Cysteine, sodium metabisulfite, and glutathione enhance crosslinking between proteins during high moisture meat analog extrusion processing and may improve the fibrousness of the products.

BACKGROUND: High moisture meat analog (HMMA) products processed using extrusion have become increasingly popular in the last few years. Because the formation of disulfide bonds is believed to play a critical role in the texturization mechanism, this study aimed to understand how chemical compounds capable of reducing disulfide bonds, specifically cysteine, sodium metabisulfite, and glutathione, affect the texture and the chemical interactions between the proteins. METHOD: Wheat protein blended with cysteine, sodium metabisulfite, or glutathione at levels of 0, 0.5, 1.0, 2.5, 5.0, and 7.5 g kg-1 was extruded at three different temperatures (115, 140, and 165 °C) using a co-rotating twin-screw extruder. The feed rate (85 g min-1), the moisture content (600 g kg-1), and the screw speed (300 rpm) were kept constant. Unextruded and extruded material was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, polymeric protein fractionation, and sulfhydryl group/disulfide bond analysis. Extruded samples were further analyzed for their hardness and their anisotropic index. RESULTS: The inclusion of reductants significantly affected the structure of the obtained extrudates. Although reducing agents had a relatively small impact on the total amount of disulfide bonds, their action significantly enhanced crosslinking between the proteins. At select conditions, samples with high fibrousness were specifically obtained when cysteine or sodium metabisulfite was included at levels of 5.0 g kg-1. DISCUSSION: In the presence of reducing agents, it is believed that disulfide bonds are split earlier during the process without binding to them, giving the protein strands more time to unravel and align, leading to a better flow behavior and more fibrous products. © 2024 Society of Chemical Industry.

Talc and calcium carbonate inclusions in direct expanded pea starch extrudates exhibit different behavior under increasing screw speeds.

The ability to modulate direct expanded product structures improves the versatility and range of product applications. The effect of nucleating agents, namely, talc and calcium carbonate (CC), on the expansion characteristics of pea starch extrudates as impacted by screw speed was explored. Pea starch blends with increasing levels of nucleating agents (0.25%, 1%, and 2%) at 18% moisture (w.b.) were extruded across a range of screw speeds (150, 250, 350, and 450 rpm). The water absorption index, water solubility index (WSI), expansion ratio (ER), unit density, and cell count were determined to evaluate the performance of nucleating agents. The nucleating efficiency of CC, as assessed by cell count, improved with increasing screw speeds. In contrast, the nucleating efficiency of talc was influenced by inclusion levels irrespective of screw speed. ER values ranged from 2.10 to 2.88, where higher nucleating agent inclusions and screw speeds corresponded with lower ER values. Increased nucleating agents and screw speeds corresponded to higher WSI values suggesting the nucleating agents promoted starch degradation. The nucleating agents appeared to promote flow instabilities indicated upon assessment of the extrudate surface. PRACTICAL APPLICATION: This study provides helpful information on the expanded extrudate structure of pea starch as influenced by screw speed and nucleating agents. These findings may help the food industry select processing parameters and appropriate nucleating agent inclusion levels when producing new expanded products with unique textures.

The interaction between wheat and pea protein influences the final chemical and sensory characteristics of extruded high moisture meat analogs.

Plant-based meat analog products, including those produced by extrusion processing, have become increasingly popular. Complete comprehension of the texturization mechanism and the formation of fibrousness would help improve existing products and extend the variety of plant sources used. Therefore, this study aimed to provide improved insight into the mechanism of texturization during the processing of high-moisture meat analog (HMMA) products. Blends with different wheat and pea protein ratios (100:0, 80:20, 60:40, 40:60, 20:80, and 0:100 wheat:pea) were extruded at a screw speed of 400 rpm, two different moisture contents (50% and 55%), and a feed rate of 90 g/min using a co-rotating twin-screw extruder. Extrudates were analyzed for their texture, free sulfhydryl groups, disulfide bonds, and solubility in different extractants relative to the raw ingredient blends. In addition, a sensory analysis was conducted using the rapid and cost-effective "rate-all-that-apply" (RATA) methodology. The interplay between the two protein types had synergistic effects on the system parameters torque, pressure, and specific mechanical energy, as well as on some textural and sensory parameters. Molecular analyses were not influenced by the interplay between wheat and pea protein as the molecular analyses followed linear trends with the pea inclusion level. Analysis of protein solubility suggests that the texturization mechanism differs slightly depending on the protein type. It is suggested that the texturization of wheat protein depends highly on disulfide bonds, whereas the texturization of pea protein relies on the combination of disulfide bonds and non-covalent interactions. Additionally, RATA was found to be a valuable tool for HMMA products.

