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Br Biotechnol J ; 2014 Jun; 4(6): 740-750
Article in English | IMSEAR | ID: sea-162473

ABSTRACT

This study was carried out after a five day germination period on TZEE*TZEEW* DEMARSCUS*TZEE-W one of the most recommended high amylolytic Nigerian maize cultivar. Purification steps comprising of fractional precipitation by ammonium sulphate, gel filtration and anion exchange chromatography, was used respectively to purify β amylase (EC 3.2.1.2) from the malt. The sensitivity of the beta amylase indicated that it has serine at its active site. An apparent 60KDa monomeric protein was detected by one dimensional sodium dodecyl sulphate- polyacrylamide gel electrophoresis (SDS-PAGE). Identity assigned to the purified protein by Matrix-assisted laser desorption ionization time –of- flight mass spectrometry ( MALDI-ToF) reference to electronic protein data base is a 58542Da high putative beta amylase – Q9AV88-ORSA from Oryza sativa. Complete primary structure thereafter deduced with the aid of MS/MS MALDI- ToF showed a signature of a highly conserved ubiquitous not yet reported beta amylase. This study paved an insight to the gene encoding the β amylase in TZEE*TZEEW* DEMARSCUS*TZEE-W and better understanding of the activity of the enzyme at molecular level.

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