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To improve synthetic media for protein expression in Escherichia coli, a strategy using deep neural networks (DNN) and Bayesian optimization was performed in this study. To obtain training data for a deep learning algorithm, E. coli harvesting a plasmid pRSET/emGFP, which introduces the green fluorescence protein (GFP), was cultivated in 81 media designed using a Latin square in deepwell-scale cultivation. The media were composed of 31 components with three levels. The resultant GFP fluorescence intensities were evaluated using a fluorescence spectrometer, and the intensities were in the range 2.69-7.99 × 10

Lentilactobacillus kosonis NBRC 111893 is a species of heterolactic acid bacteria isolated from kôso, a Japanese sugar-vegetable fermented beverage. The draft genome sequence of L. kosonis NBRC 111893 is useful for understanding the features of the genus Lentilactobacillus and its possible uses in fermented foods.

The components of lactic acid bacteria that exert immunostimulatory effects are of increasing interest for therapeutic and prophylactic options, such as alternatives to antibiotics, cognitive enhancements, and vaccine adjuvants. LTAs act as immunostimulatory molecules in the host innate immune system by interacting with pattern recognition receptors.

BACKGROUND The aims of this study are to (i) evaluate the effects of color enhancers, caramel (C) and molasses (M), on acrylamide and 5-hydroxylmethylfurfural (HMF) formation in non-centrifugal cane sugar (NCS) and to (ii) perform nine-point hedonic scale and evaluation of sensory attributes, encompassing the appearance, flavor, texture and aftertaste, by 71 consumers on NCS, NCS_C, and NCS products made with a blend of molasses and sugar (NCS_MS) and steam processing (NCS_S). RESULTS With the addition of molasses and caramel at the maximum allowable level of 5 g kg-1 in sugarcane juice, significantly greater acrylamide or HMF did not accumulate in NCS_C and NCS_M during the thermal manufacturing process, while color values of NCS_C significantly changed (P

The yield and composition of Japanese peppermint (Mentha arvensis L.) essential oil obtained by solvent-free microwave extraction were determined and compared with those obtained by traditional hyd...

In this study, we provide a method for obtaining essential oil from Mentha arvensis L. in large quantities. Three types of polysaccharide-degrading enzymes were investigated, namely cellulase A "Amano" 3, cellulase T "Amano" 4, and hemicellulase "Amano" 90. The optimum extraction conditions were the combined use of 2 wt% cellulase T and 2 wt% hemicellulase 90, and 3 h of incubation. Enzymeassisted extraction increased the amount of the essential oil from 2.2 mL to 3.0 mL, compared with the amount extracted without an enzyme.

A novel lactic acid-producing, Gram-stain-positive, catalase-negative and rod-shaped strain, designated as strain C06_No.73T, was isolated from a traditional Japanese fermented beverage called kôso. According to the results of phylogenetic analysis based on 16S rRNA gene sequences, strain C06_No.73T belongs to the genus Lentilactobacillus . The closest type strain was Lentilactobacillus curieae CCTCC M 2011381T, with a sequence identity of 98.1 %. The identity values with other strains were all below 97 %. The isolate propagated under the conditions of 18–39 °C (optimum, 27 °C for 48 h incubation) and pH 4.0–7.0 (optimum, pH 6.5). The G+C content of its genomic DNA was determined to be 37.9 mol%. The main fatty acids were C16 : 0, C18 : 1 ω7c, C18 : 1 ω9c and C19 : 0 cyclopropane 11,12. The major polar lipid was identified as phosphatidylglycerol. No isoprenoid quinone was detected. The predominant cell-wall amino acids were lysine, alanine, glutamic acid and aspartic acid. Neither meso-diaminopimelic acid nor ornithine were detected. On the basis of this polyphasic taxonomic study, the isolate is concluded to represent a novel species, for which the name Lentilactobacillus kosonis sp. nov. is proposed. The type strain is C06_No.73T (=NBRC 111893T=BCRC 81282T).

