• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.749-760
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• 2022

α-Galactosidase is a debranching enzyme widely used in the food, feed, paper, and pharmaceuticals industries and plays an important role in hemicellulose degradation. Here, T26, an aerobic bacterial strain with thermostable α-galactosidase activity, was isolated from laboratory-preserved lignocellulolytic microbial consortium TMC7, and identified as Parageobacillus thermoglucosidasius. The α-galactosidase, called T26GAL and derived from the T26 culture supernatant, exhibited a maximum enzyme activity of 0.4976 IU/ml when cultured at 60℃ and 180 rpm for 2 days. Bioinformatics analysis revealed that the α-galactosidase T26GAL belongs to the GH36 family. Subsequently, the pET-26 vector was used for the heterologous expression of the T26 α-galactosidase gene in Escherichia coli BL21 (DE3). The optimum pH for α-galactosidase T26GAL was determined to be 8.0, while the optimum temperature was 60℃. In addition, T26GAL demonstrated a remarkable thermostability with more than 93% enzyme activity, even at a high temperature of 90℃. Furthermore, Ca²⁺ and Mg²⁺ promoted the activity of T26GAL while Zn²⁺ and Cu²⁺ inhibited it. The substrate specificity studies revealed that T26GAL efficiently degraded raffinose, stachyose, and guar gum, but not locust bean gum. This study thus facilitated the discovery of an effective heat-resistant α-galactosidase with potent industrial application. Meanwhile, as part of our research on lignocellulose degradation by a microbial consortium, the present work provides an important basis for encouraging further investigation into this enzyme complex.

• Journal of Microbiology and Biotechnology
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• v.32 no.6
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• pp.800-807
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• 2022

Four aprE genes encoding alkaline serine proteases from B. subtilis strains were used as template genes for family gene shuffling. Shuffled genes obtained by DNase I digestion followed by consecutive primerless and regular PCR reactions were ligated with pHY300PLK, an E. coli-Bacillus shuttle vector. The ligation mixture was introduced into B. subtilis WB600 and one transformant (FSM4) showed higher fibrinolytic activity. DNA sequencing confirmed that the shuffled gene (aprEFSM4) consisted of DNA mostly originated from either aprEJS2 or aprE176 in addition to some DNA from either aprE3-5 or aprESJ4. Mature AprEFSM4 (275 amino acids) was different from mature AprEJS2 in 4 amino acids and mature AprE176 in 2 amino acids. aprEFSM4 was overexpressed in E. coli BL21 (DE3) by using pET26b(+) and recombinant AprEFSM4 was purified. The optimal temperature and pH of AprEFSM4 were similar to those of parental enzymes. However, AprEFM4 showed better thermostability and fibrinogen hydrolytic activity than the parental enzymes. The results indicated that DNA shuffling could be used to improve fibrinolytic enzymes from Bacillus sp. for industrial applications.

• The Korea Journal of Herbology
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• v.29 no.5
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• pp.1-8
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• 2014

Objectives : To prove the channel-tropism theory of herbal medicines on diabetes mellitus as emaciation-thirst disease in Korean Medicine Theory, we investigated the selective therapeutic effects of Mori Cortex Radidus (MCR), Schisandrae Fructus (SF), Anemarrhenae Rhizoma (AR) for the upper emaciation on different organs in high fat and high sucrose (HF/HS) feeding-induced prediabetic mice. Methods : Diabetes in C57BL/6 mice was induced by the administration of high fat (45 kal%) and high sucrose (32 kal%) for 8 weeks, and them treated with each extract at 250 or 500 mg/kg body weight for 4 weeks (once a day). Oral glucose tolerance test and body weight was measured once a week. Insulin, total cholesterol, triglyceride, ${\gamma}GTP$, GOT and GPT were measured in the sera of all mice. Histopathological changes of different organs, lung, heart, pancreas, stomach, liver, and kidney were observed by H&E staining. Results : The results revealed that MCR extract inhibited the impaired glucose tolerance and lung damage, and increased serum insulin levels in HF/HS-induced prediabetic mice. SF extract inhibited the impaired glucose tolerance and lung damage, increased serum insulin levels, and decreased serum triglycerige levels. Meanwhile, AR extract inhibited the impaired glucose tolerance and lung damage, and decreased serum levels of insulin, total cholesterol and triglycerige levels. Conclusions : These results demonstrated that MCR, SF, and AR extract as the upper emaciation herbal medicines were followed their channel-tropism theory like a lung, and may have a selective therapeutic potential for control of diabetic stage.

