Macular pigment-enriched oil production from genome-edited microalgae.

BACKGROUND: The photosynthetic microorganism Chlamydomonas reinhardtii has been approved as generally recognized as safe (GRAS) recently, this can excessively produce carotenoid pigments and fatty acids. Zeaxanthin epoxidase (ZEP), which converts zeaxanthin to violaxanthin, and ADP-glucose pyrophosphorylase (AGP). These are key regulating genes for the xanthophyll and starch pathways in C. reinhardtii respectively. In this study, to produce macular pigment-enriched microalgal oil, we attempted to edit the AGP gene as an additional knock-out target in the zep mutant as a parental strain. RESULTS: Using a sequential CRISPR-Cas9 RNP-mediated knock-out method, we generated double knock-out mutants (dZAs), in which both the ZEP and AGP genes were deleted. In dZA1, lutein (2.93 ± 0.22 mg g-1 DCW: dried cell weight), zeaxanthin (3.12 ± 0.30 mg g-1 DCW), and lipids (450.09 ± 25.48 mg g-1 DCW) were highly accumulated in N-deprivation condition. Optimization of the culture medium and process made it possible to produce pigments and oil via one-step cultivation. This optimization process enabled dZAs to achieve 81% higher oil productivity along with similar macular pigment productivity, than the conventional two-step process. The hexane/isopropanol extraction method was developed for the use of macular pigment-enriched microalgal oil for food. As a result, 196 ± 20.1 mg g-1 DCW of edible microalgal oil containing 8.42 ± 0.92 mg g-1 lutein of oil and 7.69 ± 1.03 mg g-1 zeaxanthin of oil was produced. CONCLUSION: Our research showed that lipids and pigments are simultaneously induced in the dZA strain. Since dZAs are generated by introducing pre-assembled sgRNA and Cas9-protein into cells, antibiotic resistance genes or selective markers are not inserted into the genome of dZA, which is advantageous for applying dZA mutant to food. Therefore, the enriched macular pigment oil extracted from improved strains (dZAs) can be further applied to various food products and nutraceuticals.

Effects of textured food masticatory performance in older people with different dental conditions.

BACKGROUND: Physiological deterioration (aging, poor dental status, and reduced tongue pressure) makes chewing difficult. This study aimed to investigate the chewing patterns of older people with or without dentures, evaluate the textural and masticatory properties of texture-modified radish Kimchi, and investigate the correlation between dental status and tongue pressure. Additionally, differences in the subjective-objective concordance of texture-modified Kimchi were investigated using the preference test. METHODS: This study included 32 Korean women aged between 65 and 85 years. Masticatory behavior was recorded by electromyography, and tongue pressure was measured using the Iowa Oral Performance Instrument. A preference test, with hardness as the relevant textural property, determined the participants' preferences among the test samples (food with a homogeneous structure-radish Kimchi). To assess preference differences, a questionnaire suitable for older people was designed. The preference for cooked radish Kimchi with various blanching times based on overall acceptability and self-reporting of preference was investigated to develop elderly-friendly food. The subjective scores indicated whether the sample (radish Kimchi) was hard or soft based on the chewing ability of the participants. Dental status, muscle activities, and tongue pressure were considered for the food design with optimized texture. The relationship between subject score and mastication properties were examined using multiple regression analysis. RESULTS: The number of chews and chewing time increased with hardness, significantly activating the masseter and temporalis muscles. The evaluation of masseter muscle activity, particularly for level-6 radish Kimchi, showed that older people with complete dentures chewed less actively than those with natural teeth (p < 0.05). The older people with natural teeth (18.94 ± 10.27 kPa) exhibited higher tongue pressure than those with complete dentures (10.81 ± 62.93 kPa), and the difference was statistically significant (p < 0.01). Older people preferred food with familiar tastes and textures. An association was found between the subjective hardness score and the objective hardness level. The perceived hardness intensity was linked to the chewing ability of the participant. Denture wearers exhibited a lower chewing ability, and at level 6, they perceived greater hardness of food than those with natural teeth. CONCLUSIONS: Developing food with a modified texture can bridge the gap between physiological and psychological aspects of food texture; texture-modified radish Kimchi, with limited blanching time, may be favorable for older people.

