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http://dx.doi.org/10.4014/jmb.1407.07063

Molecular and Biochemical Characteristics of ${\beta}$-Propeller Phytase from Marine Pseudomonas sp. BS10-3 and Its Potential Application for Animal Feed Additives  

Nam, Seung-Jeung (Lee Gil Ya Cancer and Diabetes Institute, Gachon University Graduate School of Medicine)
Kim, Young-Ok (Biotechnology Research Center, National Fisheries Research and Development Institute)
Ko, Tea-Kyung (Lee Gil Ya Cancer and Diabetes Institute, Gachon University Graduate School of Medicine)
Kang, Jin-Ku (Lee Gil Ya Cancer and Diabetes Institute, Gachon University Graduate School of Medicine)
Chun, Kwang-Hoon (College of Pharmacy, Gachon University)
Auh, Joong-Hyuck (Department of Food Science and Technology, Chung-Ang University)
Lee, Chul-Soon (Lee Gil Ya Cancer and Diabetes Institute, Gachon University Graduate School of Medicine)
Lee, In-Kyu (Department of Biomedical Science, Graduate School of Medicine, Kyungpook National University)
Park, Sunghoon (Lee Gil Ya Cancer and Diabetes Institute, Gachon University Graduate School of Medicine)
Oh, Byung-Chul (Lee Gil Ya Cancer and Diabetes Institute, Gachon University Graduate School of Medicine)
Publication Information
Journal of Microbiology and Biotechnology / v.24, no.10, 2014 , pp. 1413-1420 More about this Journal
Abstract
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.
Keywords
${\beta}$-Propeller phytase; Pseudomonas sp.; $Ca^{2+}$-phytate salts;
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