Effects of molecular weight fraction on antioxidation capacity of rice protein hydrolysates.

Rice protein was used as a starting material to provide rice protein hydrolysates (RPH) through enzyme-assisted extraction. RPH was further fractionated using ultrafiltration membrane (UF) and classified by molecular weight (MW; MW < 1 kDa, MW 1-10 kDa, and MW > 10 kDa). Peptides with MW < 1 kDa possessed superior antioxidant properties (p < 0.05). Therefore, UF demonstrated great efficacy in selectively separating antioxidant peptides. A Pearson correlation analysis revealed that the total phenolic concentration was correlated with oxygen radical absorbance capacity (ORAC; r = 0.999, p < 0.05). Amino acid contents had negative correlations with the scavenging activity (specifically, IC50) of 1,1-diphenyl-2-picrylhydrazyl and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) radicals (r = - 0.986 to - 1.000). Reducing power was related to aromatic amino acid contents (r = 0.997, p < 0.05). In this study, enzymatic hydrolysis was discovered to be an effective method of extracting and isolating natural antioxidant proteins from broken rice, thus preserving the nutritional quality of rice and making those proteins more accessible in future applications.

Mulberry Leaf (Morus alba L.) Extracts and Its Chlorogenic Acid Isomer Component Improve Glucolipotoxicity-Induced Hepatic Lipid Accumulation via Downregulating miR-34a and Decreased Inflammation.

Mulberry leaf (Morus alba L.) is used as a traditional medicine and potential health food to treat various metabolic diseases, such as hypertension, diabetes, and hyperlipidemia. However, we sought the mechanisms by which functional components of mulberry leaves mediate diabetic steatohepatitis. We applied an in vitro model of HepG2 cells induced by glucolipotoxicity and evaluated the effects of MLE and its major components nCGA, Crp, and CGA. The results showed that MLE and nCGA reduced liver fat accumulation by inhibiting SREBP-1/FASN, SREBP-2/HMG-CoAR, and activating PPARα/CPT-1. Additionally, MLE and nCGA decreased inflammatory responses associated with NF-κB, TNF-α, and IL-6 to alleviate steatohepatitis. Furthermore, we showed that MLE and nCGA exerted anti-glucolipotoxicity effects by downregulating miR-34a, thus activating SIRT1/AMPK signaling, and subsequently suppressing hepatic lipid accumulation.

Evaluating the Antioxidants, Whitening and Antiaging Properties of Rice Protein Hydrolysates.

Plant-derived protein hydrolysates have potential applications in nutrition. Rice protein hydrolysates (RPHs), an excellent source of proteins, have attracted attention for the development of cosmeceuticals. However, few studies have reported the potential application of RPH in analysis, and this study examined their antioxidant activities and the inhibitory activities of skin aging enzymes. The results indicated that the total phenolic and flavonoid concentrations were 2.06 ± 0.13 mg gallic acid equivalent/g RPHs and 25.96 ± 0.52 µg quercetin equivalent/g RPHs, respectively. RPHs demonstrated dose-dependent activity for scavenging free radicals from 1,1-diphenyl-2-picrylhydrazyl [half-maximal inhibitory concentration (IC50) = 42.58 ± 2.1 mg/g RPHs] and 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (IC50 = 2.11 ± 0.88 mg/g RPHs), dose-dependent reduction capacity (6.95 ± 1.40 mg vitamin C equivalent/g RPHs) and oxygen radical absorbance capacity (473 µmol Trolox equivalent/g RPHs). The concentrations of the RPH solution required to achieve 50% inhibition of hyaluronidase and tyrosinase activities were determined to be 8.91 and 107.6 mg/mL, respectively. This study demonstrated that RPHs have antioxidant, antihyaluronidase, and antityrosinase activities for future cosmetic applications.

Implementation of food safety management systems that meets ISO 22000:2018 and HACCP: A case study of capsule biotechnology products of chaga mushroom.

