Immunostimulatory potential of extruded plant-based meat: effect of extrusion moisture level on macrophage activation.

In this study, the effect of different moisture levels in extruded plant-based meat on macrophage immunostimulation, and the potential of this meat as a protein source and a solution to environmental and economic challenges associated with conventional meat was investigated. To determine the effects of the extruded plant-based meat, cell viability assay, enzyme-linked immunosorbent assay, flow cytometry, and western blotting were performed. Low-moisture (LMME) and high-moisture meat extracts (HMME) showed higher potential to activate macrophages and regulate cytokine production than raw material extract. Treatment with LMME and HMME resulted in increased expression of CD80, CD86, and MHC class I/II proteins, indicating their potential to activate macrophages. Western blotting suggested that the immune activation observed in a previous study of macrophages was because of the phosphorylation of MAPKs and NF-κB. These findings suggest that extruded plant-based meat can potentially be used as an immunostimulatory food ingredient.

Effects of Process Variables on the Physicochemical, Textural, and Structural Properties of an Isolated Pea Protein-Based High-Moisture Meat Analog.

This study investigated the optimal extrusion conditions required to produce an isolated pea protein (IPP)-based meat analog. High-moisture extrusion cooking (HMEC) was performed. The effects of the moisture content (55 and 60%), barrel temperature (165 and 175 °C), and screw speed (150 and 200 rpm) on the physicochemical, textural, and structural properties of the high-moisture meat analog (HMMA) were determined. The results showed that the moisture content had a significant effect (p < 0.05) on the physicochemical and textural properties of the HMMA. A lower moisture content had significant impact (p < 0.05) on enhancing the texturization of the HMMA and the formation of fibrous structures, thereby increasing the texture profile analysis (TPA) and cutting strength of the HMMA. Protein denaturation during HMEC resulted in a lower protein solubility of the meat analog than the raw material. The content of ß-sheets and ß-turns in the meat analogs were higher than that in the raw material, while the content of random coils and α-helices is inversely proportional. The process variables had no significant (p > 0.05) effect on the secondary structures. In conclusion, the moisture content is the most important factor affecting the properties of HMMAs. The extrusion process variables for HMMAs are a moisture content of 55%, a barrel temperature of 175 °C, and a screw speed of 200 rpm.

Investigating the Potential of Full-Fat Soy as an Alternative Ingredient in the Manufacture of Low- and High-Moisture Meat Analogs.

The increase in meat consumption could adversely affect the environment. Thus, there is growing interest in meat analogs. Soy protein isolate is the most common primary material to produce low- and high-moisture meat analogs (LMMA and HMMA), and full-fat soy (FFS) is another promising ingredient for LMMA and HMMA. Therefore, in this study, LMMA and HMMA with FFS were manufactured, and then their physicochemical properties were investigated. The water holding capacity, springiness, and cohesiveness of LMMA decreased with increasing FFS contents, whereas the integrity index, chewiness, cutting strength, degree of texturization, DPPH free radical scavenging activity, and total phenolic content of LMMA increased when FFS contents increased. While the physical properties of HMMA decreased with the increasing FFS content, its DPPH free radical scavenging activity and total phenolic contents increased. In conclusion, when full-fat soy content increased from 0% to 30%, there was a positive influence on the fibrous structure of LMMA. On the other hand, the HMMA process requires additional research to improve the fibrous structure with FFS.

Effects of mealworm larva composition and selected process parameters on the physicochemical properties of extruded meat analog.

Mealworm larva (Tenebrio molitor), the most promising edible insect species with low cost and less environmental pollution, can fulfill the flavor and nutrition which are deficient in soy-based meat analog. Consumers who might have the sensitivity and reluctance to the intact form of edible insect could be offered the high-quality extruded meat analog. Therefore, this study aimed to investigate the effects of different mealworm larva contents and extrusion process parameters using twin-screw extruder on the physicochemical properties of the extruded meat analog. Mealworm larva was added to the base formulation at the rate of 0, 15, and 30%. Extrusion process parameters were varied as die temperature (140°C and 150°C) and moisture content (40% and 50%). The integrity index, texture profile analysis, and oxidation activity (TBARS) of extruded meat analog decreased with increase in mealworm larva content. However, water holding capacity, nitrogen solubility index, protein digestibility, and DPPH radical scavenging activity significantly increased (p < .05) as the mealworm content increased. Lower die temperature and higher moisture content enhanced the textural properties, but reduced nitrogen solubility index and protein digestibility. At higher die temperature, DPPH activity increased but TBARS showed the opposite result. After extrusion cooking, the total amino acid levels of extruded meat analog were 585.21 g/kg in 30% mealworm larva content that level was lower than 591.43 g/kg in raw mixture while the sulfur-containing amino acids and glutamic acid were higher than that of raw mixture. In conclusion, addition of mealworm larva could improve the nutritional value, antioxidant functionality, and taste of extruded meat analog under controlled extrusion process conditions.

