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Ultrasonics Sonochemistry Jun 2024This study aimed to elucidate the impact of ultrasound-assisted cellulase (UC) pretreatment on nutrients, phytic acid, and the bioavailability of phenolics during brown...
This study aimed to elucidate the impact of ultrasound-assisted cellulase (UC) pretreatment on nutrients, phytic acid, and the bioavailability of phenolics during brown rice sprouting. It sought to unveil the underlying mechanisms by quantifying the activity of key enzymes implicated in these processes. The sprouted brown rice (SBR) surface structure was harmed by the UC pretreatment, which also increased the amount of γ-oryzanol and antioxidant activity in the SBR. Concurrently, the UC pretreatment boosted the activity of phytase, glutamate decarboxylase, succinate semialdehyde dehydrogenase, Gamma-aminobutyric acid (GABA) transaminase, chalcone isomerase, and phenylalanine ammonia lyase, thereby decreasing the phytic acid content and increasing the GABA, flavonoid, and phenolic content in SBR. In addition, UC-pretreated SBR showed increased phenolic release and bioaccessibility during in vitro digestion when compared to the treated group. These findings might offer theoretical direction for using SBR to maximize value.
Topics: Oryza; Phenols; Phytic Acid; Cellulase; Ultrasonic Waves; Antioxidants; Nutrients; Biological Availability
PubMed: 38669797
DOI: 10.1016/j.ultsonch.2024.106878 -
Biochemistry May 2024As a key component for NADPH oxidase 2 (NOX2) activation, the peripheral membrane protein p47 translocates a cytosolic activating complex to the membrane through its PX...
As a key component for NADPH oxidase 2 (NOX2) activation, the peripheral membrane protein p47 translocates a cytosolic activating complex to the membrane through its PX domain. This study elucidates a potential regulatory mechanism of p47 recruitment and NOX2 activation by inositol hexaphosphate (IP6). Through NMR, fluorescence polarization, and FRET experimental results, IP6 is shown to be capable of breaking the lipid binding and membrane anchoring events of p47-PX with low micromolar potency. Other phosphorylated inositol species such as IP5(1,3,4,5,6), IP4(1,3,4,5), and IP3(1,3,4) show weaker binding and no ability to inhibit lipid interactions in physiological concentration ranges. The low micromolar potency of IP6 inhibition of the p47 membrane anchoring suggests that physiologically relevant concentrations of IP6 serve as regulators, as seen in other membrane anchoring domains. The PX domain of p47 is known to be promiscuous to a variety of phosphatidylinositol phosphate (PIP) lipids, and this regulation may help target the domain only to the membranes most highly enriched with the highest affinity PIPs, such as the phagosomal membrane, while preventing aberrant binding to other membranes with high and heterogeneous PIP content, such as the plasma membrane. This study provides insight into a potential novel regulatory mechanism behind NOX2 activation and reveals a role for small-molecule regulation in this important NOX2 activator.
Topics: Phytic Acid; NADPH Oxidases; Humans; Cell Membrane; NADPH Oxidase 2; Phosphatidylinositol Phosphates
PubMed: 38669178
DOI: 10.1021/acs.biochem.4c00117 -
Current Issues in Molecular Biology Apr 2024phytase (AppA) is widely used as an exogenous enzyme in monogastric animal feed mainly because of its ability to degrade phytic acid or its salt (phytate), a natural...
phytase (AppA) is widely used as an exogenous enzyme in monogastric animal feed mainly because of its ability to degrade phytic acid or its salt (phytate), a natural source of phosphorus. Currently, successful recombinant production of soluble AppA has been achieved by gene overexpression using both bacterial and yeast systems. However, some methods for the biomembrane immobilization of phytases (including AppA), such as surface display on yeast cells and bacterial spores, have been investigated to avoid expensive enzyme purification processes. This study explored a homologous protein production approach for displaying AppA on the cell surface of by engineering its outer membrane (OM) for extracellular expression. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of total bacterial lysates and immunofluorescence microscopy of non-permeabilized cells revealed protein expression, whereas activity assays using whole cells or OM fractions indicated functional enzyme display, as evidenced by consistent hydrolytic rates on typical substrates (i.e., p-nitrophenyl phosphate and phytic acid). Furthermore, the in vitro results obtained using a simple method to simulate the gastrointestinal tract of poultry suggest that the whole-cell biocatalyst has potential as a feed additive. Overall, our findings support the notion that biomembrane-immobilized enzymes are reliable for the hydrolysis of poorly digestible substrates relevant to animal nutrition.
PubMed: 38666945
DOI: 10.3390/cimb46040215 -
Animal : An International Journal of... May 2024There is a gap in the understanding of the relationship between dietary phytate levels and the relative efficacy of phytase to improve amino acid (AA) digestibility in...