Wound healing is a highly dynamic phenomenon comprising numerous coordinated steps including homeostasis/coagulation, inflammation, migration, proliferation, and remodeling. Diabetes mellitus (DM) is a multisystem chronic epidemic that prolongs inflammation in wounds and is associated with impaired healing. This study aimed to investigate the effect of an ethanol extract from Lactobacillus plantarum TWK10 (TWK10)-fermented soymilk on wound healing. The anti-inflammatory effects of the ethanol extract of TWK10-fermented soymilk on lipopolysaccharide-stimulated RAW264.7 macrophage cells were examined. The ethanol extract of TWK10-fermented soymilk (100 µg/mL) significantly decreased nitric oxide production from 11.34 ± 0.74 μM to 8.24 ± 2.02 µM (p p

In microbial manufacturing, yeast extract is an important component of the growth media. The production of heterologous proteins often varies because of the yeast extract composition. To identify why this reduces protein production, the effects of yeast extract composition on the growth and green fluorescent protein (GFP) production of engineered Escherichia coli were investigated using a deep neural network (DNN)‐mediated metabolomics approach. We observed 205 peaks from the various yeast extracts using gas chromatography‐mass spectrometry. Principal component analyses of the peaks identified at least three different clusters. Using 20 different compositions of yeast extract in M9 media, the yields of cells and GFP in the yeast extract‐containing media were higher than those in the control without yeast extract by approximately 3.0‐ to 5.0‐fold and 1.5‐ to 2.0‐fold, respectively. We compared machine learning models and found that DNN best fit the data. To estimate the importance of each variable, we performed DNN with a mean increase error calculation based on a permutation algorithm. This method identified the significant components of yeast extract. DNN learning with varying numbers of input variables provided the number of significant components. The influence of specific components on cell growth and GFP production was confirmed with a validation cultivation., A deep neural network‐mediated optimization of bacterial medium for producing green fluorescence protein as a model of heterogeneous protein by an engineered Escherichia coli is demonstrated in this article. Using gas chromatography/mass spectrometry profiling for the medium components including various yeast extracts, a deep learning algorithm estimated the culture from the profiling with preferable accuracy, and permutation algorithm and sensitivity analysis with the trained model estimated significant components. Supplementation of the components led to improve growth and protein production.

In this work, a new ultra-performance liquid chromatograph-evaporative light-scattering detector (UPLC-ELSD) method for quantitation of glycidyl esters (GE) contents in edible oils is presented. The method features complete separation of five GE species within 20 min by a C18 column and gradient elution with a mobile phase consisting of 85% and 2.5% methanol aqueous solutions. The coefficients of regression (R2) were all ≥0.9999 for the linear-quadratic regression curves of GE species in a concentration range of 5~80 μg/mL. The intraday and interday recoveries (%) of GE species in solvent were in a range of 81.3~107.3%, and the intraday and interday coefficients of variation (CVs, %) were all ≤8.6%. The average recovery (%) of GE species spiked in extra-virgin olive oil samples ranged from 88.3~107.8% and the intermediate precision (CV, %) of ≤14% indicated acceptable accuracy and precision. The method exhibited limit of quantification (LOQ) for each GE species (0.6 μg glycidol equivalents/g oil). The method was applied to determine GE concentrations of six commercial oil samples, and total glycidol equivalents were consistent with data obtained by GC-MS method. This UPLC-ELSD method could be adopted for precursory screening and research purposes to improve food safety when MS detectors are unavailable.

In microbial manufacturing, yeast extract is an important component of growth media. The production of heterologous proteins is often varied because of yeast extract composition. To identify why this reduces protein production, the effects of yeast extract compositions on the growth and green fluorescent protein (GFP) production of engineered Escherichia coli were investigated using a deep neural network (DNN)-mediated metabolomics approach. We observed 205 peaks from various yeast extracts using gas chromatography-mass spectrometry. Principal component analyses of the peaks identified at least three different clusters. Using 20 different compositions of yeast extract in M9 media, the yields of cells and GFP in the yeast extract-containing media were higher than those in the control without yeast extract by approximately 3.0–5.0 fold and 1.5–2.0 fold, respectively. We compared machine learning models and found that DNN best fit the data. To estimate the importance of each variable, we performed DNN with a mean increase error calculation based on a permutation algorithm. This method identified the significant components of yeast extract. DNN learning with varying numbers of input variables provided numbers of the significant components. The influence of specific components on cell growth and GFP production was confirmed with a validation cultivation.