Influence of legume-derived proteins with varying solubility on the direct expansion of corn starch during twin-screw extrusion processing.

This study analyzed the effect of the inclusion of legume-derived proteins, specifically pea and fava bean protein, with varying solubility levels on the expansion of corn starch. Three different proteins exhibiting low, medium, or high solubility were mixed with corn starch to obtain blends containing 15%, 25%, and 35% (w/w) of the protein. Extrusion was performed on a twin-screw extruder at three different screw speeds (200, 400, and 600 rpm), a moisture content of 16% (w.b.), and a die temperature of 140°C. Obtained extrudates were analyzed for their expansion, unit density, and hydration properties, namely, water solubility index (WSI) and water absorption index (WAI). Extrudates containing the protein with the highest solubility showed different patterns than those that had proteins with low or medium solubility. Expansion ratio (ER) increased from a maximum of 3.55 ± 0.24 for pure corn starch up to 5.45 ± 0.43 when incorporating 35% of the protein with medium solubility but significantly decreased down to 1.24 ± 0.08 when incorporating 35% of the most soluble protein. The influence on the system parameters, as well as on the hydration properties, was also greatest for the blends containing the protein with the highest solubility. Even though significant Pearson correlations were observed between protein solubility and ER (r = -0.579), unity density (r = 0.614), WSI (r = -0.634), torque (r = -0.612), as well as specific mechanical energy (r = -0.451), further research is needed to evaluate if the solubility is indeed the reason for certain behaviors or if other protein characteristics are more critical for expansion. PRACTICAL APPLICATION: This manuscript provides practical information on the influence of the addition of legume-derived proteins with different solubility levels on direct expansion. The obtained results may help the industry with the selection of the appropriate proteins for inclusion levels in producing high protein direct-expanded extruded food products.

High methoxyl pectin can improve the extrusion characteristics and increase the dietary fiber content of starch-cellulose extrudates.

Improving total dietary fiber content while maintaining the texture/expansion of extruded products is a challenge. Pectin has a dual function; it is a source of dietary fiber and it also functions as a hydrocolloid, which could improve the texture of high-fiber extruded foods. The objective of this study was to evaluate the impacts of pectin types from citrus peel on the expansion characteristics of starch-cellulose extrudates. High and low methoxyl pectin (HMP and LMP) was added to the starch-cellulose mixtures and extruded using a twin-screw extruder. The pasting properties of raw mixtures, extrusion properties, microstructure, and dietary fiber contents of the extrudates were studied. The inclusion of HMP in raw material improved the peak viscosity (629.7 ± 8.1 to 754.7 ± 80.1 mPa s) and maintained the final viscosity compared to the control (starch-cellulose mixture alone), unlike LMP. HMP relatively maintained the extrusion process parameters such as torque, back pressure, and specific mechanical energy as the control. Interestingly, the addition of 7% of HMP had a similar expansion ratio (3.41 ± 0.08 to 2.35 ± 0.06) compared to the control (3.46 ± 0.08 to 2.32 ± 0.09) under the extrusion conditions studied. The total dietary fiber content improved from 12.22 ± 0.01% to 18.26 ± 0.63% (w/w). HMP maintained the expansion characteristic of starch-cellulose extrudates and improved its total dietary fiber content relative to LMP. Adding HMP to the mixtures improved the extensibility of the melt, favoring bubble growth and expansion of the starch-cellulose extrudates. Fourier transform infrared spectroscopy data suggested that there could be intermolecular interactions between starch, cellulose, and pectin, but the nature of these interactions needs further investigation. PRACTICAL APPLICATION: The study provides practical information on the influence of the addition of high and low methoxyl pectin on starch-cellulose extrudates. The results can help the industry to produce snack products that are more nutritious but are still well accepted by the consumers.

