Combination of Milk and Plant Proteins to Develop Novel Food Systems: What Are the Limits?

In the context of a diet transition from animal protein to plant protein, both for sustainable and healthy scopes, innovative plant-based foods are being developing. A combination with milk proteins has been proposed as a strategy to overcome the scarce functional and sensorial properties of plant proteins. Based on this mixture were designed several colloidal systems such as suspensions, gels, emulsions, and foams which can be found in many food products. This review aims to give profound scientific insights on the challenges and opportunities of developing such binary systems which could soon open a new market category in the food industry. The recent trends in the formulation of each colloidal system, as well as their limits and advantages are here considered. Lastly, new approaches to improve the coexistence of both milk and plant proteins and how they affect the sensorial profile of food products are discussed.

Edible insect as an alternative protein source: a review on the chemistry and functionalities of proteins under different processing methods.

The consumption of edible insects can be anadvantageous alternative to the conventional food supply chain, which involves global water waste, land deficit, undernutrition, and starvation. Besides the nutritional aspects, insect proteins have demonstrated a wide range of functional properties such as foamability, emulsifying and gelling abilities. The protein content and amino acid profile of some insects have revealed a good nutritional value and interesting functional properties. However, it is crucial to comprehend how the protein quality is affected by insect feeding, drying, and defatting. There is a knowledge gap about the impact of industrial treatment, such as pH, ionic strength, and heat treatment, on insect proteins' functional properties. In this review, we have aimed to highlight the potential application of insect proteins as a nutritional source and their promising technological applications. The study reported the principal insect protein characterization methodologies that have been investigated in the literature aiming to correlate the physicochemical parameters to possible protein functionalities. The research on the functional properties of insect proteins is at the exploratory level. Further detailed studies are needed to clarify the structure-function relation of insect proteins and how these functionalities and insect processing can increase consumer acceptance.

An Easy and Cheap Kiwi-Based Preparation as Vegetable Milk Coagulant: Preliminary Study at the Laboratory Scale.

In the present study, a kiwifruit aqueous extract was developed and used as a coagulant enzyme in cheesemaking. In detail, polyacrylamide gel electrophoresis (SDS-PAGE) was used to investigate the presence of actinidin, the kiwifruit enzyme involved in κ-casein hydrolysis, in different tissues (pulp, peel, and whole fruit) of ripe and unripe kiwifruits. Data revealed the presence of the enzyme both in the peel and in the pulp of the fruit. Although the aqueous extract obtained from the kiwifruit peel was able to hydrolyze semi-skimmed milk, it did not break down κ-casein. The aqueous extract obtained from the pulp showed a hydrolytic activity toward both κ-casein and semi-skimmed milk. The values for milk-clotting and proteolytic activity of the kiwifruit pulp extract were evaluated at different temperatures and pH parameters in order to obtain a high value of the MCA/PA ratio; we found that a temperature of 40 °C in combination with a pH value of 5.5 allowed us to obtain the best performance. In addition, the data revealed a higher hydrolytic activity of the enzymatic preparation from ripe kiwifruits than that from unripe ones, suggesting the use of the extract from pulp of ripe kiwifruits in the laboratory-scale cheesemaking. The data showed that 3% (v/v) of the ripe kiwifruit pulp extract determined a curd yield of 20.27%, comparable to chymosin yield. In conclusion, the extraction procedure for kiwifruit aqueous extract proposed in the present study was shown to be a fast, cheap, chemical-free, and ecofriendly technology as a plant coagulant for cheese manufacturing.

Physical properties of UHT light cream: impact of the high-pressure homogenization and addition of hydrocolloids.

The beneficial effects of a healthy diet on the quality of life have prompted the food industry to develop low-fat variants, but fat content directly affects the physicochemical and sensory properties of food products. The utilization of high-pressure homogenization (HP) and incorporation of hydrocolloids have been suggested as strategies to improve the physical stability and rheological properties of light cream. Thus, this study aims to analyze the associated effect of high-pressure homogenization (80 MPa) and three different hydrocolloids: microcrystalline cellulose, locust bean gum and xanthan gum, on emulsion stability and rheological properties of ultra-high-temperature (UHT) light cream (ULC) with a 15% w/w fat content. The stability of ULC was determined by the ζ potential of oil droplets and emulsion stability percentage. Rheological characterization was based on flow behavior tests and dynamic oscillatory measurements, which were carried out in a rheometer. Results showed that the high-pressure homogenization process did not influence the emulsion stability of the treatments. Moreover, the hydrocolloids added to systems present weak interactions with milk proteins since all ULC showed macroscopical phase separation. The samples presented the same rheological behavior and were classified as pseudoplastic fluids (n < 1). ULC treated at 80 MPa was significantly (P ≤ 0.05) more consistent than the treatments at 20 MPa. All ULC showed a predominant elastic behavior (G' > G″), and a remarkable increase in both G' and G″ at 80 MPa. The results presented in this study highlight the potential of HP for altering some rheological characteristics of UHT light cream, for example, to increase its consistency. These results are important for the dairy industry and ingredient suppliers, in the standardization of UHT light cream and/or to develop low-fat products.

