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 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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Acknowledgements
Facilities provided by Aligarh Muslim University are gratefully acknowledged. P.G. was Senior Research Fellow of the Council of Industrial Research, New Delhi, India. The authors are also thankful to Department of Science and Technology for providing lab facilities under its FIST programme and to the University Grants Commission for the DRS to the Department of Biochemistry.
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Gupta, P., Saleemuddin, M. Bioaffinity Based Oriented Immobilization of Stem Bromelain. Biotechnol Lett 28, 917–922 (2006). https://doi.org/10.1007/s10529-006-9015-9
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DOI: https://doi.org/10.1007/s10529-006-9015-9