• Journal of Microbiology and Biotechnology
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• v.33 no.12
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• pp.1648-1656
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• 2023

We have previously observed that feeding with single-cell hemoprotein (heme-SCP) in dogs (1 g/day for 6 days) and broiler chickens (1 ppm for 32 days) increased the proportion of lactic acid bacteria in the gut while reducing their body weights by approximately 1~2%. To define the roles of heme-SCP in modulating body weight and gut microbiota, obese C57BL/6N mice were administered varied heme-SCP concentrations (0, 0.05, and 0.5% heme-SCP in high fat diet) for 28 days. The heme-SCP diet seemed to restrain weight gain till day 14, but the mice gained weight again later, showing no significant differences in weight. However, the heme-SCP-fed mice had stiffer and oilier bodies compared with those of the control mice, which had flabby bodies and dull coats. When mice were dissected at day 10, the obese mice fed with heme-SCP exhibited a reduction in subcutaneous fat with an increase in muscle mass. The effect of heme-SCP on the obesity-associated dyslipidemia tended to be corroborated by the blood parameters (triglyceride, total cholesterol, and C-reactive protein) at day 10, though the correlation was not clear at day 28. Notably, the heme-SCP diet altered gut microbiota, leading to the proliferation of known anti-obesity biomarkers such as Akkermansia, Alistipes, Oscillibacter, Ruminococcus, Roseburia, and Faecalibacterium. This study suggests the potential of heme-SCP as an anti-obesity supplement, which modulates serum biochemistry and gut microbiota in high-fat diet-induced obese mice.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.50 no.6
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• pp.428-434
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• 2005

For the purpose to verify the grain and evaluate milling properties of Korean waxy wheat, com­parison analysis between waxy wheat lines and their respective maternal parents were performed. The waxy lines showed various grain yields of 4.76${\~}$5.79 t/ha depending on parentages, which were corresponding to $80{\~}96\%$ levels of their respective maternal parents. One thousand grain weights of waxy lines were also lighter than its respective part in its parentage by exhibiting 32.8${\~}$34.6 g compared to 32.9${\~}$45.2 g of their parents. Test weights of waxy lines and their parents were 720${\~}$798 g/l and 786${\~}$797 g/l, respectively. The proportions of the grains above 2.5 mm in width were higher in order of Keumgang, Olgeuru, Geuru, SW97134, Suwon 292, Woori, SW97105, and SW97110. Waxy lines exhibited low milling properties by showing the straight flour yields ranging from $61.8\%$ to $67.1\%$ compared with the yields of their parents ranging from $66.1\%$ to $72.5\%$; the waxy lines were significantly lower in first break flour (Bl) and first reduction flour (Rl) yields in the Buller test mill, while significantly higher in the yields of second and third reduction flour (R2 and R3) than the respective ones of their parent wheat.

• Journal of Food Hygiene and Safety
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• v.32 no.1
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• pp.50-56
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• 2017