Impact of tamarind seed gum on the viscosity behavior, thermal properties, and extrusion characteristics of native corn starch.

Tamarind seed gum (TSG) is a cold-swelling hydrocolloid with remarkable processing stability and starch synergy. Its use in direct expanded extruded foods has not been documented. The thermal and pasting viscosity properties of six TSG (0%, 0.5%, 1.0%, 2.5%, 5.0%, and 7.5% TSG) and native corn starch blends were characterized by differential scanning calorimetry and ViscoQuick, respectively. These same blends were extruded using a corotating twin-screw extruder at four screw speeds (SSs) (150, 300, 450, and 600 rpm). System back pressure, motor torque, and specific mechanical energy (SME) were measured. Extrudate quality metrics, such as expansion ratio (ER), water absorption index (WAI), and water solubility index (WSI), were also measured. The pasting viscosities indicated that TSG inclusion increases viscosity but also makes the starch-gum paste more susceptible to permanent shear degradation. The thermal analysis indicated that TSG inclusion narrowed the melting endotherms and lowered the energy required for melting (p < 0.05) at higher inclusion levels. Extruder back pressure, motor torque, and SME decreased with increasing TSG levels (p < 0.05) as the TSG effectively lowered the melt viscosity at high usage rates. The ER reached a maximum of 3.73 with a 2.5% TSG level extruded at 150 rpm (p < 0.05). The WAI of extrudates increased with TSG inclusion rate at equivalent SSs, whereas WSI behaved oppositely (p < 0.05). Small inclusions of TSG can improve the expansion properties of starch, whereas larger inclusions result in a lubrication effect that mitigates the shear-induced depolymerization of starch. PRACTICAL APPLICATION: The impact of cold-water soluble hydrocolloids, including tamarind seed gum, on the extrusion process, is poorly understood. From this work, tamarind seed gum effectively modifies the viscoelastic and thermal characteristics of corn starch in a way that enhances the direct expansion characteristics of the starch during extrusion processing. The effect is more beneficial at lower gum inclusion levels as higher levels result in reduced capabilities to translate shear from the extruder into useful transformations of the starch polymers during processing. Small amounts of tamarind seed gum could be used to improve the quality of extruded starch puff snacks.

Extrusion characteristics of ten novel quinoa breeding lines.

Ten novel breeding lines of quinoa (Chenopodium quinoa Willd) suitable to be grown in the Pacific Northwest of the United States were developed and utilized for extrusion processing. Understanding how a particular breeding line performs during food processing and which properties determine its performance can promote the use of quinoa as an ingredient in value-added products, such as extruded snacks. In this study, extrusion characteristics of the whole seed flours of the novel quinoa breeding lines were evaluated using a co-rotating twin-screw extruder at two temperatures (110°C and 125°C), three screw speeds (200, 350, and 500 rpm), and a moisture content of 18% (w.b.). The expansion ratio (ER) ranged from 1.15 to 2.33 and was negatively influenced by the fat content in the flours. Breeding line 11WAQ-104.88 (WAQ10) exhibited the greatest potential for use in direct expanded foods as it had the highest ER for all conditions studied. Strong correlations were found between ER and pasting properties of the flours, proving the usefulness of the pasting test for flour quality evaluation before extrusion processing. The results demonstrate the potential of using quinoa for producing direct expanded food products and highlight the importance of selecting specific breeding lines for desired product characteristics. PRACTICAL APPLICATION: This study provides the extrusion processing characteristics of 10 new quinoa breeding lines. Based on the information gained, it will be easier for the food industry, including breeders as well as processors, to select the right quinoa variety based on their requirements, and may further help to enhance the use of quinoa.