Influences of extrusion parameters on physicochemical properties of textured vegetable proteins and its meatless burger patty.

Isolated soy protein, wheat gluten, and starch at ratio 5:4:1 were texturized under different moisture contents (40 and 50%) and die temperature (130 and 150 °C) by the twin-screw extruder. Physicochemical properties were firstly studied. These textured vegetable proteins (TVPs) were used to form 100% plant-based burger patties. Cooking and textural features were secondly investigated. TVP at 50% moisture content and 130 °C die temperature represented the highest water absorption capacity and integrity index but the lowest solubility among TVPs. Cooking loss and shrinkage in diameter and thickness, cohesiveness, chewiness, hardness, and cutting strength of TVP meatless burger patties were significantly lower than that commercial meat patty, while moisture retention and springiness of TVP meatless burger patties were higher (p < 0.05). Our results found that the texture of patty made with TVP at 50% moisture content and 130 °C die temperature was the most similarity to commercial meat patty.

Fermentation of texturized vegetable proteins extruded at different moisture contents: effect on physicochemical, structural, and microbial properties.

To investigate the effect of fermentation on texturized vegetable protein (TVP), TVPs extruded at 40 and 50% feed moisture contents (MC) were fermented using Bacillus subtilis at 37 °C. Physicochemical, structural and microbial properties of TVPs were determined at 0, 12, 24, 36, 48, and 60 h after fermentation. Integrity index of fermented TVPs did not change significantly until 24-36 h after fermentation. Springiness and cohesiveness remained stable after fermentation in all samples. Significant total color change was observed during fermentation. The pH values dropped initially and rose again with a coincident increase in nitrogen solubility index and viable cell count of B. subtilis. During fermentation, TVP extruded at 50% MC maintained higher chewiness, hardness, integrity index, and layered structure than that extruded at 40% MC. The study demonstrated that fermenting the TVP extruded at 50% MC has potential to produce a new type of TVP based B. subtilis fermented food.

Effects of extrusion types, screw speed and addition of wheat gluten on physicochemical characteristics and cooking stability of meat analogues.

BACKGROUND: Plant protein-based products such as meat analogues have been receiving attention over the years. However, comparisons of product properties and mechanisms applied in the production of low- and high-moisture meat analogues have not been reported. In this study, the effects of extrusion types (low- and high-moisture extrusion cooking), absence or presence of added wheat gluten, as well as screw speed (150 and 200 rpm) on the physicochemical properties of meat analogues were evaluated. The mechanism of protein texturization of low- and high-moisture meat analogues was studied. RESULTS: Extrusion types and addition of wheat gluten had a major influence on physicochemical characteristics which were critical in controlling the fibrous texture of the final product, while screw speed had a minor impact on springiness only (P < 0.001). All high-moisture meat analogues (HMMAs) were associated with a higher integrity index and greater stability of springiness and cutting strength than low-moisture meat analogues (LMMAs) using the same formula and screw speed, while the nitrogen solubility index of HMMAs was lower. Based on the physicochemical properties determined, the higher cross-link formation in HMMAs is proposed to occur in the cooling die section. CONCLUSION: Our findings show that the utilization of high-moisture extrusion cooking and the incorporation of wheat gluten into the formula at 400 g kg-1 could impart a fibrous and compact structure to extrudates similar to that of actual muscle meat, with a greater integrity index and texture stability. © 2019 Society of Chemical Industry.

A comparison of physicochemical characteristics, texture, and structure of meat analogue and meats.

BACKGROUND: The continuous increase of global consumer's dietary behaviors towards reducing meat consumption due to health benefits, ecological, ethical, and social aspects. Texturized vegetable protein (TVP) transformed from plant protein-based ingredient can impart fibrous structure like muscle meat. However, there is limited information on a comparison of product properties of TVP and meats. In this study, the comparison of physicochemical properties, texture, and structure of TVP and different types of meats (beef, pork, and chicken) were investigated. RESULTS: The nitrogen solubility and integrity index, chewiness and cutting strength of TVP were similar to that of the chicken sample compared to other meats. However, water absorption capacity of TVP (2173.84 g kg-1 ) was significantly higher than all meat samples (1095.37 to 1653.52 g kg-1 ). The color and amino acids of TVP were different from meat samples due to the difference in protein types. TVP showed a fibrous structure with non-uniform air cells. CONCLUSION: The study demonstrated that some textural and chemical characteristics of texturized vegetable protein under intermediate moisture extrusion exhibited the most similarity to chicken meat. © 2018 Society of Chemical Industry.

