The widening gap between current supply of meat and its future demand has increased the need to produce plant-based meat analogs. Despite ongoing technical developments, one of the unresolved ...challenges of plant-based meat analogs is to safely and effectively imitate the appearance of raw and cooked animal-based meat, especially the color. This study aimed to develop a more effective and safe browning system for beet red (BR) in plant-based meat analog patties using laccase (LC) and sugar beet pectin (SBP). First, we investigated the synergistic effects of SBP and LC on BR decolorization of meat analog patties. We discovered that the red tones of LC-treated patties containing BR and SBP were remarkably browned after grilling, compared to patties that did not contain SBP. Notably, this color change by LC + SBP was similar to that of beef patties. Additionally, the hardness of LC-treated meat analog patties containing BR was higher than those that did not contain BR. Interestingly, the presence of SBP and LC enhanced the browning reaction and functional properties of meat analogs containing BR. This is the first report on a browning system for meat analogs containing BR using enzymatic methods to the best of our knowledge.
Abstract
The gap between the current supply and future demand of meat has increased the need to produce plant-based meat analogs. Methylcellulose (MC) is used in most commercial products. Consumers ...and manufacturers require the development of other novel binding systems, as MC is not chemical-free. We aimed to develop a novel chemical-free binding system for meat analogs. First, we found that laccase (LC) synergistically crosslinks proteins and sugar beet pectin (SBP). To investigate the ability of these SBP-protein crosslinks, textured vegetable protein (TVP) was used. The presence of LC and SBP improved the moldability and binding ability of patties, regardless of the type, shape, and size of TVPs. The hardness of LC-treated patties with SBP reached 32.2 N, which was 1.7- and 7.9-fold higher than that of patties with MC and transglutaminase-treated patties. Additionally, the cooking loss and water/oil-holding capacity of LC-treated patties with SBP improved by up to 8.9–9.4% and 5.8–11.3%, compared with patties with MC. Moreover, after gastrointestinal digestion, free amino nitrogen released from LC-treated patties with SBP was 2.3-fold higher than that released from patties with MC. This is the first study to report protein-SBP crosslinks by LC as chemical-free novel binding systems for meat analogs.
The widening gap between the supply and demand for meat products has increased the need to produce plant-based meat analogs as protein sources. Meat analogs are principally composed of soy-based ...textured vegetable proteins. Despite ongoing technical developments, one of the unresolved challenges for plant-based meat analogs is the off-flavor from soy, which limits their consumer acceptability. Among the various methods developed for overcoming this challenge, masking the beany flavors with cyclodextrins (CDs) is an attractive, cost-effective, and safe strategy. However, the current established CD treatment method does not meet the requirement for a clean-label. This study aimed to develop more acceptable off-flavor-masking technologies for plant-based patties for modern clean-label preferences using enzymatic methods. We used the cyclodextrin glucanotransferase (CGT), "Amano," as a commercially available food-grade CGT. The CGT-catalyzed reaction in plant-based patties yielded 17.1 g/L CD. As CGT could yield sufficient CD in the patties, we investigated whether CDs produced by CGT could mask the off-flavors released from the plant-based patties. The CGT-treated patties had significantly lower volatilization amounts of the known beany off-flavor-generating compounds compared to the non-treated patties. Moreover, CGT treatment improved the texture of the patties and increased their water- and oil-holding capacity. As CGT is rendered inactive after cooking, it would not be considered an additive. These findings indicated that CDs produced by the CGT reaction could effectively mask off-flavors of meat analogs and improve their physical properties while meeting clean-label requirements.
Despite considerable research and developments in the color properties of plant-based meat analog (PBMA) products, their color changes during the grilling process are one of the main unsolved ...challenges. Recently, it was reported that a novel browning system for PBMA products containing a red pigment (betanidin) and a multicopper oxidase (laccase; LC) can simulate the color changes (red to brown) of animal meat. However, the reaction products in PBMA using this system have not been analyzed in detail. Thus, this study aimed to clarify the reaction products of betanidin oxidized via LC-catalyzed reaction. It was found that LC-catalyzed oxidation in the PBMA patties degraded betanidin to betalamic acid and cyclodopa, subsequently polymerizing these degradation products to produce unknown brown polymers. These findings provide suggestions for the browning mechanism of PBMA patties containing betanidin via LC-catalyzed oxidation.