There is a gap in the understanding of the relationship between dietary phytate levels and the relative efficacy of phytase to improve amino acid (AA) digestibility in pigs and chickens. Two experiments were conducted to investigate the effect of exogenous phytase on standardized ileal digestibility (SID) of AA and the apparent ileal digestibility (AID) of P in both standard- (SP) and high-phytate (HP) diets for broilers and swine. There were either 40 cages of Cobb 500 male broilers or 10 crossbred barrows (35 kg) fitted with ileal T-cannulas. Both studies were allotted to five dietary treatments (8 replicates). Treatments consisted of four corn-soybean meal-based diets arranged in a 2 × 2 factorial of standard or high phytate and exogenous phytase at 0 or 1 000 phytase units (FYT)/kg; and one N-free diet. Birds were fed a common starter diet from d 0 to 20 and fed experimental diets from d 20 to 25. Birds were euthanized on d 25 via CO asphyxiation, and digesta were collected from the terminal ileum. Pigs were fed for a total of four 7-d periods, where digesta were collected on d 6 and 7 of each period. Diet and digesta samples were analyzed for DM, N, Ti, AA, and P to determine AA and P digestibility. The SID of AA was determined by correcting the AID of AA for the basal endogenous losses estimated using the N-free diet. Main effects of the diet type (standard or HP) and phytase (0 or 1 000 FYT/kg), and the interaction of diet type and phytase were evaluated. For both experiments, the HP diets produced lower SID of AA compared to the SP (P < 0.001). For broilers, there was a phytase effect (P < 0.001) for the SID of all AAs evaluated regardless of the diet type. For pigs, phytase improved (P < 0.05) the SID of Met, Lys, Cys, Glu and Ser and tended to improve (P < 0.10) Arg, Leu, Thr, and Tyr. There were no significant interactions for either experiment. For both experiments, AID of P was lower for the HP diets (P < 0.01), and phytase produced greater AID of P for both diet types (P < 0.01). These data indicate that phytase greatly improves the digestibility of P for broilers and pigs and has the ability to significantly increase the digestibility of amino acids for these animals, regardless of the dietary phytate P.
Topics: Animals; 6-Phytase; Chickens; Animal Feed; Phytic Acid; Male; Digestion; Diet; Animal Nutritional Physiological Phenomena; Ileum; Swine; Amino Acids; Dietary Supplements
PubMed: 38636148
DOI: 10.1016/j.animal.2024.101135 -
Food Chemistry Aug 2024Lentils have a valuable physicochemical profile, which can be affected by the presence of antinutrients that may impair the benefits arising from their consumption....
Lentils have a valuable physicochemical profile, which can be affected by the presence of antinutrients that may impair the benefits arising from their consumption. Different treatments can be used to reduce these undesirable compounds, although they can also affect the general composition and behaviour of the lentils. Thus, the effect of different processing methods on the physicochemical and techno-functional properties, as well as on the antinutritional factors of different lentil varieties was studied. Phytic acid was eliminated during germination, while tannins and trypsin inhibitors are mostly affected by cooking. Functional properties were also altered by processing, these being dependent on the concentration of different nutrients in lentils. All the studied treatments affected the physicochemical profile of lentils and their functional properties. Cooking and germination appear to be the most effective in reducing antinutritional factors and improving the physicochemical profile of the lentils, meeting the current nutritional demands of today's society.
Topics: Lens Plant; Seeds; Germination; Cooking; Nutritive Value; Phytic Acid; Tannins; Trypsin Inhibitors; Food Handling
PubMed: 38631207
DOI: 10.1016/j.foodchem.2024.139293 -
Food Chemistry: X Jun 2024The purpose of the study was to explore effect of four different strains on quality characteristics of soy yogurt. The results showed that four strains were all related...
The purpose of the study was to explore effect of four different strains on quality characteristics of soy yogurt. The results showed that four strains were all related to the genus and N1 was , N2 was , N3 was , and N4 was . The result analysis of biochemical, sensory, nutritional, functional and safety properties of fermentation process and end products showed that the soy yogurt fermented with L. N1 had the highest isoflavone content and the lowest phytic acid content; the soy yogurt fermented with L. N2 had the highest content of free amino acids and oligosaccharides, the lowest content of trypsin inhibitors; the soy yogurt fermented with N3 had the lowest oil content; the soy yogurt fermented with L. N4 had optimal functional properties. In summary, N4 was suitable as a fermentation strain for soymilk.
PubMed: 38623511
DOI: 10.1016/j.fochx.2024.101359 -
Food Chemistry Aug 2024This study investigated the effect of low-intensity pulsed electric field (PEF) (0.3-0.7 kV/cm) and/or germination (0-72 h, 20 °C) on faba beans prior to flour- and...
Low-intensity pulsed electric field processing prior to germination improves in vitro digestibility of faba bean (Vicia faba L.) flour and its derived products: A case study on legume-enriched wheat bread.
This study investigated the effect of low-intensity pulsed electric field (PEF) (0.3-0.7 kV/cm) and/or germination (0-72 h, 20 °C) on faba beans prior to flour- and breadmaking. PEF (0.5 and 0.7 kV/cm) had no significant effect on the germination performance of faba bean but had a positive effect on in vitro starch and protein hydrolysis of PEF-treated beans germinated for 72 h. The incorporation of flour from soaked, germinated, PEF-treated, and PEF-treated+germinated faba beans into wheat bread, at 30% mass level, improved the nutritional composition (total starch and protein contents) and protein digestibility but it reduced the specific volume and increased the density, brownness, and hardness of the bread. This finding shows for the first time that PEF-treatment (<0.7 kV/cm) of faba beans followed by germination (72 h) improved in vitro starch and protein hydrolysis of its flour and the protein digestibility at gastric phase of its enriched wheat bread.
