Genome-wide identification, characterization, and expression analysis of MIPS family genes in legume species.

BACKGROUND: Evolutionarily conserved in plants, the enzyme D-myo-inositol-3-phosphate synthase (MIPS; EC 5.5.1.4) regulates the initial, rate-limiting reaction in the phytic acid biosynthetic pathway. They are reported to be transcriptional regulators involved in various physiological functions in the plants, growth, and biotic/abiotic stress responses. Even though the genomes of most legumes are fully sequenced and available, an all-inclusive study of the MIPS family members in legumes is still ongoing. RESULTS: We found 24 MIPS genes in ten legumes: Arachis hypogea, Cicer arietinum, Cajanus cajan, Glycine max, Lablab purpureus, Medicago truncatula, Pisum sativum, Phaseolus vulgaris, Trifolium pratense and Vigna unguiculata. The total number of MIPS genes found in each species ranged from two to three. The MIPS genes were classified into five clades based on their evolutionary relationships with Arabidopsis genes. The structural patterns of intron/exon and the protein motifs that were conserved in each gene were highly group-specific. In legumes, MIPS genes were inconsistently distributed across their genomes. A comparison of genomes and gene sequences showed that this family was subjected to purifying selection and the gene expansion in MIPS family in legumes was mainly caused by segmental duplication. Through quantitative PCR, expression patterns of MIPS in response to various abiotic stresses, in the vegetative tissues of various legumes were studied. Expression pattern shows that MIPS genes control the development and differentiation of various organs, and have significant responses to salinity and drought stress. CONCLUSION: The MIPS genes in the genomes of legumes have been identified, characterized and their expression was analysed. The findings pave way for understanding their molecular functions and evolution, and lead to identify the putative MIPS genes associated with different cell and tissue development.

Soybean glycinin impaired immune function and caused inflammation associated with PKC-ζ/NF-κb and mTORC1 signaling in the intestine of juvenile grass carp (Ctenopharyngodon idella).

Glycinin is a major protein and antinutritional factor of soybean. However, how dietary glycinin affect intestinal immune function of fish were largely unknown. In this study, we used juvenile grass carp as a model to investigate the impacts of glycinin on intestinal immune function of fish and involved mechanisms. We set three treatments including control, glycinin and glycinin + glutamine in this trial. For immune components, results revealed that compared with control group, glycinin group had lower acid phosphatase activities in the foregut, midgut and hindgut, lower C3 and C4 content, and lower mRNA abundances of IgM, IgZ, hepcidin, LEAP-2A, LEAP-2B and ß-defensin-1 in the midgut and hindgut rather than foregut of grass carp. For pro-inflammatory cytokines and relevant signaling, glycinin elevated mRNA abundances of IL-1ß, IL-8, IL-12p35, IL-12p40 and IL-17D in the midgut and IL-1ß, IFN-γ2, IL-6, IL-8, IL-12p35, IL-12p40 and IL-17D in the hindgut, and increased protein abundances of PKC-ζ and nuclear NF-κB p65 in the midgut and hindgut in comparison to control. For anti-inflammatory cytokines and relevant signaling, glycinin reduced mRNA abundances of TGF-ß1, TGF-ß2, IL-4/13B (rather than IL-4/13A), IL-10 and IL-11 in the midgut and hindgut, and reduced p-mTOR (Ser 2448), p-S6K1 (Thr 389) and p-4EBP1 (Thr 37/46) protein abundances in the midgut and hindgut rather than foregut. Co-administration of glutamine with glycinin could partially enhance intestinal function and reduce intestinal inflammation compared with glycinin treatment. Concluded, glycinin decreased intestinal immune components and caused intestinal inflammation associated with PKC-ζ/NF-κB and mTORC1 signaling.

Effects of gossypol from cottonseed cake on the blood profile in sheep.

Cottonseed cake contains gossypol, a potentially toxic compound that, when consumed by sheep, can affect reproduction, the immune system, and the liver. Changes in hematologic and serum biochemical parameters were monitored for 63 days in 12 Santa Inês ewes, six of which received ration containing 400 g kg(-1) of cottonseed cake. Blood samples were collected at the start of the experiment and weekly thereafter for hematologic assessment and determination of serum urea, creatinine, total protein, and albumin concentrations and for measurement of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatase, and γ-glutamyl transferase activities. No clinical signs of toxicity were observed. Evaluation of the erythron showed that sheep consuming cottonseed cake had an increased packed cell volume (p < 0.05) and increased erythrocyte counts and hemoglobin concentrations (p < 0.05) in the leukogram and serum biochemistry panel. In conclusion, consumption of 400 g kg(-1) cottonseed cake by sheep for 63 days may induce changes in the erythron but no consistent changes in serum biochemical parameters, indicating no damage to the liver or kidneys.

Optimization of soybean processing into kinema, a Bacillus-fermented alkaline food, with respect to a minimum level of antinutrients.

