1 Amino acid sequences of phytase from Pseudomonas syringae and gene sequences of it thereof

摘要

PURPOSE: Amino acid sequences of phytase derived from Pseudomonas syringae MOK1 and nucleotide sequences encoding the same are provided, thereby hydrolyzing phytate in plant fodder to separate inorganic phosphate, improving the use of phosphate, and decreasing the amount of phosphate in the feces of domestic animals in order to minimize environmental pollution. CONSTITUTION: Phytase derived from Pseudomonas syringae MOK1 (KFCC-11192) has the amino acid sequence of SEQ ID NO: 2. A gene encoding the phytase derived from Pseudomonas syringae MOK1 (KFCC-11192) has the nucleotide sequence of SEQ ID NO: 1. A method for sequencing the amino acid sequence of the phytase derived from Pseudomonas syringae MOK1 (KFCC-11192) comprises the steps of: extracting and purifying total DNA from Pseudomonas syringae MOK1 (KFCC-11192); analyzing the internal amino acid sequence from the extracted and purified MOK1 phytase; constructing degenerate primers A and B using the N-terminal and internal amino acid sequence of the MOK1 phytase; carrying out PCR using the degenerate primers A and B to obtain a DNA fragment having 691 kb; cloning the 691 kb DNA fragment to pGEM-T easy vector and sequencing it; constructing inverse PCR primers A and B; and carrying out inverse PCR using the inverse PCR primers A and B to obtain the total amino acid sequence of the MOK1.