Topics: Vicia faba; Germination; Flour; Bread; Triticum; Digestion; Food Handling; Seeds; Starch; Electricity; Plant Proteins; Hydrolysis
PubMed: 38615637
DOI: 10.1016/j.foodchem.2024.139321 -
International Journal of Molecular... Mar 2024As compounds of natural origin enter human body, it is necessary to investigate their possible interactions with the metabolism of drugs and xenobiotics in general,...
As compounds of natural origin enter human body, it is necessary to investigate their possible interactions with the metabolism of drugs and xenobiotics in general, namely with the cytochrome P450 (CYP) system. Phytic acid (myo-inositol hexaphosphoric acid, IP6) is mainly present in plants but is also an endogenous compound present in mammalian cells and tissues. It has been shown to exhibit protective effect in many pathological conditions. For this paper, its interaction with CYPs was studied using human liver microsomes, primary human hepatocytes, the HepG2 cell line, and molecular docking. Docking experiments and absorption spectra demonstrated the weak ability of IP6 to interact in the heme active site of CYP1A. Molecular docking suggested that IP6 preferentially binds to the protein surface, whereas binding to the active site of CYP1A2 was found to be less probable. Subsequently, we investigated the ability of IP6 to modulate the metabolism of xenobiotics for both the mRNA expression and enzymatic activity of CYP1A enzymes. Our findings revealed that IP6 can slightly modulate the mRNA levels and enzyme activity of CYP1A. However, thanks to the relatively weak interactions of IP6 with CYPs, the chances of the mechanisms of clinically important drug-drug interactions involving IP6 are low.
Topics: Humans; Animals; Phytic Acid; Molecular Docking Simulation; Xenobiotics; Cytochrome P-450 Enzyme System; RNA, Messenger; Mammals
PubMed: 38612422
DOI: 10.3390/ijms25073610 -
Plants (Basel, Switzerland) Mar 2024Lupin species provide essential nutrients and bioactive compounds. Within pulses, they have one of the highest contents of proteins and fibers and are among the poorest...
Lupin species provide essential nutrients and bioactive compounds. Within pulses, they have one of the highest contents of proteins and fibers and are among the poorest in carbohydrates. The Mediterranean region is an important cradle area of the origin and domestication of cultivated white lupin ( L.). In this work, we present the characterization of 19 white lupin landraces collected from several sites in southern Italy, characterized by different pedoclimatic conditions. The protein contents and electrophoretic patterns, total polyphenols, phytic acid, lipids and phosphorous content, and reducing and anti-tryptic activities have been determined for each landrace. The relationships of the compositional characteristics, the area of origin of landraces and between compositional characteristics and thermo-pluviometric trends that occurred in the genotype comparison field during the two-year period between 2019 and 2020 are compared and discussed. From a nutritional point of view, some of the analyzed landraces differ from the commercial reference. The panel of molecular analyses performed can help in building an identity card for the grain to rapidly identify those varieties with the desired characteristics.
PubMed: 38592821
DOI: 10.3390/plants13060785 -
Journal of Microbiology and... May 2024Phytase increases the availability of phosphate and trace elements by hydrolyzing the phosphomonoester bond in phytate present in animal feed. It is also an important...
Phytase increases the availability of phosphate and trace elements by hydrolyzing the phosphomonoester bond in phytate present in animal feed. It is also an important enzyme from an environmental perspective because it not only promotes the growth of livestocks but also prevents phosphorus contamination released into the environment. Here we present a novel phytase derived from , TmPhy, which has distinctive structure and properties compared to other previously known phytases. TmPhy gene expressed in the system was confirmed to be 41 kDa in size and was used in purified form to evaluate optimal conditions for maximum activity. TmPhy has a dual optimum pH at pH3 and pH6.8 and exhibited the highest activity at 70°C. However, the heat tolerance of the wildtype was not satisfactory for feed application. Therefore, random mutation, disulfide bond introduction, and N-terminal mutation were performed to improve the thermostability of the TmPhy. Random mutation resulted in TmPhyM with about 45% improvement in stability at 60°C. Through further improvements, a total of three mutants were screened and their heat tolerance was evaluated. As a result, we obtained TmPhyMD1 with 46.5% residual activity, TmPhyMD2 with 74.1%, and TmPhyMD3 with 66.8% at 80°C heat treatment without significant loss of or with increased activity.
Topics: 6-Phytase; Enzyme Stability; Hydrogen-Ion Concentration; Hot Temperature; Mutation; Pichia; Temperature; Animal Feed; Kinetics; Recombinant Proteins
PubMed: 38563103
DOI: 10.4014/jmb.2311.11044