• Journal of Microbiology and Biotechnology
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• v.30 no.11
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• pp.1720-1728
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• 2020

We have previously characterized AprESJ4, the major fibrinolytic enzyme from Bacillus subtilis SJ4 (Yao et al., 2019). During that study, we observed a 68 kDa protein with fibrinolytic activity. In this study, we cloned the gene (vprSJ4) encoding the 68 kDa protein, a mature Vpr and minor protease secreted by Bacillus species. vprSJ4 encodes a preproenzyme consisting of 810 amino acids (aa) including signal sequence (28 aa) and prosequence (132 aa). The mature enzyme (650 aa) has a predicted molecular weight of 68,467.35. Unlike Vprs from other B. subtilis strains, VprSJ4 has 4 additional amino acids (DEFA) at the C-terminus. vprSJ4 was overexpressed in Escherichia coli. PreproVprSJ4 was localized in inclusion bodies, and subjected to in vitro renaturation and purification by an affinity column. SDS-PAGE and western blot showed that autoprocessing of preproVprSJ4 occurred and 68 kDa and smaller proteins were produced. The optimum pH and temperature of the recombinant VprSJ4 were pH 7.0 and 40℃, respectively. Kinetic parameters of recombinant VprSJ4 were measured by using an artificial substrate, N-succinyl-ala-ala-pro-phe-p-nitroanilide. Coexpression of vprSJ4 and aprESJ4 using pHY300PLK increased the fibrinolytic activity a further 117% when compared with aprESJ4 single expression using the same vector in B. subtilis WB600.

• Korean Chemical Engineering Research
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• v.49 no.4
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• pp.449-455
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• 2011

This study was performed to develop antimicrobial films using polyvinyl alcohol and methyl cellulose. Methyl cellulose and polyvinyl alcohol films plasticized with PEG(polyethylene glycol) were prepared by solvent casting process under addition of 0.025~1.0 wt% ampicillin and 0.1~1.0 wt% streptomycin as an antimicrobial agent. The mechanical properties of prepared films were examined by universal testing machine(UTM). Tensile strength of methyl cellulose films was 15.44~21.70 $N/mm^2$. Tensile strength of PVA(15 wt%) film was 20.2~51.5 $N/mm^2$, and the tensile strength of the antimicrobial films were decreased linearly with increasing the antibiotic loading amount up to 1 wt%. Antimicrobial activities of PVA and methyl cellulose films containing ampicillin and streptomycin through the disc diffusion test for the Staphylococcus aureus and Escherichia coli. The antimicrobial activity of methyl cellulose films and PVA containing ampicillin were higher than that of containing streptomycin methyl cellulose films. The results indicate the films may be a proper materials for antimicrobial packing applications.

• Bulletin of the Korean Chemical Society
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• v.34 no.8
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• pp.2419-2424
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• 2013

Guanosine-5'-diphosphate 3'-diphosphate (ppGpp) serves as alarmone in bacterial stringent responses. In this study, an affinity column was constructed by immobilizing ppGpp to NHS-Sepharose for isolating ppGpp-binding proteins. A novel ppGpp-binding protein, YjgA, was isolated and characterized by MALDI-TOF MS (matrix-assisted laser desorption ionization-time-of-flight mass spectrometry) coupled with two-dimensional gel electrophoresis. YjgA and truncated forms of YjgA were cloned and over-expressed in BL21 (DE3). The binding affinity of YjgA to ppGpp was determined by equilibrium dialysis. The interaction of YjgA with ppGpp was very specific, considering that the dissociation constant of YjgA with ppGpp was measured as $5.2{\pm}2.0{\mu}M$, while the affinities to GTP and GDP were about 60 and 30 times weaker than ppGpp. Expression of yjgA gene in Escherichia coli K-12 MG1655 was examined by reverse transcription polymerase chain reaction (RT-PCR). RT-PCR results revealed that yjgA was expressed from early to late stationary phase. The yjgA deletion mutant exhibited decreased cell number at stationary phase compared to parent strain and the over-expression of YjgA increased the cell number. These results suggested that YjgA might stimulate cell division under stationary phase. In most prokaryotic genome, about half of the protein candidates are hypothetical, that are expected to be expressed but there is no experimental report on their functions. The approach utilized in this study may serve as an effective mean to probe the functions of hypothetical proteins.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.23-37
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• 2022

