β-propeller phytase hydrolyzes insoluble Ca2+-phytate salts and completely abrogates the ability of phytate to chelate metal ions

Ok Hee Kim, Young Ok Kim, Jae Hoon Shim, Yun Shin Jung, Woo Jin Jung, Won Chan Choi, Heeseob Lee, Sang Jun Lee, Kyung Kil Kim, Joong Huck Auh, Hyeonjin Kim, Jung Wan Kim, Tae Kwang Oh, Byung Chul Oh

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32 Citations (Scopus)

Abstract

Phytate is an antinutritional factor that influences the bioavailability of essential minerals by forming complexes with them and converting them into insoluble salts. To further our understanding of the chemistry of phytate's binding interactions with biologically important metal cations, we determined the stoichiometry, affinity, and thermodynamics of these interactions by isothermal titration calorimetry. The results suggest that phytate has multiple Ca2+-binding sites and forms insoluble tricalcium- or tetracalcium-phytate salts over a wide pH range (pH 3.0-9.0). We overexpressed the β-propeller phytase from Hahella chejuensis (HcBPP) that hydrolyzes insoluble Ca2+-phytate salts. Structure-based sequence alignments indicated that the active site of HcBPP may contain multiple calcium-binding sites that provide a favorable electrostatic environment for the binding of Ca2+-phytate salts. Biochemical and kinetic studies further confirmed that HcBPP preferentially recognizes its substrate and selectively hydrolyzes insoluble Ca2+-phytate salts at three phosphate group sites, yielding the final product, myo-inositol 2,4,6-trisphosphate. More importantly, ITC analysis of this final product with several cations revealed that HcBPP efficiently eliminates the ability of phytate to chelate several divalent cations strongly and thereby provides free minerals and phosphate ions as nutrients for the growth of bacteria. Collectively, our results provide significant new insights into the potential application of HcBPP in enhancing the bioavailability and absorption of divalent cations.

Original languageEnglish
Pages (from-to)10216-10227
Number of pages12
JournalBiochemistry
Volume49
Issue number47
DOIs
StatePublished - 30 Nov 2010

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6-Phytase
Phytic Acid
Propellers
Metal ions
Salts
Metals
Ions
Divalent Cations
Biological Availability
Minerals
Cations
Phosphates
Binding Sites
Calorimetry
Sequence Alignment
Static Electricity
Titration
Thermodynamics
Stoichiometry
Nutrients

Cite this

Kim, O. H., Kim, Y. O., Shim, J. H., Jung, Y. S., Jung, W. J., Choi, W. C., ... Oh, B. C. (2010). β-propeller phytase hydrolyzes insoluble Ca2+-phytate salts and completely abrogates the ability of phytate to chelate metal ions. Biochemistry, 49(47), 10216-10227. https://doi.org/10.1021/bi1010249
Kim, Ok Hee ; Kim, Young Ok ; Shim, Jae Hoon ; Jung, Yun Shin ; Jung, Woo Jin ; Choi, Won Chan ; Lee, Heeseob ; Lee, Sang Jun ; Kim, Kyung Kil ; Auh, Joong Huck ; Kim, Hyeonjin ; Kim, Jung Wan ; Oh, Tae Kwang ; Oh, Byung Chul. / β-propeller phytase hydrolyzes insoluble Ca2+-phytate salts and completely abrogates the ability of phytate to chelate metal ions. In: Biochemistry. 2010 ; Vol. 49, No. 47. pp. 10216-10227.
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title = "β-propeller phytase hydrolyzes insoluble Ca2+-phytate salts and completely abrogates the ability of phytate to chelate metal ions",
abstract = "Phytate is an antinutritional factor that influences the bioavailability of essential minerals by forming complexes with them and converting them into insoluble salts. To further our understanding of the chemistry of phytate's binding interactions with biologically important metal cations, we determined the stoichiometry, affinity, and thermodynamics of these interactions by isothermal titration calorimetry. The results suggest that phytate has multiple Ca2+-binding sites and forms insoluble tricalcium- or tetracalcium-phytate salts over a wide pH range (pH 3.0-9.0). We overexpressed the β-propeller phytase from Hahella chejuensis (HcBPP) that hydrolyzes insoluble Ca2+-phytate salts. Structure-based sequence alignments indicated that the active site of HcBPP may contain multiple calcium-binding sites that provide a favorable electrostatic environment for the binding of Ca2+-phytate salts. Biochemical and kinetic studies further confirmed that HcBPP preferentially recognizes its substrate and selectively hydrolyzes insoluble Ca2+-phytate salts at three phosphate group sites, yielding the final product, myo-inositol 2,4,6-trisphosphate. More importantly, ITC analysis of this final product with several cations revealed that HcBPP efficiently eliminates the ability of phytate to chelate several divalent cations strongly and thereby provides free minerals and phosphate ions as nutrients for the growth of bacteria. Collectively, our results provide significant new insights into the potential application of HcBPP in enhancing the bioavailability and absorption of divalent cations.",
author = "Kim, {Ok Hee} and Kim, {Young Ok} and Shim, {Jae Hoon} and Jung, {Yun Shin} and Jung, {Woo Jin} and Choi, {Won Chan} and Heeseob Lee and Lee, {Sang Jun} and Kim, {Kyung Kil} and Auh, {Joong Huck} and Hyeonjin Kim and Kim, {Jung Wan} and Oh, {Tae Kwang} and Oh, {Byung Chul}",
year = "2010",
month = "11",
day = "30",
doi = "10.1021/bi1010249",
language = "English",
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Kim, OH, Kim, YO, Shim, JH, Jung, YS, Jung, WJ, Choi, WC, Lee, H, Lee, SJ, Kim, KK, Auh, JH, Kim, H, Kim, JW, Oh, TK & Oh, BC 2010, 'β-propeller phytase hydrolyzes insoluble Ca2+-phytate salts and completely abrogates the ability of phytate to chelate metal ions', Biochemistry, vol. 49, no. 47, pp. 10216-10227. https://doi.org/10.1021/bi1010249

