Cell viability studies and to evaluate anti-cancerous activity of recombinant clone carrying Methionine-?-lyase (MGL) of Brevibacterium linens

Order of Publishing in Issue: 
4
Volume :8
Issue :4
October, 2014
Page No: 
372-377
Authors: 
[*]Kolasani Pavani[1] and S. Vijaya Saradhi[2]
Address: 
[1]Department of Biotechnology, Acharya Nagarjuna University, Nagarjuna Nagar, Guntur, Andhra Pradesh-522 510, India
Address: 
[2]Department of Biotechnology, KL University, Vaddeswaram, Guntur, Andhra Pradesh-522 303, India

Brevibacterium linens is a normal flora present in the whey of curd, which is a rich source of L-Methionine γ-lyase (EC4.4.1.11;MGL), also known as methionase, L-methionine γ- demethiolase. This enzyme can be utilized as a therapeutic agent in treating cancer. The MGL gene was cloned into pRSET-A vector and expressed in E.coliDH5α. In light of the importance of MGL of Brevibacterium linens in anti-cancerous activity, an attempt was made to analyze its anti-cancerous activity by studying the viability of cancerous cell lines and by studying the toxicity studies on the cancerous cell lines by treating with cloned bacterial extracts. Dried bacterial extract powder was obtained by freeze drying. The percentage viability of HCT 15 and T47D cancer cells carried out by using Trypan blue staining were 71.48%, 79.83% respectively which are most suitable to perform cytotoxicity studies. With the increase in the concentration of the bacterial extract (20 to 120 μl/ml acetone, methanol and ethyl acetate bacterial extracts); the percent viability of cells tends to decrease.

Keywords: 
Brevibacterium linens, Cell viability, Trypan blue staining, MTT assay.
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