Reduction in the antigenicity of beta-lactoglobulin in whole milk powder via supercritical CO2 treatment.

Cow milk allergy is a common phenomenon experienced in early childhood (<5 yr of age) with an average occurrence rate of roughly 2.5%. The most prevalent allergen in cow milk is believed to be ß-LG. The objective of this study was to evaluate the use of hydrophobic supercritical CO2 (ScCO2) to modify the chemical structure ß-LG, thus impairing its recognition by antibodies. Whole milk powder (WMP) was selected because of its closest compositional resemblance to bovine fluid milk and its applications in reconstitution and in the beverage (infant, toddler, and adult), confectionary, bakery, and meat industries. For this study, WMP was treated with food-grade CO2 at temperatures of 50, 63, and 75°C under operating pressures of 100, 150, 200, 250, and 300 bar. Proteins in WMP were examined using SDS-PAGE, western blot, and ELISA. Orbitrap Fusion liquid chromatography-tandem MS (LC-MS/MS) and periodic staining was performed to confirm post-translational modifications in ß-LG. Functional properties of WMP before and after treatment were assessed by its solubility index, oil holding capacity, emulsion capacity and stability, zeta potential, particle size, and color analysis. SDS-PAGE of treated samples yielded fuzzy bands (variable mobility of molecules due to different molecular weights results in ill-defined bands) indicative of an increase in molecular weight, presumably due to chemical change in the protein, and demonstrated a maximum of 71.13 ± 0.29% decrease in the band intensity of ß-LG under treatment conditions of 75°C/300 bar for 30 min. These changes were small with samples treated with heat only. Lighter, diffused bands were observed using western blot analysis. The ELISA tests proved that ScCO2 treatment specifically and significantly affected the antigenicity of ß-LG with a reduction of 42.9 ± 2.83% and 54.75 ± 2.43% at 63°C/200 bar and 75°C/300 bar, respectively. Orbitrap fusion detected the presence of fatty acids and sugar moieties bound to ß-LG and the latter was confirmed by periodic staining. Functional properties of ScCO2-treated milk powder yielded a decrease in solubility index and an increase in emulsion capacity of WMP was observed under ScCO2 treatment at 75°C/300 bar, with small and insignificant changes at other treatments producing a decrease in antigenicity. Color changes were small for most samples, except at 63°C/200 bar, where a significant increase in yellowness was observed. Zeta potential and particle size measurements indicated that most changes were temperature driven. This study demonstrates 2 approaches to mitigate ß-LG antigenicity via fatty acid binding and lactosylation using hydrophobic ScCO2.

The relevance of a potential bioactive ingredient: The milk fat globule membrane.

The milk fat globule membrane (MFGM) is the surrounding membrane of the triacyl glyceride-dense fat globules in milk. The native MFGM is a tri-layer structure formed during the biosynthesis of lipids in the rough endoplasmic reticulum of lactocytes within the mammary gland. It is composed of a phospholipid tri-layer, cholesterol, many proteins, glycosylated proteins, and various lipids (Brink and Lönnerdal, 2020). The 2 main sources of MFGM are cream derived during butter production and whey derived from cheese manufacturing. The use of MFGM ingredients in nutritional products, especially in infant nutrition, has received great attention in recent years. Today, there is a range of commercially available MFGM-enriched ingredients from various suppliers across the globe. The basic components of the tri-layer, phospholipids, and proteins have been shown to exert bioactive effects in various models including in humans. It is not yet known how the differences in composition of such complex ingredients impact the bioactive potential of MFGM within the dairy matrix. The MFGM becomes more complex as a result of processing as it loses its native tri-layer structure and MFGM fragments are formed. This review aims to bring to light the underlying mechanisms for the bioactive effects of MFGM ingredients on human health.

Evaluation of kefir grain microbiota, grain viability, and bioactivity from fermenting dairy processing by-products.

