title: Cloning, Sequencing, and In Silico Analysis of β-Propeller Phytase Bacillus licheniformis Strain PB-13.
abstract: β -Propeller phytases (BPPhy) are widely distributed in nature and play a major role in phytate-phosphorus cycling. In the present study, a BPPhy gene from Bacillus licheniformis strain was expressed in E. coli with a phytase activity of 1.15 U/mL and specific activity of 0.92 U/mg proteins. The expressed enzyme represented a full length ORF "PhyPB13" of 381 amino acid residues and differs by 3 residues from the closest similar existing BPPhy sequences. The PhyPB13 sequence was characterized in silico using various bioinformatic tools to better understand structural, functional, and evolutionary aspects of BPPhy class by multiple sequence alignment and homology search, phylogenetic tree construction, variation in biochemical features, and distribution of motifs and superfamilies. In all sequences, conserved sites were observed toward their N-terminus and C-terminus. Cysteine was not present in the sequence. Overall, three major clusters were observed in phylogenetic tree with variation in biophysical characteristics. A total of 10 motifs were reported with motif "1" observed in all 44 protein sequences and might be used for diversity and expression analysis of BPPhy enzymes. This study revealed important sequence features of BPPhy and pave a way for determining catalytic mechanism and selection of phytase with desirable characteristics.
authors.affiliation: Department of Biochemistry, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar 263145, India ; Akal School of Biotechnology, Eternal University, Baru Sahib, Sirmour 173101, India.
authors.forename: Vinod
authors.identifier: 0000-0002-1583-7749
authors.initials: V
authors.lastname: Kumar
authors.affiliation: Department of Biochemistry, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar 263145, India.
authors.forename: Gopal
authors.identifier: 0000-0002-2113-4230
authors.initials: G
authors.lastname: Singh
authors.affiliation: Department of Biochemistry, C. C. S. Haryana Agricultural University, Hisar 125001, India.
authors.forename: Punesh
authors.initials: P
authors.lastname: Sangwan
authors.affiliation: Department of Biochemistry, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar 263145, India.
authors.forename: A K
authors.initials: AK
authors.lastname: Verma
authors.affiliation: Department of Biochemistry, College of Basic Sciences and Humanities, G. B. Pant University of Agriculture and Technology, Pantnagar 263145, India.
authors.forename: Sanjeev
authors.initials: S
authors.lastname: Agrawal