Nutritional attributes and microbial metagenomic profile during solid-state fermentation of soybean meal inoculated with Lactobacillus acidophilus under non-sterile conditions.

BACKGROUND: Soybean meal (SBM) is used widely in animal feed but it contains anti-nutritional factors (ANFs) such as protease inhibitors - immunogenic proteins that limit its utilization. Fermentative processes could help to reduce these ANFs. The aim of this study was to evaluate the nutritional attributes, bacterial community dynamics, and microbial metagenomic profile during the solid-state fermentation of SBM using a strain of the bacterium Lactobacillus acidophilus with or without pre-autoclaving treatment. RESULTS: Following fermentation, there was a reduction in the pH and a concurrent increase in the population of lactic acid bacteria. Fermentation also resulted in an increase in both crude and soluble protein levels. Trypsin inhibitor levels decreased after fermentation, particularly in fermented SBM that had not been pre-autoclaved, with an inactivation rate higher than 90%. Moreover, high-molecular-weight peptides (44-158 kDa), specifically some polypeptides from the soybean immunogen glycinin and ß-conglycinin, underwent degradation during the fermentation process. Bacterial community analysis revealed the dominance of the Lactobacillus genus in all samples, regardless of the treatments applied. Metagenomic profiling identified L. acidophilus as the dominant species in inoculated SBM, irrespective of whether pre-autoclaving was conducted or not. CONCLUSION: This study demonstrates the feasibility of solid-state fermentation with L. acidophilus under non-sterile conditions to inactivate trypsin inhibitor and increase protein concentration and hydrolysate immunogen proteins into low-molecular-weight peptides in SBM. Lactobacillus acidophilus inoculum also inhibited the growth of undesirable bacteria. This knowledge contributes to our understanding of the potential applications of solid-state fermentation with L. acidophilus in improving the nutritional quality of SBM. © 2024 Society of Chemical Industry.

Bioactive peptides produced by engineered probiotics and other food-grade bacteria: A review.

Synthetic biology is employed for the study and design of engineered microbes with new and improved therapeutic functions. The main advantage of synthetic biology is the selective genetic manipulation of living organisms with desirable beneficial effects such as probiotics. Engineering technologies have contributed to the edition of metabolic processes involved in the mechanisms of action of probiotics, such as the generation of bioactive peptides. Hence, current information related to bioactive peptides, produced by different engineering probiotics, with antimicrobial, antiviral, antidiabetic, and antihypertensive activities, as well as their potential use as functional ingredients, is discussed here. Besides, the effectiveness and safety aspects of these bioactive peptides were also described.

Effect of oral supplementation of Lactobacillus reuteri in reduction of intestinal absorption of aflatoxin B(1) in rats.

The goals of this work were to assess the ability of Lactobacillus reuteri to bind aflatoxin B(1) in the intestinal tract and determine its effect on intestinal absorption of the toxin dispensed in either single or multiple doses in a murine model. Male Wistar rats were used, and two experiments were conducted after bacteria were implanted. Experiment one involved a single-oral dose of toxin, and the subsequent flow cytometric analysis of bacteria isolated from the small intestine and treated with specific FITC-labeled AFB(1) antibodies. The second experiment was carried out supplying the toxin in 7 oral sub-doses, and the later quantification of AFB(1)-Lys adducts in blood samples by ELISA assay. The results demonstrated that L. reuteri was able to bind AFB(1) in the intestinal tract, mostly in the duodenum. Furthermore, the AFB(1)-Lys adducts were present at significantly lower levels in those animals receiving AFB(1) plus bacteria than in those receiving only AFB(1). Our findings confirm that probiotic bacteria could act as biological barriers in normal intestinal conditions thereby reducing the bioavailability of AFB(1) ingested orally in a single or multiple doses, thus avoiding its toxic effects.

[Bioactive peptides derived from milk proteins]. / Péptidos bioactivos derivados de las proteínas de la leche.

Milk proteins are known for having a wide range of nutritional, functional and biological properties that make them important ingredients in functional or health promoting foods. These properties are partly attributed to bioactive peptides coded in the different milk proteins. Bioactive peptides are inactive within the protein sequence but may be released by the action of native proteolitic enzymes from milk, enzymes from lactic acid bacteria or from exogenous sources or may be produced during gastrointestinal digestion or processing of foods. Peptides derived from caseins and whey proteins were shown to present several bioactive properties such as opioid, antihypertensive, antimicrobial, immunodulatory, mineral carrier and antithrombotic. This overview presents a perspective of the importance of dairy proteins in the production of bioactive peptides and their biological activities, as well as the main analytical tecniques that have been used for the isolation and identification of these peptides.

Micropreparative separation, fractionation, and peptide mapping of beta-lactoglobulin A and B variants by capillary electrophoresis.

