Separation of phytic acid and other related inositol phosphates by high-performance ion chromatography and its applications

Chen, Qingchuan; Li, Betty W.

doi:10.1016/j.chroma.2003.08.040

Cited by 72 publications

(83 citation statements)

References 35 publications

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“…As shown, there is one major and two minor degradation pathways for wheat phytase: the major pathway proceeds via d-I(1,2,3,5,6)P 5 , d-I(1,2,3,6)P 4 , and d-I(1,2,5,6)P 4 , I(1,2,3)P 3 -d-I(1,2,6)P 3 , d/l-I(1,2)P 2 , and finally into I(2)P 1 and d/l-I(1)P 1 . This pathway of degradation is consistent with published results (Chen and Li, 2003;Nakano et al, 2000). One minor route proceeds via d/l-I(1,2,4,5,6)P 5, d/l-I(1,2,5,6)P 4 , d/l-I(1,2,6)P 3 , d/l-I(1,2)P 2 , I(2)P 1 , and then to d/l-I(1)P 1 , similar to our previous study (Wu et al, 2015).…”

Section: Major and Minor Pathways Of Phytate Degradationsupporting

confidence: 82%

“…High-performance ion chromatography, the most appropriate technique to separate various inositol phosphate isomers, was used following the method of Chen and Li (2003). Briefly, a 1-mL collected sample was freeze-dried and redissolved in 0.5 mL of H 2 O, filtered through a 0.2-mm filter disk, and then analyzed immediately.…”

Section: High-performance Ion Chromatography Analysismentioning

confidence: 99%

“…After post-column reaction with an Fe solution, the separated components were detected at 295 nm. An in-house reference standard was also prepared according to the Chen and Li (2003) method to identify different IP x . Furthermore, a series of commercial IP x standards (see above) was used to identity and quantify the degradation products.…”

Section: High-performance Ion Chromatography Analysismentioning

confidence: 99%

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Phytate Degradation by Different Phosphohydrolase Enzymes: Contrasting Kinetics, Decay Rates, Pathways, and Isotope Effects

Sun

Alikhani

Massoudieh

et al. 2017

Soil Science Soc of Amer J

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Phytate (IP 6 ) is often the most common organic P compound particularly in agricultural soils. understanding the fate of inositol phosphate (IP x ) in the environment in terms of isomeric composition and concentration and assessing relative resistance to (or preference for) degradation is essential to estimate the potential role of IP x in generating inorganic P (P i ) as well as overall P cycling in the environment. In this study, we analyzed IP 6 degradation by four common phosphohydrolase enzymes (phytase from wheat [Triticum aestivum l.] and Aspergillus niger and acid phosphatase from wheat germ and potato [Solanum tuberosum l.]), with particular focus on degradation pathways, isomer kinetic decay rate, and isotope effects using a combination of high-performance ion chromatography, nuclear magnetic resonance, stable isotopes, and process-based modeling techniques. our results show that the degradation pathways are often distinct among enzymes. The process-based Bayesian inverse modeling was used to capture the trend and magnitude of the measured concentrations for each IP x isomer and to determine the decay constants. Furthermore, o isotope ratios (d 18 o P ) of released P i enabled the identification of isotopically identical phosphate moieties in phytate derived from natural sources. Distinctly different fractionation factors, degradation pathways, and kinetic decay rate coefficients among the enzymes studied could lead to potential discrimination and tracking of phytate sources and products as well as active enzymes present in the environment.Abbreviations: HPIC, high-performance ion chromatography; IP, inositol phosphate; NMR, nuclear magnetic resonance; P i , inorganic phosphorus; pNPP, para-nitrophenyl phosphate.I nositol phosphates are a group of organic P compounds widely present in the natural environment (Turner et al., 2002). Phytate (the salt of myo-inositol 1,2,3,4,5,6-hexakisphosphate or IP 6 ) is a P storage molecule in cereals and grains and represents between 60 and 80% of P in mature seeds (Raboy, 1997). Since it is reported that ?51 million tonnes of phytate is formed in commercially produced fruits and crop seeds every year (Lott et al., 2000), a good fraction of phytate in seeds and grains is released to the soil environment as plant residues and animal manures (Dao, 2007;Gerke, 2015). Phytate readily sorbs onto minerals or precipitates with soil cations and organic matter, and then accumulates to constitute an often dominant class of organic P (Celi and Barberis, 2007;Giles and Cade-Menun, 2014). Although sorption and precipitation immobilizes a large fraction of phytate in soil, there is a potential for its transfer to water bodies with soil particulates and colloids (Turner et al., 2002;Turner and Newman, 2005 Core Ideas• Phytate is degraded through distinct pathways for a particular enzyme.• oxygen isotope ratios of phosphate moieties in phytate are isotopically identical.• These findings bring new insights into tracking phytate sources in the environment.

