Purification of active myrosinase from plants by aqueous two-phase counter-current chromatography.
Phytochem Anal
; 26(1): 47-53, 2015.
Article
em En
| MEDLINE
| ID: mdl-25130502
ABSTRACT
INTRODUCTION:
Myrosinase (thioglucoside glucohydrolase; E.C. 3.2.1.147), is a plant enzyme of increasing interest and importance to the biomedical community. Myrosinase catalyses the formation of isothiocyanates such as sulforaphane (from broccoli) and 4-(α-l-rhamnopyranosyloxy)benzyl isothiocyanate (from moringa), which are potent inducers of the cytoprotective phase-2 response in humans, by hydrolysis of their abundant glucosinolate (ß-thioglucoside N-hydroxysulphate) precursors.OBJECTIVE:
To develop an aqueous two-phase counter-current chromatography (CCC) system for the rapid, three-step purification of catalytically active myrosinase.METHODS:
A high-concentration potassium phosphate and polyethylene glycol biphasic aqueous two-phase system (ATPS) is used with a newly developed CCC configuration that utilises spiral-wound, flat-twisted tubing (with an ovoid cross-section).RESULTS:
Making the initial crude plant extract directly in the ATPS and injecting only the lower phase permitted highly selective partitioning of the myrosinase complex before a short chromatography on a spiral disk CCC. Optimum phase retention and separation of myrosinase from other plant proteins afforded a 60-fold purification.CONCLUSION:
Catalytically active myrosinase is purified from 3-day broccoli sprouts, 7-day daikon sprouts, mustard seeds and the leaves of field-grown moringa trees, in a CCC system that is predictably scalable.Palavras-chave
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Base de dados:
MEDLINE
Assunto principal:
Brassica
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Isotiocianatos
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Moringa
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Raphanus
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Glicosídeo Hidrolases
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Mostardeira
Idioma:
En
Ano de publicação:
2015
Tipo de documento:
Article