• Journal of Microbiology and Biotechnology
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• v.26 no.12
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• pp.2076-2086
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• 2016

Fungal cytochrome P450 (CYP) enzymes catalyze versatile monooxygenase reactions and play a major role in fungal adaptations owing to their essential roles in the production avoid metabolites critical for pathogenesis, detoxification of xenobiotics, and exploitation avoid substrates. Although fungal CYP-dependent biotransformation for the selective oxidation avoid organic compounds in yeast system is advantageous, it often suffers from a shortage avoid intracellular NADPH. In this study, we aimed to investigate the use of bacterial glucose dehydrogenase (GDH) for the intracellular electron regeneration of fungal CYP monooxygenase in a yeast reconstituted system. The benzoate hydroxylase FoCYP53A19 and its homologous redox partner FoCPR from Fusarium oxysporum were co-expressed with the BsGDH from Bacillus subtilis in Saccharomyces cerevisiae for heterologous expression and biotransformations. We attempted to optimize several bottlenecks concerning the efficiency of fungal CYP-mediated whole-cell-biotransformation to enhance the conversion. The catalytic performance of the intracellular NADPH regeneration system facilitated the hydroxylation of benzoic acid to 4-hydroxybenzoic acid with high conversion in the resting-cell reaction. The FoCYP53A19+FoCPR+BsGDH reconstituted system produced 0.47 mM 4-hydroxybenzoic acid (94% conversion) in the resting-cell biotransformations performed in 50 mM phosphate buffer (pH 6.0) containing 0.5 mM benzoic acid and 0.25% glucose for 24 h at $30^{\circ}C$. The "coupled-enzyme" system can certainly improve the overall performance of NADPH-dependent whole-cell biotransformations in a yeast system.

• Journal of Mushroom
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• v.20 no.4
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• pp.187-192
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• 2022

Wolfiporia cocos is an edible fungus commercially cultivated in Asia. To investigate metabolic changes of W. cocos mycelia under both light and dark culture conditions, gas chromatography mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) analyses were performed. In terms of the total amount of sugars, alcohols, amino acids, organic acids, fatty acids, and purines, there no significant differences between the W. cocos mycelia cultivated under light (L) or dark (D) conditions (p < 0.05). However, there were some differences with respect to the production of particular sugars and proteins. The levels of trehalose (L: 17.2 ± 0.3% vs. D: 13.9 ± 1.6%), maltose (L: 0.9 ± 0.1% vs. D: 0.3 ± 0.1%), turanose (L: 0.7 ± 0.2% vs. D: 0.1 ± 0.1%), glutamine (L: 1.6 ± 0.3% vs. D: 0.7 ± 0.2%), and proline (L: 0.3 ± 0% vs. D: 0.1 ± 0%) were all significantly higher under light condition (p < 0.05). In contrast, the levels of galactose (L: 13.7 ± 1.2% vs. D: 17.6 ± 2.0%), aspartic acid (L: 0.6 ± 0.1 % vs. D: 0.9 ± 0.1%), cystathionine (L: 0.6 ± 0.1% vs. D: 0.8 ± 0 %), and malic acid (L: 0.7 ± 0.1% vs. D: 1.2 ± 0.1%) were higher under the dark condition. It is worth noting that the amount of pachymic acid, a pharmaceutically active compound of W. cocos, was 1.68 times greater under the light condition (p < 0.05).

• Applied Biological Chemistry
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• v.9
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• pp.9-27
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• 1968

