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Myrosinase activity in different plant samples; optimisation of measurement conditions for spectrophotometric and pH-stat methods
Abstract
Myrosinase found in Brassicaceae plants, is the enzyme responsible for hydrolysis of glucosinolates. As a result a variety of biologically active metabolites are liberated, whose importance in crop protection and especially in cancer chemoprevention is rapidly gaining recognition. The growing practical application of glucosinolate degradation products requires that sensitive and reliable methods of myrosinase activity determination in different types of plant samples are established. With the use of commercial myrosinase prep, we systematically optimised conditions of measurement of this enzyme activity by spectrophotometric and pH-stat methods. The parameters evaluated included: sample preparation, choice of substrate, its concentration, reaction temperature and detection wavelength. Two substrates with different spectral properties were chosen: sinigrin (SIN) and glucotropaeolin (GTL). For both substrates, the best reliability was achieved at reaction temperature of 37*C and substrate concentration of 0,2mM and 5mM for spectrophotometric and pH-stat methods, respectively. GTL exhibiting higher absortpion at the recommended detection wavelenght of 230 nm ensured greater sensitivity of spectrophotometric determination of myrosinase activity in the case of transparent plan samples. GTL seemed to increase also the sensitivity of pH-stat method, however, in this casehomogenisation of plant samples turned out to be most important. The optimised conditions were then verified for a range of plant samples. Based on these results, the optimised protocols of myrosinase activity determination for both methods are proposed.
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- Category:
- Articles
- Type:
- artykuł w czasopiśmie wyróżnionym w JCR
- Published in:
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INDUSTRIAL CROPS AND PRODUCTS
no. 50,
pages 58 - 67,
ISSN: 0926-6690 - Language:
- English
- Publication year:
- 2013
- Bibliographic description:
- Piekarska A., Kusznierewicz B., Meller M., Dziedziul K., Namieśnik J., Bartoszek-Pączkowska A.: Myrosinase activity in different plant samples; optimisation of measurement conditions for spectrophotometric and pH-stat methods// INDUSTRIAL CROPS AND PRODUCTS. -Vol. 50, (2013), s.58-67
- DOI:
- Digital Object Identifier (open in new tab) 10.1016/j.indcrop.2013.06.048
- Verified by:
- Gdańsk University of Technology
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