Purification and characterization of a novel neutral and heat-tolerant phytase from a newly isolated strain Bacillus nealsonii ZJ0702.
BACKGROUND
Phytic acid and phytates can interact with biomolecules, such as proteins and carbohydrates, and are anti-nutritional factors found in food and feed. Therefore, it is necessary to remove these compounds in food and feed processing. Phytase can hydrolyze phytic acid and phytates to release a series of lower phosphate esters of myoinositol and orthophosphate. Thus, the purification and characterization of novel phytases that can be used in food and feed processing is of particular interest to the food and feed industries.


RESULTS
A novel neutral and heat-tolerant phytase from a newly isolated strain Bacillus nealsonii ZJ0702 was purified to homogeneity with a yield of 5.7% and a purification fold of 44. The molecular weight of the purified phytase obtained by SDS-PAGE was 43 kDa. The homology analysis based on N-terminal amino acid and DNA sequencing indicated that the purified phytase was different from other known phytases. The optimal thermal and pH activity of the phytase was observed at 55°C and 7.5, respectively. Seventy-three percent of the original activity of the phytase was maintained following incubation at 90°C for 10 min. The phytase was stable within a pH range of 6.0-8.0 and showed high substrate specificity for sodium phytate. Cu²⁺, Co²⁺, Zn²⁺, Mn²⁺, Ba²⁺ and Ni²⁺ ions were found to inhibit the activity of the phytase.


CONCLUSIONS
A novel phytase purified from B. nealsonii ZJ0702 was identified. The phytase was found to be thermally stable over a wide temperature range at neutral pH. These properties suggest that this phytase is a suitable alternative to fungal phytases for the hydrolysis of phytic acid and phytates in food and feed processing industries.
College of Food Science and Biotechnology, Zhejiang Gongshang University, 149 Jiaogong Road, Hangzhou 310035, Zhejiang Province, PR China. yup9202@gmail.com.
Ping
P
Yu
Yirun
Y
Chen
D010832:6-Phytase / Q000737:chemistry / Q000302:isolation & purification*; D001407:Bacillus / Q000145:classification / Q000201:enzymology*; D004591:Electrophoresis, Polyacrylamide Gel; D006358:Hot Temperature; D006863:Hydrogen-Ion Concentration; D008970:Molecular Weight; D010710:Phosphates / Q000032:analysis; D010833:Phytic Acid / Q000032:analysis; D017422:Sequence Analysis, DNA; D013379:Substrate Specificity
D010710:Phosphates; D010833:Phytic Acid; D010832:6-Phytase