Physical Stability and Antioxidant Ability of a Sustainable Oil-in-Water Emulsion Containing Perilla Skin Extract and Upcycled Aquasoya Powder.

In response to environmental issues, upcycling has become a growing trend in the food industry. Aquasoya is a promising method to upcycle by-product from soybean processing due to its high protein contents and excellent emulsifying ability. In the present research, Aquasoya powder was used an emulsifier to incorporate the antioxidant compounds from perilla skin extract (PSE), namely rosmarinic acid, into oil-in-water (O/W) emulsion system and its physochemical stability was assessed. As a result, droplet size of the emulsion was smaller in PSE-incorporated emulsion (PO, 350.57 ± 9.60 b nm) than the emulsion without PSE (PX, 1045.37 ± 142.63 a nm). Centrifugal photosedimentometry analysis also revealed that the physical stability was significantly improved in PO, and the stability was maintained over 30 d of storage. Furthermore, as PO had a higher ABTS radical scavenging ability and showed slower initial lipid oxidation, it was concluded that PO has a higher antioxidant ability than PX. Conclusively, Aquasoya can be considered as an emulsifier in O/W emulsion with PSE because it can effectively integrate and stabilize the antioxidant substance derived from perilla skin.

Impact of varying food hardness on mastication/swallowing.

The purpose of this study was to analyze mastication and swallowing in the elderly and confirm the association with dysphagia characteristics. A questionnaire was developed to evaluate the masticating and swallowing functions of the elderly. Mastication was analyzed using electromyography, and tongue/lip pressures were measured using Iowa Oral Performance Instrument. The results of the questionnaire showed that statistical difference in the number of teeth between the group without and with, decreased ability to swallow, and there was a correlation with lip pressure. Additionally, the higher number of teeth, the higher muscle activity, and there is a positive correlation between the number of chews and the lip pressure. Consequently, our findings suggested oral health parameters are closely associated with mastication/swallowing ability. Finally, based on the results obtained for different foods tested, we suggested that texture-modified foods are necessary to enhance swallowing ability.

Modified Korean MIND Diet: A Nutritional Intervention for Improved Cognitive Function in Elderly Women through Mitochondrial Respiration, Inflammation Suppression, and Amino Acid Metabolism Regulation.

SCOPE: Mild cognitive impairment is associated with a high prevalence of dementia. The study examines the benefits of a modified Korean MIND (K-MIND) diet and explores biomarkers using multi-omics analysis. METHODS AND RESULTS: The K-MIND diet, tailored to the elderly Korean population, includes perilla oil, milk, or fermented milk, and avoids alcohol consumption. As a result, the K-MIND diet significantly improves subjects "orientation to place" in the Korean version of the Mini-Mental State Examination, 2nd edition test. According to multi-omics analysis, the K-MIND diet upregulates genes associated with mitochondrial respiration, including ubiquinone oxidoreductase, cytochrome C oxidase, and ATP synthase, and immune system processes, and downregulates genes related to nuclear factor kappa B activity and inflammatory responses. In addition, K-MIND affects the metabolic pathways of glycine, serine, threonine, tryptophan, and sphingolipids, which are closely linked to cognitive function through synthesis of neurotransmitters and structures of brain cell membranes. CONCLUSION: The findings imply that the K-MIND diet improves cognitive function by upregulating key genes involved in oxidative phosphorylation and downregulating pro-inflammatory cytokines.

Clinical Practice Guidelines for Oropharyngeal Dysphagia.