The methodological aspects for the separation, fractionation, and peptide mapping by free zone capillary electrophoresis (CZE) of beta-lactoglobulin (beta-Lg) variants A and B were established. First, beta-Lg variants A or B were separated and fractionated by CZE. Then, the collected protein fraction was subjected to off-line tryptic digestion. Second, peptide mapping of the tryptic hydrolysates and peptide fraction collection were carried out by CZE. beta-Lg variants were separated and collected using an uncoated capillary (72 cm x 75 microm i.d.) in 0.05 M borate buffer containing 0.1% Tween 20 at pH 8.0 by applying 20 kV. By subjecting the capillary under pressure after a delay time of 15%, the protein was collected in a microvial containing digestion buffer. The most suitable conditions for the tryptic digestion of beta-Lg were established by monitoring the reaction products with CZE. A tryptic hydrolysis with an enzyme-to-substrate ratio (E/S) of 1/20 and incubation for 20 hr at 37 degrees C was found to result in the most suitable conditions. Peptides were separated and collected using an uncoated capillary (120 cm x 75 microm i.d.) in 0.15 M formic acid at pH 2.3 by applying 28 kV. Peptide maps were highly reproducible as shown by coefficients of variation of less than 0.89 and 5.42% for migration times and peak areas, respectively. Moreover, very good resolution of the peptide maps revealed the region in which the aberrant peptides of the beta-Lg variants may be located.

Péptidos bioactivos derivados de las proteínas de la leche / Bioactive peptides derived from milk proteins

Las proteínas lácteas son conocidas por tener propiedades nutricionales, funcionales y biológicas que hacen de ellas ingredientes importantes en alimentos funcionales promotores de la salud. Estas propiedades son parcialmente atribuidas a los péptidos bioactivos codificados en las diferentes proteínas de la leche. Los péptidos bioactivos, son inactivos dentro de la secuencia de la proteína intacta y pueden ser liberados por acción de enzimas proteolíticas nativas de la leche, enzimas de bacterias ácido lácticas o de fuentes exógenas, durante la digestión gastrointestinal o durante el proceso del alimento. Los péptidos derivados de las proteínas caseicas y séricas han demostrado poseer varias propiedades bioactivas como lo son: opioide, antihipertensiva, antimicrobial, inmunomodulatoria, transporte de minerales y antitrombótica. Esta revisión presenta una perspectiva de la importancia de las proteínas lácteas en la producción de péptidos bioactivos y sus actividades biológicas, así como de las principales técnicas utilizadas para el aislamiento e identificación de estos péptidos

Tequila volatile characterization and ethyl ester determination by solid phase microextraction gas chromatography/mass spectrometry analysis.

Solid phase microextraction (SPME) and gas chromatography were used for tequila volatile characterization and ethyl ester quantitation. Several factors determined the differences in tequila volatile profiles obtained by the SPME technique, namely, sampling mode, fiber coating, and fiber exposure time. Each of these factors determined the most suitable conditions for the analysis of volatile profiles in tequila. Volatile extraction consisted of placing 40 mL of tequila in a sealed vial kept at 40 degrees C. A poly(dimethylsiloxane) fiber was immersed in the liquid for 60 min and desorbed for 5 min into the gas chromatograph. The identified volatiles by mass spectrometry were mainly alcohols, esters, and ketones. The calibration curves for ethyl hexanoate, octanoate, and decanoate followed linear relationships with highly significant (p < 0.001) determination coefficients (R2 = 0.99). The coefficients of variation of less than 10% for ethyl ester concentrations indicated that the technique was reproducible. The limits of quantitation for ethyl esters were 0.05 parts per million, which were below the concentration range (0.27-15.03 ppm) found for different tequila samples. Quantitative differences in ethyl esters were found for the four most commonly known tequila types: silver, gold, aged, and extra-aged.

Detection and prediction of hydrolytic rancidity in milk by multiple regression analysis of short-chain free Fatty acids determined by solid phase microextraction gas chromatography and quantitative flavor intensity assessment.

The objective was to establish a method for detecting and predicting hydrolytic rancidity in milk by correlating quantitative sensory data with individual short-chain free fatty acids (FFA) (C(4)-C(12)) in milk determined by solid phase microextraction and gas chromatography (SPME-GC). A FFA-based equation for determining rancid flavor intensities in milk was derived by stepwise regression analysis. A highly significant (p < 0.001) correlation coefficient (R (2)) of 0.84 indicated that rancidity scores were dependent on FFA obtained by SPME-GC and that a good proportion of the variation in the rancidity scores was explained by the model. When rancidity scores were predicted for 19 commercial milks, one sample was found to be distinctly rancid by the statistical model and by the trained sensory panel. The rest of the samples were found to be nonrancid by either method. Thus, the predicting power of the model was shown because there was 100% correct flavor classification for the samples tested.