Anti-cancerous/Anti-bacterial Activities of Allicin Generated In situ from Diastereo Pure Alliins by Alliinase.
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Date
2015
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Abstract
Aims: To facilitate allicin generation in-situ from pure diastereomers of alliin by enzymatic reaction
of alliinase and assess its anti-cancerous/anti-bacterial activities.
Study Design: Chemical synthesis and in-vitro assay of anti-cancerous/anti-bacterial activities.
Place and Duration of Study: Protein Research Laboratory, Research Resources Center,
University of Illinois at Chicago, between February 2014 and February 2015.
Methodology: Cancer cell viability assay MTT assay, bacterial plate-diffusion growth inhibition
assay, and flow cytometry cell cycle analysis have been used to demonstrate the anticancerous/
anti-pathogen activities of the in-situ allicin. Diastereomers of alliin are produced by
H2O2 oxidation of deoxyalliin, which is prepared by mixing L-cysteine and allyl bromide. Deoxyalliin
and diastereomers of alliin are purified to high purity with repeated fractional crystallization. In addition, fluorenylmethyloxycarbonyl (Fmoc) protected alliin and alliin methyl ester are synthesized
and purified with RP-HPLC to test the importance of amino and carboxyl groups of alliin in alliinase
enzymatic reaction. Alliinase is produced by a simple and effective method from an aqueous garlic
extract
Results: Results from spectrophotometric alliinase activity assay indicate that (+)-L-alliin is more
reactive toward alliinase than (-)-L-alliin, and both amino and carboxyl groups of the cysteine
portion of alliin are critical in alliinase enzymatic reaction. Results from cancer cell viability assay
MTT assay, bacterial plate-diffusion growth inhibition, and flow cytometry cell cycle analysis confirm
that the in-situ allicin is as active as allicin purified from aqueous garlic extract or allicin synthesized
chemically in a dose-dependent manner.
Conclusion: We describe here facile pathways to synthesize diastereomerically pure alliins and
isolate allinase. The in-situ allicin conversed from alliin by allinase is very active. The data obtained
here provide useful information on the design of the in-situ allicin strategy.
Description
Keywords
Allium sativum, alliin, allicin, alliinase, anti-cancerous, anti-bacterial
Citation
Gupta Shalini, Jayathilaka Lasanthi, Huang Jin-Sheng, Lee Jenny, Lee Bao-Shiang. Anti-cancerous/Anti-bacterial Activities of Allicin Generated In situ from Diastereo Pure Alliins by Alliinase. International Journal of Biochemistry Research & Review 2015 8(2): 1-16.