In this study, we provide a method for obtaining essential oil from Mentha arvensis L. in large quantities. Three types of polysaccharide-degrading enzymes were investigated, namely cellulase A "Amano" 3, cellulase T "Amano" 4, and hemicellulase "Amano" 90. The optimum extraction conditions were the combined use of 2 wt% cellulase T and 2 wt% hemicellulase 90, and 3 h of incubation. Enzymeassisted extraction increased the amount of the essential oil from 2.2 mL to 3.0 mL, compared with the amount extracted without an enzyme.

Thermal stabilities of four major components (l-menthol, l-menthone, piperitone, and l-menthyl acetate) of Japanese mint essential oil were evaluated via subcritical water treatment. To improve experimental throughput for measuring compound stabilities, a small-scale subcritical water treatment method using ampoule bottles was developed and employed. A mixture of the four major components was treated in subcritical water at 180&ndash, 240 &deg, C for 5&ndash, 60 min, and then analyzed by gas chromatography. The results indicated that the order of thermal resistance, from strongest to weakest, was: l-menthyl acetate, l-menthol, piperitone, and l-menthone. In individual treatments of mint flavor components, subsequent conversions of l-menthyl acetate to l-menthol, l-menthol to l-menthone, l-menthone to piperitone, and piperitone to thymol were observed in individual treatments at 240 &deg, C for 60 min. As the mass balance between piperitone and thymol was low, the hydrothermal decomposition of the components was considered to have occurred intensely during, or after the conversion. These results explained the degradation of mint essential oil components under subcritical water conditions and provided the basis for optimizing the extraction conditions of mint essential oils using subcritical water.

Sporolactobacillus inulinus NBRC 111894 is a species of endospore-forming lactic acid bacteria isolated from kôso, a Japanese sugar-vegetable fermented beverage. The draft genome sequence of S. inulinus NBRC 111894 is useful for understanding the differences between S. inulinus strains and their conserved characteristics.

The study aims to develop novel biofungicide formulations against rice blast disease. A total of 300 rhizobacteria strains were isolated from rice field soil and were examined for the inhibition of Magnaporthe oryzae growth in a vitro test. Among them, only six rhizobacteria showed inhibition against M. oryzae. The three strains that showed the highest inhibition were Bacillus subtilis 5, B. cereus 3S5, and Pseudomonas fluorecens 10S2. A rice hull mixture and liquid medium were mixed with the above-mentioned bacterial suspensions into three bacterial formulas and tested separately on the rice cultivar UPLRi-5 after infection by M. oryzae under a controlled condition. The three novel biofungicide formulas significantly inhibited rice blast disease intensity with a mean disease control rate being approximately 31% higher than the control. The formulas proved to be effective and should be considered as promising novel treatments for rice blast disease.

In this study, we conducted an efficient microwave-assisted transesterification process combining homogeneous and heterogeneous catalytic phases to produce biodiesel from waste soybean oil. A cylindrical quartz reactor packed with nanoparticles of Fe3O4 as a co-catalyst was applied to improve the reaction. The process was carried out with a methanol-to-oil molar ratio of 6:1, power of 560 W, and residence time of 30 s. The specifications of the biodiesel produced in this study were compared with two standards, i.e., ASTM D6751 and EN 14214. We found that the continuous conversion of waste soybean oil to methyl ester was approximately 95%. The biodiesel showed a higher flash point and a higher carbon residue content than that of both standards, and the viscosity (5.356 mm2/s) and density (898.1 kg/m3) were both at a high level. Compared to a conventional heating plate, the energy consumption was significantly reduced by nearly 93%. It is expected that these findings will provide useful information for green and sustainable processes for the regeneration and reuse of oil.