AIMS: Optimization of traditional processing of soybeans using response surface methodology (RSM) to achieve a minimum level of antinutritional factors (ANFs) in kinema. METHODS AND RESULTS: Central composite rotatable designs were used to optimize the processing stages of kinema preparation. In each stage, the linear or quadratic effects of independent variables were significant in minimizing ANF levels. The predicted optimum condition for soaking was when the raw beans-water ratio was 1 : 10, and the soaking temperature, time and pH were 10°C, 20 h and 8·0 respectively. Here, tannins content (TC), phytic acid content (PAC) and trypsin inhibitor activity (TIA) decreased (P < 0·05). While haemagglutinating activity (HA) level remained unchanged (P < 0·05), total biogenic amines content (TBAC) increased. The optimum condition for cooking was optimally soaked beans-water ratio of 1 : 5, and cooking pressure and time were 1·10 kg cm(-2) and 20 min respectively. Here, TC, PAC, TIA and HA decreased (P < 0·05), but TBAC remained unchanged compared to optimally soaked beans. TC and HA went below the level of detection. The optimum condition for fermentation was obtained when inoculum load was 10(3) total cells g(-1) grits, and fermentation temperature and time were 37°C and 48 h respectively. Fermentation of optimally cooked beans caused a reduction (P < 0·05) of PAC. While TIA remained unchanged (P < 0·05), TBAC increased. In kinema, TC, PAC, TIA and HA decreased (P < 0·05) over raw beans by 100, 61, 71 and 100% respectively. Good agreement was observed between predicted values and experimental values. CONCLUSIONS: The processing treatments significantly minimized the level of ANFs in soybeans. SIGNIFICANCE AND IMPACT OF THE STUDY: RSM was successfully deployed to obtain the optimum condition for kinema-making with a minimum level of ANFs without impairing sensory attributes of the product. The results are useful for commercial production of kinema.

Bile enhances glucose uptake, reduces permeability, and modulates effects of lectins, trypsin inhibitors and saponins on intestinal tissue.

Antinutritional factors (ANFs) can disrupt digestive and other intestinal functions. ANFs in soybean meal (SBM) are implicated in proliferative and inflammatory responses in the intestine of various (functionally) monogastric animals, including Atlantic salmon (Salmo salar L.). The goal of the current study was to investigate the effect of ex vivo exposure of mid and distal intestinal tissue of salmon to soybean saponins (SAP), lectin (LEC) and Kunitz' trypsin inhibitor (KTI), singly and in combination, on epithelial function, as assessed by measuring in vitro glucose uptake pathways along a glucose concentration gradient. As solubilization of SAP in the calcium-containing Ringer's solution was problematic but resolved with the addition of a physiological concentration of bile collected from the gall bladder of salmon, an evaluation of bile effects became an added element. Results indicated that bile increased baseline glucose absorption and possibly transport, and also had a protective effect on the epithelial barrier, at least partially due to taurocholate. Compared to controls, tissues exposed to LEC+bile, KTI+bile and LEC+KTI+bile exhibited increased glucose uptake at the higher glucose concentrations, apparently due to markedly increased tissue permeability. Addition of SAP, however, attenuated the response, possibly by binding bile components. SAP+bile, also in combination with LEC and/or KTI, as well as LEC, KTI and LEC+KTI without bile often reduced transcellular glucose uptake pathways, while maintaining low tissue permeability. SAP+LEC+KTI+bile, LEC and KTI caused the most marked reductions. The distal intestine was more affected, reflecting the restriction of in vivo SBM-induced inflammatory changes to this region.

Characterization of anti-soybean agglutinin (SBA) IgY antibodies: a new strategy for neutralization of the detrimental biological activity of SBA.

Anti-soybean agglutinin (SBA) IgY was produced, and its potential to neutralize the haemagglutinating activity of SBA in vitro was tested. Thirty-five-week-old hens [treatment (n = 5) and control (n = 5)] were immunized with SBA or injected with saline 4 times every 15 days. Eggs were collected after the last immunization, and IgY was extracted using the polyethylene glycol (PEG) method. Serum anti-SBA IgY titres in immunized hens increased after the first immunization and reached a plateau between days 45 and 60. In contrast, specific IgY titres in the control group remained at basal levels throughout the evaluation. Average IgY titres were significantly higher in the treatment group on days 15, 30, 45, and 60. Total IgY content in the egg yolk extract was 38.7 ± 1.6 and 37.7 ± 1.5 mg/ml for the treatment and control groups, respectively. The specific anti-SBA IgY titer detected in the egg yolk extract was significantly higher (p < 0.001) for hens in the treatment group compared to the control group, with OD450nm values of 0.98 ± 0.05 and 0.058 ± 0.02, respectively. The specificity of anti-SBA IgY was confirmed by the Western blotting, and the inhibition of SBA-induced haemagglutination in vitro was compared with D-galactose, a known molecule that binds to SBA and blocks its binding to erythrocytes. The inhibition of SBA-induced haemagglutination by the anti-SBA IgY reached 512 units of haemagglutination inhibition (UHI), compared to 8 or 256 UHI, respectively, when IgY from control chickens or D-galactose was used. Thus, anti-SBA IgY antibodies were efficiently produced in large quantities and effectively inhibited SBA-induced haemagglutination in vitro.

