International Journal of Pharma and Bio Sciences
ijpbs.net
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10.22376/ijpbs.2019.10.1.p1-12
Volume 5 Issue 1
2014 (January - March)
ISOLATION, PURIFICATION AND QUANTITATIVE ANALYSIS OF CYSTEINE PROTEASE, BROMELAIN FROM ANANAS COMOSUS (PINEAPPLE)
Proteases were extracted and assayed from the fruit of lessThan i greaterThan Ananas comosus lessThan /i greaterThan (Pineapple). They were precipitated utilizing variable concentrations of acetone (40%,60%,80%,100%), ammonium sulphate (45%,65%) and Jello (40%,60%,80%,100%). The extracted enzymes exhibited proteolytic activity which was determined using other assay techniques. Activity determined for lessThan i greaterThan Ananas comosus lessThan /i greaterThan was 2U/mL. The enzymes were, in their crude state, analyzed using SDS-AGE. Bands were observed between the molecular weight range of 24-45 KDa. The enzymes were purified from the extract by anion exchange chromatography using Silica Gel Column (pH-8.0, Sodium Phosphate Buffer). The elution of lessThan i greaterThan Ananas comosus lessThan /i greaterThan extract resulted in two bound fractions. Since the second fraction of the lessThan i greaterThan Ananas comosus lessThan /i greaterThan extract exhibited no protease activity, it was ignored. The purified samples were analyzed using SDS-AGE, the purified extract of lessThan i greaterThan Ananas comosus lessThan /i greaterThan displays a single band (characteristic of bromelain, a monomeric molecule).
HEMANT KR. SHARMA, RICHA SRIVASTAVA AND SHUBHANGI SHUKLA
Protease, SDS-PAGE, Silica Gel, Ananas comosus
429-437