β-propeller phytase hydrolyzes insoluble Ca2+-phytate salts and completely abrogates the ability of phytate to chelate metal ions. / Kim, Ok Hee; Kim, Young Ok; Shim, Jae Hoon; Jung, Yun Shin; Jung, Woo Jin; Choi, Won Chan; Lee, Heeseob; Lee, Sang Jun; Kim, Kyung Kil; Auh, Joong Huck; Kim, Hyeonjin; Kim, Jung Wan; Oh, Tae Kwang; Oh, Byung Chul.

In: Biochemistry, Vol. 49, No. 47, 30.11.2010, p. 10216-10227.

Research output: Contribution to journalArticleResearchpeer-review

TY - JOUR

T1 - β-propeller phytase hydrolyzes insoluble Ca2+-phytate salts and completely abrogates the ability of phytate to chelate metal ions

AU - Kim, Ok Hee

AU - Kim, Young Ok

AU - Shim, Jae Hoon

AU - Jung, Yun Shin

AU - Jung, Woo Jin

AU - Choi, Won Chan

AU - Lee, Heeseob

AU - Lee, Sang Jun

AU - Kim, Kyung Kil

AU - Auh, Joong Huck

AU - Kim, Hyeonjin

AU - Kim, Jung Wan

AU - Oh, Tae Kwang

AU - Oh, Byung Chul

PY - 2010/11/30

Y1 - 2010/11/30

N2 - Phytate is an antinutritional factor that influences the bioavailability of essential minerals by forming complexes with them and converting them into insoluble salts. To further our understanding of the chemistry of phytate's binding interactions with biologically important metal cations, we determined the stoichiometry, affinity, and thermodynamics of these interactions by isothermal titration calorimetry. The results suggest that phytate has multiple Ca2+-binding sites and forms insoluble tricalcium- or tetracalcium-phytate salts over a wide pH range (pH 3.0-9.0). We overexpressed the β-propeller phytase from Hahella chejuensis (HcBPP) that hydrolyzes insoluble Ca2+-phytate salts. Structure-based sequence alignments indicated that the active site of HcBPP may contain multiple calcium-binding sites that provide a favorable electrostatic environment for the binding of Ca2+-phytate salts. Biochemical and kinetic studies further confirmed that HcBPP preferentially recognizes its substrate and selectively hydrolyzes insoluble Ca2+-phytate salts at three phosphate group sites, yielding the final product, myo-inositol 2,4,6-trisphosphate. More importantly, ITC analysis of this final product with several cations revealed that HcBPP efficiently eliminates the ability of phytate to chelate several divalent cations strongly and thereby provides free minerals and phosphate ions as nutrients for the growth of bacteria. Collectively, our results provide significant new insights into the potential application of HcBPP in enhancing the bioavailability and absorption of divalent cations.

AB - Phytate is an antinutritional factor that influences the bioavailability of essential minerals by forming complexes with them and converting them into insoluble salts. To further our understanding of the chemistry of phytate's binding interactions with biologically important metal cations, we determined the stoichiometry, affinity, and thermodynamics of these interactions by isothermal titration calorimetry. The results suggest that phytate has multiple Ca2+-binding sites and forms insoluble tricalcium- or tetracalcium-phytate salts over a wide pH range (pH 3.0-9.0). We overexpressed the β-propeller phytase from Hahella chejuensis (HcBPP) that hydrolyzes insoluble Ca2+-phytate salts. Structure-based sequence alignments indicated that the active site of HcBPP may contain multiple calcium-binding sites that provide a favorable electrostatic environment for the binding of Ca2+-phytate salts. Biochemical and kinetic studies further confirmed that HcBPP preferentially recognizes its substrate and selectively hydrolyzes insoluble Ca2+-phytate salts at three phosphate group sites, yielding the final product, myo-inositol 2,4,6-trisphosphate. More importantly, ITC analysis of this final product with several cations revealed that HcBPP efficiently eliminates the ability of phytate to chelate several divalent cations strongly and thereby provides free minerals and phosphate ions as nutrients for the growth of bacteria. Collectively, our results provide significant new insights into the potential application of HcBPP in enhancing the bioavailability and absorption of divalent cations.

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DO - 10.1021/bi1010249

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