Effects of fermented unconventional protein feed on pig production in China.

Unconventional protein feeds, characterized by low nutritional value, high variability, and poor palatability, have limited their application in swine production. Fermentation technology holds the key to addressing these shortcomings. Given the ban on antibiotics in China, the inferior quality of imported pig breeds, and long-term dependence on imported soybean, the prospects for fermented unconventional protein feeds are promising. This paper delves into the common types of fermented unconventional protein feeds, factors influencing the fermentation process, the mechanisms by which they enhance swine health, and the challenges and prospects of fermented feeds, offering theoretical insights for the future development of the feed industry.

Extrusion as a tool to enhance the nutritional and bioactive potential of cereal and legume by-products.

Cereal and legume by-products obtained from primary food production industries pose an environmental and economic problem. Nevertheless, these residues can potentially yield value-added products due to their elevated content of dietary fiber, phytochemicals, vitamins, minerals, and residual levels of proteins, which makes them a suitable and heightened option for reutilization in human consumption. Several studies identify extrusion as an innovative technology to modify the technofunctionality and nutritional properties of cereal and legume by-products, resulting in the production of improved ingredients. This review focuses on studies that evaluate the effect of extrusion to improve the nutritional and bioactive potential of cereal and legume by-products. A revision of the extrusion process parameters that improve the profile and bioavailability of dietary fiber, proteins, and phenolic compounds, and minimize antinutritional factors associated to cereal and legume by-products was done. The composition of by-products and process parameters such as feed moisture, barrel temperature and screw speed influence the resulting effect of extrusion. Studies suggest that extruding composite feedstock containing cereal or legume by-products may limit the molecular modifications that trigger the nutritional improvements. Therefore, extrusion applied as a pretreatment represents an interesting and economic alternative to improve the profile and bioavailability of the nutrients found in cereal and legume by-products which might lead to the development of functional ingredients useful to produce foods aimed to prevent chronic diseases.

Nutritional quality and physicochemical properties of biscuit from composite flour of wheat, African yam bean and tigernut.

This study investigated the effect of the supplementation of wheat flour with flour blends of African yam bean (AYB) and tigernut for the production of biscuits and evaluated the sensory characteristics. The composite flours were of ratio 70:25:5 (TWB), 70:20:10 (ATW), 70:15:15 (BTT) for wheat: AYB: tigernut, respectively, while 100 % wheat flour served as control (WTY). The composite flour samples were analyzed for proximate, functional and pasting properties. The physical and chemical properties and sensory attributes of the developed biscuits were carried out. The moisture, protein, fat, ash, crude fiber, carbohydrate, and energy contents of the composite flour ranged from 6.63 to 8.13 %, 11.22-18.36 %, 13.27-19.15 %, 0.98-0.99 %, 3.96-7.43 %, 59.97-62.55 % and 400.89 to 410.40 Kcal/100g, respectively. The results showed that protein fat, ash and crude fiber of the biscuit were improved. The water and oil absorption capacity of composite flour was low while the pasting properties of the composite flour blends reduced as the AYB flour increased. All the composite flour blend biscuit samples possessed high essential nutrients and antioxidant potential. All the biscuits samples were accepted by the panelists, however, sample BTT (70 % wheat flour+15 % AYB flour+ 15 % tigernut flour) was most accepted in appearance, aroma, taste, crispness and overall acceptability. Therefore, biscuits from the flour blends of wheat, AYB and tigernut could be nutritionally beneficial and good for adults.

Label-Free Quantification by Liquid Chromatography-Tandem Mass Spectrometry of the Kunitz Inhibitor of Trypsin KTI3 in Soy Products.

The greater awareness of consumers regarding the sustainability of food chains has shifted part of the consumption from animal protein sources to vegetable sources. Among these, of relevance both for human food use and for animal feed, is soy. However, its high protein content is unfortunately accompanied by the presence of antinutritional factors, including Kunitz's trypsin inhibitor (KTI). Now there are few analytical methods available for its direct quantification, as the inhibitory activity against trypsin is generically measured, which however can be given by many other molecules and undergo numerous interferences. Therefore, in this work, a direct label-free liquid chromatography-mass spectrometry (LC-MS) method for the identification and quantification of trypsin Kunitz inhibitor KTI3 in soybean and derivative products has been developed. The method is based on the identification and quantification of a marker peptide, specific for the protein of interest. Quantification is achieved with an external calibration curve in the matrix, and the limit of detection and the limit of quantification of the method are 0.75 and 2.51 µg/g, respectively. The results of the LC-MS method were also compared with trypsin inhibition measured spectrophotometrically, highlighting the complementarity of these two different pieces of information.