In our study, a food safety management system was developed for a chaga mushroom biotechnology product manufacturer, with the purpose of meeting the quality demands of customers and enhancing the manufacturer's reputation. The study focused on identifying the potentially significant hazards present at each stage of the production process for chaga mushroom capsule products, and on ensuring that the biotechnology company in question has fully implemented ISO22000:2018 and the HACCP methodology. The results indicate that, in the 1 year following the implementation of ISO 22000:2018, there was a statistically significant drop (P < 0.05) in the coliforms level of the tested biotechnology product samples that started in the ninth month following implementation. The rapid screening of mycotoxin, heavy metal, and pesticide residue levels also increased monitoring intensity and strengthened the periodic rotation plan, enabling control over potential problems relating to raw materials and ensuring product quality. This finding reveals the importance and necessity of rapid screening for small- and medium-sized food industry enterprises. Furthermore, 1 year after the system's implementation, the defect rate for chaga products was also observed to have declined, and the number of process flow anomalies requiring correction was also found to have decreased significantly (P < 0.05), indicating that safety and quality levels were improving and stabilizing. If implemented over a longer period of time, the food safety management system's benefits would stand out even more, and significant improvements would be observed for more monitored items. PRACTICAL APPLICATION: Few studies have touched on food safety management systems (FSMSs) developed for capsule health food products. The enterprise examined in this study had actively worked to improve its quality system and meet its customers' needs through the implementation of the FSMSs.

Effects of operating conditions on biogas production in an anaerobic digestion system of the food and beverage industry.

BACKGROUND: Food residuals (FR) were anaerobically biotransformed to produce biogases (e.g. methane and hydrogen), and different pre-treatment conditions, including particle size, oil content, pH and salt content, were controlled in this study. The bio-solids of a municipal solid waste (MSW) from a wastewater treatment plant were added to assess its effect on anaerobic transformation efficiency and gas yields. RESULTS: The breaking of FR and the application of MSW were effective in enhancing the transformation efficiency and yield of biogases. The energy transfer efficiency value of the combined FRs used in this study was probably 23%. However, it can be very cost effective to apply arbitrary proportions to treat two types of FR in the anaerobic digestion tank of a wastewater treatment plant. It was also found that the alkalinity and pH value were two major parameters that controlled the success of the transformation. About 0.16-0.17 kg of alkalinity was needed during the anaerobic digestion of 1 kg dry FR, but this requirement was decreased by the treatment applying MSW. Olive oil had higher reducing rates when used as a substitute for heat-oxidized oil to study the effect of oil content on methylation. CONCLUSION: The conditions for anaerobic digestion established in this study were practical for the digestion of FR in wastewater treatment plants in Taiwan. However, we nonetheless found that it was cost effective to use arbitrary proportions for both types of FR and integrate the anaerobic digestion process used in wastewater treatment plants. © 2020 Society of Chemical Industry.

The Influence of Submerged Fermentation of Inonotus obliquus with Control Atmosphere Treatment on Enhancing Bioactive Ingredient Contents.

This study led in the pioneering technique incubated in a bioreactor with the forced air injection system. The purpose of this study was to establish the optimal incubation conditions for this technique. The results showed that the speed at which Inonotus obliquus was incubated with the forced air injection system was superior to that with a normal bioreactor. A nitrogen to oxygen ratio of 50:50 provided the best results with the forced air injection system, including in terms of the achievement of biomass, total triterpenes, betulinic acid content, and the scavenging activities of DPPH radicals, which reached up to 21.3 g/1000 mL, 2.1 g/1000 mL, 1.9 g/1000 mL, and 87.3%, respectively. The results showed that the bioreactor with the forced air injection system could more effectively incubate I. obliquus by using less vapor while still utilizing a model close to that of a traditional bioreactor. The innovative bioreactor fermentation model was thus more economical than the traditional bioreactor model.

Crystallographic analysis of the Staphylococcus epidermidis lipase involved in esterification in aqueous solution.