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Section: Major and Minor Pathways Of Phytate Degradationsupporting

confidence: 82%

Section: High-performance Ion Chromatography Analysismentioning

confidence: 99%

Section: High-performance Ion Chromatography Analysismentioning

confidence: 99%

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Phytate Degradation by Different Phosphohydrolase Enzymes: Contrasting Kinetics, Decay Rates, Pathways, and Isotope Effects

Sun

Alikhani

Massoudieh

et al. 2017

Soil Science Soc of Amer J

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“…For the phytase of Aspergillus niger var. ficuum a dual phytate dephosphorylation pathway resulting in the generation of two myo-inositol trisphosphates was reported (4). The appearance of D-Ins(1,5,6)P 3 as one of the myo-inositol trisphosphate intermediates requires removal of the phosphate residue at position 2 of the myo-inositol ring.…”

Section: Introductionmentioning

confidence: 99%

“…Individual myo-inositol phosphate esters have been shown to have important physiological functions in man (22). Some of these compounds, in particular D-Ins(1,4,5)P 3 and D-Ins(1,3,4,5)P 4 , have been demonstrated to play an important role as intracellular second messengers (22), and several isomers of myo-inositol phosphates have shown important pharmacological effects, such as prevention of diabetes complications and antiinflammatory effects (2,5) as well as antiangiogenic and antitumour effects (17). In addition, dietary myo-inositol phosphates have been suggested to bring about benefits for human health, such as amelioration of heart disease conditions by controlling hypercholesterolemia and atheriosclerosis (14), prevention of renal stone formation (8), and protection against a variety of cancers, in particular colon cancer (31).…”

Section: Introductionmentioning

confidence: 99%

Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9

Greiner¹,

Silva

Couri

2009

Braz. J. Microbiol.

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An extracellular phytase from Aspergillus niger 11T53A9 was purified about 51-fold to apparent homogeneity with a recovery of 20.3% referred to the phytase activity in the crude extract. Purification was achieved by ammonium sulphate precipitation, ion chromataography and gel filtration. The purified enzyme behaved as a monomeric protein with a molecular mass of about 85 kDa and exhibited maximal phytate-degrading activity at pH 5.0. Optimum temperature for the degradation of phytate was 55°C. The kinetic parameters for the hydrolysis of sodium phytate were determined to be K M = 54 µmol l -1 and k cat = 190 sec -1 at pH 5.0 and 37°C.The purified enzyme was rather specific for phytate dephosphorylation. It was shown that the phytase preferably dephosphorylates myo-inositol hexakisphosphate in a stereospecific way by sequential removal of phosphate groups via D-Ins(1,2,4,5,6)P 5 , D-Ins(1,2,5,6)P 4 , D-Ins(1,2,6)P 3 , D-Ins(1,2)P 2 to finally Ins(2)P.

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Interaction of phytate with protein and minerals in a soybean–maize meal blend depends on pH and calcium addition

Pontoppidan¹,

Pettersson²,

Sandberg³

2007

J Sci Food Agric

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In order to investigate possible interactions of phytate with protein and minerals in simplified animal diets, studies were conducted on the solubility of endogenous phytate, protein and essential minerals in a soybean-maize meal blend within a physiological relevant pH range. The blend was mixed with water for 10 min and then allowed to incubate at 40• C (30 min) after adjustment of the pH. Finally, soluble phytate, protein, zinc, manganese and iron were determined. Phytate and mineral solubility was highly influenced by pH whereas protein solubility was less affected. Addition of 5 g Ca 2+ kg −1 drastically reduced the solubility of phytate, zinc, manganese and iron at pH above 4.4, indicating that the formation of insoluble phytate-mineral complexes is increased in the presence of calcium. The action of pepsin increased the solubility of protein and phytate at pH below 4, indicating that insoluble phytate-protein complexes are present at low pH. Calcium had the same solubilising effect as pepsin at pH 2-4 but to a lesser degree.

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Separation of phytic acid and other related inositol phosphates by high-performance ion chromatography and its applications

Cited by 72 publications

References 35 publications

Phytate Degradation by Different Phosphohydrolase Enzymes: Contrasting Kinetics, Decay Rates, Pathways, and Isotope Effects

Phytate Degradation by Different Phosphohydrolase Enzymes: Contrasting Kinetics, Decay Rates, Pathways, and Isotope Effects

Purification and characterisation of an extracellular phytase from Aspergillus niger 11T53A9

Interaction of phytate with protein and minerals in a soybean–maize meal blend depends on pH and calcium addition

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