Purification and some properties of a metalloprotease from sorghum malt variety KSV8-1

A metalloprotease from sorghum malt variety KSV8-I was purified by a combination of 4-M sucrose fractionation, ion-exchange chromatography on Q-Sepharose (Fast flow), gel-filtration chromatography on Sephadex G-100 and hydrophobic interaction chromatography on phenyl-Sepharose CL-4B. The enzyme was...

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Bibliographic Details
Published in:World journal of microbiology & biotechnology 2005-10, Vol.21 (6-7), p.1051-1056
Main Authors: Ogbonna, A.C, Okolo, B.N
Format: Article
Language:English
Subjects:
Bacteria
Biological and medical sciences
Biotechnology
Chromatography
enzyme activity
Enzymes
Fractionation
Fundamental and applied biological sciences. Psychology
malt
metalloproteinases
Sorghum
Sorghum bicolor
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Summary:A metalloprotease from sorghum malt variety KSV8-I was purified by a combination of 4-M sucrose fractionation, ion-exchange chromatography on Q-Sepharose (Fast flow), gel-filtration chromatography on Sephadex G-100 and hydrophobic interaction chromatography on phenyl-Sepharose CL-4B. The enzyme was purified 7.9-fold to give a 13.4% yield relative to the total activity in the crude extract and a final specific activity of 2128.7 U mg^sup -1^ protein. SDS-PAGE revealed a single migrating protein band corresponding to a relative molecular mass of 35 kDa. The purified enzyme had optimal activity at 60 °C and maximal temperature stability between 40 and 60 °C but retained over 77% of its initial activity after incubation at 70 °C for 30 min. Both pH optimum and maximal stability were at 7.0 but 60% of the activity remained after 24 h between pH 5.0 and 8.0. Using 0.2 ml of 5 mM solution of each metal ion, the purified protease was slightly (P
ISSN:0959-3993
1573-0972
DOI:10.1007/s11274-004-8026-8