OBJECTIVE: Dysphagia is a common clinical condition characterized by difficulty in swallowing. It is sub-classified into oropharyngeal dysphagia, which refers to problems in the mouth and pharynx, and esophageal dysphagia, which refers to problems in the esophageal body and esophagogastric junction. Dysphagia can have a significant negative impact one's physical health and quality of life as its severity increases. Therefore, proper assessment and management of dysphagia are critical for improving swallowing function and preventing complications. Thus a guideline was developed to provide evidence-based recommendations for assessment and management in patients with dysphagia. METHODS: Nineteen key questions on dysphagia were developed. These questions dealt with various aspects of problems related to dysphagia, including assessment, management, and complications. A literature search for relevant articles was conducted using Pubmed, Embase, the Cochrane Library, and one domestic database of KoreaMed, until April 2021. The level of evidence and recommendation grade were established according to the Grading of Recommendation Assessment, Development and Evaluation methodology. RESULTS: Early screening and assessment of videofluoroscopic swallowing were recommended for assessing the presence of dysphagia. Therapeutic methods, such as tongue and pharyngeal muscle strengthening exercises and neuromuscular electrical stimulation with swallowing therapy, were effective in improving swallowing function and quality of life in patients with dysphagia. Nutritional intervention and an oral care program were also recommended. CONCLUSION: This guideline presents recommendations for the assessment and management of patients with oropharyngeal dysphagia, including rehabilitative strategies.

Evaluation of the Physicochemical and Functional Properties of Aquasoya (Glycine max Merr.) Powder for Vegan Muffin Preparation.

Recent concerns on health and sustainability have prompted the use of legumes as a source of plant-based proteins, resulting in the application of their cooking water as a substitute for egg whites. In this study, the cooking water of yellow soybeans was powdered, and, subsequently, the nutritional and functional characteristics of powders from yellow soybeans (YSP), chickpeas (CHP), and egg whites (EWP) were compared. The main components of these powders (total polyphenol, total carbohydrate, and protein), along with their hydration properties (hygroscopicity, water solubility index, and water/oil holding capacities), and emulsifying and foaming properties, were identified. The muffins prepared with YSP, CHP, and EWP were analyzed to determine their basic characteristics, such as volume, baking loss, and sensory attributes. The results of the powder analyses indicated that YSP was significantly superior to CHP and EWP, particularly in terms of holding capacities, and emulsion and foam stabilities. The sensory evaluation results showed that there was no statistically significant difference in overall acceptance among the muffin samples. Therefore, YSP can be used as an alternative to CHP or EWP, and applied as a novel ingredient in various vegan products.

Structural and functional modification of proteins from black soybean Aquasoya via ultrasonication.

Plant-based proteins obtained from agricultural by-products have garnered growing interest in response to consumer awareness of health and environmental issues. This study aimed to improve the functionalities of the proteins recovered from black soybean Aquasoya (PBSA) by modifying their structure via ultrasonication. PBSA was ultrasonicated with a frequency of 40 kHz at 350 W for different time periods (0, 20, 40, and 60 min), and its structural characteristics, physicochemical properties, and functional properties were investigated. Ultrasonication left the primary structure intact but altered the secondary and tertiary structures of the PBSA; α-helix and ß-sheet contents decreased, random coil contents increased, and buried non-polar amino acid residues were exposed. Moreover, ultrasound promoted an increase in free sulfhydryl content and a reduction in particle size. Consequently, functional properties, such as solubility, emulsion stability, and foaming performance were improved by modifying the structure and physicochemical properties of PBSA. This work demonstrates the potential of ultrasound, which is favorable for modifying the spatial conformation and related functionalities of proteins, thus meeting the needs of manufacturers to use function-enhanced proteins as food additives.

Formation of a novel coating material containing lutein and zeaxanthin via a Maillard reaction between bovine serum albumin and fucoidan.

The effective delivery of bioactive compounds has recently been receiving attention. In this study, a conjugate with BSA and fucoidan synthesized via the Maillard reaction was confirmed through electrophoresis, the o-phthalaldehyde assay, and through changes in absorbance. Two moles of fucoidan were glycated with one mole of BSA at 60 °C and 79% relative humidity for 4 days. The droplet coated with B-F conjugate remained stable during storage at 4 and 25 °C and slightly increased only at 55 °C however, the droplet coated with intact BSA and B/F mixture significantly increased. L/Z were degraded about 82, 79, and 36% for 4, 25, and 55 °C, respectively, regardless of the type of emulsifier. Although the conjugates could not prevent the degradation of lutein and zeaxanthin during storage, they improved the stability of the emulsion and showed 4.20-fold and 1.32-fold higher bioaccessibility than intact BSA and B/F mixtures, respectively.