The purpose of this study was to investigate the microbiological contamination of water and drinking cups in springs and to estimate the toxin gene, enterotoxin production ability and antibiotic susceptibility of foodborne pathogens. Ten spring water and 34 drinking cups were tested. The average number of total aerobic bacteria and coliform bacteria in spring water were 1.8 log CFU/mL and 1.2 log CFU/mL, and in drinking cups were $4.7log\;CFU/100cm^2$ and $1.7log\;CFU/100cm^2$. Salmonella spp., Staphylococcus aureus, E. coli O157:H7, Listeria monocytogenes, Vibrio parahaemolyticus, Yersinia enterocolitica were not isolated from all of samples but Bacillus cereus was detected in 5 (14.7%) of 34 drinking cups. The nheA and entFM genes were major enterotoxin genes in B. cereus isolated from drinking cups. All of B. cereus tested in this study produce non-heamolytic enterotoxin but only 2 isolates possessed heamolysin BL enterotoxin producing ability. B. cereus was resistant to ${\beta}-lactam$ antibiotics. These results revealed that the sanitary conditions of drinking cups in spring should be improved promptly. The substitution carrying a personal drinking cup for the public drinking cups equipped in springs is suggested to prevent food-borne illness.

• The Korea Journal of Herbology
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• v.32 no.1
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• pp.25-31
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• 2017

Objectives : Mori Radicis Cortex (MRC), the root epidermis of Morus alba L., has been traditionally used to treat lung-related diseases in Korean Medicine. The common of MRC is Mulberry bark Morus bark, and it's pharmaceutical properties and taste are known as sweet and cold, and it promotes urination and reduce edema by reducing heat from the lungs and soothe asthma. In the present study, anti-apoptotic mechanism of MRC in middle cerebral artery occlusion (MCAO) model in mice. Methods : Two-hundred grams of MRC was extracted with methanol at room temperature for 5 days, and this was repeated one time. After filtration, the methanol was removed using vacuum evaporator, then stored at $-20^{\circ}C$ until use. C57BL/6 male mice were housed in an environment with controlled humidity, temperature, and light cycle. In order to determine beneficial effects of MRC on ischemia induced brain damage, infarct volume, neurological deficit scores, activities of several apoptosis-related proteins such as caspase-8, -9, Bcl-xL in MCAO-induced brains of mice were analyzed. Mice in MRC-treated groups were orally administered 30, 100, or 300 mg/kg of body weight for three consecutive days before commencing the MCAO procedure. Results : Pre-treatment of MRC significantly reduced infarct volume in MCAO subjected mice applied with 300 mg/kg of MRC methanol extract, and MRC effectively inhibited Bcl-xL reduction and caspase-9 activation caused by MCAO-induced brain damage. Conclusions : MRC showed neuro-protective effects by regulating apoptosis-related protein signals, and it can be a potential candidate for the therapy of ischemia-induced brain damage.

• Journal of Korean Medicine Rehabilitation
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• v.32 no.2
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• pp.19-36
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• 2022

Objectives This study is to investigate the effects and mechanisms of Imyo-san (IMS) on the obese mice model induced by high-fat diet. Methods Antioxidative capacity was measured by in vitro method. C57BL/6 mice were randomly assigned into 5 groups (n=7). Normal group was fed general diet (Normal). The other 4 groups were fed high fat diet (HFD) with water (Control), with Garcinia gummi-gutta (GG, Garcinia gummi-gutta 200 mg/kg), with low-dose IMS (IMSL, Imyo-san 0.54 g/kg) and with high-dose IMS (IMSH, Imyo-san 1.08 g/kg). Results IMS showed high radical scavenging activity. After 6 week experiment, body weight, food intake, food efficiency ratio (FER), epididymal fat and liver weight, triglyceride (TG), total cholesterol (TC), high density lipoprotein (HDL) cholesterol, low density lipoprotein (LDL) cholesterol, very low density lipoprotein (VLDL) cholesterol, sterol regulatory element-binding protein-1 (SREBP-1), phospho-acetyl-CoA carboxylase (p-ACC), fatty acid synthase (FAS), stearoyl-CoA desaturase-1 (SCD-1), SREBP-2, 3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR), phospho-liver kinase B1 (p-LKB1), phospho-AMP-activated protein kinase (p-AMPK), peroxisome proliferator-activated receptor 𝛼 (PPAR𝛼), peroxisome proliferator-activated receptor 𝛾 coactivator-1𝛼 (PGC-1𝛼), uncoupling protein-2 (UCP-2), carnitine palmitoyltransferase 1A (CPT-1A), and histology of liver and epididymal fat were measured and analysed. Body weight gain, FER, liver and epididymal fat weight of IMS groups were significantly decreased. There were significant improvements in blood lipids with less TG, TC, LDL-cholesterol, VLDL-cholesterol and more HDL-cholesterol. Proteins associated with lipid synthesis (SREBP-1, p-ACC, FAS, SCD-1) and cholesterol (SREBP-2, HMGCR) was improved. Factors regulating lipid synthesis and lipid catabolism (p-LKBI, p-AMPK, PPARα, PGC-1α, UCP-2, CPT-1A) were increased. In histological examinations, IMS group had smaller fat droplets than control group. All results increased depending on concentration. Conclusions It can be suggested that IMS has anti-obesity effects with improving lipid metabolism.

