%0 Journal Article
%T Cloning and Expression of a Novel Phytase Gene (phyMS) from <em>Mycobacterium smegmatis</em>
%A Tamrin Nuge
%A Yumi Zuhanis Has-Yun Hashim
%A Abd-El Aziem Farouk
%A Hamzah Mohd Salleh
%J Advances in Enzyme Research
%P 27-38
%@ 2328-4854
%D 2014
%I Scientific Research Publishing
%R 10.4236/aer.2014.21003
%X Phytase, also known as phytate-degrading enzyme, catalyzes the hydrolysis of phytate (inositol hexakisphosphate) with sequential release of phosphate and lower inositol phosphate. We report here a new plasmid construct designated as pMSuia from pBAD-TOPO that harbors a 1.1 kb phytase gene <em>(phyMS) </em>from <em>Mycobacterium smegmatis</em>, and expression as well as characterization of the purified recombinant <em>M. smegmatis</em> phytase. DNA sequencing analysis and multiple alignment exercise indicated that the <em>M. smegmatis phytase</em> is different from both known acid and alkaline phytases. The active ~45 kDa recombinant enzyme was expressed and confirmed by enzyme assay and Western blot analyses. Ni-NTA affinity purified recombinant <em>M. smegmatis</em> phytase exhibited specific activity of 233.51 U/mg, optimal pH of 3 and 7 and optimal temperature of 60¡ãC. The purified enzyme retains almost 30% of the initial activity after incubation at 90¡ãC for 60 min. The enzyme showed broad substrate specificity with K<sub>m</sub> and V<sub>max</sub> of the recombinant enzyme for sodium phytate substrate of 0.105 ¡À 0.016 mM and 26.93 ¡À 1.21 mM min<sup>-1</sup>, respectively.
%K Mycobacterium smegmatis
%K Thermostability
%K Phytase
%K Broad pH Activity
%K Broad Substrate
%U http://www.scirp.org/journal/PaperInformation.aspx?PaperID=43904