Four dairy foods processing by-products (acid whey permeate [AWP], buttermilk [BM], sweet whey permeate [SWP], and sweet whey permeate with added milk fat globule ingredient [SWP+MFGM]) were fermented for 4 wk and compared with traditional kefir milks for production of novel kefir-like dairy products. Sweet whey permeates and SWP supplemented with 1.5% milk fat globule membrane (MFGM) proved to be the most viable by-products for kefir grain fermentation, exhibiting diverse abundance of traditional kefir microorganisms and positive indicators of bioactive properties. Grain viability was assessed with shotgun metagenomics, texture profile analysis, live cell counts, and scanning electron microscopy. Assessed bioactivities of the kefir-like products included antibacterial, antioxidant, potential anticancerogenic properties, and membrane barrier effects on human colorectal adenocarcinoma Caco-2 cells. All kefir grains were most abundant in Lactobacillus kefiranofaciens when analyzed with shotgun metagenomics. When analyzed with live cell counts on selective media, AWP kefir-like product had no countable Lactococcus spp., indicating suboptimal conditions for kefir grain microbiota survival and application for fermented dairy starter culture bacterium. Live cell counts were affirmed with kefir grain surface scanning electron microscopy images. The SWP treatment had the most adhesive kefir grain surface, and SWP+MFGM had the largest exopolysaccharide yield from grain extraction. All kefir and kefir-like products were able to achieve a 6-log reduction against Listeria innocua and Escherichia coli. Traditional milk kefirs had the highest antioxidant capacity for 2,2-diphenyl-1-picrylhydrazyl (DPPH) and the 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid; ABTS) assay. The AWP formulation had a significantly higher DPPH antioxidant activity compared with the other kefir and kefir-like products, and SWP had the lowest Trolox equivalence concentration in the ABTS assay. Sweet whey and supplemented milk fat sweet whey had upregulation of Cldn-1 and Ocln-1 gene expression, which correspond with a significant increase in transepithelial electrical resistance.

Development of probiotic yogurt products incorporated with Lactobacillus kefiranofaciens OSU-BSGOA1 in mono- and coculture with Kluyveromyces marxianus.

The bacterium Lactobacillus kefiranofaciens OSU-BDGOA1 and yeast Kluyveromyces marxianus bdgo-ym6 were previously isolated from kefir grains and have shown probiotic traits in mono- and coculture. This research evaluates the effect of introducing probiotic kefir microorganisms in monoculture and in coculture alongside yogurt starter cultures on the physicochemical and rheological properties, volatile flavor compounds, survival of the microorganisms during simulated digestion, and sensory attributes of the final fermented products. The incorporation of L. kefiranofaciens OSU-BDGOA1 in monoculture showed promising outcomes, resulting in a final product showing more solid-like characteristics and potentially improving the texture of the product. There was also a significant increase in the concentration of desirable volatile flavor compounds in the yogurt with the monoculture, particularly 2,3-butanedione, displaying a positive correlation with buttery flavor in the sensory analysis. The inclusion of L. kefiranofaciens in monoculture also promoted better sensory attributes and was significantly better than the yogurt with the coculture with the yeast, showing promising results for the incorporation of this probiotic bacterium into functional fermented dairy products.

Chemical transesterification of coconut and corn oils using different metal hydroxides as catalysts to determine the chemical and physiochemical changes to the oils.

