ISOLATION, KINETICS AND IN VITRO ANTI-LEUKEMIC ACTIVITY OF CAPSICUM ANNUM L-GLUTAMINASE
Mohannad Idress, Mohammed Salim, Asia El-Hassan
Sudan Academy of Sciences, Khartoum, Sudan. Biotechnology Park, Africa City of Technology, Khartoum, Sudan.Department of Biochemistry, School of Medicine, Ahfad University for Women.
Keywords: Chilies, in vitro anti-leukemic activity, L-glutaminase, Capsicum annum
Abstract

This study is aimed at exploring the potential of developing naturally occurring L-glutaminase isolated from green chilies as anti-leukemic drug which selectively targets cancer cell metabolism unlike other non-selective chemotherapeutics. This was done by determination of its specific activity and optimum condition for L-glutaminase activity followed by partial purification and in vitro cytotoxicity assay. Activity of L-glutaminase was measured in the aqueous extract of fruits by Nesslerization method after homogenization in liquid nitrogen and extraction with 0.05 M sodium borate buffer (pH 8.5). Results showed that L-glutaminase activity per milligram of total protein was 18.7 U/mg. Optimum conditions for the catalytic activity of crude L-glutaminase extracted from chili fruits were studied. Results showed maximum activity of L-glutaminase was achieved when the enzyme was incubated with 250 mM of l-glutamine at 37°C for 30 minutes in the presence of phosphate buffered saline at pH 7.2. The maximum velocity (Vmax) and affinity constant (Km) of L-gutaminase were 14.1 mM and 90.2 mM respectively. Crude L-glutaminase was purified by salting out using seven different concentrations of ammonium sulfate ranging from 20% to 80% saturation. Specific activity of purified L-glutaminase was increased from 18.7 to 98.5 U/mg at when salted out in 50% ammonium sulfate solution. This result indicated the high efficiency of ammonium sulfate precipitation as purification technique. Assessment of anti-leukemic property was done by comparing the cytotixicity of L-glutaminase with doxorubicin and combination of them against THP-1 cell line by MTT assay. Results showed significant anti-cancer activity with inhibition percent of 72.75 % in case of combination of the enzyme with doxorubicin when compared to the enzyme alone (60.28%). These findings concluded that L-glutaminase of green chilies could be developed as anti-leukemic agent owing to its high content in chilies, ease of purification and signicant cytotoxicity against human monocytic leukemia cells.

Article Information

Identifiers and Pagination:
Year:2015
Volume:7
First Page:140
Last Page:151
Publisher Id:19204159.7:3.2015
Article History:
Received:May 15, 2015
Accepted:June 2, 2015
Collection year:2015
First Published:July 1,2016


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Editor in Chief
Prof. Dr. Cornelia M. Keck (Philipps-Universität Marburg)
Marburg, Germany

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Welcome to the research group of Prof. Dr. Cornelia M. Keck in Marburg. Cornelia M. Keck is a pharmacist and obtained her PhD in 2006 from the Freie Universität (FU) in Berlin. In 2009 she was appointed as Adjunct Professor for Pharmaceutical and Nutritional Nanotechnology at the University Putra Malaysia (UPM) and in 2011 she obtained her Venia legendi (Habilitation) at the Freie Universität Berlin and was appointed as a Professor for Pharmacology and Pharmaceutics at the University of Applied Sciences Kaiserslautern. Since 2016 she is Professor of Pharmaceutics and Biopharmaceutics at the Philipps-Universität Marburg. Her field of research is the development and characterization of innovative nanocarriers for improved delivery of poorly soluble actives for healthcare and cosmetics. Prof. Keck is executive board member of the German Association of Nanotechnology (Deutscher Verband Nanotechnologie), Vize-chairman of the unit „Dermocosmetics“ at the German Society of Dermopharmacy, active member in many pharmaceutical societies and member of the BfR Committee for Cosmetics at the Federal Institute for Risk Assessment (BfR).

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Abbreviation: J App Pharm
doi: http://dx.doi.org/10.21065/19204159
Frequency: Annual 
Current Volume: 9 (2017)
Next scheduled volume: December, 2018 (Volume 10)
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Starting year: 2009
Nature: Online 
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