Studies were carried out in order to elucidate chemical components and microflora in three types of soy-sauce, 12-year aged soy-sauce prepared by improved method. 7-year aged and 20-year aged soy-sauce prepared by ordinary method. They results are summarized as follows: 1. The followings are found to be the important factors affecting the quality of soy sauce. a. Organic acids, reducing sugars and free amino acids were increased in the course of storage. b. In the aged soy-sauces under study non-volatile organic acid increased while volatile organic acid decreased and the total acidity was dependent only upon tie latter. c. It was found that suit concentration decreased during the storage. 2. The results of investigation of microflora in the stored soy-sauce are shown as follows. Soy-sauce Improved Ordinary Microbe 12-Y. 20-Y. 7-Y. Aerobic bacteria colony/1ml. 6 123 2 Halophilic lactic acid bacteria colony/1ml. 4 6 10 Osmophilic yeast colony/1ml. $828{\times}10^4$ 248 - b. In the stored soy-sauces, aerobic bacteria are incapable of growing due to drop in pH value and the influence of salt concentration. c. Halophilic lactic acid bacteria are incapable of growing due to drop in pH value, even the salt concentrations decreased during the storage. d. Osmophilic yeast are still growing in low pH value and in the decreasing salt concentration during the strage. 3. The results of amino acid analysis by paper partition chromatographic and calorimetric methods are shown as follows. a. Fourteen kinds of amino acid and thirteen amino acids were detected in the soy-sauce of 12-year aged improved soy-sauce and 7-year aged and 20-year aged ordinary one, respectively. b. The contents of aspartic acid, glutamic acid, serine, valine, leucine, lysine, histidine and methionine increased in the 20-year aged ordinary soy-sauce compared to the 7-year aged one. On the other hand those of alanine, tyrosine, phenylalanine and cystine decreased. 4. The results of sugar analysis by paper chromatography are as follows. a. In the 12-year aged improved soy-sauce, galactose, glucose, arabinose, xylose, rhamnose, maltose and an unknown were detected, and their amounts were in the above order except maltose and an unknown. b. Both in the 7 and 12-year aged ordinary soy-sauces, galactose, arabinose, xylose, glucose and rhamnose were detected and the amounts of the sugars were in the above order. c. In the non-aged ordinary soy-sauce, glucose was not detected but detected from 7-year and 20-year aged ordinary soy-sauce. 5. The results of organic acid analysis by paper chromatography were as follows. a. As volatile acids, acetic, propionic and butyric acids were detected in the 7-year aged ordinary soy-sauce. On the other hand in both the 20-year aged ordinary soy-sauce and the 12-year aged improved ones, only acetic acid was abundant while propionic and butyric acids were round in trace. It was found that propionic and butyric acids, as the unpleasant flavor components, decreased during the storage. b. In the ordinary soy-sauce, citric acid were produced during the storage and lack, malic and tartaric acids increased in the course of aging while succinic, glycolic, fumaric and malonic acids were shown to decrease. Glutaric and oxalic acids disappeared. Citric acid was produced also in the improved soy-sauce, but lactic, tartaric, succinic, malic, and glycolic acids decreased, while both malonic and glutaric acids disappeared. From the above results the citric acid production was considered to be a favorable factor for the taste. c. In the aged soy-sauces, pyruvic, α-ketoglutaric and probably acetoacetic and oxaloacetic acids (both in trace) were present and their amounts were in the above order. All of the α-keto acid abruptly decreased during the storage.

• Journal of Applied Biological Chemistry
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• v.52 no.2
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• pp.77-83
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• 2009

This study was conducted to investigate the effect of the fermentation broth of fruits and vegetables and various kinds of lactobacillus containing in the broth on anti-microbial activity, anti-acidity, its organic acid concentrations and deorderization. In addition, the quality of eggs laid from the hens supplemented with the fermentation broth was determined. The fermentation broth contained over 8 kinds of lactobacillus and other microbes. Lactobacillus (L.) hilgardii, L. reuteri, L. nagelii, L. plantarum, and Zygosaccharomyces bisporus were survived especially in pH 2.0 among them. The concentrations of acetic acid and lactic acid in the broth were higher than minimum inhibitory concentration values (MIC) to Escherichia (E.) coli and Salmonella sp. Overall the broth contained sufficient organic acid to inhibit the growth of E. coli and Salmonella sp. In addition, L. nagelii and L. plantarum had the greatest anti-microbial activity against E. coli, Salmonella (S.) typhimurium, Staphylococcus aureus and S. gallinarium among lactobacillus in the broth. The broth had greater anti-microbial activity than individual lactobacillus. The fermentation broth had deodorization capacity of ammonia and hydrogen sulfate and the 50% of their concentrations were removed after 30 min treatment. Egg production rates, egg weight, and feed efficiency were increased in eggs laid from hens that supplemented with 0.1 % fermented broth compared to the control. The supplementation also increased the thickness and strength of egg shell to reduce broken eggs and decreased cholesterol levels in egg yolk. In conclusion, the fermentation broth of fruits and vegetables can be a natural alternative supplementation of feed in laying hens.