BACKGROUND: The transesterification of butteroil has been shown to alter its lipid chemistry and thus alter the crystallization of the fat. The reaction kinetics and resulting crystallization of the butteroil differ depending on the nature of the catalyst used. Modeling the reaction with vegetable oils is a simpler method for the analysis of resulting products to understand the chemical and physiochemical changes that occur based on catalyst selection. The objective of this work is to perform a chemical transesterification of coconut and corn oil using monovalent and divalent catalysts to investigate the chemical and crystal changes that occur. RESULTS: Coconut and corn oil were subjected to chemical transesterification using both Ca(OH)2 and KOH as catalysts. In both the coconut and corn oil samples, transesterification caused monoglycerides (MAGs) and diacylglycerides (DAGs) to form from the most abundant fatty acid found in each sample. Coconut oil's melting temperature, solid fat content (SFC), and storage modulus decreased as a result of the transesterification, and crystals began to form in the corn oil causing melting thermograms to be evident, higher SFC, and a more viscous oil as a result. Using Ca(OH)2 as a catalyst resulted in more MAG formation, and a higher SFC and melting temperature than when KOH was used as a catalyst. CONCLUSION: The results demonstrate that the chemical changes that result from transesterification of plant-based oils change the crystallization behavior of the oils and can therefore be used for different applications in the food industry. © 2024 The Author(s). Journal of The Science of Food and Agriculture published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Invited review: Properties of ß-galactosidases derived from Lactobacillaceae species and their capacity for galacto-oligosaccharide production.

ß-galactosidase (enzymatic class 3.2.1.23) is one of the dairy industry's most important and widely used enzymes. The enzyme is part of a large family known to catalyze hydrolysis and transglycosylation reactions. Its hydrolytic activity is commonly used to decrease lactose content in dairy products, while its transglycosylase activity has recently been used to synthesize galacto-oligosaccharides (GOS). During the past couple of years, researchers have focused on studying ß-galactosidase isolated and purified from lactic acid bacteria. This review will focus on ß-galactosidase purified and characterized from what used to be the Lactobacillus genera. Furthermore, particular emphasis is given to its kinetics, biochemical characteristics, GOS production, market, and utilization by Lactobacilllaceae species.

Partition of milk phospholipids during ice cream manufacturing.

The distribution of phospholipids (PL) within the fat and serum phase of ice cream manufacturing was evaluated through partition coefficients (KPL) after mixing, pasteurization, freezing, and hardening. Ice creams containing about 40.41 ± 3.45 (± standard deviation; control formulation) and 112.29 ± 9.06 (enriched PL formulation) mg of PL per g of fat were formulated with nonfat dry milk and ß-serum, respectively. Overall, the KPL were lower than 1, indicating that the PL were predominantly found in the fat phase, and only a small amount was left in the serum and sediment. Confocal micrographs visually confirmed this generalization. The addition of PL significantly increased the viscosity of the mixes between 4- and 9-fold, depending on the shear rate. Additionally, mixes containing high PL exhibited higher yield stress than those formulated with low PL (0.15 ± 0.09 and 0.016 ± 0.08 Pa, respectively). Ice creams with high PL delayed the onset of meltdown and exhibited a slower rate of a meltdown than low-PL ice creams (18.53 ± 0.57 and 14.83 ± 0.85 min, and 1.01 ± 0.05 and 0.71 ± 0.04% min-1, respectively). This study provides useful guidelines for manufacturing ice cream enriched in milk PL. Additionally, the use of ß-serum, a byproduct stream, as a source of PL is illustrated. The development will require studying the sensorial description of the product as well as consumer acceptance.

Efficient in vitro digestion of lipids and proteins in bovine milk fat globule membrane ingredient (MFGMi) and whey-casein infant formula with added MFGMi.