Effects of extrusion with CO2 injection on physical and antioxidant properties of cornmeal-based extrudates with carrot powder.

Carrot powder and cornmeal were extruded at ratios of 0:100, 10:90, and 20:80 with and without CO2 injection at die temperatures of 80, 100, and 120 °C. The effects of the composition of the extrudate, die temperature, and CO2 injection on physicochemical and antioxidant properties of extruded products were studied. The results showed that die temperature had a significant effect on expansion ratio (ER), specific length, piece density, color, water absorption index (WAI), and water solubility index (WSI) (p < 0.05). The injection of CO2 significantly affected the ER, WAI, WSI, lightness, redness, microstructure, total phenolic content, and the 2,2-diphenyl-1-picrylhydrazyl radical-scavenging activity of extrudates (p < 0.05). Increasing the proportion of carrot powder in extrudates resulted in better antioxidant properties and higher levels of crude ash, crude fat, crude protein, and redness; however, it resulted in lower WAI, lightness, and yellowness (p < 0.05). The study demonstrated that extrusion with CO2 injection and addition of carrot powder may improve the nutritional quality and structure-forming ability of extrudates.

The Effects of CO2 Injection and Barrel Temperatures on the Physiochemical and Antioxidant Properties of Extruded Cereals.

The effects of CO2 injection and barrel temperatures on the physiochemical and antioxidant properties of extruded cereals (sorghum, barley, oats, and millet) were studied. Extrusion was carried out using a twin-screw extruder at different barrel temperatures (80, 110, and 140°C), CO2 injection (0 and 500 mL/min), screw speed of 200 rpm, and moisture content of 25%. Extrusion significantly increased the total flavonoid content (TFC) of extruded oats, and ß-glucan and protein digestibility (PD) of extruded barley and oats. In contrast, there were significant reductions in 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, PD of extruded sorghum and millet, as well as resistant starch (RS) of extruded sorghum and barley, and total phenolic content (TPC) of all extrudates, except extruded millet. At a barrel temperature of 140°C, TPC in extruded barley was significantly increased, and there was also an increase in DPPH and PD in extruded millet with or without CO2 injection. In contrast, at a barrel temperature of 140°C, the TPC of extruded sorghum decreased, TFC of extruded oats decreased, and at a barrel temperature of 110°C, PD of extruded sorghum without CO2 decreased. Some physical properties [expansion ratio (ER), specific length, piece density, color, and water absorption index] of the extrudates were significantly affected by the increase in barrel temperature. The CO2 injection significantly affected some physical properties (ER, specific length, piece density, water solubility index, and water absorption index), TPC, DPPH, ß-glucan, and PD. In conclusion, extruded barley and millet had higher potential for making value added cereal-based foods than the other cereals.

The Effect of Extrusion Conditions on Water-extractable Arabinoxylans from Corn Fiber.

The effect of feed moisture contents (30%, 40%, and 50%) and screw speed (200 rpm, 250 rpm, and 300 rpm) on the corn fiber gum (CFG) yield and soluble arabinoxylans (SAX) content of destarched corn fiber was investigated. The CFG yields and SAX contents of extruded, destarched corn fiber were higher than that of destarched corn fiber. In extruded, destarched corn fiber, increased screw speed and decreased feed moisture contents resulted in a higher SAX contents. The maximum yields of CFG obtained from extruded, destarched corn fiber were 79.1±19.0 g/kg (30% feed moisture content) and 82.3±11.30 g/kg (300 rpm screw speed). The highest SAX content was also observed at a screw speed of 300 rpm. The results of the present study show that water extraction and extrusion combined have the potential to increase CFG and SAX yields from corn fiber.

Effects of extrusion cooking on physicochemical properties of white and red ginseng (powder).