Thirty-six weaned piglets with an initial body weight (BW) of 8.43 ± 0.40 kg (28 days of age, ([Landrace × Yorkshire] × Duroc) were randomly assigned to 6 treatments for a 2-week feeding trial to determine the effects of different inorganic zinc (IZ), organic zinc (OZ) or combination of low crude protein diet (LP) and Mixed feed additive (MFA) on diarrhea score, nutrient digestibility, zinc utilization, blood profiles, organ weight, and fecal microflora in weaned piglet diet. The pigs were individually placed in 45 × 55 × 45 cm stainless steel metabolism cages in an environmentally controlled room (30 ± 1℃). The dietary treatments included a negative control (NC), positive control (PC; zinc oxide, 1,000 mg/kg), T1 (IZ : OZ, 850 : 150), T2 (IZ : OZ 700 : 300), T3 (IZ : OZ, 500 : 500), and T4 (LP + MFA [0.1% Essential oils + 0.08% Protease + 0.02% Xylanase]). The daily feed allowance was adjusted to 2.7 times the maintenance requirement for digestible energy (2.7 × 110 kcal of DE/kg BW^0.75). This allowance was divided into two equal parts, and the piglets were fed at 08 : 30 and 17 : 30 each day. Water was provided ad libitum through a drinking nipple. The diarrhea score was significantly increased (p < 0.05) in NC treatment compared with other treatments. The apparent total tract digestibility (ATTD) of dry matter (DM), nitrogen (N), and gross energy (GE) was significantly increased (p < 0.05) in the T2 treatment compared with the PC and NC treatments in week 1. In week 2, the ATTD of DM, N, and GE was significantly decreased (p < 0.05) in the NC treatment compared with other treatments. The T3 treatment had significantly higher (p < 0.05) ATTD and apparent ileal digestibility of zinc than the PC and T1 treatments. The Escherichia coli count in feces was significantly decreased in the T4 treatment compared with the NC and T2 treatments. The Lactobacillus count in feces was significantly increased in the T4 and T1 treatment compared with the T2 and T3 treatments. In conclusion, IZ : OZ 500 : 500 levels could improve nutrient digestibility and zinc utilization in weaned piglets, Moreover, MFA in LP diets could be used as a zinc alternative.

• Journal of Animal Science and Technology
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• v.64 no.1
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• pp.70-83
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• 2022

A set of studies was performed to determine the influence of dietary ZnO concentration and source during two phases (days 0 to 14 and days 15 to 28). Experiment 1: 168 weaned piglets were allocated to four treatment groups in six replicates. The treatments included a basal diet without ZnO supplementation (control), 2,500 mg ZnO/kg (In2500), 500 mg nano-ZnO/kg (N500), and 150 mg nano-ZnO/kg (N150). Experiment 2: 168 weaned piglets were divided into three treatment groups with eight replicates. The treatments included control, In2500, N300, and 150 mg nano-ZnO/kg (N150). An in vitro trial showed that the growth of Listeria monocytogenes, Escherichia coli, and Salmonella typhimurium was inhibited when exposed to 300 and 500 ppm of ZnO after 24 h of incubation. In experiment 1, the average daily gain (ADG) by the pigs was improved in the N500 and IN2500 treatment groups. Colonization of coliforms and Clostridium spp. significantly decreased in the pigs fed the N500 and IN2500 diets in phase 1. The total plasma antioxidant capacity was greater in the IN2500 and N500 treatment groups than in the control. Superoxide dismutase (SOD) activity was greater in pigs fed the IN2500 (phase 1) or the IN2500 and N500 (phase 2) diets than in the control and N150 treatment group. In experiment 2, pigs in the N300 treatment group showed a higher ADG and lower fecal score colonization of coliforms and Clostridium spp. compared with those in the N150 treatment group. In conclusion, nano-ZnO at a dose of 300 ppm showed the same growth as the pharmacological dose of Zn. This provides an option to the pharmacological dose.

• Food Science of Animal Resources
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• v.30 no.6
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• pp.975-988
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• 2010

