Effects of Natural Extract Mixtures on the Quality Characteristics of Sausages during Refrigerated Storage.

Owing to the residual toxicity and adverse health effects of chemical preservatives, there is an increasing demand for using natural preservatives in food. Although many natural extracts have been evaluated, research on their antibacterial effects remains insufficient. Therefore, this study aimed to explore the possibility of developing Psidium guajava, Ecklonia cava, and Paeonia japonica (Makino) Miyabe & Takeda extracts as natural food preservatives. Further, the effect of mixing these extracts on microbial growth and quality was evaluated during the refrigeration of sausages. Optimal mixing ratios were determined based on the minimum inhibitory and bactericidal concentrations of each mixed extract against the Listeria monocytogenes, Salmonella spp. and Escherichia coli. D-optimal mixing design optimization tool was further used to obtain an optimum mixing ratio of Formulation 1 (F1). The antibacterial activity of F1 increased with increasing concentration, with similar activities at 0.5% and 1%. The sausages with synthetic or natural preservatives showed significantly lower lipid oxidation than those of the control and grapefruit extract-treated sausages after 4 wk of refrigeration. Total plate counts were observed only in the control and treatment groups stored for 3 wk, and no significant effect of ascorbic acid was observed. Compared to the other samples, sausages with added natural extracts showed the highest overall acceptability scores initially and after 4 wk. Therefore, similar amounts of grapefruit seed and natural extracts had the same effect on microbiological analysis and lipid rancidity during sausage storage. Hence, this mixture can serve as a potential natural preservative in meat products.

Inhibitory effect of natural extract mixtures on microbial growth and lipid oxidation of sausages during storage.

Large amounts of additives are used during meat product processing to maintain product quality and shelf life. With the growing interest in healthy foods, natural plant-based additives are being used as alternatives to synthetic additives. In this study, six types of natural extracts with excellent antibacterial activity were selected, and their antibacterial and antioxidant activities against four types of pathogens were evaluated in various combinations. In addition, the pH, color, amount of thiobarbituric acid reactive substances (TBARS), and growth of pathogenic microorganisms were analyzed during the storage of sausages treated with various combinations of these extracts. The natural extract mixtures exhibited different antibacterial activities, depending on the combination. Compared to grapefruit seed extract, a mixture of natural extracts extracted with ethanol (M4) reduced the Escherichia coli content by more than 99.9% after 8 days of storage and slowed the growth of Listeria monocytogenes and Salmonella spp. by more than 80% after 14 days. Compared to untreated (NC) and grapefruit extract (PC)-treated sausages, sausages treated with the natural extract mixtures showed a significant decrease in CIE L* and an increase in CIE a* and CIE b* (p < 0.05). The pH value was significantly lower in sausages containing natural extract mixtures than in the NC and PC sausages (p < 0.05). The natural plant extract mixtures significantly prevented lipid oxidation (p < 0.05). In summary, different types of natural extract mixtures have a synergistic effect when used together, suggesting that natural preservatives can generally inhibit the growth of microorganisms and oxidation of processed meat.

Characterization of Lactic Acid Bacteria and Yeast from Grains as Starter Cultures for Gluten-Free Sourdough.

With the increasing number of people affected by gluten consumption-related diseases, adhering to a gluten-free (GF) diet is the most effective preventive measure. Herein, we aimed to isolate and characterize the functional properties of autochthonous lactic acid bacteria (LAB) and yeast from various GF sourdoughs to determine their suitability in starter cultures for sourdough preparation. Three LAB, Weissella confusa BAQ2, Lactobacillus brevis AQ2, Leuconostoc citreum YC2, and Saccharomyces cerevisiae BW1, were identified. The isolated LAB exhibited greater TTA, faster acidification rates, and higher acid tolerance than commercial LAB. W. confusa BAQ2 exhibited the highest EPS production, W. confusa BAQ2 and L. brevis AQ2 showed high maltose utilization, and S. cerevisiae BW1 exhibited the highest CO2 production rate. Accordingly, all four microbial strains were mixed for the starter culture. The sourdough prepared with starter cultures exhibited differences in gas production depending on fermentation time, which influenced the volume of GF bread dough. GF bread prepared with fermented sourdough exhibited a 16% higher specific volume and enhanced crumb firmness and elasticity than that prepared using non-fermented sourdough. Thus, autochthonous LAB strains isolated from various GF sourdoughs can be used together to improve the quality of sourdough bread, demonstrating their potential for use in starter cultures for GF sourdough production.