Milk quality, production process and physicochemical characteristics of Porungo, an artisanal cheese from the state of Sao Paulo, Brazil.

Porungo is a traditional pasta filata cheese produced using raw milk throughout the southwest region of the state of São Paulo, Brazil. The objectives of this Research Communication were to evaluate the quality of raw milk used to make Porungo cheese, to characterize its production process, and to determine its chemical composition. The results showed that the raw milk met both Brazilian and international quality requirements. Chemically, Porungo can be classified as a medium to full fat semi-hard fresh cheese. Our study has allowed the first standards and regulations for Porungo to be established in Brazil. By virtue of this, the local producers are able to formalize their activity while consumers can have access to a safe and certified product.

5-Hydroxymethylfurfural formation and color change in lactose-hydrolyzed Dulce de leche.

The work described in this Research Communication concerns the production of Dulce de leche (DL), that is a traditional product from South America obtained by concentration. Maillard reaction (MR) products are mainly responsible for the formation of color and flavor in this product. Lactose-hydrolyzed products have been developed to supply consumer demand, but this hydrolysis may affect the flavor, color, taste, texture and even some nutritional aspects of the product. We studied the influence of different levels of lactose-hydrolysis, sucrose addition and initial pH on the development of MR, appraised by the determination of 5-hydroxymethylfurfural (HMF). A process simulator with multi-monitoring system was used to produce 15 DL. Box-Behnken 33 experimental design was applied for the three factors: pH, lactose-hydrolysis level and sucrose concentration. Lipids, protein, ashes, carbohydrates, water activity, dissolved solids, colorimetric analysis and HMF (free and total) are among the physicochemical attributes and MR indicators analyzed in this work. The products showed significant differences in composition but all the values were in agreement with the literature. Moreover, higher levels of lactose hydrolysis and higher pH presented a direct relation with the development of MR, observed by an increase in coloration (lower luminosity) and more formation of HMF, both free and total. The present study expands the knowledge about DL spread made of lactose-hydrolyzed milk, allowing the food industries to produce a lactose free DL with nutritional and sensory characteristics closer to the traditional product.

Impact of co-cultivation with Enterococcus faecalis over growth, enterotoxin production and gene expression of Staphylococcus aureus in broth and fresh cheeses.

Biocontrol of Staphylococcus aureus by lactic acid bacteria can be considered as a feasible alternative to the use of chemicals in foods, but the mechanisms underlying this antagonistic interaction remains poorly understood. This study aimed to evaluate the impact of a LAB species (Enterococcus faecalis) over the growth, enterotoxin production and gene expression of S. aureus under experimental conditions. E. faecalis 41FL1 and S. aureus ATCC 29213 were inoculated isolated and together in brain heart infusion (BHI) at 30 °C and in a fresh cheese model at 15 °C: microbial populations were monitored by culture plating, production of classical staphylococcal enterotoxins (SEs) was verified by an ELISA assay, expression of S. aureus genes (virulence, transcriptional regulation and central carbon metabolism) was investigated by quantitative real-time PCR, and pH and contents of water-soluble metabolites in both matrices were measured. S. aureus growth was inhibited in co-cultures assays, with a 2.02-log reduction in BHI and a 3.39-log reduction in cheese model compared to respective single cultures. Classical SEs were detected in S. aureus single culture assays (BHI and cheese), in BHI inoculated with both strains after 48 h of incubation, but not detected in co-inoculated cheeses. pH in all matrices containing E. faecalis reached lower values than in matrices containing S. aureus alone, due to lactate production by E. faecalis. Expression of genes coding for transcription regulators (ccpA and rex) and enzymes involved in central carbon metabolism (alsD and citZ) was mostly upregulated in co-inoculated cheeses, whereas expression of several virulence determinants (agrC, hld, hla, entA and spa) was strongly downregulated. This study provides relevant data on the behaviour of S. aureus in the presence of competing microbiota and support the use of controlled population dominance by LAB as an effective biopreservation strategy to ensuring food safety.