Effect of the emulsifier type on the physicochemical stability and in vitro digestibility of a lutein/zeaxanthin-enriched emulsion.

Lutein (L) and zeaxanthin (Z), as macular pigments, are water-insoluble, chemically unstable, and have low bioaccessibilities; they are often emulsified to overcome these limitations. This study investigated the impact of various emulsifiers (ethyl lauroyl arginate (LAE); Tween 80; and sodium dodecyl sulfate (SDS)) on the physicochemical properties and in vitro digestibilities of L/Z-fortified oil-in-water emulsions. Droplet aggregation and creaming extents were dependent on the emulsifier type. The ζ-potentials of emulsions stabilized by LAE, Tween 80, and SDS were + 87, - 26, and - 95 mV, respectively. SDS-stabilized emulsion had the smallest particles, while the particle sizes for the LAE- and Tween 80-stabilized emulsions were larger and not significantly different. The rates of L/Z degradation were sensitive to the emulsifier type and to heat, not to light. The L/Z bioaccessibility was the highest for the Tween 80 emulsion. Surfactants should therefore be carefully selected to optimize L/Z physicochemical stability and bioaccessibility in emulsions.

The particle size of rice flour greatly affects the structural, textural and masticatory properties of steamed rice cake (Baekseolgi).

In this study, steamed rice cakes prepared with different particle sized flour were used to investigate how the structures of rice cakes affected masticatory properties and bolus starch hydrolysis. Decreasing the particle size increased the surface areas requiring hydration, resulting in a loose structure and fluffy starch network during gelatinization. Increasing the particle size led to a tight and firm network, but was easily melted in the oral cavity. The chewing cycle and time differed among the samples. The conditions of inter-individual salivary flow rate and salivary α-amylase were diverse. The oral carbohydrate hydrolysis in the bolus before swallowing showed no significant differences in reducing sugar levels among particle sizes. However, salivary concentration was related to initial starch hydrolysis in the oral cavity, indicating the food structures affected mastication factors and physiological conditions. This study provides food structure and physiological factor information that could help design customized foods in industry.

Revalorization of the Cooking Water (Aquafaba) from Soybean Varieties Generated as a By-Product of Food Manufacturing in Korea.

Concerns regarding sustainability have prompted the search of value in the by-products of food manufacturing. Such is the case of the cooking water (CW) of chickpeas, which has shown its potential as a vegan egg white replacement. This study aimed to characterize and compare the CW from three novel legumes (black soybeans, BSB; yellow soybeans, YSB; and small black beans, SBB) obtained from the processing of Korean soybean foods, and the widely used CW from chickpeas (CH), with regard to total polyphenol, total carbohydrate, and protein contents, and further compare their foaming and emulsifying abilities and stabilities. Compositional analysis revealed that all the studied legumes possessed higher values than CH for all parameters. Furthermore, the CW from these legumes exhibited enhanced functional properties, particularly foaming capacity and stability. Taken together, our results suggest that the CW from BSB, YSB, and SBB, sourced from the manufacturing of legume food products, has the potential of being revalorized as a plant-based functional ingredient for vegan product development.

Aquafaba from Korean Soybean I: A Functional Vegan Food Additive.

The substitution of animal-based foods (meat, eggs, and milk) with plant-based products can increase the global food supply. Recently, pulse cooking water (a.k.a. aquafaba) was described as a cost-effective alternative to the egg in gluten-free, vegan cooking and baking applications. Aquafaba (AQ) forms stable edible foams and emulsions with functional properties that are like those produced by whole egg and egg white. However, the functional ingredients of AQ are usually discarded during food preparation. In this study, Korean-grown soy (ver. Backtae, Seoritae, and Jwinunikong) and chickpea were used to produce AQ. Two approaches were compared. In the first, seed was cooked at an elevated pressure without presoaking. In the second, seed was soaked, then, the soaking water was discarded, and soaked seed was cooked at an elevated pressure. Both approaches produced a useful emulsifier, but the latter, with presoaking, produced a superior product. This approach could lead to a process that involves a small number of efficient steps to recover an effective oil emulsifier, produces no waste, and is cost-effective. The AQ product from Backtae (yellow soybean) produced emulsions with better properties (90%) than AQ produced from other cultivars and produced more stable food oil emulsions. This study will potentially lead to gluten-free, vegan products for vegetarians and consumers with animal protein allergies. This is the first report of the efficient production of AQ, an egg white substitute derived from cooked soybean of known cultivars.