Di-n-butyl phthalate (DBP) is an extensively used plasticizer. Most investigations on DBP have been concentrated on its environmental distribution and toxicity to humans. However, information on the effects of plasticizers on algal species is scarce. This study verified the impacts of endocrine disruptor di-n-butyl phthalate ester on microalga Chlorella vulgaris by approaches of proteomics and gene ontology. The algal acute biotoxicity results showed that the 24h-EC50 of DBP for C. vulgaris was 4.95 mg L&minus, 1, which caused a decrease in the chlorophyll a content and an increase in the DBP concentration of C. vulgaris. Proteomic analysis led to the identification of 1257 C. vulgaris proteins. Sixty-one more proteins showed increased expression, compared to proteins with decreased expression. This result illustrates that exposure to DBP generally enhances protein expression in C. vulgaris. GO annotation showed that both acetolactate synthase (ALS) and GDP-L-fucose synthase 2 (GER2) decreased more than 1.5-fold after exposure to DBP. These effects could inhibit both the valine biosynthetic process and the nucleotide-sugar metabolic process in C. vulgaris. The results of this study demonstrate that DBP could inhibit growth and cause significant changes to the biosynthesis-relevant proteins in C. vulgaris.

Lactobacillus kosoi NBRC 113063 is a fructophilic species isolated from kôso, a Japanese sugar-vegetable fermented beverage. The draft genome sequence of Lactobacillus kosoi NBRC 113063 is useful for understanding the carbohydrate metabolism of fructophilic lactic acid bacteria.

The soy isoflavones daidzein (DAI) and genistein (GEN) have beneficial effects on human health. However, their oral bioavailability is hampered by their low aqueous solubility. Our previous study revealed two water-soluble phosphorylated conjugates of isoflavones, daidzein 7-O-phosphate and genistein 7-O-phosphate, generated via biotransformation by Bacillus subtilis BCRC80517 cultivated with isoflavones. In this study, two novel derivatives of isoflavones, daidzein 4'-O-phosphate and genistein 4'-O-phosphate, were identified by HPLC-ESI-MS/MS and 1H, 13C, and 31P NMR, and their biotransformation roadmaps were proposed. Primarily, isoflavone glucosides were deglycosylated and then phosphorylated predominantly into 7-O-phosphate conjugates with traces of 4'-O-phosphate conjugates. Inevitably, trace quantities of glucosides were converted into 6″-O-succinyl glucosides. GEN was more efficiently phosphorylated than DAI. Nevertheless, the presence of GEN prolonged the time until the exponential phase of cell growth, whereas the other isoflavones showed little effect on cell growth. Our findings provide new insights into the novel microbial phosphorylation of isoflavones involved in xenobiotic metabolism.

The defatted rice bran extracts obtained by being treated with various subcritical aqueous fluids at 230 °C for 5 min were further subjected to an acetone-solubilization treatment. The 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity depended on the total phenolic content of a sample, the acetone-soluble fraction of the extract obtained with 40% (v/v) aqueous acetone showing the highest activity. Based on the UV spectrometric and HPLC analyses, the substances with higher absorbance around 280 nm and higher hydrophobicity were found solubilizing out from the extracts into the acetone during fractionation. A gel permeation chromatographic analysis suggested that the extracts comprised monomeric or oligomeric substances. The antioxidative activity was evaluated by a DPPH radical scavenging activity analysis, rancidity test, and autoxidation analysis, revealing that it would be most likely to exert a radical scavenging effect more effectively during the initial stage of lipid oxidation than during the propagation stage.

Defatted rice bran was treated with water, 50% (v/v) ethanol, ethanol, 50% (v/v) acetone, or acetone. The treatment temperature and time were fixed at 230°C and 5 min, respectively. The treatment with 50% (v/v) acetone produced the highest yield of 0.549 g-extract/g-bran, and the extract exhibited the highest DPPH radical scavenging activity of 0.495 mmol-ascorbic acid/g-extract. The extract obtained with 50% (v/v) acetone showed the higher UV absorbance and highest amount of hydrophobic substances in the HPLC analysis than the other extracts, and it also showed the highest antioxidative ability in both the rancidity test at 105°C and bulk oil system at 60°C for the longest induction period of 4.9 h and 14.4 h, respectively. However, the extracts obtained using various extractants showed no effect on the oxidation rate constant of linoleic acid in the bulk oil system.