The Staphylococcus epidermidis lipase (SeLip, GehC) can be used in flavour-compound production via esterification in aqueous solution. This study reports the crystallization and crystallographic analysis of recombinant GehC (rGehC; Lys303-Lys688) with a molecular weight of 43 kDa. rGehC was crystallized at 293 K using PEG 10 000 as a precipitant, and a 99.9% complete native data set was collected from a cooled crystal at 77 K to a resolution of 1.9 Šwith an overall Rmerge value of 7.3%. The crystals were orthorhombic and belonged to space group P212121, with unit-cell parameters a = 42.07, b = 59.31, c = 171.30 Å, α = ß = γ = 90°. Solvent-content calculations suggest that there is likely to be one lipase subunit in the asymmetric unit.

A 90-Day Subchronic Toxicity Study of Submerged Mycelial Culture of Cordyceps cicadae (Ascomycetes) in Rats.

Cordyceps cicadae is a parasitic fungus that hibernates inside a host (Cicada flammata Dist.) and then grows its fruiting body on the surface of the insect. The complete insect/fungus combination of C. cicadae has been widely applied in Chinese traditional medicine. Recent studies have demonstrated that the medicinal benefits of cultured mycelia are as effective as those found in the wild. However, toxicological information regarding the chronic consumption of C. cicadae mycelia culture is not available. This study was conducted to evaluate the possible toxicity arising from repeated exposure to freeze-dried submerged mycelial culture of C. cicadae for 90 days. A total of eighty 8-week-old Sprague-Dawley rats were divided into 4 groups (10 males and 10 females in each group). C. cicadae was administered daily to animals by gavage at doses of 0, 500, 1000, and 2000 mg/kg body weight for 90 days. No animal deaths occurred and no treatment-related clinical signs were observed during the study period. No statistical differences in body weight gain, relative organ weight, hematology, serum chemistry, and urinalysis were observed. Gross necropsy and histopathological findings indicated that there was no treatment-related abnormality. Based on the results, the no observed adverse effect level of C. cicadae whole broth is determined to be > 2000 mg/kg for male and female Sprague-Dawley rats. The results of this study provides support for the use of C. cicadae fermentation product as a safe agent in functional food.

Evaluation of the toxicological safety of erinacine A-enriched Hericium erinaceus in a 28-day oral feeding study in Sprague-Dawley rats.

Natural products have attained great importance as they are believed to be the new alternative medicines for conventional therapy. As numerous studies have proved the tremendous medicinal values of Hericium erinaceus, it is necessary to take into account its safety as well as its risk for the recipient. However, mushroom mycelium has an identity distinct from mushrooms, as two specific classes of compounds, hericenones and erinacines, can only be extracted from both the fruit body and the cultured mycelium, respectively. Therefore, this is the first report on the evaluation of the toxicity of H.erinaceus mycelium, enriched with 5mg/g erinacine A, by a 28-day repeated oral administration study in Sprague-Dawley rats. Three doses of 1 (Low), 2 (Mid) and 3 (High) g/kg body weight/day were selected for the study while distilled water served as control. All animals survived to the end of the study. No abnormal changes were observed in clinical signs. No adverse or test article-related differences were found in urinalysis, haematology and serum biochemistry parameters, between the treatment and control groups. No gross pathological findings and histopathological differences were seen. Therefore, the no-observed-adverse-effect level of erinacine A-enriched H.erinaceus is greater than 3g/kgbody weight/day.

Simultaneous production of fatty acid methyl esters and diglycerides by four recombinant Candida rugosa lipase's isozymes.

In this study, the catalytic efficiency of four recombinant CRL (Candida rugosa lipase) isozymes (LIP1-LIP4) towards the production of fatty acid methyl ester (FAME) was compared and evaluated as an alternative green method for industrial applications. The results indicated that the recombinant C. rugosa LIP1 enzyme exhibited the highest catalytic efficiency for FAME production compared to the recombinant C. rugosa LIP2-LIP4 enzymes. The optimal conditions were as follows: pH 7.0, methanol/soybean oil molar ratio: 3/1, enzyme amount: 2U (1.6 µL), reaction temperature: 20°C, 22 h of reaction time, and 3 times of methanol addition (1 mol/6h), and resulted in 61.5 ± 1.5 wt.% of FAME conversion. The reaction product contained also 10 wt.% of DAG with a ratio of 1,3-DAG to 1,2-DAG of approximately 4:6, and can be potentially used in industrial applications as a food emulsifier.