The relative immaturity of the infant digestive system has the potential to affect the bioavailability of dietary lipids, proteins, and their digested products. We performed a lipidomic analysis of a commercial bovine milk fat globule membrane ingredient (MFGMi) and determined the profile of lipids and proteins in the bioaccessible fraction after in vitro digestion of both the ingredient and whey-casein-based infant formula without and with MFGMi. Test materials were digested using a static 2-phase in vitro model, with conditions simulating those in the infant gut. The extent of digestion and the bioaccessibility of various classes of neutral and polar lipids were monitored by measuring a wide targeted lipid profile using direct infusion-mass spectrometry. Digestion of abundant proteins in the ingredient and whey-casein infant formula containing the ingredient was determined by denaturing PAGE with imaging of Coomassie Brilliant Blue stained bands. Cholesterol esters, diacylglycerides, triacylglycerides, phosphatidylcholines, and phosphatidylethanolamines in MFGMi were hydrolyzed readily during in vitro digestion, which resulted in marked increases in the amounts of free fatty acids and lyso-phospholipids in the bioaccessible fraction. In contrast, sphingomyelins, ceramides, and gangliosides were largely resistant to simulated digestion. Proteins in MFGMi and the infant formulas also were hydrolyzed efficiently. The results suggest that neutral lipids, cholesterol esters, phospholipids, and proteins in MFGMi are digested efficiently during conditions that simulate the prandial lumen of the stomach and small intestine of infants. Also, supplementation of whey-casein-based infant formula with MFGMi did not appear to alter the profiles of lipids and proteins in the bioaccessible fraction after digestion.

Effects of Fructose and Palmitic Acid on Gene Expression in Drosophila melanogaster Larvae: Implications for Neurodegenerative Diseases.

One of the largest health problems worldwide is the development of chronic noncommunicable diseases due to the consumption of hypercaloric diets. Among the most common alterations are cardiovascular diseases, and a high correlation between overnutrition and neurodegenerative diseases has also been found. The urgency in the study of specific damage to tissues such as the brain and intestine led us to use Drosophila melanogaster to study the metabolic effects caused by the consumption of fructose and palmitic acid in specific tissues. Thus, third instar larvae (96 ± 4 h) of the wild Canton-S strain of D. melanogaster were used to perform transcriptomic profiling in brain and midgut tissues to test for the potential metabolic effects of a diet supplemented with fructose and palmitic acid. Our data infer that this diet can alter the biosynthesis of proteins at the mRNA level that participate in the synthesis of amino acids, as well as fundamental enzymes for the dopaminergic and GABAergic systems in the midgut and brain. These also demonstrated alterations in the tissues of flies that may help explain the development of various reported human diseases associated with the consumption of fructose and palmitic acid in humans. These studies will not only help to better understand the mechanisms by which the consumption of these alimentary products is related to the development of neuronal diseases but may also contribute to the prevention of these conditions.

Monitoring the ripening attributes of Turkish white cheese using miniaturized vibrational spectrometers.

Monitoring the ripening process by prevalent analytic methods is laborious, expensive, and time consuming. Our objective was to develop a rapid and simple method based on vibrational spectroscopic techniques to understand the biochemical changes occurring during the ripening process of Turkish white cheese and to generate predictive algorithms for the determination of the content of key cheese quality and ripening indicator compounds. Turkish white cheese samples were produced in a pilot plant scale and ripened for 100 d, and samples were analyzed at 20 d intervals during storage. The collected spectra (Fourier-transform infrared, Raman, and near-infrared) correlated with major composition characteristics (fat, protein, and moisture) and primary products of the ripening process and analyzed by pattern recognition to generate prediction (partial least squares regression) and classification (soft independent analysis of class analogy) models. The soft independent analysis of class analogy models classified cheese samples based on the unique biochemical changes taking place during the ripening process. partial least squares regression models showed good correlation (RPre = 0.87 to 0.98) between the predicted values by vibrational spectroscopy and the reference values, giving low standard errors of prediction (0.01 to 0.57). Portable and handheld vibrational spectroscopy units can be used as a rapid, simple, and in situ technique for monitoring the quality of cheese during aging and provide real-time tools for addressing deviations in manufacturing.

Use of casein micelles to improve the solubility of hydrophobic pea proteins in aqueous solutions via low-temperature homogenization.

