Research Article | Volume: 6, Issue: 7, July, 2016

Purification and characterization of a novel thermo stable L-methioninase from Streptomyces sp. DMMMH4 and its evaluation for anticancer activity

Mohsen Helmy Selim Husein Hosny Elshikh Moataza Mahmoud Saad Elsayed Eliwa Mostafa Mohamed Abdelraof Mahmoud   

Open Access   

Published:  Jul 28, 2016

DOI: 10.7324/JAPS.2016.60708
Abstract

L-methioninase has been purified 2.55-fold from the crude extract of Streptomyces sp. DMMMH4. The purification procedure was carried out by heat treatment and gel filtration on Sephadex G-200 column chromatography. SDS-PAGE electrophoresis showed a migrating protein band molecular mass of 47‚ÄČkDa. The kinetic properties determined for the purified enzyme displayed optimum activity at 70OC and thermal stability were 70OC for 30 min. The enzyme showed maximum activity at pH 6 using acetate buffer 0.05M and was relatively stable across a broad range of pH values (5.5-8 pH). The enzyme strongly inhibited by Cr+2, Fe+2, Ni+2, Cd+2, PMSF, β-mercaptoethanol and SDS while Hg+2,Cu+2 and iodoacetate completely inhibited the enzyme activity at a final concentration of 10mM. The purified enzyme exhibited a Km of 0.7, 0.15 and 0.25 mM for L-methionine, DL-ethionine and L-cystine respectively. Cytotoxicity test demonstrate that enzyme was active against liver HepG2, breast MCF-7, lung A549, prostate PC3 and colon HCT116 cancer cell lines and has negligible toxicity toward a normal melanocyte cell line HFB4.


Keyword:     L-methioninase Streptomyces sp DMMMH4 Purification Thermo stable anticancer.


Citation:

Selim MH, Elshikh HH, Saad MM, Eliwa E, Abdelraof M. Purification and characterization of a novel thermo stable Lmethioninase from Streptomyces sp. DMMMH4 and its evaluation for anticancer activity. J App Pharm Sci, 2016; 6 (07): 053-060.

Copyright:The Author(s). This is an open access article distributed under the Creative Commons Attribution Non-Commercial License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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