An unresolved challenge for plant-based meat analogs (PBMAs) is their lack of juiciness. Saturated fats significantly contribute to the juiciness of PBMAs, but there are concerns about the ...undesirable health effects related to saturated fats; thus, demand for their replacement with vegetable unsaturated oils has increased. Although many food additives are used to reduce the leakage of unsaturated oils, this solution cannot meet the clean-label requirements that have been trending in recent years. In this study, we aimed to develop better consumer-acceptable methods using protein-glutaminase (PG) to improve the juiciness of PBMA patties to meet clean-label trends. We found no significant difference between the visual surface of control and PG-treated textured vegetable proteins (TVPs). However, the microstructure of PG-treated TVP had a more rounded shape than that of the control TVP as observed under a scanning electron microscope. After grilling process, the PBMA patties composed of PG-treated TVP showed significantly higher liquid-holding capacities (a juiciness indicator) than the control patties. This suggested that PG treatment could potentially produce PBMA patties with increased juiciness. Interestingly, after the PG-treated TVP underwent the wash process, we found that PG treatment of TVP easily reduced the various beany off-flavor compounds by 58-85%. Moreover, the results of the in vitro protein digestion test showed that the amounts of free amino nitrogen released from PBMA patties composed of PG-treated TVP were 1.5- and 1.7-fold higher than those from control patties in the gastric and intestinal phases, respectively. These findings indicate that PG treatment of TVP could enhance the physical, sensory, and nutritional properties of PBMA patties and meet the clean-label requirements.
The gap between the current supply of meat and its predicted future demand is widening, increasing the need to produce plant-based meat analogs. Despite ongoing technical developments, one of the ...unresolved challenges of plant-based meat analogs is to safely and effectively decolor plant proteins that originally exhibit yellow-brown or strong brown color. This study aimed to develop an effective and safe decoloring system for soy-based protein products using food-grade hydrogen peroxide and catalase. First, soy-based protein isolate (PI) and textured vegetable protein (TVP) were treated with hydrogen peroxide, and then the residual hydrogen peroxide was degraded using catalase. This process caused notable decolorization of PI and TVP, and residual hydrogen peroxide was not detected in these products. These findings indicate that this process could safely and effectively decolorize soy-based proteins. Interestingly, this decoloring process enhanced the solubility, water- and oil-holding capacities, foaming capacity, and emulsifying stability of decolored soy-based PI. Additionally, cooking loss and juiciness of decolored TVP-based foods were improved compared to those of non-treated foods. These findings indicate that the decoloring process also enhances the physical properties of soy-based protein products.
The activity of a self-sufficient cytochrome P450 enzyme, CYP505D6, from the lignin-degrading basidiomycete
was characterized. Recombinant CYP505D6 was produced in
and purified. In the presence of ...NADPH, CYP505D6 used a series of saturated fatty alcohols with C
carbon chain lengths as the substrates. Hydroxylation occurred at the ω-1 to ω-6 positions of such substrates with C
carbon chain lengths, except for 1-dodecanol, which was hydroxylated at the ω-1 to ω-7 positions. Fatty acids were also substrates of CYP505D6. Based on the sequence alignment, the corresponding amino acid of Tyr51, which is located at the entrance to the active-site pocket in CYP102A1, was Val51 in CYP505D6. To understand the diverse hydroxylation mechanism, wild-type CYP505D6 and its V51Y variant and wild-type CYP102A1 and its Y51V variant were generated, and the products of their reaction with dodecanoic acid were analyzed. Compared with wild-type CYP505D6, its V51Y variant generated few products hydroxylated at the ω-4 to ω-6 positions. The products generated by wild-type CYP102A1 were hydroxylated at the ω-1 to ω-4 positions, whereas its Y51V variant generated ω-1 to ω-7 hydroxydodecanoic acids. These observations indicated that Val51 plays an important role in determining the regiospecificity of fatty acid hydroxylation, at least that at the ω-4 to ω-6 positions. Aromatic compounds, such as naphthalene and 1-naphthol, were also hydroxylated by CYP505D6. These findings highlight a unique broad substrate spectrum of CYP505D6, rendering it an attractive candidate enzyme for the biotechnological industry.
is a white-rot fungus whose metabolism of lignin, aromatic pollutants, and lipids has been most extensively studied. This fungus harbors 154 cytochrome P450-encoding genes in the genome. As evidenced in this study,
CYP505D6, a fused protein of P450 and its reductase, hydroxylates fatty alcohols (C
) and fatty acids (C
) at the ω-1 to ω-7 or ω-1 to ω-6 positions, respectively. Naphthalene and 1-naphthol were also hydroxylated, indicating that the substrate specificity of CYP505D6 is broader than those of the known fused proteins CYP102A1 and CYP505A1. The substrate versatility of CYP505D6 makes this enzyme an attractive candidate for biotechnological applications.
•A pectate lyase, AsPelA, from Aspergillus luchuensis var. saitoi was identified.•AsPelA gene was cloned and expressed and the enzyme was biochemically characterized.•AsPelA was thermostable and ...stable and active over a neutral-to-alkaline pH range.•AsPelA was more thermostable than AkPelA, which differs from AsPelA by 2 amino acids.