The dairy industry struggles to maintain consumer attention in the midst of declining fluid milk sales. Current trends create an opportunity to incorporate plant-based proteins with milk to produce a high-protein, multisourced, functional food product. Plant-based proteins, such as those in peas, can be challenging to use in food systems because of their low solubility and undesirable off-flavors. Casein micelles have unique structural properties that allow for interactions with small ions and larger macromolecules that aid in their noteworthy ability as a nanovehicle for hydrophobic compounds. The objective of this study was to use the inherent structure of the casein micelle along with common dairy processing equipment to create a stable colloidal dispersion of casein micelles with pea protein to improve its solubility in aqueous solutions. We created 3 blends with varying ratios of casein-to-pea protein (90:10, 80:20, 50:50). We subjected the mixtures to 3 cycles of homogenization using a bench-top GEA 2-stage homogenizer at 27,580 kPa maintained at 4°C, followed by pasteurization at 63°C for 30 min. The resulting blends were homogeneous liquids with increased stability due to the lack of protein precipitation. Further protein analysis by HPLC and AA sequencing revealed that vicilin, an insoluble storage protein, was the main pea protein incorporated within the casein micelle structure. These results supported our hypothesis that low-temperature homogenization can successfully be used to create a colloidal dispersion with increased stability, in which insoluble plant-based proteins may be incorporated with casein micelles in an aqueous solution. Additionally, 3-dimensional microscope images of the blends indicated a noticeable difference between the surface roughness upon addition of pea protein to the casein micelle matrix. This research highlights a promising application for other plant-based proteins to be used within the dairy industry to help drive future product innovation while also meeting current processing conditions and consumer demands.

Invited review: Milk kefir microbiota-Direct and indirect antimicrobial effects.

Kefir is a fermented dairy product with well recognized probiotic properties. Recently, consumer interest in fermented products with probiotic microorganisms has increased due to the accumulating evidence of the effects of kefir microorganisms on the modulation of gut microbiota and their antimicrobial activity. Although the health properties of kefir have been reviewed in other works, the present review addresses the antimicrobial effects of kefir microbiota and associated compounds. The antimicrobial activity of kefir microorganisms could derive from different mechanisms. The microorganisms' capacity to adhere to the intestinal epithelium, preventing the adhesion of pathogens, and their immunomodulation properties are among the mechanisms suggested. Bacteria and yeast isolated from kefir have been shown to have in vivo and in vitro antimicrobial activity against enteropathogenic bacteria and spoilage fungi. However, most reports have focused their approach on single-strain antimicrobial properties; evaluation of antimicrobial activity of cocultures of kefir microbiota and their potential mechanisms of action has been neglected. Kefir microbiota and associated compounds have shown promising antimicrobial effects; however, more research needs to be done to discern the mechanisms of action.

Effects of pressure, shear, temperature, and their interactions on selected milk quality attributes.

The effects of pressure, temperature, shear, and their interactions on selected quality attributes and stability of milk during ultra-shear technology (UST) were investigated. The UST experiments include pressure (400 MPa) treatment of the milk sample preconditioned at 2 different initial temperatures (25°C and 15°C) and subsequently depressurizing it via a shear valve at 2 flow rates (low: 0.15-0.36 g/s; high: 1.11-1.22 g/s). Raw milk, high-pressure processed (HPP; 400 MPa, ~40°C for 0 and 3 min) and thermal treated (72°C for 15 s) milk samples served as the controls. The effect of different process parameters on milk quality attributes were evaluated using particle size, zeta potential, viscosity, pH, creaming, lipase activity, and protein profile. The HPP treatment did not cause apparent particle size reduction but increased the sample viscosity up to 3.08 mPa·s compared with 2.68 mPa·s for raw milk. Moreover, it produced varied effects on creaming and lipase activity depending on hold time. Thermal treatment induced slight reduction in particle size and creaming as compared with raw milk. The UST treatment at 35°C reduced the effective diameter of sample particles from 3,511.76 nm (raw milk) to 291.45 nm. This treatment also showed minimum relative lipase activity (29.93%) and kept milk stable by preventing creaming. The differential effects of pressure, shear, temperature, and their interactions were evident, which would be useful information for equipment developers and food processors interested in developing improved food processes for dairy beverages.