Genotoxicity profile of erinacine A-enriched Hericium erinaceus mycelium.

Hericium erinaceus (H. erinaceus) has a long history of usage in traditional Chinese medicine for the treatment of gastric disorders. Recently, it has become a well-established candidate in causing positive brain and nerve health-related activities by inducing nerve growth factor (NGF) from its bioactive ingredient, erinacine A. This active compound, which exists only in fermented mycelium but not in its fruiting body, increases NGF levels in astroglial cells in vitro as well as catecholamine and NGF levels in vivo. With increasing recognition of erinacine A in H. erinaceus (EAHE) mycelium improving neurodegenerative diseases, numerous products are being marketed based on these functional claims. To our knowledge, there have been no reports on the mutagenicity of EAHE prior to this paper. Hence, the present study was undertaken to determine the mutagenicity and genotoxicity effects of EAHE mycelium conducted in three standard battery of tests (reverse mutation, chromosomal aberration, and micronuclei tests) according to the latest guidelines in order to meet all international regulatory requirements and provide information on the safety of this new and promising natural remedy. Our results have indicated that EAHE mycelium did not significantly increase the number of revertant colonies in the bacterial reverse mutation test nor induce higher frequency of aberrations in the chromosome aberration test. Moreover, no statistically significant EAHE mycelium-related increase was observed in the incidence of reticulocytes per 1000 red blood cells and micronucleated reticulocytes per 1000 reticulocytes. In conclusion, the three standard battery of tests suggested that EAHE mycelium was devoid of mutagenicity and genotoxicity in the tested doses and experimental conditions.

Enhancement of the immune response against Salmonella infection of mice by heat-killed multispecies combinations of lactic acid bacteria.

Heat-killed lactic acid bacteria (LAB) has advantages over live LAB in that it has a long shelf-life and is therefore easy to store and transport. From four LAB strains selected by immunomodulatory activity and adherent properties, we prepared the heat-killed multispecies combination of LAB (MLAB) and the cell walls from MLAB under two conditions (100 °C for 30 min and 121 °C for 15 min). Different effects on the adherent properties of these four LAB strains were observed, depending on the heating conditions. With mouse macrophage cells, the two heat-killed MLABs (HMLABs) showed significantly higher induction activities on the production of interleukin 12 (IL-12) than their individual strains did. Heat-killed MLABs and cell-wall preparations were able to reduce the Salmonella invasion of Caco-2 and mouse macrophage cells. Feeding mice with HMLAB could inhibit the Salmonella invasion of mice significantly. For these mice, the expression level of pro-inflammatory cytokines, such as TNF-α and IL-6, in mouse serum was reduced while that of the anti-inflammatory cytokine, i.e. IL-10, was enhanced. The HMLABs developed in this study showed higher protective effect against Salmonella invasion either of Caco-2 cells or of mice, relative to the heat-killed lactobacilli, which consisted of Lactobacillus acidophilus strains selected at random. In conclusion, the HMLABs were potentially useful for the protection of mice against Salmonella infection and the induced inflammation.

Integrated biocatalytic process for trehalose production and separation from rice hydrolysate using a bioreactor system.

Rice starch can be hydrolyzed into maltose for trehalose bioconversion by enzymatic methods. In this study, we have successfully established an efficient production system for our recombinant PTTS in large scale. Three bio-treatments were developed to simplify the separation and purification of trehalose from complex rice saccharified liquid. The trehalose conversion rate of 64.63±4.05% at 30 °C can be reached using rice hydrolysate as the substrate in a 5l fermentor system. By 1% of raw material koji fermentation, the highest concentration of bioethanol (3.61±0.07%) was obtained at 30 °C for 36 h. After 12h of reaction time, the gluconic acid (24.47±0.33 mM) was successfully produced by glucose oxidase (40 U/g rice) using residual glucose as a substrate. After the batch/continuous ionic exchange process, the trehalose can be successfully separated, crystallized and identified as 92.6±0.02% purity and 94.2% of the recovery yield, respectively.