Characterization of chitosan extracted from Mealworm Beetle (Tenebrio molitor, Zophobas morio) and Rhinoceros Beetle (Allomyrina dichotoma) and their antibacterial activities.

Chitin was isolated from the larvae, adult, and superworm of Mealworm Beetle and the larvae, pupa, and adult of Rhinoceros Beetle by deproteinization and demineralization. Chitin yields were 4.60, 8.40, and 3.90% in the larvae, adult, and superworm of Mealworm Beetle and 10.53, 12.70, and 14.20% in the larvae, pupa, and adult of Rhinoceros Beetle, respectively. Subsequently, chitosan was obtained by deacetylation of chitin extractions from all stages of both beetle species, and its yields from chitin were 80.00, 78.33, and 83.33% in the larvae, adult, and superworm of Mealworm Beetle, and 83.37, 83.37, and 75.00% in larvae, pupa, and adult of Rhinoceros Beetle, respectively. The Fourier transform infrared spectroscopy (FTIR) and X-ray diffraction (XRD) analysis of the chitosan from the two insect species showed the similar structural characteristics as those of commercial chitosan. Chitosan from the Mealworm Beetle showed about 1 mm inhibition zones against Bacillus cereus, Listeria monocytogenes, and Escherischia coli, and a 2 mm inhibition zone against Staphylococcus aureus in antimicrobial activity test.

Characteristics of cricket (Gryllus bimaculatus) chitosan and chitosan-based nanoparticles.

The field cricket (Gryllus bimaculatus) is commonly consumed as food in different parts of the world. This study was performed to characterize the chitosan extracted from crickets and to assess its potential use to the growing functional market. The degree of deacetylation (DA), Fourier-transform infrared spectra, X-ray diffraction patterns, molecular mass, scanning electron microscopy spectra, and color were measured. Cricket chitosan nanoparticles were prepared, and the optimal conditions were identified. The molecular mass of the cricket chitosan was lower than that of commercial chitosan; however, the DA, FTIR, and XRD spectra were similar. The particle size (208.27 ± 3.47 nm), zeta potential (35.72 ± 1.29 mV), and polydispersity index (PDI: 0.27 ± 0.03) of the cricket chitosan NPs were superior to the commercial. Addition of NaCl reduced the cricket chitosan NPs size up to 15.5%. This finding is a novel trial to prove the availability of the insect chitosan with a low molecular mass as an active carrier source.

Fucoidan improves the structural integrity and the molecular stability of ß-lactoglobulin.

ß-lactoglobulin (ß-lg) was covalently bonded with fucoidan through Maillard reaction at 60 °C for 96 h under 79% RH condition. The molecular characters of the conjugate were investigated using fourier transform infrared spectroscopy (FT-IR), atomic force microscopy (AFM), and circular dichroism (CD) spectroscopy. And, its thermal properties, surface activity, and zeta-potential were compared with intact ß-lg, ß-lg-fucoidan mixture, and fucoidan under different pH conditions. AFM indicated that the conjugate was nano-structured, regular spherical-shaped and generally large sized compared to ß-lg-fucoidan mixture. CD spectra and FT-IR showed that tertiary structure of ß-lg slightly unfolded, but little change in secondary structure occurred. This explained that glycation under Maillard condition resulted in a molten globule state of ß-lg. Differential scanning calorimetry (DSC) data exhibited that fucoidan shifted the temperature of phase transition and improved thermal stability of ß-lg molecule. In addition, the conjugate prominently decreased the surface tension with pH-dependency.

Quantitative analysis of phosphinothricin-N-acetyltransferase in genetically modified herbicide tolerant pepper by an enzyme-linked immunosorbent assay.