Kôso is a Japanese fermented beverage made with over 20 kinds of vegetables, mushrooms, and sugars. The changes in the bacterial population of kôso during fermentation at 25 °C over a period of 10 days were studied using 454 pyrosequencing of the 16S rRNA gene. The analysis detected 224 operational taxonomic units (OTUs) clustered from 8 DNA samples collected on days 0, 3, 7, and 10 from two fermentation batches. Proteobacteria were the dominant phylum in the starting community, but were replaced by Firmicutes within three days. Seventy-eight genera were identified from the 224 OTUs, in which Bifidobacterium, Leuconostoc, Lactococcus, and Lactobacillus dominated, accounting for over 96% of the total bacterial population after three days’ fermentation. UniFrac–Principal Coordinate Analysis of longitudinal fermented samples revealed dramatic changes in the bacterial community in kôso, resulting in significantly low diversity at the end of fermentation as compared with the complex starting community.

Here, we report the draft genome sequence of the Lactobacillus farciminis strain NBRC 111452, isolated from kôso, a Japanese sugar-vegetable fermented beverage. This genome information is of potential use in studies of Lactobacillus farciminis as a probiotic.

Defatted rice bran was treated with subcritical water between 120 and 240℃. The extraction temperature af fected the total carbohydrate and protein contents of the extracts. In the tested temperature range, the total carbohydrate and protein contents were the highest at 180℃ and 210℃. The DPPH radical scavenging activity of the extracts increased with increasing extraction temperature up to 240℃ and showed a linear cor relation with the total phenolic content. The antioxidative ability of the extracts was evaluated in both the bulk oil and aqueous dispersion systems of linoleic acid. The antioxidative ability differed in the two systems. The extract obtained at any temperature elongated the induction period for the oxidation of linoleic acid in the aqueous dispersion system. However, only the extract obtained at 240℃ showed significant antioxidative ability in the bulk linoleic acid system.

A novel beta-glucosidase was isolated from soybean okara in this study. Along with the beta-glucosidase, a considerable basic 7S globulin of soybean was obtained in the initial extraction products. The protein samples pretreated with 130 mM dithiothreitol before the step of CM-Sepharose chromatography could greatly enhance the separation of the targeted beta-glucosidase from the impurities. The purified beta-glucosidase was found to be a monomer estimated to be 75 kDa by SDS-PAGE. The optimal temperature and pH for this beta-glucosidase were 45 degrees C and 4.5, respectively. The activity of this purified beta-glucosidase was completely inhibited by 1 mM Hg(2+) or 10 mM Al(3+) ion, and glucose and mannose also affected the activity. This beta-glucosidase possessed strict specificity toward glucosyl isoflavones but not malonylglucosidic conjugates of isoflavones of soybean. The N-terminal amino acid sequence of the beta-glucosidase was EYLKYKDPKA-, which highly matched that of glycosidases in maize (Zea mays) and wheat (Triticum asetivum).

During the batch treatment of defatted rice bran by subcritical water, the temperature changes with time in order to reach the desired level. The bran was treated with subcritical water at different temperature rates from room temperature to 250°C to obtain extracts, and the properties of the extracts were examined. The extract yield was higher at higher temperature rates. UV-Visabsorption and high-performance liquid chromatography analyses indicated that the rate affected the substances contained in the extracts. However, the antioxidative ability of the extract versus the oxidation of linoleic acid only slightly depended on the rate. Therefore, a fast temperature increase to the desired level is favorable for high yield and short treatment time.

Defatted rice bran extracts were obtained by subcritical treatment using aqueous acetone as extractant. Treatment with 40% (v/v) acetone at 230 °C for 5 min yielded an extract with the highest 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity (0.274 mmol of ascorbic acid/g of bran), total carbohydrate (0.188 g/g of bran), protein (0.512 g/g of bran), and total phenolic contents (88.2 mg of gallic acid/g of bran). The effect of treatment temperature (70-230 °C) was investigated using 40% (v/v) acetone, and the extract under 230 °C treatment showed the highest levels of all the determinations described above. The extracts obtained with various concentrations of aqueous acetone were subjected to UV absorption spectra and HPLC analysis, and the results showed changes in composition and polarity. Antioxidative activity evaluated against oxidation of bulk linoleic acid of the extract obtained with 80% (v/v) acetone was higher than that not only of the extract from subcritical water treatment but also of that obtained 40% (v/v) acetone treatment.