Simultaneous production of trehalose, bioethanol, and high-protein product from rice by an enzymatic process.

Rice is a starch-rich raw material that can be used for trehalose production. It can be hydrolyzed with alpha-amylase, beta-amylase, and pullulanase to produce high-maltose content of rice saccharified solution for bioconversion of maltose into trehalose by trehalose synthase (TSase). For this purpose, an efficient enzymatic procedure has been successfully developed to simultaneously produce value-added trehalose, bioethanol, and high-protein product from rice as substrate. The highest maltose yield produced from the liquefied rice starch hydrolysate was 82.4 +/- 2.8% at 50 degrees C and pH 5.0 for 21-22 h. The trehalose conversion rate can reach at least 50% at 50 degrees C and pH 5.0 for 20-24 h by a novel thermostable recombinant Picrophilus torridus trehalose synthase (PTTS). All residual sugar, except trehalose, can be fully hydrolyzed by glucoamylase into glucose for further bioethanol production. The insoluble byproduct containing high yields of protein (75.99%) and dietary fiber (14.01%) can be processed as breakfast cereal product, health food, animal forage, etc. The conversion yield of bioethanol was about 98% after 64 h of fermentation time by Saccharomyces cerevisiae without any artificial culture solution addition. Ethanol can easily be separated from trehalose by distillation with a high recovery yield and purity of crystalline trehalose of 92.5 +/- 8.7% and 92.3%, respectively.

High dose vitamin C supplementation increases the Th1/Th2 cytokine secretion ratio, but decreases eosinophilic infiltration in bronchoalveolar lavage fluid of ovalbumin-sensitized and challenged mice.

Vitamin C is traditionally regarded to be beneficial for asthma, however the benefit is still controversial. In the present study, high dose vitamin C was supplemented to ovalbumin (OVA)-sensitized and challenged mice to evaluate the effects of dietary vitamin C on allergic asthma. In this study, the experimental mice were divided into four groups, including nonsensitized control, dietary control, positive control (cured ip with dexamethasone), and high dose vitamin C supplementation (130 mg of vitamin C/kg bw/day by gavage for 5 weeks). Differential leukocyte counts, levels of inflammatory mediators, as well as type 1 T-helper lymphocytes (Th1)-type and type 2 T-helper lymphocytes (Th2)-type cytokines in the bronchoalveolar lavage fluid (BALF) were determined. The results showed that both high dose vitamin C supplementation and dexamethasone treatments significantly (P < 0.05) decreased eosinophilic infiltration into BALF. High dose vitamin C supplementation significantly increased the secretion ratio of interferon (IFN)-gamma/interleukin (IL)-5 cytokines. This study suggests that high dose vitamin C supplementation might attenuate allergic inflammation in vivo via modulating the Th1/Th2 balance toward the Th1 pole during the Th2-skewed allergic airway inflammation and decreasing eosinophilic infiltration into BALF.

Dietary perilla oil lowers serum lipids and ovalbumin-specific IgG1, but increases total IgE levels in ovalbumin-challenged mice.

