Gene Cloning Studies of a Novel Phytase from Bacillus megaterium EBD 9-1 Strain

Abstract

Phytases are hydrolytic enzymes which release inorganic phosphate by cleaving phosphate groups of phytate and attracted attention because of their usage in many industries providing benefit in eco-friendly way. Therefore, in this study we tried to clone the phytase gene from newly isolated Bacillus megaterium EBD 9-1 strain into the E. coli DH5α strain. The entire genome sequences of Bacillus megaterium DSM319, Bacillus megaterium QM B1551 and Bacillus megaterium WSH-002 strains are available in the GenBank database. However, a phytase sequence was not found in any of them. Thence, phytase sequences of different Bacillus species were aligned, and a degenerate primer pair was prepared. We tried to clone the phytase gene of the Bacillus megaterium EBD 9-1 strain using degenerate primers.We were able to amplify, clone and sequence 3 amplification products, but the sequencing results indicated that we could not clone the phytase gene. This study is important for literature because of being the first report on cloning studies of phytase from Bacillus megaterium.