This paper should be cited as: Nazemi, A. Vaseghi, N. Khataminejad, MR. Nasrollahi Omran, A. Eskandari, M.
Expression of Bacillus Species
Isolated Phytase Gene from Soil by PCR Method
Nazemi, A. (PhD)1, Vaseghi, N. (MSc)2*, Khataminejad,
MR. (PhD)3, Nasrollahi Omran,
A. (PhD)4, Eskandari, M. (MSc)5
1.Assistant professor of Genetics,Faculty of Biology Sciences,Islamic
Azad University,Tonekabon Branch,Tonekabon,
Iran
2.MSc of Microbiology, Faculty of Biology Sciences, Islamic Azad University, Tonekabon Branch, Tonekabon, Iran
3.PhD of Microbiology,Department
of Microbiology, Faculty of Biology Sciences, Islamic Azad University,Tonekabon
Branch, Tonekabon, Iran
4.Associate Professor of
Microbiology, Department of Microbiology,
Faculty of Biology Sciences, Islamic Azad University,Tonekabon
Branch, Tonekabon, Iran
5.MSc of Cellular and Molecular
Biology, Faculty of Sciences, Gilan University,
Rasht, Iran
Abstract
Background
and Objective: Recognizing and using of isolated phytase
in the soil microorganisms are paramount importance to produce the Phytase enzyme utilized commercially in different
industries. This study was conducted to recognize different bacillus species
which are Phytase producers and detection of the gene
that can produce this enzyme.
Material
and Methods: Soil samples were gathered through different parts of mountainous
areas. The early isolation of bacillus was carried out in Bacillus Medium Agar.
After isolating the bacteria and genome extraction, the responsible gene of
enzyme producer recognized and amplified by PCR method. The size of this
protein and the optimal production situation in supplemental exploitation such
as SDS-PAGE and the enzymatic activity of its size were evaluated.
Results:
Of 40 samples, one bacterium secreting Phytase
enzyme was isolated. This bacterium was sequenced and recognized Bacillus Sobtlis species that is classified in STR Genus. The size
of protein phytase produced by this gene was about 45
KD and the enzyme activity at 55 degrees was measured about 5.65 in wavelength
of 415 NM. The phytase gene with the size of 1200 bp was propagated.
Conclusion: the
microorganisms, in natural conditions, produce Phytase
enzyme in limited amount and with the quality appropriate to microorganisms.
Thus, isolating the bacilli producing Phytase enzyme
and purifying this protein are highly significant.
Keywords:BacillusSubtilis;Phytase;SDS-PAGE;Enzymatic Activity;Polymerization Chain Reaction
Corresponding Author: Vaseghi, N.
Email: NVasegi@ yahoo.
Com
Received 18 Jul 2012 Revised 30 Aug 2013
Accepted 4 Sep 2013