Potential interactions between starch and fruit pomace may impact the expansion ratio of direct expanded extrudates.

Due to their dense characteristics, direct-expanded products fortified with insoluble fiber are generally not well accepted. Understanding the interactions between starch and fiber could help to effectively choose and modify ingredients to produce products containing high amounts of fiber. Therefore, this study aims to explain the interplay between two starches (native and waxy corn) and two pomace types (blueberry and cranberry). Blends up to 100% of pomace were extruded using a co-rotating twin-screw extruder. Raw material and milled extrudates were analyzed for their pasting and hydration properties. Fourier-transform infrared (FTIR) and solid-state nuclear magnetic resonance (NMR) spectroscopy were conducted to observe molecular changes. The expansion ratio (ER) significantly decreased as pomace was added and ranged from 3.85 for pure waxy corn starch to approximately 1 for blends that contained 80% pomace. Distinctions between the blends were observed. Particularly, at 20% of pomace inclusion, native corn starch with cranberry pomace showed a significantly higher ER. Different behaviors were also detected during the physicochemical analyses. A nonlinear trend between pomace level and water solubility as well as absorption was observed for native corn starch blends, suggesting that molecular interactions between the biopolymers occur. FTIR and NMR results give no evidence for new covalent bonds; hence, the most likely interactions occurring are hydrogen bonds. In addition to the dilution effect of pomace addition, the enhancement or weakening of such interactions between starch molecules by pomace compounds may reduce the ER.

Optical Modulation of Antibiotic Resistance by Photoswitchable Cystobactamids.

The rise of antibiotic resistance causes a serious health care problem, and its counterfeit demands novel, innovative concepts. The combination of photopharmacology, enabling a light-controlled reversible modulation of drug activity, with antibiotic drug design has led to first photoswitchable antibiotic compounds derived from established scaffolds. In this study, we converted cystobactamids, gyrase-inhibiting natural products with an oligoaryl scaffold and highly potent antibacterial activities, into photoswitchable agents by inserting azobenzene in the N-terminal part and/or an acylhydrazone moiety near the C-terminus, yielding twenty analogs that contain mono- as well as double-switches. Antibiotic and gyrase inhibition properties could be modulated 3.4-fold and 5-fold by light, respectively. Notably, the sensitivity of photoswitchable cystobactamids towards two known resistance factors, the peptidase AlbD and the scavenger protein AlbA, was light-dependent. While irradiation of an analog with an N-terminal azobenzene with 365 nm light led to less degradation by AlbD, the AlbA-mediated inactivation was induced. This provides a proof-of-principle that resistance towards photoswitchable antibiotics can be optically controlled.

Heat resistance comparison of Salmonella and Enterococcus faecium in cornmeal at different moisture levels.

Adequate surrogate identification is critical for validating in-plant thermal process controls for Salmonella inactivation in different food matrices. This study compared the thermal inactivation parameters (D- and z-values) and evaluated the heat resistance of Enterococcus faecium (8459) as a surrogate for a 5-serovar Salmonella cocktail in cornmeal. The cornmeal was spray inoculated with the respective bacteria to achieve ~9 log CFU/g population and set to the desired moisture contents (16, 22, and 28% w.b.). The inoculated cornmeal was then heat-treated at pre-determined temperatures (60, 64, and 68 °C) in sealed aluminum thermal-death-time disks in hot water baths for pre-determined time intervals. Injury-recovery media [brain heart infusion (BHI) agar overlaid with xylose lysine deoxycholate (XLD) agar for Salmonella or BHI agar overlaid m-enterococcus agar for E. faecium] were used for microbial enumeration to account for thermally injured bacterial cells. The D-values of Salmonella in cornmeal at 16, 22, and 28% moisture content were 37.5, 8.4, and 2.4 min at 60 °C, 19.9, 3.5, and 1.1 min at 64 °C, and 10.1, 1.4, and 0.5 min at 68 °C, respectively. The D-values of E. faecium in cornmeal at 16, 22, and 28% moisture content were 140.4, 18.9, and 3.3 min at 60 °C, 78.4, 7.1, and 1.6 min at 64 °C, and 37.3, 2.8, and 0.8 min at 68 °C, respectively. The z-values of E. faecium and Salmonella in cornmeal at 16, 22, and 28% moisture content were 13.9, 9.7, and 12.5 °C, and 14.0, 10.4, and 11.7 °C, respectively. These results indicated similar or higher thermal resistance (D-values) and equivalent thermal sensitivity (z-values) of E. faecium compared to Salmonella at different moisture contents and respective temperatures (P ≤ 0.05). Therefore, E. faecium could be used as a surrogate for Salmonella during thermal process validation of cornmeal processing.

