• Korean Journal of Food Science and Technology
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• v.22 no.7
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• pp.780-785
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• 1990

In order to establish the effective extraction and purification process of rapeseed protein, the extraction solvents were compared with one another ; and the residues of glucosinolate and phytate and the extraction yield of protein, which had been extracted by 1% sodium hexa mata-phosphate(SHMP) and purified through isoelectric precipitation, acid-washing and UF concentration, were investigated. As for the condition for extraction of rapeseed proteins, the solvent of 1% SHMP(pH 8.0) turned out the most appropriate ; so far as the purification process for the elimination of glucosinolate and phytate was concerned, the acid-washing twice or the process of the acid-washing once and UF concentration was considered the most effective. The yield and content of rapeseed protein were 37.1% and 75.3% respectively in the case of the acid-washing twice, 42.1% and 72.4% respectively in the case of the acid-washing once and UF concentraction, Consequently, with the elimination effects of glucosinolate and phytate put into consideration, the process of isoelectric precipitation, acid-washing once(pH 3.5), neutralizing(pH 7.5), UF concentration and then freeze drying proved the most effective purification process.

• Microbiology and Biotechnology Letters
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• v.30 no.1
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• pp.1-7
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• 2002

Phytase from Escherichia coli WC7 was purified from cell extracts and its molecular mass was estimated to be 45 kDa by SDS-PAGE. Its optimum temperature and pH for phytate hydrolysis was 6$0^{\circ}C$ and pH 5.0, respectively. The enzyme was stable up to 6$0^{\circ}C$ and over broad pH range (pH 2-12). The enzyme had higher affinity for sodium phytate than p-nitrophenylphosphate (pNPP). That is, the apparent Km value for sodium phytate and pNPP were $0.15\pm$0.02 mM and 2.82$\pm$0.05 mM, respectively. The gene encoding the phytase was cloned in E. coli XL1-Blue. Sequence analysis showed an open reading frame of 1241 Up encoding a signal peptide (22 aa) and a mature enzyme (410 aa). WC7 phytase was expressed up to 17.5 U/ml in the transformed E. coli XL1-Blue/pUEP, which was 23-fold higher than the activity from wild strain.

• Journal of Food Hygiene and Safety
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• v.22 no.1
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• pp.37-44
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• 2007

The approval of use of certain food-grade phosphates as food additives in a wide variety of meat products greatly stimulated research on the applications of phosphates in foods. Although phosphates have never been classified as antimicrobial agents, a number of investigators have reported that phosphates have antimicrobial activities. Phytic acid is a natural plant inositol hexaphosphate constituting 1-5% of most cereals, nuts, legumes, oil seeds, pollen, and spores. In this study, we investigated antibacterial activities of sodium phytate(SPT), sodium pyrophosphate (SPP), sodium tripolyphosphate (STPP) on Escherichia coli O157:H7 on tryptic soy broth and in beef, pork and chicken. In tryptic soy broth, SPT, SPP and STPP at the concentrations of 0.05, 0.1, and 0.5% effectively inhibited the growth of Escherichia coli O157:H7 in a concentration-dependent manner. The bactericidal activity of SPT was the stronger than that of SPP or STPP at the same concentrations. In addition, the antibacterial effects of SPT, SPP and STPP at the concentrations of 0.05, 0.1, 0.3, and 0.5% on Escherichia coli O157:H7 were also investigated in raw or cooked meats including beef, pork and chicken. SPT, SPP and STPP significantly inhibited the bacterial growth in a dose-dependant manner (p<0.05). The bactericidal effect of SPT was stronger than that of SPP or STPP. The addition of SPT, SPP and STPP in meats increased meat pHs. SPP and STPP also increased the levels of soluble orthophosphate in meats but STP did not. These results indicate that SPT is very effective for inhibition of bacterial growth and that can be used as a muscle food additive for increasing functions of meats.

• Journal of the Korean Society of Food Science and Nutrition
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• v.38 no.7
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• pp.870-878
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• 2009

This study was conducted to select the soybean cultivar to minimize the formation of calcium oxalate, and investigate the ingredients for total oxalate (Ox), phytate ($InsP_6$) and minerals such as calcium (Ca), magnesium (Mg), sodium (Na), zinc (Zn), and potassium (K) in 113 recommended soybean cultivars in Korea. Ca content ranged from 0.586 mg/g in Saealkong to 3.177 mg/g in Daolkong, and Mg content ranged from 0.559 mg/g in Taekwankong to 3.085 mg/g of dry seed in Seonheukkong. The total oxalate content ranged from 1.24 mg/g in Seonheukkong to 3.81 mg/g in Ilmikong, and InsP6 content ranged from 0.43 mg/g in Mailikong to 4.72 mg/g of dry seed in Dagikong. In the cross-correlation analysis for the contents of Ca, Mg, Ox and $InsP_6$, Seonheukkong and Danmi2 were selected to minimize the formation of calcium oxalate because the contents of Ca and $InsP_6$ were much higher than the content of Ox. These cultivars could be useful for producing soy foods beneficial to populations at risk for calcium oxalate kidney stones and for improved mineral bioavailability.