A systematic comparison of the physicochemical properties of white ginseng (WG), extruded white ginseng (EWG), red ginseng (RG), and extruded red ginseng (ERG) was performed. The aim of the present study was to identify the effects of the physicochemical properties of ginseng by extrusion cooking. The highest value of the water absorption index (WAI) was 3.64 g/g obtained from EWG, and the highest value of the water solubility index (WSI) was 45.27% obtained from ERG. The ERG had a better dispersibility compared with other samples. Extrusion cooking led to a significant increase in acidic polysaccharide and total sugar content but resulted in a decrease in crude fat and reducing sugar contents. Enzyme treatment led to a sharp increase in acidic polysaccharide content, especially the cellulose enzyme. Extrusion cooking led to a significant increase in 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and reducing power, and the increases in WG and RG were 13.56% (0.038) and 3.56% (0.026), respectively. The data of this study provide valuable information about the effects of extrusion on quality changes of EWG and ERG.

Effect of thermostable α-amylase injection on mechanical and physiochemical properties for saccharification of extruded corn starch.

BACKGROUND: In industry, a jet cooker is used to gelatinize starch by mixing the starch slurry with steam under pressure at 100-175 °C. A higher degree of starch hydrolysis in an extruder is possible with glucoamylase. Unfortunately, it is difficult to carry out liquefaction and saccharification in parallel, because the temperature of gelatinization will be too high and will inactivate glucoamylase. Since the temperature for liquefaction and saccharification is different, it is hard to change the temperature from high (required for liquefaction) to low (required for saccharification). The industrial gelatinization process is usually carried out with 30-35% (w/w) dry solids starch slurry. Conventional jet cookers cannot be used any more at high substrate concentrations owing to higher viscosity. In this study, therefore, corn starch was extruded at different melt temperatures to overcome these limitations and to produce the highest enzyme-accessible starch extrudates. RESULTS: Significant effects on physical properties (water solubility index, water absorption index and color) and chemical properties (reducing sugar and % increase in reducing sugar after saccharification) were achieved by addition of thermostable α-amylase at melt temperatures of 115 and 135 °C. However, there was no significant effect on % increase in reducing sugar of extruded corn starch at 95 °C. CONCLUSION: The results show the great potential of extrusion with thermostable α-amylase injection at 115 and 135 °C as an effective pretreatment for breaking down starch granules, because of the significant increase (P < 0.05) in % reducing sugar and enzyme-accessible extrudates for saccharification yield.

Influence of germination and extrusion with CO(2) injection on physicochemical properties of wheat extrudates.

Whole wheat and germinated wheat flour were extruded in a laboratory co-rotating twin screw extruder with die temperatures (90 and 130°C), screw speeds (150 and 200rpm) and CO2 injection. The effects of germination and extrusion process on specific mechanical energy (SME) input, expansion ratio, specific length, piece density, elastic modulus, breaking strength, colour, water solubility index (WSI), water absorption index (WAI) and microstructure were determined. The study showed that the use of germinated wheat flour increased the specific length, lightness and the WSI. When CO2 was injected, the expansion ratios (only 90°C die temperature for extruded germinated wheat) and lightness were significantly increased (p<0.05). The chemical properties (crude protein, fat, ash, reducing sugar, γ-aminobutyric acid, soluble arabinoxylans, ß-glucan and phytic acid) were also investigated. The germination step and extrusion process mainly affected the chemical properties. However, the difference of die temperatures, screw speed and CO2 injection had slight effect on the chemical properties.

The effect of feed moisture and temperature on tannin content, antioxidant and antimicrobial activities of extruded chestnuts.

This study focuses on the effect of extrusion processing on tannin reduction, phenolic content, flavonoid content, antioxidant and anitimicrobial activity. Extrusion temperature (120 and 140 °C) and feed moisture (25% and 28%) were used on the tannin content, antioxidant and antimicrobial activities. Extrusion cooking reduced tannin content up to 78%, and improved antioxidant activity from 12.89% to 21.17% in a concentration dependant manner without affecting its antimicrobial activity that varied from 250 to 500 mg. The time-kill assay confirmed the ability of extruded chestnut to reduce Pseudomonas aeruginosa count below detectable limit that reduced the original inoculum by 3log10 CFU/mL. Overall, the results showed that extrusion cooking might serve as a tool for tannin reduction and could improve the antioxidant and antimicrobial properties of chestnut, which might be helpful for chestnut related products in the food industry.

The effect of extrusion conditions on the acidic polysaccharide, ginsenoside contents and antioxidant properties of extruded Korean red ginseng.