Temperature modulates the production and activity of a metalloprotease from Pseudomonas fluorescens 07A in milk.

This work evaluated the expression and activity of a metalloprotease released by Pseudomonas fluorescens 07A in milk. Low relative expression of the protease by the strain was observed after incubation for 12 h at 25°C while the strain was in the logarithmic growth phase. After 24 h, protease production significantly increased and remained constant for up to 48 h, a time range during which the strain remained in the stationary phase. Conversely, at refrigeration temperatures, at 12 h the strain was still in the lag phase and expressed the protease at higher levels than when the logarithmic phase was reached. Casein fractions were highly degraded by P. fluorescens 07A, the purified protease, and the bacterial pellet on d 7 of incubation at 25°C and to a lesser extent at 10°C for the sample incubated with the bacterium. Heat treatment at 90°C for 5 min completely inactivated the proteolytic activity of the purified protease and the bacterial pellet. This work contributes to the knowledge about the conditions of milk storage that influence the production and activity of this extracellular metalloprotease. The results demonstrate the need to find alternative strategies to control the synthesis and activity of proteolytic enzymes in the dairy industry to ensure the quality of processed products.

Characterization of a heat-resistant extracellular protease from Pseudomonas fluorescens 07A shows that low temperature treatments are more effective in deactivating its proteolytic activity.

This work discusses the biological and biochemical characterization of an extracellular protease produced by Pseudomonas fluorescens. The enzyme has a molecular weight of 49.486 kDa and hydrolyzes gelatin, casein, and azocasein, but not BSA. Its maximum activity is found at 37°C and pH 7.5, but it retained almost 70% activity at pH 10.0. It was shown to be a metalloprotease inhibited by Cu(2+), Ni(2+), Zn(2+), Hg(2+), Fe(2+), and Mg(2+), but induced by Mn(2+). After incubation at 100°C for 5min, the enzyme presented over 40% activity, but only 14 to 30% when submitted to milder heat treatments. This behavior may cause significant problems under conditions commonly used for the processing and storage of milk and dairy products, particularly UHT milk. A specific peptide sequenced by mass spectrometer analysis allowed the identification of gene that encodes this extracellular protease in the genome of Pseudomonas fluorescens 07A strain. The enzyme has 477 AA and highly conserved Ca(2+)- and Zn(2+)-binding domains, indicating that Ca(2+), the main ion in milk, is also a cofactor. This work contributes to the understanding of the biochemical aspects of enzyme activity and associates them with its sequence and structure. These findings are essential for the full understanding and control of these enzymes and the technological problems they cause in the dairy industry.

Evaluation of the synergistic effects of milk proteins in a rapid viscosity analyzer.

Protein systems (PS) are routinely used by companies from Brazil and around the globe to improve the texture, yield, and palatability of processed foods. Understanding the synergistic behavior among the different protein structures of these systems during thermal treatment under the influence of pH can help to better define optimum conditions for products and processes. The interpretation of the reactions and interactions that occur simultaneously among the protein constituents of these systems as dispersions during thermal processing is still a major challenge. Here, using a rapid viscosity analyzer, we observed the rheological changes in the startup viscosities of 5 PS obtained by combining varying proportions of milk protein concentrate and whey protein concentrate under different conditions of pH (5.0, 6.5, and 7.0) and heat processing (85°C/15min and 95°C/5min). The solutions were standardized to 25% of total solids and 17% of protein. Ten analytical parameters were used to characterize each of the startup-viscosity ramps for 35 experiments conducted in a 2×3 × 5 mixed planning matrix, using principal component analysis to interpret behavioral similarities. The study showed the clear influence of pH 5.5 in the elevation of the initial temperature of the PS startup viscosity by at least 5°C, as well as the effect of different milk protein concentrate:whey protein concentrate ratios above 15:85 at pH 7.0 on the viscographic profile curves. These results suggested that the primary agent driving the changes was the synergism among the reactions and interactions of casein with whey proteins during processing. This study reinforces the importance of the rapid viscosity analyzer as an analytical tool for the simulation of industrial processes involving PS, and the use of the startup viscosity ramp as a means of interpreting the interactions of system components with respect to changes related to the treatment temperature.