• Animal Bioscience
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• v.35 no.10
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• pp.1592-1605
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• 2022

Objective: The objective of this study was to select an effective in vitro digestion-fermentation model to estimate the effect of decreasing dietary crude protein (CP) on odor emission during pig production and to suggest potential prediction markers through in vitro and in vivo experiments. Methods: In the in vitro experiment, three diet formulations with different CP contents (170 g/kg, 150 g/kg, and 130 g/kg) but containing the same standardized ileal digestible essential amino acids (SID-EAA) were assessed. Each diet was evaluated by two different in vitro gastric-intestinal phase digestion methods (flask and dialysis), combined with fresh pig feces-ferment inoculation. Eighteen growing barrows (31.9±1.6 kg) were divided into three groups: control diet (180 g CP/kg, without SID-EAA adjustment), 170 g CP/kg diet, and 150 g CP/kg diet for 4 weeks. Results: The in vitro digestion results indicated that in vitro digestibility was affected by the gastric-intestinal phase digestion method and dietary CP level. According to the gas kinetic and digestibility results, the dialysis method showed greater distinguishability for dietary CP level adjustment. Nitrogen-related odor compounds (NH₃-N, indole, p-cresol, and skatole) were highly correlated with urease and protease activity. The feeding study indicated that both EAA-adjusted diets resulted in a lower odor emission especially in p-cresol and skatole. Both protease and urease activity in feces were also closely related to odor emissions from nitrogen metabolism compounds. Conclusion: Dialysis digestion in the gastric-intestinal phase followed by fresh fecal inoculation fermentation is suitable for in vitro diet evaluation. The enzyme activity in the fermentation and the fecal samples might provide a simple and effective estimation tool for nitrogen-related odor emission prediction in both in vitro and in vivo experiments.

• Genomics & Informatics
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• v.15 no.4
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• pp.142-146
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• 2017

More effective production of human insulin is important, because insulin is the main medication that is used to treat multiple types of diabetes and because many people are suffering from diabetes. The current system of insulin production is based on recombinant DNA technology, and the expression vector is composed of a preproinsulin sequence that is a fused form of an artificial leader peptide and the native proinsulin. It has been reported that the sequence of the leader peptide affects the production of insulin. To analyze how the leader peptide affects the maturation of insulin structurally, we adapted several in silico simulations using 13 artificial proinsulin sequences. Three-dimensional structures of models were predicted and compared. Although their sequences had few differences, the predicted structures were somewhat different. The structures were refined by molecular dynamics simulation, and the energy of each model was estimated. Then, protein-protein docking between the models and trypsin was carried out to compare how efficiently the protease could access the cleavage sites of the proinsulin models. The results showed some concordance with experimental results that have been reported; so, we expect our analysis will be used to predict the optimized sequence of artificial proinsulin for more effective production.

• 한국생물공학회:학술대회논문집
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• 2003.10a
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• pp.351-355
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• 2003

Response surface methodology was employed to study the interactive effect of sucrose, nitrogen, temperature and to optimize their levels to enhance the production of human granulocyte-macrophage colony-stimulation factor from Nicotiana tabacum cell suspension cultures. A 15-runs Box-Behnken design including three center points was the response surface method selected for the initial set of experiments. The analysis of the data from the Box-Behnken experiments showed interactive effects of sucrose:nitrogen, sucrose:temperature and nitrogen:temperature. The optimal combinations of sucrose, nitrogen and temperature for hGM-CSF production from surface plot were sucrose 90 g/L, nitrogen 41 mM and 22$^{\circ}C$, respectively. The optimization of there factors enhanced the hGM-CSF production by 2 times because high sucrose concentration stimulated the secretion of hGM-CSF and low temperature prevented hGM-CSF degradation in media by pretenses.