Natural Extracts as Inhibitors of Microorganisms and Lipid Oxidation in Emulsion Sausage during Storage.

Food additives are required to maintain the freshness and quality of foods, particularly meats. However, chemical additives may not be preferred by consumers, and natural materials with antimicrobial and antioxidant effects may be used as replacements for common chemical additives. Accordingly, in this study, we compared the antimicrobial and antioxidant activities of natural compounds extracted with ethanol and hot water, and emulsion sausage prepared with natural ethanol extracts was analyzed for pH, color, thiobarbituric acid reactive substances (TBARS), and Clostridium perfringens growth during storage. The antimicrobial activities of 49 natural extract candidates against Listeria monocytogenes, C. perfringens, Salmonella spp., and Escherichia coli were analyzed, and six natural materials with excellent antibacterial activities, i.e., Elaeagnus umbellata Thunb. f. nakaiana (Araki) H. Ohba, Punica granatum L., Ecklonia cava, Nelumbo nucifera Gaertner, and Schisandra chinensis (Turcz.) Baill., and Rubus coreanus Miq. were evaluated to determine their total polyphenol contents and DPPH radical scavenging activities. The total polyphenol contents of ethanol extracts were higher than those of hot water extracts, whereas DPPH radical scavenging activity was found to be higher in hot water extracts. The TBARS values of emulsion sausages were significantly increased as storage time increased, and the TBARS values of emulsion sausages prepared with natural extracts were lower than those of control sausages. Natural extract-treated emulsion sausages showed a 99% reduction in bacterial contents compared with untreated sausages on day 2, with greater than 99.9% reduction after day 3. Thus, these results demonstrated that natural extracts could have applications as natural preservatives in meat products.

Changes in physicochemical properties of rice flour by fermentation with koji and its potential use in gluten-free noodles.

To use rice flour as an ingredient in gluten-free noodles, improved texture properties such as increased hardness and reduced adhesiveness are required. We investigated the physicochemical characteristics of rice flour obtained by fermenting japonica rice with koji, determined the pasting and textural properties of the resulting gel, and suggested a method for producing gluten-free noodles. Koji-fermented rice flour was obtained by fermenting rice grains for 24 and 48 h. Koji fermentation reduced the protein and ash content of the rice and lowered the pH. The change in amylose content was not significant, but the short-to-long-chain ratio of amylopectin increased. Changes in the structural and compositional characteristics facilitated swelling of the rice flour and starch leaching. Variations in the gelatinization and hydration properties of the rice flour increased its peak viscosity and gel hardness, and reduced its gel adhesiveness. Noodles made from koji-fermented rice flour have improved physical features, such as modified textural properties resulting from a gel texture and increased whiteness, indicating that koji-fermented rice flour is a desirable noodle ingredient for gluten-free foods.

Development of Freeze-Thaw Stable Starch through Enzymatic Modification.

The use of unmodified starch in frozen foods can cause extremely undesirable textural changes after the freeze-thaw process. In this study, using cyclodextrin glucanotransferase (CGTase) and branching enzymes, an amylopectin cluster with high freeze-thaw stability was produced, and was named CBAC. It was found to have a water solubility seven times higher, and a molecular weight 77 times lower, than corn starch. According to the results of a differential scanning calorimetry (DSC) analysis, dough containing 5% CBAC lost 19% less water than a control dough after three freeze-thaw cycles. During storage for 7 days at 4 °C, bread produced using CBAC-treated dough exhibited a 14% smaller retrogradation peak and 37% less hardness than a control dough, suggesting that CBAC could be a potential candidate for clean label starch, providing high-level food stability under repeated freeze-thaw conditions.