Invited review: Acid whey trends and health benefits.

In recent years, acid whey production has increased due to a growing demand for Greek yogurt and acid-coagulated cheeses. Acid whey is a dairy by-product for which the industry has long struggled to find a sustainable application. Bulk amounts of acid whey associated with the dairy industry have led to increasing research on ways to valorize it. Industry players are finding ways to use acid whey on-site with ultrafiltration techniques and biodigesters, to reduce transportation costs and provide energy for the facility. Academia has sought to further investigate practical uses and benefits of this by-product. Although modern research has shown many other possible applications for acid whey, no comprehensive review yet exists about its composition, utilization, and health benefits. In this review, the industrial trends, the applications and uses, and the potential health benefits associated with the consumption of acid whey are discussed. The proximal composition of acid whey is discussed in depth. In addition, the potential applications of acid whey, such as its use as a starting material in the production of fermented beverages, as growth medium for cultivation of lactic acid bacteria in replacement of commercial media, and as a substrate for the isolation of lactose and minerals, are reviewed. Finally, the potential health benefits of the major protein constituents of acid whey, bioactive phospholipids, and organic acids such as lactic acid are described. Acid whey has promising applications related to potential health benefits, ranging from antibacterial effects to cognitive development for babies to human gut health.

The interactions between anthocyanin and whey protein: A review.

Anthocyanins (ACN) are natural pigments that produce bright red, blue, and purple colors in plants and can be used to color food products. However, ACN sensitivity to different factors limits their applications in the food industry. Whey protein (WP), a functional nutritional additive, has been shown to interact with ACN and improve the color, stability, antioxidant capacity, bioavailability, and other functional properties of the ACN-WP complex. The WP's secondary structure is expected to unfold due to heat treatment, which may increase its binding affinity with ACN. Different ACN structures will also have different binding affinity with WP and their interaction mechanism may also be different. Circular dichroism (CD) spectroscopy and Fourier transform infrared (FTIR) spectroscopy show that the WP secondary structure changes after binding with ACN. Fluorescence spectroscopy shows that the WP maximum fluorescence emission wavelength shifts, and the fluorescence intensity decreases after interaction with ACN. Moreover, thermodynamic analysis suggests that the ACN-WP binding forces are mainly hydrophobic interactions, although there is also evidence of electrostatic interactions and hydrogen bonding between ACN and WP. In this review, we summarize the information available on ACN-WP interactions under different conditions and discuss the impact of different ACN chemical structures and of WP conformation changes on the affinity between ACN and WP. This summary helps improve our understanding of WP protection of ACN against color degradation, thus providing new tools to improve ACN color stability and expanding the applications of ACN and WP in the food and pharmacy industries.

Technically relevant enzymes and proteins produced by LAB suitable for industrial and biological activity.

Lactic acid bacteria (LAB) are a unique subset of microorganisms that have co-evolved with humans since the beginning of agricultural practices and animal domestication and throughout our never-ending quest for food preservation, digestibility, and flavor enhancement. LAB have historically played a preponderant role in our foods. In this review, we focus on the enzymatic activities and current or potential applications of LAB in our lives. A description of each of the enzymatic systems in LAB is included. Glycosidases, which hydrolyze the most abundant food molecules and as sources of carbon, sustain the lives of organisms on Earth as well as ensure microbial innocuity by the production of lactic acid from the uniquely mammalian carbohydrate, lactose. Lipases and proteases or proteinases are of fundamental importance in food fermentations and in dairy foods for flavor development. Bacteriocins and peptidoglycan hydrolases are part of the enzymatic system of LAB that has evolved to make these bacteria fierce competitors in various microbiomes, which are highly important for the human gut. In this review, we also present an explanation on how the versatility of the genetics of LAB can adapt to the matrix where they are placed with the advantage of not having any toxicity to humans. The systematic study of LAB enzymes has allowed for some unique applications in foods and biopharmaceutical industries. Here, we summarize how different enzyme systems in LAB are classified, and thus, facilitate much-needed further studies to understand the fundamentals and translate them into applications to improve our lives.