• 한국균학회소식:학술대회논문집
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• 2015.11a
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• pp.39-39
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• 2015

Korean traditional Nuruk has been developed with various materials and shapes according to geographical environments and climates of their origins. Nuruk is also known as kokja in Korea, reflecting the understanding that microorganisms such as wild fungi, yeasts, and lactobacillus bacteria are naturally inoculated and reproduced. The objective of this study is to identify the characteristics of traditional Nuruk through recreating traditional production methods detailed in ancient Korean documents. In the present study, a total of 58 different kinds of Korean traditional Nuruk were prepared, including 46 kinds of recreated products. Each Nuruk sample was evaluated for its enzymatic activities, including glucoamylase, protease, and glucanase. Their suitability for alcoholic beverage production were compared to each other. To isolate valuable microorganisms from Nuruk samples, alcoholic beverages produced using each sample were subjected to sensory evaluation to determine their taste. In addition, metabolite changes in traditional alcoholic beverages fermented with different kinds of Nuruk were analyzed through mass-based metabolomics approach. This study presents, for the first time, the traditional production methods written in ancient Korean documents using workable production methods supported by modern technologies. In addition, this study analyzed the characteristics of reproduced Nuruk. It could be utilized as a basis for studying traditional Korean traditional alcoholic beverages and their valuable microorganisms.

• The Korean Journal of Food And Nutrition
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• v.7 no.2
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• pp.151-158
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• 1994

This study was carried out to clear up the cause of low-acid producing phenomenon occurred In non-fat dry milk during liquid-yoghurt fermentation by Lactobacillus case, and to present its improving methods. All samples of non-fat dry milks which were low in TCA-soluble peptides showed low-acid production, but those high in TCA-soluble peptides showed high-acid production. The addition of trypsin-hydrolysate of Na-caseinate to non-fat dry milk showed some improving effect on acid production but that of papain-hydrolysate did not show any improving effect and that of bacterical neutral protease-hydrolysate showed some inhibitory effect. The improving effects on growth and acid production of lactic acid bacteria were more prominent when the trypsin-hydrolysate of Na-caseinate was added. to such fermenting system in which the levels of TCA-soluble peptides and the proteolytic ability of starter bacteria were abnormally low. The liquid-yoghurt made with non-hydrolysed Na-caseinate and defective non-fat dry milk showed precipitate occurrence but that with trypsin-hydrolysate of Na-caseinate and defective non-fat dry milk did not make any precipitate during storage as with normal non-fat dry milk.

• Preventive Nutrition and Food Science
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• v.2 no.2
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• pp.101-108
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• 1997

A lactic acid bacterium LAB3113, isolated from traditionally fermented Kimchi was found to produce bacteriocin whose activity was very specific toward lactobacilli and not effective against any other bacteria. Lactobacilli affected by the inhibitory substance included Lactobacillus delbrueckii-lactis, L. johnsonii, L. gsseri, and L. curvatus. Based upon biochemical and physiological characteristics, LAB3113 was classified as Lactococcus lactis, and its bacteriocin was named as lactococcin K3113. Lactococcus lactis. LAB3113 produced bacteriocin at th early stage of growth and the concentration of the bacteriocin did not decrease even after alt stationalry phase. Optimal temperature of bacteriocin production was $25^{\circ}C$ at the initial pH 7.0. Partially purified lactococcin K3113 was completely inactivated by protease, but not affected by lipase, lysozyme and RNase. The bacteriocin was very heat-stable even after autoclaving for 20 min. It was also stable in pH changes, an was not affected by th presence of solvents. lacotococcin K3113 appeared to act in bactericidal mode against L. delbrueckii-lactis ATCC4797. Molecular weight of lactococcin K3113 was calibrated as 10,500 dal by SDS-PAGE an activity staining. Lactococcus lactis LAB3113 had four residential plasmids of 3.7kb, 11.2kb, 15.5kb, and 48kh in molecular sizes. Plasmid profile analysis of mutant strain revealed that 15.5 kb plasmid was re-sponsible for the production of lactococcin K3113 and its immunity to the bacteriocin.

• Korean Journal of Food Science and Technology
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• v.47 no.4
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• pp.438-445
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• 2015

The production of GABA was optimized by co-cultivation of immobilized Lactobacillus plantarum K154 (ILK) with Ceriporia lacerata cultures. The mycelial culture of C. lacerata was performed in a defined medium containing 3% glucose, 3% soybean flour, and 0.15% $MgSO_4$ in a submerged condition for 7 days at $25^{\circ}C$, resulting in the production of 29.7 g/L mycelia, 3.1 g/L exopolysaccharides, 2% (w/w) ${\beta}$-glucan, 68.96 unit/mL protease, and 10.37 unit/mL ${\alpha}$-amylase. ILK in C. lacerata culture showed viable cell counts of $3.13{\time}10^9CFU/mL$ for immobilized cells and $1.48{\time}10^8CFU/mL$ for free cells after 1 day. GABA production in the free and immobilized cells was 9.96 mg/mL and 6.30 mg/mL, respectively, after 7 days. A recycling test of ILK in the co-fermentation was consequently performed five times at $30^{\circ}C$ for 15 days, resulting in the highest production of GABA. GABA could also be efficiently overproduced by co-cultivation with the produced polysaccharides, ${\beta}$-glucan, peptides, and probiotics.

