• Journal of Microbiology and Biotechnology
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• v.24 no.10
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• pp.1413-1420
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• 2014

Phytate is an antinutritional factor that impacts the bioavailability of essential minerals such as $Ca^{2+}$, $Mg^{2+}$, $Mn^{2+}$, $Zn^{2+}$, and $Fe^{2+}$ by forming insoluble mineral-phytate salts. These insoluble mineral-phytate salts are hydrolyzed rarely by monogastric animals, because they lack the hydrolyzing phytases and thus excrete the majority of them. The ${\beta}$-propeller phytases (BPPs) hydrolyze these insoluble mineral-phytate salts efficiently. In this study, we cloned a novel BPP gene from a marine Pseudomonas sp. This Pseudomonas BPP gene (PsBPP) had low sequence identity with other known phytases and contained an extra internal repeat domain (residues 24-279) and a typical BPP domain (residues 280-634) at the C-terminus. Structure-based sequence alignment suggested that the N-terminal repeat domain did not possess the active-site residues, whereas the C-terminal BPP domain contained multiple calcium-binding sites, which provide a favorable electrostatic environment for substrate binding and catalytic activity. Thus, we overexpressed the BPP domain from Pseudomonas sp. to potentially hydrolyze insoluble mineral-phytate salts. Purified recombinant PsBPP required $Ca^{2+}$ or $Fe^{2+}$ for phytase activity, indicating that PsBPP hydrolyzes insoluble $Fe^{2+}$-phytate or $Ca^{2+}$-phytate salts. The optimal temperature and pH for the hydrolysis of $Ca^{2+}$-phytate by PsBPP were $50^{\circ}C$ and 6.0, respectively. Biochemical and kinetic studies clearly showed that PsBPP efficiently hydrolyzed $Ca^{2+}$-phytate salts and yielded myo-inositol 2,4,6-trisphosphate and three phosphate groups as final products. Finally, we showed that PsBPP was highly effective for hydrolyzing rice bran with high phytate content. Taken together, our results suggest that PsBPP has great potential in the animal feed industry for reducing phytates.

• Applied Biological Chemistry
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• v.29 no.2
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• pp.212-218
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• 1986

Proteins in Korean rapeseeds, as in many other plantseeds, are usually bound to phytate molecules. These phytate-bound proteins are of little value as foodstuffs because of their poor solubility in digestive systems. Therefore it is necessary to remove phytates from proteins in order to convert these proteins io a useful foodstuff. In the work, an efficient procedure for removal of phytates from defatted Korean rapeseed was found. The influence of pH on the solubility of protein and phytate of rapeseed flour showed that the former was the lowest at pH 5.0 and began to increase as pH further raised. Meanwhile, the latter was the highest at pH 6.0, however, it was decreased abruptly at alkaline pH, especially to content of 1.3% at pH 11.5. The solubility cf protein was relatively high in NaCl aqueous solution at $pH\;6.0{\sim}8.0$, and did not male any noticeable difference depending on NaCl concentration. On the other hand, the solubility of phytate was high at pH of below 6.0 showing an abrupt decrease at pH of above 6.0. The solubility of protein in $CaCl_2$ aqueous solution was highest at $pH\;6.0{\sim}8.0$, however, there was no significant change at the whole range of tested pH of the solution. A maximum solubility of phytate was shown at $pH\;3.0{\sim}4.0$. And it was decreased abruptly at a higher pH of the above range and also decreased at a lower pH with higher $CaCl_2$ concentration. The solubility of phytate in $Na_2SO_3$ aqueous solution was highest at $pH\;5.0{\sim}8.0$. As the concentration goes up the maximum value of solubility was found to move to higher pHs. Depending on the concentration of $Na_2SO_3$, the decreasing pattern was changed in an alkaline solution. The solubility of phytate in the solution containing low concentration of $Ca^{2+}$ ion was low in all treatments at pH of above 7.0 and showed the maximum value at low pH as $Ca^{2+}$ ion concentration increases. The solubility of protein at pH 11.5 showed the highest value in $1mM\;Ca^{2+}$ ion solution.