Purification and characterization of a phospholipid-hydrolyzing phosphoesterase produced by Pediococcus acidilactici isolated from Gouda cheese.

Lipolysis occurs during ripening of dairy products as a result of esterase or lipase activity. Lactic acid bacteria (LAB) are considered to be weakly lipolytic bacteria compared with other species. In cheeses with extended ripening periods, lipolytic LAB may have several advantages. Pediococcus acidilactici is a LAB frequently found in fermented dairy products, but no previous reports exist on their production of esterases or lipases. Our interest in the relationship of LAB and enzymatic characterization is due to the multiple reports of the benefits of LAB in the gut microbiome, particularly at the intestinal membrane. Pediococci have been characterized as probiotic and especially active in membrane interactions. The aim of this project was to purify, characterize, and identify the phosphoesterase produced by P. acidilactici originally isolated from Gouda cheese and determine its phospholipid (PL) hydrolysis profile, with a focus on increased absorption of these compounds in the human gut. Native zymograms were performed to identify a protein with lipolytic activity in the intracellular fraction of P. acidilactici. The enzyme was purified via size-exclusion HPLC, concentrated via ultrafiltration, and identified using sequence analysis in liquid chromatography (LC)-MS/MS. The purified fraction was subjected to biochemical characterization as a function of pH, temperature, ion concentration, hydrolysis of different substrates, and PL. A single protein with a molecular weight of 86 kDa and esterase activity was detected by zymography. Analysis of the LC-MS/MS results identified a putative metallophosphoesterase with a calculated molecular weight of 45.5 kDa, suggesting that this protein is active as a homodimer. The pure protein showed an optimal activity between pH 8.0 to 9.0. The optimal temperature for activity was 37°C, and the enzyme lost 15% of activity after incubation at 90°C for 1 h. This enzyme showed activity on short-chain fatty acids and exhibited high hydrolysis of phosphatidylinositol. It also hydrolyzed phosphatidylserine, phosphatidylcholine, and sphingomyelin. Phosphatidylethanolamine was hydrolyzed but with less efficiency. The characteristics and lipolytic actions exerted by this protein obtained from LAB hold promise for a potential strain of esterase or lipase that may exert human health benefits through increased digestibility and absorption of nutrients found in dairy products.

Growth of lactic acid bacteria in milk phospholipids enhances their adhesion to Caco-2 cells.

The mechanisms of bacterial adhesion to human cells involve several complex reactions and activation of genes and proteins. It has been reported that the food components in dairy matrices, such as sugar or salt, can decrease bacterial adhesion to Caco-2 cells. However, it has not been evaluated whether the bacteria grown in media supplemented with milk phospholipids (MPL) can increase or decrease the adhesion of these cells. The objective of this work was to evaluate the effects of MPL on the kinetic growth of lactic acid bacteria (LAB) and their functional characteristics as probiotics, expression of surface protein genes, and adherence to Caco-2 cells. Seven LAB strains isolated from various dairy products were characterized. Five of the tested LAB strains were able to grow in a chemically defined medium supplemented with MPL. Lactobacillus reuteri OSU-PECh-48 showed the highest growth rate and the greatest optical density. All of the strains tested showed tolerance to acidic conditions at pH 3.0 and to bile salts at 0.5 and 1% concentrations. Auto-aggregation and cell surface hydrophobicity ability were evaluated, with nonsignificant differences between the strains grown in MPL and without MPL. Gene expression of 6 surface proteins was evaluated in the presence or absence of MPL. Pediococcus acidilactici OSU-PECh-L and OSU-PECh-48 were the strains with highest relative expression of 5 of the 6 genes evaluated. Lactobacillus paracasei OSU-PECh-BA was the strain with the lowest level of expression of surface protein genes. Most of the bacteria tested had increased adhesion to Caco-2 cells after growth in MPL. The bacteria with the highest degrees of adhesion observed were Lactobacillus paracasei OSU-PECh-3B, Pediococcus acidilactici OSU-PECh-L, and Lactobacillus reuteri OSU-PECh-48. The genes Cnb and EF-Tu increased in expression in the presence of MPL in most of the LAB tested. The results obtained in this work demonstrate the high potential of these LAB strains for use as starters or beneficial cultures in fermentation of not only dairy products but also other food fermentation processes, with promising ability to increase residence time in the gut, modify the microbiome, and improve human health.