Our previous studies indicated that alpha-linolenic acid (ALA)-rich perilla oil might alleviate bronchoalveolar inflammation. However, it failed to modulate the Th1/Th2 balance toward the Th1 pole during Th2-skewed allergic airway inflammation in mice. This study attempts to further investigate the effects of dietary perilla oil on serum lipids and immunoglobulin profiles using an ovalbumin (OVA)-challenged mouse model. The inbred female BALB/c mice were randomly divided into four groups and fed different AIN-76 feeds containing 5% corn oil (rich in linoleic acid, 18:2n-6 polyunsaturated fatty acids (PUFA), as a control diet), 5% perilla oil (rich in alpha-linolenic acid, 18:3n-3 PUFA) or 5% compound oil containing 50% corn oil and 50% perilla oil, respectively, for 35 consecutive days ad libitum. Experimental mice were sensitized by an intraperitoneal injection of alum-precipitated antigen containing ovalbumin on 7, 14 and 21 days after supply of the specified experimental diets. One week later, the mice were then challenged by aerosolized OVA. The results showed that dietary perilla oil administration significantly (P < 0.05) decreased the relative liver tissue weight (RTW) and serum lipid levels including triglycerides, total cholesterol, HDL- and LDL-cholesterol. However, the HDL/LDL ratio was also significantly lowered by dietary perilla oil. Dietary perilla oil markedly decreased serum OVA-specific IgG1 level and total IgA antibodies (Th2 antibodies). Unfortunately, it also increased non-specific serum IgE (Th2 antibody) levels. The results suggest that dietary perilla oil might have a moderately beneficial effect on asthmatic allergy via lowering serum lipids and OVA-specific IgG1, as well as total IgA levels. However, it failed to obviously modulate Th1/Th2 antibody levels via isotype switching of B cells from Th2 antibody to Th1 antibody.

Effect of microbial fermentation on content of statin, GABA, and polyphenols in Pu-Erh tea.

Besides cancer prevention, the hypolipidemic effects of tea have been well studied in animals and humans. Recently, statin has been identified in Pu-erh tea extract. Clinical trials have confirmed that statin decreases the incidence of major coronary and cerebrovascular events and this may be due to its hypolipidemic and antiinflammatory effects. Since a good Pu-erh tea needs longer storage (10 years or more) of fermentation to enhance the flavor and fragrance, we screened microorganisms from two Pu-erh teas, 20 and 25 years old. Species of fungi and bacteria strains that contributed to a good taste of Pu-erh tea were isolated. The effect of fermentation was investigated by inoculating fresh tea leaves with individual strains of isolated microorganisms. Results showed that statin, total polyphenol content, and the scavenging activities of alpha,alpha-diphenyl-beta-picrylhydrazyl (DPPH) radicals increased during fermentation. Tea leaves inoculated with Streptomyces bacillaris strain R9 had the highest polyphenol content (3.3 mg/100 g) and scavenging ability to DPPH radicals (92%). Streptomyces cinereus strain Y11 was equally good for polyphenol content but yielded the highest amount of statin (1012 ng/g) after 42 days of fermentation. Interestingly, the statin content of fresh tea leaves fermented with strain R9 or Y11 after 180 days was much higher (4- and 8-fold, respectively) than that of the 25-year-old Pu-erh tea (513 ng/g) as measured by the HPLC method. Similarly, these two strains also increased the content of gamma-aminobutyric acid (GABA) 5.7- and 4.7-fold in tea fermented for 180 days as compared with the fresh leaves (1270 microg/g) and that were higher than that of the Pu-erh tea (4900 microg/g). Taken together, the present results indicate that tea short-term fermented with S. bacillaris or S. cinereus enhances the color and content of statin, GABA, and polyphenols.

An antifungal chitinase produced by Bacillus cereus with shrimp and crab shell powder as a carbon source.

The production of inexpensive chitinolytic enzymes is an element in the utilization of shellfish processing wastes. In this study, shrimp and crab shell powder prepared by treating shrimp and crab processing wastes with boiling and crushing was used as a substrate for the isolation of an antifungal chitinase-producing microorganism. Bacillus cereus YQ 308, a strain isolated from the soil samples, excreted one chitinase when cultured in a medium containing 2% (wt/vol) shrimp and crab shell powder as major carbon source. The chitinase, purified by sequential chromatography, had an Mr of 48 kDa and pI of 5.2. The purified chitinase (2 mg/ml) inhibited the hyphal extension of the fungi Fusarium oxysporum and Pythium ultimum.