• Journal of Korean Medicine Rehabilitation
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• v.32 no.4
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• pp.1-8
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• 2022

Objectives This study is to investigate the effects and mechanisms of Gastrodia elata extract (GEE) on the high-fat diet-induced obesity model. Methods C57BL/6 mice were randomly assigned into 5 groups (n=10). Control group was fed normal diet (ND). Obesity group was fed 60% high fat diet (HFD). The other three groups were fed HFD with 100, 200, 500 mg/kg GEE. After five weeks, body weight, liver and epididymal fat weight, triglyceride concentration in liver and serum, sterol regulatory element-binding protein-1 (SREBP-1), acetyl-CoA carboxylase (ACC), fatty acid synthase, peroxisome proliferator-activated receptor 𝛾 (PPAR-𝛾), CCAAT/enhancer binding protein 𝛼 (C/EBP-𝛼) expression level, insulin concentration in serum were measured. Results The GEE (100, 200, and 500 mg/kg)-treated animals exhibited substantial decreases in body mass, liver weight and epididymal white adipose tissue collate to the HFD-fed group. GEE treatment also reduced hepatic and serum triglyceride level. Furthermore, GEE treatment significantly inhibited adipogenesis in the GEE group by reducing the protein expression of SREBP-1, ACC and the messenger RNA expression of PPAR𝛾, C/EBP-𝛼, which are adipocyte differentiation-related genes. Conclusions These research outcomes recommend that GEE is possibly valuable for the prevention of HFD-induced obesity via modification of various pathways related with adipogenesis and adipocyte differentiation.

• THE JOURNAL OF KOREAN ORIENTAL ONCOLOGY
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• v.2 no.1
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• pp.57-73
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• 1996

These studies were consist of two sub-experiment. In order to study the effect of Banhahubaktang on the Cell-cytotoxicity In vitro. We had put through MTT Assay. In order to investigate the effects of Banhahubaktang on the ICR mice which had Abdominal tumor induced by Sarcoma-180 cell line, C57BL/6 mice which had pulmonary melanoma induced by B16 cell line. After Sarcoma-180 cell line and B16 cell line were transplanted, the extract of Banhahubaktang was orally administered to the mice to observe the extension of survival time of the mice, inhibition of solid tumor, inhibition of pulmonary melanoma metastasis. productivity of Interleukin-2, NK-Activity. The results were summarized as follows: 1. On the MTT assay, in case of $100{\mu}g/ml$ and $10{\mu}g/ml$ of Banhahubaktang concentration were inhibited cell viability significantly. But $1{\mu}g/ml$ of Banhahubaktang was tended to inhibit cell viability with no significance. 2. In the effect of life extension, Banhahubaktang treated group appeared to survive longer than the control group, but which were not significant. 3. In the effect of inhibit solid tumor, Banhahubaktang treated group appeared to decrease than the control group, but which were not significant. 4. In the effect of inhibit melanoma pulmonary metastasis. Banhahubaktang treated group appeared to inhibit than the control group significantly. 5. In the productivity of Interleukin-2, on 7 and 14 day, Banhahubaktang treated group increased than control group, which were significant. But on 21 day, test group and control group were much in common. 6. In the NK-Activity, Banhahubaktang treated group and control group were much in common.