Solid state fermentation of de-oiled rice bran: Effect on in vitro protein digestibility, fatty acid profile and anti-nutritional factors.

An experiment was conducted to study the effect of solid state fermentation of de-oiled rice bran (DORB) with Rhizopus oryzae on in vitro protein digestibility, anti-nutritional factors and fatty acid profile. The fermentation of DORB with Rhizopus oryzae significantly reduced the in vitro protein digestibility of DORB (p < .05). The fermentation of DORB with Rhizopus oryzae increased the saturated fatty acid (SFA) content by 46.83%, while decreased the MUFA and PUFA contents by 14.01 and 8.76%, respectively. The n-6 fatty acid content of FDORB increased by 6.19%, while n-3 fatty acid content decreased by 53.92%. The fermentation of DORB resulted in significant reduction in phytate and trypsin inhibitor activity (p < .05). Based on the present result it is concluded that the fermentation of DORB with Rhizopus oryzae improves the n-6 fatty acid profile and brings significant reduction in the phyate and trypsin inhibitor content but fails to improve the in vitro protein digestibility and hence cannot be recommended as a suitable microbe for DORB fermentation.

ß-propeller phytase hydrolyzes insoluble Ca(2+)-phytate salts and completely abrogates the ability of phytate to chelate metal ions.

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 Ca(2+)-binding sites and forms insoluble tricalcium- or tetracalcium-phytate salts over a wide pH range (pH 3.0-9.0). We overexpressed the ß-propeller phytase from Hahella chejuensis (HcBPP) that hydrolyzes insoluble Ca(2+)-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 Ca(2+)-phytate salts. Biochemical and kinetic studies further confirmed that HcBPP preferentially recognizes its substrate and selectively hydrolyzes insoluble Ca(2+)-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.

Value addition to finger millet (Eleusine coracana) by germination and fermentation with Monascus purpureus.

The present investigation has been carried out to highlight the importance of germination and fermentation of finger millet with Monascus purpureus. Finger millet was subjected to (i) germination, (ii) to fermentation with M. purpureus, and (iii) germination followed by fermentation with M. purpureus. The results of this experiment suggest that the germinated (72 h) finger millet fermented (10 days) with M. purpureus showed reduction in phytic acid and tannin contents by 88.8% and 90.1%, respectively, with an increase of 61.5% HCl-extractable minerals, reducing sugars and soluble proteins thereby supporting the production of antihypercholesterolemic metabolite, statin.

Inhibition of PCR amplification by phytic acid, and treatment of bovine fecal specimens with phytase to reduce inhibition.

Development of effective polymerase chain reaction (PCR)-based diagnostic tests using ruminant fecal specimens has been thwarted by excessive inhibition. A PCR system based on amplification of 1000 copies of bacteriophage lambda-DNA was used as a model to evaluate inhibition levels in bovine feces. Dilution experiments using a bovine fecal specimen suggested that as little as 40 microg of feces (in a 100-microl PCR) affected the efficiency of amplification. It was discovered that phytic acid (the hexaphosphoric ester of inositol) is a powerful inhibitor of PCR. Above 0.3 mM phytate, the PCR is completely inhibited. In a very narrow range around 0.2 mM target-specific amplification proceeds efficiently. At concentrations between 10 and 100 microM, phytate nonspecific amplification (e.g., primer-dimer formation) is dominant. Below 10 microM, phytate target-specific amplification proceeds efficiently. A simple processing procedure using 50 units/ml of Aspergillus niger 3-phytase [E.C. 3.1.3.8] was developed that reduced PCR inhibition levels in bovine fecal specimens by approximately 500-fold.

Beta-carotene and inhibitors of iron absorption modify iron uptake by Caco-2 cells.

A National fortification program instituted in Venezuela in 1993 reduced iron deficiency and anemia by half in only 1 y. The fortification mixture contained ferrous fumarate, vitamin A and other vitamins. We conducted experiments to characterize ferrous fumarate uptake by Caco-2 cells. Increasing amounts of ferrous fumarate, vitamin A, phytate, tannic acid and beta-carotene were added to incubation mixtures using a range of concentrations that included the molar ratios used in the Venezuelan fortification program. Cells were incubated for 1 h at 37 degrees C with 37 kBq (59)Fe and the compound to be evaluated. They were then rinsed, trypsinized and counted to measure uptake. Effects of ascorbic acid, days in culture and use of flasks or inserts were also evaluated. Optimal conditions for uptake experiments were pH 5.5, in the presence of ascorbic acid and at 16 d in culture. Use of flasks or inserts did not affect uptake. Vitamin A did not significantly increase iron uptake under the experimental conditions employed. However, beta-carotene (6 micromol/L) significantly increased iron uptake compared to no beta-carotene addition (114.9 +/- 6.3 and 47.2 +/- 5.9 pmol/mg cell protein, respectively). Moreover, in the presence of phytates or tannic acid, beta-carotene generally overcame the inhibitory effects of both compounds depending on their concentrations. We conclude that beta-carotene improves iron uptake and overcomes the inhibition by potent inhibitors of iron absorption. These experiments also validated the usefulness of Caco-2 cell model system to evaluate iron metabolism.

Iron absorption in man: ascorbic acid and dose-dependent inhibition by phytate.

The dose-dependent inhibitory effect of sodium phytate on iron absorption was studied in man by serving wheat rolls containing no phytates and rolls to which various amounts (seven dose levels between 2 and 250 mg expressed as phytate phosphorus) were added just before serving. Fe in the two kinds of rolls was labeled with two radioisotopes of Fe (55Fe, 59Fe) and the rolls were served on alternate days. The inhibition of Fe absorption was strongly related to the amount of phytate added; 2 mg inhibited absorption by 18%, (p less than 0.001), 25 mg by 64% (p less than 0.001), and 250 mg by 82% (p less than 0.001). The addition of ascorbic acid significantly counteracted the inhibition whereas the corresponding effect of meat was less well defined and only seen at the highest phytate level. The marked inhibition of Fe absorption by phytates and the significant counteracting effect of ascorbic acid have wide nutritional implications.

Regulation of avian erythrocyte AMP-deaminase.

1. Kinetic data for avian erythrocyte AMP-deaminase in lysate supernatants and 2000-fold purified enzyme were consistent with an allosteric model having four binding sites for substrate. 2. Relative to the purified enzyme, AMP-deaminase in lysate supernatants exhibited a greater S0.5 and enhanced sensitivity toward phytic acid, but was far less sensitive toward potassium ion. 3. In the absence of potassium chloride, the enzymatic activity in lysates exhibited hysteresis at subsaturating 5'-AMP. This response was modified reversibly by allosteric ligands. 4. It is concluded that the characteristics of avian RBC AMP-deaminase, as expressed in lysates, may reflect important intermolecular interactions and better represent the regulatory properties of this enzyme in erythrocytes.