Two experiments were conducted to determine whether water extracts from Artemisia capillaries (A. capillaries) and Camellia sinensis (C. sinensis) could be used as alternatives to antibiotic growth promoters in broiler feed. The experiment 1 was verified their chemical composition, extracts yields, total phenolic compounds concentration, antioxidant activity, antimicrobial activity, and chicken splenocytes proliferation through in vitro test. The extract yields of A. capillaries and C. sinensis were 26.5 and 16.8%, respectively. Total phenolic compounds concentrations of them expressed as gallic acid equivalent were 15.28 and 26.74 mg/mL, respectively. Electron donating abilities of them expressed as $SC_{50}$ showing 50% DPPH radical scavenging were 0.30 and 0.06 mg, respectively. Bacterial inhibitory rates of them against Escherichia coli, Staphylococcus aureus, and Salmonella Typhimurium were ranged from 42.1 to 52.3% and from 21.6 to 33.7%, respectively. And, these extracts increased proliferation of chicken splenocytes. Especially, A. capillaris was more excellent than Echinacea and Concanavalin A known as T-cell stimulator. The experiment 2 was investigated their effects on growth performance, relative organ weight, cecal microflora, blood biochemical parameters, and splenic cytokines mRNA expression in broiler chicks. Four hundred eighty 1-day-old male broiler chicks (Ross 308) were divided in to 4 treatment groups with 4 replicates of 30 birds in each group: NC (control, no antibiotics), PC (avilamycin, 10 ppm; salinomycin, 60 ppm), AC (A. capillaries, 100 ppm), and CS (C. sinensis, 100 ppm); treatments were administered through water supplementation. Final body weight was significantly higher in all treated groups than in NC (p<0.05). Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in NC (p<0.05). The relative weights and lengths of the small intestine were more significantly decreased in the PC and AC groups than in the other groups. Cecal Salmonella numbers were significantly or somewhat decreased in all treated groups than in the NC group (p<0.05). The contents of total cholesterol, aspatate aminotransferase, and alanine aminotransferase in blood serum were more significantly decreased in all treated groups than in NC (p<0.05). In conclusion, these results suggested the possibility that these extracts could serve as alternatives for antibiotic growth promoters.

• Food Science and Preservation
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• v.12 no.6
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• pp.558-564
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• 2005

This study was carried out to investigate the cleaning effect of sesame leaf, the sterilization effect and physicochemical properties, treated with various electrolyzed water. Initial physicochemical properties could be kept more than 1 month in electrolyzed oxidizing water(EW-1) of diaphragm type and 15 days in electrolyzed water(EW-2 and EW-3) of non-diaphragm system, there was no significant difference by storage temperature. 4 kinds of microorganism (initial total counts, $10^7\~10^9$ CFU/mL) were sterilized within $0.5\~1$ minutes by electrolyzed water. In fresh sesame leaves, total viable cell count and coliform group in the treatment of electolyzed water were decreased to about $2\~3$ log scale comparing non-treated ones. Especially Bacillus cereus was not detected until 13th day when treated with EW-l. Decaying ratio of sesame leaf appears on day 6 of storage in the untreated but the treatments of electrolyzed water has no sign until day 10 of storage. Change in color difference(${\Delta}E$) during storage was observed the treatments of electrolyzed low-alkaline water(EW-2) and electrolyzed neutral water(EW-3) were very desirable at the level $1\~2$ after day 13 of storage comparative to the untreated Change of Chlorophyll content was biggest decreased to 6.8 $mg\%$ on the untreated and decreased least to 8.35 $mg\%$ on EW-3 treated group on 13th day from initial value of $9.0\~10.3\;mg\%$ The overall sensory evaluation appeared most acceptable in the treatments of EW-2 and EW-3.

• Journal of Food Hygiene and Safety
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• v.15 no.4
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• pp.328-333
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• 2000

This study was carried out to changes in chemical and microbiological properties of spring waters in Tongyeoung area. In this paper, ninety spring water samples were collected from 9 station for 11 month to evaluated chemical and bacteriological water quality. Range and mean values of constituents of the samples are as followed; water temperature 5.2~25.8$^{\circ}C$, 16.3$^{\circ}C$, pH 6.0~7.2, 6.7, total residue 33.6~210 mg/1, 90.6 mg/1, turbidity 0.35~5.48, 1.45NTU, KMnO4 consumed 0.51~4.21 mg/1, 1.39 mg/1, chloride ion 6.23~42.5, 16.7 mg/l, phosphate-phosphorus ND-0.04, 0.02 mg/1, nitrite-nitrogen ND~0.02, 0.01 mg/1, nitrate-nitrogen ND~3.56, 1.42 mg/1, ammonia-nitrogen ND~0.20, 0.14 mg/1, dissolved total nitrogen ND~3.78, 1.57 mg/1, iron 0.04~0.28, 0.13ppm, zinc 0.03~0.66, 0.20ppm, mangan ND~0.01, allumium 0.14~0.58, 0.39ppm, copper ND~0.01, 0.01, lead ND~0.01, 0.01ppm, Arsenic ND~0.01, 0.01ppm, mercury ND~0.02, chrome not detected, cadmium not detetced respectively. The viable cell counts of the spring waters ranged 5.0~760/m1(means 130/m1). Range and mean value of total coliform and focal coliform MPN's of the spring waters were 0~2,400MPN/100 ml, 73MPN/100 ml and 0~540MPN/100 ml, 21MPN/100 ml. Spring water quality was usually poor with viable cell counts exceeding 130 CFU/liter and the coliform counts in spring waters of 73 MPN/liter. Composition of coliform by IMViC reaction was 33.3% E. coli, 15.6% Citrobacter freundii, 35.6% Klebsiella aerogenes and others.