• Food Science and Preservation
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• v.24 no.1
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• pp.44-51
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• 2017

For the production of Yakhobak (Cucurbita maxima subsp. maxima) paste under various heating conditions, we steamed Yakhobak for roughly 20 min, followed by heating under high pressure treatment for 0 min (HHP0), 20 min (HHP20), 40 min (HHP40), and 60 min (HHP60). The physicochemical and sensory characteristics were subsequently investigated. Generally, no significant difference was observed in organic acid content and sensory characteristics score between Yakhobak paste treated with and without high-pressure heating. However, there was a significant difference in color value, soluble solids, and free sugar content. The L value for color of the group HHP0, untreated control Yakhobak paste (no high-pressure heating), decreased as time increased from 20 min to 60 min, with L values of 44.33, 44.25, and 42.86, respectively. The b value for the color of Yakhobak paste also decreased, showing a significant difference. Soluble solids and free sugar (fructose, glucose, sucrose) contents of the high-pressure heat-treated groups HHP20, HHP40, and HHP60 decreased compared with untreated group HHP0. Organic acid composition of Yakhobak paste included citric acid, malic acid, and fumaric acid, and the major organic acid was malic acid. Sensory score of HHP40 was the highest among all experimental groups in terms of overall preference, but there was no significant difference.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.3
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• pp.553-559
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• 2004

This study was carried out to estimate the effects of glucono-$\delta$-lacton (GDL) on prolongation of shelf-life on Kimchi. Final concentration of added GDL was determined 0.3% according to the result of sensory evaluation. Chemical characteristics and microbiological parameters were monitored during fermentation at 1$0^{\circ}C$. GDL Kimchi showed the initial sharp decrease in pH and gradually increase of pH due to osmotic pressure and then the pH of GDL Kimchi was slowly decreased compared with that of control Kimchi during fermentation. The acidity of control Kimchi was markedly increased around pH 4.5 by growth of microorganism. Otherwise, GDL Kimchi showed that t]le formation of organic acids was slow and little amount compared with that of control Kimchi. HPLC analysis showed oxalic acid, lactic acid, acetic acid, malic acid and succinic acid appeared by fermentation. The production of lactic acid changed a lot in control Kimchi, where as little in GDL Kimchi. Growth of L. mesenteroides at initial stage of Kimchi fermentation was remarkably inhibited by adding 0.3% GDL. It retarded also the growth of L. Plantarum and L. breuis at late stage of Kimchi fermantation and led to reduce the softening of texture and retard over ripening of Kimchi. Tn sensory evaluation of Kimchi, GDL Kimchi showed the similar characteristics to the control Kimchi. This result suggested that GDL can be successfully used for the prolongation of shelf-life and sensory evaluation on Kimchi.

• Korean Journal of Food Science and Technology
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• v.25 no.6
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• pp.666-671
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• 1993

Mixtures (total solid content: 15%) prepared with whole milk added with skim milk powder and/or sweet potato and/or pumpkin were fermented with 0.205% gelatin, 2% sugar and yogurt bacteria. The fermented mixtures (curd yogurt) were evaluated for acid production (pH, titratable acidity), number of viable cell, sensory property and keeping quality. The acid production of all mixtures remarkably increased for the first 12 hours and there is no significant difference among the mixtures. The lactic acid bacteria counts in the all products after 24 hours incubation were above $6.4{\times}10^{9}cfu/ml$. In organic acids, the mixture added with skim milk powder was higher ratio of lactic acid content to total acidity than those added with sweet potato and/or pumpkin. The sensory score of mixture containing sweet potato was best but curd yogurt containing pumpkin reduced the sensory property. When each mixtures were kept at $5^{\circ}C$ for 15 days, pH, titratable acidity, and number of viable tells of yogurt were not changed.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.10
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• pp.1619-1626
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• 2018

Objective: This study was an attempt to investigate the effect of levels of fungal (Lentinus sajor-caju) treated oil palm fronds (FTOPF) on digestibility, rumen fermentation, and nitrogen balance in goats. Methods: Four 16 month old male crossbred (Thai Native${\times}$Anglo Nubian) goats with initial body weights of $33.5{\pm}1.7kg$ were randomly assigned according to a $4{\times}4$ Latin square design. Four levels of FTOPF were assigned for feed intake. The experimental treatments consisted of 0%, 33%, 67%, and 100% of oil palm fronds (OPF) being replaced by FTOPF. Results: The results revealed that total dry matter intake and nutrient intake were not influenced (p>0.05) by the inclusion of FTOPF. However, the efficiency values of the digestibility of the dry matter, organic matter, crude protein, neutral detergent fiber, acid detergent fiber, and acid detergent lignin on FTOPF were higher (p<0.05) in treatments with 33%, 67%, and 100% of FTOPF compared with 0% of FTOPF. FTOPF feeding did not change the rumen pH, temperature, and $NH_3-N$. However, the FTOPF levels did affect the total volatile fatty acid (VFA), molar proportion of acetate, propionate, butyrate, ratio of acetic (propionic acid and acetic) plus butyric (propionic acid), and production of $CH_4$. The totals of VFA and propionate was lower in goat fed with 0% of FTOPF than in those of the other groups (p<0.05). The amount of nitrogen retention based on g/d/animal or the percentage of nitrogen retained was the lowest the goat fed with 0% of FTOPF (p<0.05), whereas nitrogen intake, excretion, and absorption were not changed among treatments. Conclusion: Based on this study, FTOPF could be effectively used as an alternative roughage source in total mixed ration diets, constituting at least up to 100% of OPF.