• Journal of Animal Science and Technology
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• v.51 no.2
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• pp.171-176
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• 2009

A bacterial strain producing intracellular phytase was isolated from cultivable soil near cowsheds and identified as a fluorescent Pseudomonas sp. BUN1. The BUN1 phytase, partially purified by cation and anion exchange chromatography, exhibited its optimal activity at $40^{\circ}C$ and pH 5.5. As for substrate specificity, it was very specific for phytate and showed little activity on other phosphorylated conjugates. Its activity was greatly inhibited by metal ions such as $Cu^{2+}$, $Cd^{2+}$, and $Zn^{2+}$. Addition of corn starch to PSM (phytasesynthetic medium) [0.5% sodium phytate, 0.5% $(NH_4)_2SO_4$, 0.5% KCl, 0.01% $MgSO_4\cdot7H_2O$, 0.01% $CaCl_2\cdot2H_2O$, 0.01% NaCl, 0.001% $FeSO_4\cdot7H_2O$, 0.001% $MnSO_4\cdot4H_2O$; pH 6.5] for the phytase production significantly induced its enzyme activity in comparison with other carbon sources tested.

• Journal of Animal Science and Technology
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• v.61 no.2
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• pp.87-97
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• 2019

Phytate induced excessive mineral excretion through poultry litter leads to poor performance and environmental pollution. Exogenous microbial phytase supplementation to poultry diets reduce the environmental excretion of nutrient and improve bird's performance. However, excessive dietary sodium (Na) level may hinder the phytase-mediated phytate hydrolysis and negate the beneficial effects of phytase. Therefore, this experiment was conducted to investigate the effects of different concentration dietary Na on phytase activity and subsequent impact on broiler performance, bone mineralisation and nutrient utilisation. In this study, six experimental diets, consisting of three different levels of Na (1.5, 2.5, or 3.5 g/kg) and two levels of microbial phytase (0 or 500 U/kg) were formulated by using $3{\times}2$ factorial design. The six experimental diets were offered to 360 day-old Ross 306 male chicks for 35 days, where, each experimental diet consisted of 6 replicates groups with 10 birds. Along with growth performance, nutrient utilization, intestinal enzyme activity, dry matter (DM) content of litter and mineral status in bone were analysed. Dietary Na and phytase had no effect on bode weight gain and feed intake. Birds on the low Na diet showed higher (p < 0.05) feed conversion ratio (FCR) than the mid-Na diets. High dietary Na adversely affected (p < 0.001) excreta DM content. Phytase supplementation to the high-Na diet increased (p < 0.01) the litter ammonia content. High dietary Na with phytase supplementation improved ($Na{\times}phytase$, p < 0.05) the AME value and ileal digestibility of Ca and Mg. The total tract retention of Ca, P, and Mg was reduced with high Na diet, which was counteracted by phytase supplementation ($Na{\times}phytase$, p < 0.001). The diets containing mid-level of Na improved (p < 0.001) the function of Na-K-ATPase and Mg-ATPase in the jejunum. The overall results indicate that high dietary Na did not affect phytase activity but influenced the nutrient utilization of birds, which was not reflected in bird overall performance.

• Journal of the Korean Society of Food Science and Nutrition
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• v.24 no.4
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• pp.613-618
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• 1995

This study was carried out to investigate the effect of extracting pH on the components and color of seasame protein concentrate(SPC). The protein contents of SPC by extracted at pH 2.0, 7.0, 9.0 and 11.0 were 60.57, 67.72, 79.50 and 83.44%, respectively. Most of the phytates were removed in SPC extracted at pH 7.0, 9.0 and 11.0, but the phytate content of SPC extracted at pH 2.0 was about the same as that of defatted sesame flour. The highest decrease of phytate was found in SPC extracted at pH 11.0(94.80%). SPC extracted at pH 2.0 contained the highest amount of Ca, Mg, Fe and Zn than those in other SPC prepared, while highest amount of Cu was found in SPC extracted at pH 11.0. Sodium content was similar among all the SPC prepared. SPC extracted at pH 7.0 resulted in brighter clor, but SPC extracted at pH 11.0 showed a little darker in appearance.