This study was conducted to investigate the effect of extrusion conditions (moisture content 20% and 30%, screw speed 200 and 250 rpm, barrel temperature 115℃ and 130℃) on the acidic polysaccharide, ginsenoside contents and antioxidant properties of extruded Korean red ginseng (KRG). Extruded KRGs showed relatively higher amounts of acidic polysaccharide (6.80% to 9.34%) than nonextruded KRG (4.34%). Increased barrel temperature and screw speed significantly increased the content of acidic polysaccharide. The major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, Rg2s, Rg3s, Rh1, and Rg3r) of KRG increased through extrusion, while the ginsenoside (Rg1) decreased. The EX8 (moisture 30%, screw speed 250 rpm, and temperature 130℃) had more total phenolics and had a better scavenging effect on 2,2-diphenyl-1-picrylhydrazyl radicals than those of extruded KRG samples. The extrusion cooking showed a significant increase (6.8% to 20.9%) in reducing power. Increased barrel temperature significantly increased the values of reducing power, the highest value was 1.152 obtained from EX4 (feed moisture 20%, screw speed 250 rpm, and temperature 130℃). These results suggest that extrusion conditions can be optimized to retain the health promoting compounds in KRG products.

Influence of twin-screw extrusion on soluble arabinoxylans and corn fiber gum from corn fiber.

BACKGROUND: The effect of feed moisture content and screw speed in the extrusion process with and without chemical pretreatment of corn fiber was investigated. Different chemical pretreatment methods (NaOH and H2 SO4 solution) were compared. The improvement of reducing sugar, soluble arabinoxylans (SAX) content and the yield of corn fiber gum was measured. RESULTS: A high reducing sugar content was obtained in the filtrate fraction from the extruded destarched corn fiber (EDCF) with H2SO4 pretreatment. Feed moisture content most effectively improved both reducing sugar and SAX content of filtrate. Increasing feed moisture content and screw speed resulted in a higher SAX content in the filtrate of the EDCF with NaOH pretreatment. The SAX content of the residual solid from the EDCF with NaOH pretreatment was higher compared to H2SO4 pretreated and unpretreated samples and significantly increased with decreasing feed moisture content. The screw speed did not have a major impact after enzyme hydrolysis. The yield of corn fiber gum was increased by 12% using NaOH pretreatment combined with extrusion process as compared to the destarched corn fiber. CONCLUSION: The results show the great potential of the extrusion process as an effective pretreatment for disruption the lignocelluloses of corn fiber, leading to conversion of cellulose to glucose and hemicelluloses to SAX and isolation of corn fiber gum.

Effects of extrusion conditions on the physicochemical properties of extruded red ginseng.

The effects of variable moisture content, screw speed and barrel temperature on the physicochemical properties of red ginseng powder extrudates were investigated. The raw red ginseng powders were processed in a co-rotating intermeshing twin-screw extruder. Primary extrusion variables were feed moisture content (20 and 30%), screw speed (200 and 250 rpm) and barrel temperature (115 and 130°C). Extruded red ginseng showed higher crude saponin contents (6.72~7.18%) than raw red ginseng (5.50%). Tested extrusion conditions did not significantly affect the crude saponin content of extrudates. Increased feed moisture content resulted in increased bulk density, specific length, water absorption index (WAI), breaking strength, elastic modulus and crude protein content and decreased water solubility index (WSI) and expansion (p<0.05). Increased barrel temperature resulted in increased total sugar content, but decreased reducing sugar content in the extrudate (p<0.05). Furthermore, increased barrel temperature resulted in increased amino acid content and specific length and decreased expansion and bulk density of extrudates only at a higher feed moisture content. The physicochemical properties of extrudates were mainly dependent on the feed moisture content and barrel temperature, whereas the screw speed showed a lesser effect. These results will be used to help define optimized process conditions for controlling and predicting qualities and characteristics of extruded red ginseng.

Identification of new microsatellite markers in Panax ginseng.

Microsatellites, also called simple sequence repeats (SSR), are very useful molecular genetic markers commonly used in crop breeding, species identification and linkage analysis. In the present study, we constructed a microsatellite-enriched genomic library of Panax ginseng, and identified 251 novel microsatellite sequences. Tri-nt repeat units were the most abundant (46.6%), followed by di-nt repeats (35.5%). The (AG)n motif was most common (23.1%), followed by the (AAC)n motif (22.3%). From the genotyping of 94 microsatellites using marker-specific primer sets, we identified 11 intraspecific polymorphic markers as well as 14 possible interspecific polymorphic markers differing between P. ginseng and P. quinquefolius. The exact allele structures of the polymorphic markers were determined and the alleles were named. This study represents the first report of the bulk isolation of microsatellites by screening a microsatellite-enriched genomic library in P. ginseng. The microsatellite markers could be useful for linkage analysis, genetic breeding and authentication of Panax species.