Selectivity of enzymes involved in the formation of opposite enantiomeric series of p-menthane monoterpenoids in peppermint and Japanese catnip.

Peppermint (Mentha x piperita L.) and Japanese catnip (Schizonepeta tenuifolia (Benth.) Briq.) accumulate p-menthane monoterpenoids with identical functionalization patterns but opposite stereochemistry. In the present study, we investigate the enantioselectivity of multiple enzymes involved in monoterpenoid biosynthesis in these species. Based on kinetic assays, mint limonene synthase, limonene 3-hydroxylase, isopiperitenol dehydrogenase, isopiperitenone reductase, and menthone reductase exhibited significant enantioselectivity toward intermediates of the pathway that proceeds through (-)-4S-limonene. Limonene synthase, isopiperitenol dehydrogenase and isopiperitenone reductase of Japanese catnip preferred intermediates of the pathway that involves (+)-4R-limonene, whereas limonene 3-hydroxylase was not enantioselective, and the activities of pulegone reductase and menthone reductase were too low to acquire meaningful kinetic data. Molecular modeling studies with docked ligands generally supported the experimental data obtained with peppermint enzymes, indicating that the preferred enantiomer was aligned well with the requisite cofactor and amino acid residues implicated in catalysis. A striking example for enantioselectivity was peppermint (-)-menthone reductase, which binds (-)-menthone with exquisite affinity but was predicted to bind (+)-menthone in a non-productive orientation that positions its carbonyl functional group at considerable distance to the NADPH cofactor. The work presented here lays the groundwork for structure-function studies aimed at unraveling how enantioselectivity evolved in closely related species of the Lamiaceae and beyond.

Quantitative in situ13C NMR studies of the electro-catalytic oxidation of ethanol.

The identification and quantification of reaction intermediates and products in a time-resolved fashion is of great value for the understanding of mechanisms underlying the electro-catalytic oxidation of alcohols. Using a newly developed in situ cell, the identification of different intermediates, products, and side products became possible by monitoring the ethanol electro-oxidation in operando by 13C NMR spectroscopy.

Comparison of different extraction methods for the determination of essential oils and related compounds from aromatic plants and optimization of solid-phase microextraction/gas chromatography.

Different extraction methods for the subsequent gas chromatographic determination of the composition of essential oils and related compounds from marjoram (Origanum majorana L.), caraway (Carum carvi L.), sage (Salvia officinalis L.), and thyme (Thymus vulgaris L.) have been compared. The comparison was also discussed with regard to transformation processes of genuine compounds, particularly in terms of expenditure of time. Hydrodistillation is the method of choice for the determination of the essential oil content of plants. For investigating the composition of genuine essential oils and related, aroma-active compounds, hydrodistillation is not very useful, because of discrimination and transformation processes due to high temperatures and acidic conditions. With cold solvent extraction, accelerated solvent extraction, and supercritical fluid extraction, discrimination of high and non-volatile aroma-active components as well as transformation processes can be diminished, but non-aroma-active fats, waxes, or pigments are often extracted, too. As solid-phase microextraction is a solvent-free fully automizable sample preparation technique, this was the most sparing to sensitive components and the most time-saving method for the rapid determination of the aroma compounds composition in marjoram, caraway, sage, and thyme. Finally, solid-phase microextraction could be successfully optimized for the extraction of the aroma components from the plants for their subsequent gas chromatographic determination.