• Journal of Microbiology and Biotechnology
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• v.14 no.5
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• pp.1004-1008
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• 2004

An extracellular acid phytase from Pseudomonas fragi Y9451 was purified to homogeneity from the culture supernatant by salting-out, DEAE-Sepharose column chromatography, CM-Sepharose column chromatography, and Sephacryl S-300 gel filtration. The molecular weight of the purified enzyme was estimated to be 74 kDa on gel filtration and 54 kDa and 25 kDa on SDS-PAGE, suggesting that the native enzyme was a heterodimeric protein. The purified enzyme was most active at pH 4.5 and $70^{\circ}C$ and fairly stable from pH 4.0- 6.0. It was specific for phytate and exhibited a $K_{m}$ value of 27 mM (sodium phytate, pH 4.5, $50^{\circ}C$). The phytase activity was strongly inhibited (at maximum by 87%) by $Fe^{3+},\;Cu^{2+},\;Fe^{2+}$, and $Zn^{2+}$ at 5 mM concentration, and greatly inhibited by $Ca^{2+}$ at 10 mM concentration. However, EDTA notably stimulated the phytase activity at 10 mM concentration. With optimum pH and stability, Pseudomonas fragi phytase could be a potential candidate for animal feed applications.

• Journal of Microbiology and Biotechnology
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• v.9 no.2
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• pp.223-226
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• 1999

A bacterial strain producing high level of an extracellular phytase was isolated from cooked rice and identified as a strain of Bacillus sp. and designated as Bacillus sp. KHU-10. Optimum culture conditions were investigated for the maximum productivity of phytase by Bacillus sp. KHU-10. 1.0% Maltose and 1.0% peptone with 0.5% beef extract were the best carbon source and nitrogen source, respectively. The addition of $CaCl_2$, stimulated the enzyme productivity with concentration between 0.01% and 0.2%, in the medium. Although sodium phosphate increased the cell mass, the enzyme activity decreased. Calcium phytate and wheat bran containing phytate did not enhance the enzyme production. Under the optimum medium, the production of the phytase reached the highest level of 0.2 unit/ml after 4 days of incubation.

• Applied Biological Chemistry
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• v.10
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• pp.1-13
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• 1968

Phytin is a salt(mainly calcium and magnesium) of phytic acid and its purity and molecular formula can be determined by assaying the contents of phosporus, calcium and magnesium in phytin. In order to devise a new method for the quantitative analysis of the three elements in phytin, the chelatometric method was developed as follows: 1) As the pretreatment for phytin analysis, it was ashfied st $550{\sim}600^{\circ}C$ in the presence of concentrated nitric acid. This dry process is more accurate than the wet process. 2) Phosphorus, calcium and megnesium were analyzed by the conventional and the new method described here, for the phytin sample decomposed by the dry process. The ashfied phytin solution in hydrochloric acid was partitioned into cation and anion fractions by means of a ration exchange resin. A portion of the ration fraction was adjusted to pH 7.0, followed by readjustment to pH 10 and titrated with standard EDTA solution using the BT [Eriochrome black T] indicator to obtain the combined value of calcium and magnesium. Another portion of the ration fraction was made to pH 7.0, and a small volume of standard EDTA solution was added to it. pH was adjusted to $12{\sim}13$ with 8 N KOH and it was titrate by a standard EDTA solution in the presence of N-N[2-Hydroxy-1-(2-hydroxy-4-sulfo-1-naphytate)-3-naphthoic acid] diluted powder indicator in order to obtain the calcium content. Magnesium content was calculated from the difference between the two values. From the anion fraction the magnesium ammonium phosphate precipitate was obtained. The precipitate was dissolved in hydrochloric acid, and a standard EDTA solution was added to it. The solution was adjusted to pH 7.0 and then readjusted to pH 10.0 by a buffer solution and titrated with a standard magnesium sulfate solution in the presence of BT indicator to obtain the phosphorus content. The analytical data for phosphorus, calcium and magnesium were 98.9%, 97.1% and 99.1% respectively, in reference to the theoretical values for the formula $C_6H_6O_{24}P_6Mg_4CaNa_2{\cdot}5H_2O$. Statical analysis indicated a good coincidence of the theoretical and experimental values. On the other hand, the observed values for the three elements by the conventional method were 92.4%, 86.8% and 93.8%, respectively, revealing a remarkable difference from the theoretical. 3) When sodium phytate was admixed with starch and subjected to the analysis of phosphorus, calcium and magnesium by the chelatometric method, their recovery was almost 100% 4) In order to confirm the accuracy of this method, phytic acid was reacted with calcium chloride and magnesium chloride in the molar ratio of phytic: calcium chloride: magnesium chloride=1 : 5 : 20 to obtain sodium phytate containing one calcium atom and four magnesium atoms per molecule of sodium phytate. The analytical data for phosporus, calcium and magnesium were coincident with those as determine d by the aforementioned method. The new method employing the dry process, ion exchange resin and chelatometric assay of phosphorus, calcium and magnesium is considered accurate and rapid for the determination of phytin.