Isolation and purification of proteins from Tinospora cordifolia against Pseudomonas aeruginosa
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Date
2023-05-14
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College of Horticulture and Forestry Dr YSP UHF, Neri, Hamirpur(H.P.)
Abstract
The present investigation on the “Isolation and purification of proteins from Tinospora cordifoliaagainst Pseudomonas aeruginosa.” were carried out in the Department of Biotechnology, College of Horticulture and Forestry (Dr. Y.S. Parmar University of Horticulture and Forestry), Neri Hamirpur (HP) during 2021-2022. Tinospora cordifolia is a deciduous shrub. It is a well-recognized for its medicinal properties in Indian Ayurveda system. It is one of the most versatile shrub commonly known as “giloy”. The plant gained attention due to its various biological activities such as anti-diabetic, anti-microbial, anti-allergic, anti-oxidant and anti-cancer activities. The plant parts contains various chemical constituents such as alkaloids, steroids, glycosides and polysaccharides. The study was
undertaken for isolation and purification of antibacterial protein of Tinospora cordifolia collected from Hamirpur district (Neri), Himachal Pradesh. Analysis was done by stem ethanolic, methanolic, extract of samples taken from Neri village. It was found that methanol act as better solvent for extraction of proteins. Ammonium sulphate precipitation of methaolic crude extract showed that 60-70% precipitation was found best for precipitation of proteins. Further, 60-70% precipitates was subject for dialysis followed by ion exchange chromatography. The fractions obtained after ion exchange chromatography was assayed by antibacterial activity against test organism. The active fractions were pooled together and carried out for SDS-PAGE. The total protein content present in methanolic crude extract was 2.056 mg/ml. The ammonium sulphate precipitation showed 10.833±0.928 mm zone of inhibition and after purification the antibacterial activity increased to 15±0.289 mm. The
partial characterization of proteins revealed that the antibacterial proteins was stable at optimal pH 8, heat stable at 40 ℃ temperature and stable at low salt concentration. For future aspects, the stem extract and protein can be further exploited for its other properties such as its sequencing.