Journal of Pure and Applied MicrobiologyVol. 11 No. 4

Isolation, Identification and Molecular Characterization of Phytase Producing Bacteria, Pseudomonas Sp. Aazad

Ch. Shalu Singh1, Sarita Singh1, P.K. Sharma2 and Deepak Chand Sharma3*

1Department of Microbiology, Ch. Charan Singh University, Meerut-250004, India. 2Department of Genetics and Plant Breeding, Ch. Charan Singh University, Meerut-250004, India. 3Department of Microbiology, Dr. Shakuntala Misra National Rehabilitation University, Lucknow. India.

Received on 03 October 2017 and accepted on 06 November 2017

 

ABSTRACT

Phytases are enzyme capable of hydrolysing phytic acid to myo-inositol and inorganic phosphorus. The aim of the present study was the screening and isolation of efficient phytase producing bacteria from soil samples. 14 bacterial isolates were found to produce clear zone of hydrolysis on PSM containing 0.5% ca-phytate. Out of all phytase positive isolates, five bacterial isolates 100A, 9H, 14A, 4D, and 2B showed significant phytase production in submerged fermentation. The isolate 14A was found to be the best among all with the maximum production of 89.09 U/ml after 72 h of incubation at 30 C. Colony morphology and microscopic examination (Grams staining) of maximal phytase producer suggested that the isolate 14A is gram negative rod. Further on molecular analysis the bacterial isolate showed closest similarity with Pseudomonas sp. 16S rDNA sequencing and sequence similarity search using BLAST showed that it matches 97 % with Pseudomonas pulluriniana, while 3% was found to be unique sequence and named as Pseudomonas sp. aazad. The crude phytase from the isolate showed optimum activity at pH 6 and temperature 50 C.

Keywords : Phytase, PSM, Pseudomonas sp., ca-phytate, fermentation.