Development of an enzymatic encapsulation process for a cycloamylose inclusion complex with resveratrol.

Cyclodextrin glucanotransferase (CGTase; EC 2.4.1.19) produces cycloamyloses (CAs), which are large cyclic glucans, and subsequently transforms them to α-, ß-, and γ-cyclodextrins. We developed a novel encapsulation process based on the cyclization activity of CGTase and applied it to the formation of CA inclusion complexes with resveratrol (RVT), which has limited bioavailability due to its low water solubility. The encapsulated RVT (CA-RVT) was purified using preparative high-performance liquid chromatography. The water solubility of CA-RVT was 6,000-fold higher than that of RVT. CA-RVT in water demonstrated 98% stability for 1 week at 4 °C. According to radical scavenging activity and anti-inflammatory assays, CA-RVT in aqueous solution exhibited similar activities as an equal amount of RVT in dimethyl sulfoxide, suggesting the limited solubility of RVT can be overcome through CA encapsulation by CGTase, thus enhancing its nutraceutical value as a functional ingredient in the food industry.

Properties of recombinant 4-α-glucanotransferase from Bifidobacterium longum subsp. longum JCM 1217 and its application.

To determine the physiochemical properties of the 4-α-glucanotransferase from Bifidobacterium sp., the bllj_0114 gene encoding 4-α-glucanotransferase was cloned from Bifidobacterium longum subsp. longum JCM 1217 and expressed in Escherichia coli. The amino acid sequence alignment indicated that the recombinant protein, named BL-αGTase, belongs to the glycoside hydrolase (GH) family 77. BL-αGTase was purified using nickel-nitrilotriacetic acid affinity chromatography and characterized using various substrates. The enzyme catalyzed the disproportionation activity, which transfers a glucosyl unit from oligosaccharides to acceptor molecules, and had the highest activity at 40 °C and pH 6.0. In the presence of 5 mM metal ions, in particular Cu2+, Zn2+, and Fe2+, BL-αGTase activity was reduced. To determine whether BL-αGTase can be used to generate thermoreversible gels, potato starch was treated with BL-αGTase for various reaction times. The BL-αGTase-treated starches showed sol-gel reversibility and melted at 59.6-75.7 °C.

Characterization of novel α-galactosidase in glycohydrolase family 97 from Bacteroides thetaiotaomicron and its immobilization for industrial application.

Bacteroides thetaiotaomicron (B. thetaiotaomicron), which resides in the human intestinal tract, has a number of carbohydrate enzymes, including glycoside hydrolase (GH) family 97. Only a few GH 97 enzymes have been characterized to date. In this study, a novel α-galactosidase (Bt_3294) was cloned from B. thetaiotaomicron, expressed in Escherichia coli, and purified using affinity chromatography. This novel enzyme showed optimal activity at 60 °C and pH 7.0. Enzyme activity was reduced by 94.4% and 95.7% in the presence of 5 mM Ca2+ and Fe2+, respectively. It is interesting that Bt_3294 specifically hydrolyzed shorter α-galactosyl oligosaccharides, such as melibiose and raffinose. The D-values of Bt_3294 at 40 °C and 50 °C were about 107 and 6 min, respectively. After immobilization of Bt_3294, the D-values at 40 °C and 50 °C were about 37.6 and 29.7 times higher than those of the free enzyme, respectively. As a practical application, the immobilized Bt_3294 was used to hydrolyze raffinose family oligosaccharides (RFOs) in soy milk, decreasing the RFOs by 98.9%.