• Pediatric Infection and Vaccine
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• v.25 no.2
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• pp.82-90
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• 2018

Purpose: Survival after liver transplantation (LT) has improved over the years, but infection is still a major complication. We aimed to identify the characteristics of bacterial infections in pediatric LT recipients. Methods: This study is a retrospective review of 189 consecutive children undergoing LT between 2000 and 2015 at a single center. In this study, the incidence of infection was determined for the following periods: within 1 month, between 1-5 months, and between 6-12 months. Patients who underwent liver transplants more than once or multiple organ transplants were excluded. Results: All patients had received postoperative antibiotic for 3 days. Only the maintenance immunosuppression with oral tacrolimus and steroids were performed. As a result, 132 bacterial infections developed in 87 (46.0%) patients (0.70 events per person-year). Bacterial infections occurred most frequently within the first month (n=84, 63.6%) after LT. In the pathogens, Staphylococcus aureus (15.2%), Enterococcus species (15.2%), and Klebsiella species (13.6%) were most common. Regarding the organ infected, bloodstream was most common (n=39, 29.5%), followed by peritoneum (n=28, 21.2%), urinary tract (n=25, 18.9%), and lungs (n=20, 15.2%). We changed prophylactic antibiotics from ampicillin-sulbactam to piperacillin-tazobactam at 2011, October, there were no significant effects in the prevalence of antibiotics resistant bacterial infections. The 1-year mortality was 9.0% (n=17), in which 41.2% (n=7) was attributable to bacterial infection; septicemia (n=4), pneumonia (n=2), and peritonitis (n=1). Conclusions: The incidence and type of bacterial infectious complications after LT in pediatric patients were similar to those of previous studies. Bacterial complications affecting mortality occur within 6 months after transplantation, so proper prophylaxis and treatment in this period may improve the prognosis of LT.

• Horticultural Science & Technology
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• v.28 no.5
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• pp.871-876
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• 2010

Seven native and four introduced herbs namely $Thymus$ $quinquecostatus$, $Chrysanthemum$ $zawadskii$ var. $latilobum$, $Rosmarinus$ $officinalis$, etc. were selected for analysis of the DPPH radical scavenging and anti-microbial activity of their extracts. These perennial herbs are classified as $Labiatae$ and $Compositae$ except for $Saururus$ $chinensis$ and can be propagated through seedling and cuttage. These edible herbs are used as medicinal as well as ornamental plants. Their extract has electron donating ability which ranges from 69.7 to 78.7% for native herbs and 67.4 to 75.4% for introduced herbs. Native herbs have higher (3.54%) average DPPH radical scavenging than introduced herbs. In native herbs, maximum DPPH radical scavenging activity was observed in $Agastache$ $rugosa$ (78.7%) followed by $Saururus$ $chinensis$ while $Chamaemelum$ $nobile$ showed highest activity among the introduced herbs. Many herbs viz. $Saururus$ $chinensis$, $Chrysanthemum$ $zawadskii$ var. $latilobum$ and $Solidago$ $virga-aurea$ var. $gigantean$ showed excellent anti-microbial activity against gram positive $Enterococcus$ $faecalis$, maximum (80.0%) by $Saururus$ $chinensis$. Other herbs viz. $Solidago$ $virga-aurea$ var. $gigantea$, $Chrysanthemum$ $zawadskii$ var. $latilobum$, $Salvia$Salvia $plebeia$, $Chrysanthemum$ $indicum$, $Rosmarinus$ $officinalis$, $Chamaemelum$ $nobile$ and $Lavandula$ $stoechas$ showed anti-microbial activity against gram negative $Citrobacter$ $freundii$. Especially, the inhibition of colony growth of $Citrobacter$ $freundii$ was highest in the extract of $Chrysanthemum$ $zawadskii$ var. $latilobum$, and $Chamaemelum$ $nobile$.