Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent : Allicin
Joint Authors
Source
Saudi Journal of Biological Sciences
Issue
Vol. 25, Issue 7 (30 Nov. 2018), pp.1429-1438, 10 p.
Publisher
Publication Date
2018-11-30
Country of Publication
Saudi Arabia
No. of Pages
10
Main Subjects
Topics
Abstract EN
Allicin, an extremely active constituent of freshly crushed garlic, is produced upon reaction of alliin with the enzyme alliinase (EC 4.4.1.4).
A bacterium Cupriavidus necator with the ability of alliinase production was isolated from a soil sample and was identified by morphological, biochemical and 16S rRNA sequence.
Alliinase production was optimised and it was further purified to apparent homogeneity with 103-fold purification and specific activity of 209 U/mg of protein by using DEAE Cellulose and Sephadex G-100 chromatography.
The enzyme is a homodimer of molecular weight 110 kDa with two subunits of molecular weight 55 kDa each.
The optimum activity of the purified enzyme was found at pH 7 and the optimum temperature was 35 C.
The enzyme exhibited maximum reaction rate (Vmax) at 74.65 U/mg and Michaelis–Menten constant (Km) was determined to be 0.83 mM when alliin was used as a substrate.
The cytotoxic activity of in-situ generated allicin using purified alliinase and alliin was assessed on MIA PaCa-2 cell line using MTT assay and Acridine orange–ethidium bromide staining.
This approach of in-situ allicin generation suggests a novel therapeutic strategy wherein alliin and alliinase work together synergistically to produce cytotoxic agent allicin.
American Psychological Association (APA)
Chhabria, Sagar& Desai, Krutika. 2018. Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent : Allicin. Saudi Journal of Biological Sciences،Vol. 25, no. 7, pp.1429-1438.
https://search.emarefa.net/detail/BIM-896813
Modern Language Association (MLA)
Chhabria, Sagar& Desai, Krutika. Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent : Allicin. Saudi Journal of Biological Sciences Vol. 25, no. 7 (Nov. 2018), pp.1429-1438.
https://search.emarefa.net/detail/BIM-896813
American Medical Association (AMA)
Chhabria, Sagar& Desai, Krutika. Purification and characterisation of alliinase produced by Cupriavidus necator and its application for generation of cytotoxic agent : Allicin. Saudi Journal of Biological Sciences. 2018. Vol. 25, no. 7, pp.1429-1438.
https://search.emarefa.net/detail/BIM-896813
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references : p. 1437-1438
Record ID
BIM-896813