Purification and characterization of a novel neutral and heat-tolerant phytase from a newly isolated strain Bacillus nealsoniiZJ0702

Abstract

BackgroundPhytic 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.ResultsA 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.ConclusionsA 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.

Microbial diversity of mangrove sediment in Shenzhen Bay and gene cloning, characterization of an isolated phytase-producing strain of SPC09 B. cereus

“…Effective phosphate compounds other than phytate are not hydrolyzed by the Bacillus phytases and remain available for plant uptake, particularly in nutrient-limited environments such as mangrove forests (Oh et al 2004;Yu and Chen 2013). The heterologously recombinant Bacillus phytase has been studied by several teams and was shown to exhibit low production levels and high protease activity (Guerrero-Olazarán et al 2010;Kerovuo et al 1998;Rao et al 2008); most of the enzymes originated from B. subtilis, and none were from B. cereus.…”

Purification and characterization of a novel phytase fromNocardiasp. MB 36

“…A purifi ed phytase each from Bacillus laevolacticus ), Shigella sp. CD2 (Roy et al 2012), and B. nealsonii ZJ0702 (Yu and Chen 2013) showed molecular weights of 43 kDa. However, purifi ed Bacillus sp.…”

“…ion exchange, gel fi ltration, hydrophobic interaction chromatography, etc. have widely been used for purifi cation of phytases (Hong et al 2011;Roy et al 2012;Yu and Chen 2013). An alkaline phytase was purifi ed 6.5-fold from Bacillus laevolacticus by DEAE-Sepharose and gel fi ltration chromatography .…”

“…Phytases hydrolyze phytates to simple esters of inositol (I) and phosphates (P), and can fi nd application in improving animal nutrition from phytate-rich animal diets, and for environmental protection against phosphorus-rich manures (Singh and Satyanarayana 2011;Roy et al 2012;Yu and Chen 2013). Phytate, a deposited complex of phytic acid (IP 6 ), constitutes a major storage form of phosphorus in grain, seeds, and cereals (Selle et al 2003).…”

Phytase-Producing Bacteria from Extreme Regions in Indonesia

“…In contrast to the other two phytase subfamilies, ß-propeller phytases need calcium ions for optimal stability and enzymatic activity (Powar and Jagannathan 1982;Shimizu 1992;Kerovuo et al 1998;Kim et al 1998;Choi et al 2001;Oh et al 2001;Park 2001;Shin et al 2001;Gulati et al 2007;Anis Sobirin et al 2009;El-Toukhy et al 2013;Kumar et al 2013;Yu and Chen 2013;Jain and Chauhan 2014). Maximum phytase activity of ß-propeller phytases occurs in the pH range of 6.5 to 8.0 (Choi et al 2001;Gulati et al 2007;Jain and Chauhan 2014;Kerovuo et al 1998;Kim et al 1998;Oh et al 2001;Park 2001;Powar and Jagannathan 1982;Shimizu 1992;Yu and Chen 2013). All the six Bacillus strain included in this study exhibited a broad pH profile.…”