Pectinolytic enzymes are used in diverse industrial applications. We sought to isolate a pectate lyase from Aspergillus luchuensis var. saitoi, a filamentous fungus used in traditional food and beverage preparation in Japan. The identified enzyme, named AsPelA, is orthologous to PelA from A. luchuensis mut. kawachii (AkPelA); the enzymes exhibit 99% amino acid sequence identity, with Ile140 and Val197 of AsPelA being replaced by Val and Asp in AkPelA, respectively. AsPelA activity decreased to 71%, 61%, and 46% of maximal activity after 60-min incubation at 60 °C, 70 °C, and 80 °C, whereas AkPelA activity dropped to 16%, 10%, and 8.5%, respectively, indicating that AsPelA is more thermostable than AkPelA. Furthermore, AsPelA was stable within a neutral-to-alkaline pH range, as well as in the presence of organic solvents, detergents, and metal ions. Our findings suggest that AsPelA represents a candidate pectate lyase for applications in food, paper, and textile industries.
An unresolved issue with plant-based meat analog (PBMA) products is their lower texture and juiciness than animal-derived meat products. Almost all commercial PBMA patties are constructed using ...methylcellulose (MC). Thus, this study aimed to develop better formulations of binding systems for PBMA patties that simulate the physical properties of beef patties than MC binder. We investigated the formulations of binding systems using crosslinking-catalyzed enzymes (transglutaminase TG and laccase LC) and 2–4 % carbohydrate binders (MC and sugar beet pectin SBP). Texture profile analysis using a rheometer and sensory scores by nine panelists revealed that TG-catalyzed PBMA patties with 3 % MC and LC-catalyzed PBMA patties with 4 % SBP were similar to beef patties than PBMA patties constructed with any MC concentration. These findings suggest that binding systems using crosslinking-catalyzed enzymes and carbohydrate binders could bridge the gap in physical properties between the current MC-based PBMA and beef patties.
One major challenge of reducing salt content in food is the risk of the overall taste becoming bland. Enhancing saltiness is an effective strategy for salt reduction, and the development of ...salt-reduced foods using these saltiness-enhancing flavorants as food additives is underway. However, an increasing number of consumers demand a reduction in additives in clean-label foods.IntroductionOne major challenge of reducing salt content in food is the risk of the overall taste becoming bland. Enhancing saltiness is an effective strategy for salt reduction, and the development of salt-reduced foods using these saltiness-enhancing flavorants as food additives is underway. However, an increasing number of consumers demand a reduction in additives in clean-label foods.Enzyme processing of food is an attractive strategy for developing clean-label foods because enzymes are not considered additives. We aimed to improve the saltiness and umami intensity of vegetable soups by enzyme treatment while meeting clean-label requirements. We first optimized the enzymatic reaction conditions of a protease and glutaminase blend and then investigated the synergistic effects of this enzyme blend on the taste of vegetable soup.ObjectiveEnzyme processing of food is an attractive strategy for developing clean-label foods because enzymes are not considered additives. We aimed to improve the saltiness and umami intensity of vegetable soups by enzyme treatment while meeting clean-label requirements. We first optimized the enzymatic reaction conditions of a protease and glutaminase blend and then investigated the synergistic effects of this enzyme blend on the taste of vegetable soup.Sensory evaluations indicated that the reaction products (e.g., protein hydrolysates or amino acids) could enhance the umami, kokumi, and saltiness intensity of vegetable soup supplemented with 0.5% NaCl. Notably, the saltiness intensity ratio of the enzyme-treated soup with 0.50, 0.45, and 0.40% NaCl were increased by 1.31-, 1.16-, and 0.99-fold, respectively, when this ratio for the control soup with 0.50% NaCl was set to 1.0. This indicates a 20% salt reduction rate can be achieved by enzyme treatment. Moreover, we found that these enhancements were synergically caused by enzyme-produced glutamic acid and branched-chain amino acids.ResultsSensory evaluations indicated that the reaction products (e.g., protein hydrolysates or amino acids) could enhance the umami, kokumi, and saltiness intensity of vegetable soup supplemented with 0.5% NaCl. Notably, the saltiness intensity ratio of the enzyme-treated soup with 0.50, 0.45, and 0.40% NaCl were increased by 1.31-, 1.16-, and 0.99-fold, respectively, when this ratio for the control soup with 0.50% NaCl was set to 1.0. This indicates a 20% salt reduction rate can be achieved by enzyme treatment. Moreover, we found that these enhancements were synergically caused by enzyme-produced glutamic acid and branched-chain amino acids.Our findings suggest that using enzyme blends of bacterial and fungal proteases and glutaminase is an effective approach to enhancing the saltiness levels of vegetable soups while meeting clean-label requirements.ConclusionOur findings suggest that using enzyme blends of bacterial and fungal proteases and glutaminase is an effective approach to enhancing the saltiness levels of vegetable soups while meeting clean-label requirements.
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