An immunoassay method was developed to quantitatively detect phosphinothricin-N-acetyltransferase (PAT) encoded by the Bialaphos resistance (bar) gene in genetically modified (GM) pepper. The histidine-tagged PAT was overexpressed in Escherichia coli M15 (pQE31-bar) and efficiently purified by Ni2+ affinity chromatography. A developed sandwich enzyme-linked immunosorbent assay (S-ELISA) method (detection limit: 0.01 microg/ml) was 100-fold more sensitive than a competitive indirect ELISA (CI-ELISA) method or Western blot analysis in detecting the recombinant PAT. In real sample tests, PAT in genetically modified herbicide-tolerant (GMHT) peppers was successfully quantified [4.9 +/- 0.4 microg/g of sample (n = 6)] by the S-ELISA method. The S-ELISA method developed here could be applied to other GMHT crops and vegetables producing PAT.

Functional improvements in bovine serum albumin-fucoidan conjugate through the Maillard reaction.

The solubility, thermal stability, surface activity and emulsifying properties of native bovine serum albumin (BSA), heat-treated BSA, a BSA-fucoidan mixture, and a BSA-fucoidan conjugate were assessed. Covalent linkage of BSA with fucoidan resulted in significantly (p < 0.05) high solubility after heating at 90 °C for 15 min, particularly at pH 5. The BSA-fucoidan conjugate had a high melting temperature (97.09 ± 1.45 °C), as found by differential scanning calorimetry, indicating strong heat stability and high resistance to denaturation. Although the attachment of fucoidan, a non-surface-active hydrophilic polysaccharide, gave no change in the surface activity, the emulsifying activity and the emulsion stability of the conjugate at pH 5 were superior to those of native BSA, heat-treated BSA, and the BSA-fucoidan mixture. Conclusively, fucoidan attachment enhanced the solubility, thermal stability and emulsifying properties of the protein molecules with negative charge distribution and steric stabilization.

Characterisation of bovine serum albumin-fucoidan conjugates prepared via the Maillard reaction.

Bovine serum albumin (BSA)-fucoidan conjugates were prepared by the Maillard reaction (60 °C and 79% relative humidity for 96 h), and were then identified by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and size-exclusion chromatography (SEC). Molecular characteristics of the BSA-fucoidan conjugates were investigated, using atomic force microscopy (AFM), dynamic light scattering (DLS), fluorescence spectroscopy, and circular dichroism spectroscopy. SDS-PAGE patterns provided evidence for the covalent bonding between BSA and fucoidan. SEC profiles showed that about 1.5-2.0 mol of fucoidan were covalently linked to 1 mol of BSA, resulting in high-molecular-weight compositions (conjugates). AFM images and DLS results indicated that most particles in the conjugates were nano-structured and more spherical than those of a regular BSA-fucoidan mixture. The fluorescence intensity and maximum emission wavelength of the conjugates together revealed that the BSA molecules had converted from an ordered conformation into a partially folded molten globule state.

Differences in eating behaviors and masticatory performances by gender and obesity status.

This study aimed to determine whether there might be differences in masticatory performance and eating behaviors by gender and obesity status. Forty eight (24 males; 24 females) non-obese and pre-obese young adults were matched for age, gender, and dental health. Eating behaviors were assessed using the Three-Factor Eating Questionnaire (TFEQ), and chewing performance while eating 152g of boiled rice was measured using electromyography (EMG). Compared with non-obese participants, pre-obese participants had significantly higher levels of disinhibition according to the TFEQ (P<0.05 for males; P<0.01 for females). Microstructures of chewing performance were significantly different by gender but not by obesity status. Males had a greater bite size (P<0.05), greater chewing power (P<0.001) and a faster eating rate (P<0.05) than females. Females habitually chewed more (P<0.05) and had a longer meal duration (P<0.01) than males. Eating rate was positively associated with disinhibition (r=0.490, P<0.05) score in males. Bite size (r=-0.449, P<0.05 for non-obese) and chewing power (r=-0.581, P<0.01 for non-obese and r=-0.446, P<0.05 for pre-obese) were negatively associated with disinhibition score. Results suggest that the effects of gender and, in part, obesity on eating responses may be explained as chewing performance. Therefore, gender-specific interventions and counseling aimed at slowing the rate of ingestion could be promising behavioral treatments for obese persons.