGE and HPLC. © 2019 Elsevier B.V.
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Keywords
Antigen-antibody reactions, Molar ratio, Plants (botany), Proteins, Purification, Sodium chloride, Solvent extraction, Aqueous two phase system, Aqueous two-phase extraction, Partial purification, Partitioning coefficients, Partitioning techniques, Purification factors, Selective extraction, Tie line, Molar concentration, adenosine triphosphate, citrate potassium, citrate sodium, glucose, lectin, macrogol, macrogol 6000, mannose, Pisum sativum extract, plant extract, unclassified drug, mannose binding lectin, plant lectin, Article, cultivar, hemagglutination, high performance liquid chromatography, hydrophobicity, molecular weight, nonhuman, pea, polyacrylamide gel electrophoresis, priority journal, chemistry, isolation and purification, liquid liquid extraction, pH, plant seed, procedures, Chromatography, High Pressure Liquid, Electrophoresis, Polyacrylamide Gel, Hydrogen-Ion Concentration, Liquid-Liquid Extraction, Mannose-Binding Lectins, Peas, Plant Lectins, Polyethylene Glycols, Seeds, Sodium Citrate
Citation
Journal of Chromatography B: Analytical Technologies in the Biomedical and Life Sciences, 2019, 1114-1115, , pp. 13-23