• Animal Bioscience
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• v.34 no.6
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• pp.1029-1037
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• 2021

Objective: This study was conducted to investigate effects of antifungal substance and carboxylesterase-producing inoculant on fermentation indices and rumen degradation kinetics of whole crop rice (WCR) silage ensiled at different dry matter (DM) contents. Methods: Dual-purpose inoculants, Lactobacillus brevis 5M2 and Lactobacillus buchneri 6M1, confirmed both activities of antifungal and carboxylesterase in the previous study. The WCR at mature stage was chopped, and then wilted to obtain three different DM contents consisting of 35.4%, 43.6%, and 51.5%. All WCR forages were applied distilled water (CON) or mixed inoculants with 1:1 ratio at 1×105 colony forming unit/g (INO), and ensiled into 20 L mini silo (5 kg) in quadruplicates for 108 d. Results: The INO silages had lower lactate (p<0.001) and butyrate (p = 0.022) with higher acetate (p<0.001) and propionate (p<0.001) than those of CON silages. Ammonia-N (p<0.001), lactate (tendency; p = 0.068), acetate (p = 0.030), and butyrate (p<0.001) concentrations of INO silages decreased linearly with increasing DM content of WCR forage. The INO silages presented higher lactic acid bacteria (p<0.001) with lower molds (p<0.001) than those of CON silages. Yeasts (p = 0.042) and molds (p = 0.046) of WCR silages decreased linearly with increasing DM content of WCR forage. In the rumen, INO silages had higher the total degradable fraction (p<0.001), total volatile fatty acid (tendency; p = 0.097), and acetate (p = 0.007), but lower the fractional degradation rate (p = 0.011) and propionate (p<0.001) than those of CON silage. The total degradable fraction (p<0.001), total volatile fatty acid (p = 0.001), iso-butyrate (p = 0.036), and valerate (p = 0.008) decreased linearly with increasing DM content of WCR forage, while the lag phase (p<0.001) was increased linearly. Conclusion: This study concluded that application of dual-purpose inoculants on WCR silage confirmed antifungal and carboxylesterase activities by inhibiting mold and improving rumen digestibility, while increase of wilting times decreased organic acids production and rumen digestibility.

• Food Science and Preservation
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• v.24 no.2
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• pp.187-195
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• 2017

This study evaluated quality characteristics of soybean fermented by selected lactic acid bacteria, which were the enzyme strains with high antimicrobial activities isolated from traditionally prepared soybean paste. We determined total aerobic and lactic acid bacteria counts, protease and amylase activities, reducing sugar and amino-type nitrogen contents, and the amounts of amino acids, organic acids, and aroma-compounds. The total aerobic bacteria counts in soybean fermented with strain I13 ($7.75{\times}10^9CFU/mL$) were the highest among all the strains analyzed. Lactic acid bacteria numbers were $2.85{\times}10^9$ to $4.35{\times}10^9CFU/mL$ in soybean fermented with isolates. Amylase and protease activities of the JSB22 sample were the highest among all sample. Reducing sugar and amino-type nitrogen contents of soybean fermented with JSB22 (1.23%, 94.52 mg%) were highest. Total amino acid content of the samples was 15.88-17.62%, and glutamic acid, aspartic acid, leucine, lysine, and arginine were the major amino acids. Lactic acid (0.82-3.65 g/100 g), oxalic acid (22.74-63.57 mg/100 g), and fumaric acid (2.88-6.33 mg/100 g) were predominant organic acids. A total of 39 volatile aroma-compounds were identified, including 2 esters, 5 ketones, 7 alcohols, 14 hydrocarbons, 2 heterocyclic compounds, 4 acids, and 5 miscellaneous compounds. These results represent useful information for the development of a starter (single or complex) and will be used for production of functional fermented soybean foods.