• The Korean Journal of Mycology
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• v.39 no.2
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• pp.99-104
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• 2011

Shiitake breeding requires the procedures of mating of two different parental strains and selection of hybrid strains that have good traits for the mushroom production. In this study, we tested the possibility of the use of chromogenic plate-based assay for extracellular enzyme production in order to assess and find good biochemical properties-possessed hybrid strains that were generated from genetic cross of the monokaryotic strains derived from two different parental strains of Lentinula edodes Sanjo-101ho and Sanjo-108ho. We observed that there was difference in the ability of producing ${\beta}$-glucosidase, avicelase, CM-cellulase, amylase, pectinase, xylanase, and protease among the monokaryotic strains. We could also comparatively assess that the ability of the seven extracellular enzymes production in the hybrid strains depended on the mating combination of the monokaryotic strains. Our results demonstrate that the assessment method for extracellular enzyme production using chromogenic plate assay could be usefully applied to the assessment of the hybrid strains derived from the breeding procedure of L. edodes.

• Korean Journal of Microbiology
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• v.55 no.3
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• pp.226-233
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• 2019

Ochratoxin A (OTA) that is one of mycotoxins produced mainly by Aspergillus spp. is a common contaminant of stored grains and poses health hazards to human and livestock. The aim of this study is to explore the ability of isolated bacteria Bacillus subtilis AF13 and Streptomyces shenzhenensis YR226 to inhibit growth and OTA production of 3 ochratoxigenic Aspergillus strains. The antifungal activity against mycelial growth and sporulation of Aspergillus strains was examined by coculture with AF13 and YR226 on potato dextrose agar plate. AF13 and YR226 reduced 77.58 and 78.48% of fungal colony radius, respectively, and both strains inhibited fungal sporulation up to 99% in 10 days of incubation. YR226 also reduced more than 91% of spore germination of 3 fungal strains. When Aspergillus strains were cocultured with AF13 or YR226 in yeast extract sucrose medium, mycelial growth and OTA production decreased in all three fungal strains. In particular, AF13 completely inhibited the mycelial growth of A. alutaceus and inhibited its OTA production by 99%, and YR226 also reduced mycelial growth and toxin production up to 99%, respectively. Antimicrobial substances produced by AF13 and YR226 included siderophore, chitinase, protease, ${\beta}$-1,3-glucanase and biosurfactant. These results suggest that AF13 and YR226 can be used in a biological method to prevent valuable crops against mycotoxigenic fungi, and therefore decrease economic damage in agriculture and feed industry.

• Microbiology and Biotechnology Letters
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• v.49 no.2
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• pp.255-263
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• 2021

In this study, we assessed the technological characteristics and safety of 88 Enterococcus faecium strains isolated from meju; the strains possess the glutamate decarboxylase gene gadA/B involved in γ-aminobutyric acid production. The study was conducted to evaluate the possibility of introducing E. faecium meju isolates as food fermentation starters. We observed that a NaCl concentration of 6% (w/v) facilitated the growth and acid production of all strains. At a NaCl concentration of 7%, 21 strains (24%) exhibited a low growth rate, 72 strains (82%) a weak acid production, and 16 strains (18%) showed no acid production. All strains exhibited protease activity at a NaCl concentration of 4%. At a NaCl concentration of 5%, 86 strains exhibited weak activity, and one strain showed no protease activity. We could not detect any lipase activity in the investigated strains. None of the strains exhibited an acquired antibiotic resistance to the seven antibiotics tested in the present study, namely ampicillin, chloramphenicol, ciprofloxacin, gentamicin, penicillin G, tetracycline, and vancomycin. We could identify the Enterococcus endocarditis antigen gene efaA and the tyrosine decarboxylase gene tdc contributing to tyramine production, in 88 meju isolates. We could not detect the Enterococcus surface protein gene esp, which is specifically possessed by human-originated E. faecium strains, in any of the 88 strains tested in the study.