Effect of gestational age (preterm or full term) on lipid composition of the milk fat globule and its membrane in human colostrum.

Human colostrum is the first milk secreted by the mother after birth and constitutes the ideal food for the newborn, because its chemical composition, rich in immunoglobulins, antimicrobial peptides, growth factors, bioactive lipids, and other important molecules, is perfectly adapted to the metabolic, digestive, and immunological immaturity of the newborn. An incomplete gestational period can affect the maturity of the mammary gland and its ability to secrete milk with the proper composition for the newborn's condition. Previous studies indicate that the mammary gland modulates the profiles of bioactive lipids present in the different phases of lactation from colostrum to mature milk. Given the key role played by the polar lipids (PL) (phospho- and sphingolipids) of the milk fat globule membrane (MFGM) in the immune system and cognitive development of the newborn, it is crucial to analyze whether the content and distribution of the PL are affected by gestation period. Therefore, this study aimed to determine the milk fat globule (MFG) and MFGM lipid compositions of human colostrum samples from 20 healthy preterm and full-term mothers. Lipid characterization using chromatographic techniques (gas chromatograph mass spectrometry and HPLC-evaporative light-scattering detection) revealed differences related to length of gestation in the profiles of lipid classes and fatty acid and triacylglyceride contents of colostrum. This comparative analysis leads to noteworthy outcomes about the changing roles of the PL, considering the preterm or full-term condition. We found a lack of correlation of some PL (such as phosphatidylcholine, phosphatidylinositol, and phosphatidylserine) with the delivery term; these could be denoted as structural category lipids. However, sphingomyelin and phosphatidyl-ethanolamine exhibited trends to decrease in full-term colostrum, indicating that in the final stage of pregnancy specific accretion of some PL occurs, which should be denoted as a nutritional redistribution.

Monitoring Hydroxycinnamic Acid Decarboxylation by Lactic Acid Bacteria Using High-Throughput UV-Vis Spectroscopy.

Hydroxycinnamic acid (HCA) decarboxylation by lactic acid bacteria (LAB) results in the production of 4-vinylplenols with great impact on the sensorial characteristics of foods. The determination of LAB decarboxylating capabilities is key for optimal strain selection for food production. The activity of LAB strains from the Ohio State University-Parker Endowed Chair (OSU-PECh) collection potentially capable of synthesizing phenolic acid decarboxylase was evaluated after incubation with HCAs for 36 h at 32 °C. A high-throughput method for monitoring HCAs decarboxylation was developed based on hypsochromic shifts at pH 1.0. Out of 22 strains evaluated, only Enterococcus mundtii, Lactobacillus plantarum and Pediococcus pentosaceus were capable of decarboxylating all p-coumaric, caffeic and ferulic acids. Other strains only decarboxylated p-coumaric and caffeic acid (6), only p-coumaric acid (2) or only caffeic acid (1), while 10 strains did not decarboxylate any HCA. p-Coumaric acid had the highest conversion efficiency, followed by caffeic acid and lastly ferulic acid. Results were confirmed by HPLC-DAD-ESI-MS analyses, showing the conversion of HCAs into their 4-vinylphenol derivatives. This work can help improve the sensory characteristics of HCA-rich foods where fermentation with LAB was used during processing.