Bioaffinity based oriented immobilization of stem bromelain.

Pawan Gupta; M Saleemuddin

doi:10.1007/s10529-006-9015-9

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Bioaffinity based oriented immobilization of stem bromelain.

Pawan Gupta·M Saleemuddin

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Abstract

Bromelain is a basic, 23.8 kDa thiol proteinase obtained from stem of the pineapple plant (Ananas comosus) and is unique in containing a single oligosaccharide chain attached to the polypeptide. This property allowed its affinity binding and favorable orientation on a Sepharose support pre-coupled with the lectin, concanavalin A (Con A). For comparison, bromelain was also immobilized by covalently coupling to the CNBr-activated Sepharose. The preparation obtained was more resistant to thermal inactivation as evident from the retention of over 50% activity after incubation at 60 degrees C for 100 min (as compared to 20% retained by the native enzyme and 30% retained by the covalently immobilized enzyme), exhibited a broader pH-activity profile with the enzyme retaining over 60% activity at pH 11 (as compared to over 25% retained by native and the enzyme immobilized covalently). The native, covalently-coupled and affinity-bound bromelains had apparent K (m) values of 1.1, 2 and 0.54 mg/ml, respectively using casein as the substrate. The V (max) values remained unaffected on immobilization.

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Bromelains Chromatography, Affinity Concanavalin A Enzymes, Immobilized Kinetics Plant Extracts Plant Stems Sepharose Journal Article Research Support, Non-U.S. Gov't

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Pawan Gupta, M Saleemuddin. 2006-06-08. Bioaffinity based oriented immobilization of stem bromelain.. https://doi.org/10.1007/s10529-006-9015-9

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