• KSBB Journal
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• v.23 no.3
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• pp.231-238
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• 2008

The second family member of tissue inhibitors of matrix metalloproteinases, TIMP-2, is a 21kDa protein which inhibits matrix metalloproteinases 2 (MMP-2). Expression of mammalian proteins in E. coli often forms inclusion bodies that are made up of mis-folded or insoluble protein aggregates. The requirement for the formation of 6 disulfide bonds in the process of the TIMP-2 folding is likely to be incompatible with the reducing environment of E. coli. However, this incompatibility can be often overcome by introducing a mutagenesis that could lead to enhancement of the protein solubility. In this reason, we have attempted to express the soluble TIMP-2 in E. coli by applying a modified staggered extension process (StEP), one of the in vitro PCR-based recombinant mutagenesis methods, and error-prone PCR. C-terminally located CAT fusion protein with respect to mutated TIMP-2 proteins enables us to differentiate the soluble TIMP-2 from the insoluble in E. coli by virtue of chloramphenicol resistance. According to this scheme, E. coli harboring properly-folded CAT fused to TIMP-2 protein was selected, and some of the resulting colonies exhibited an enhanced, soluble expression of TIMP-2 compared to the wild type, implying (i) the StEP technique is successfully employed to enhance the proper folding thereby increasing the solubility of TIMP-2, and (ii) the CAT dependent screening may be a simple and effective method to differentiate the soluble protein expression in E. coli.

• Journal of Life Science
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• v.24 no.10
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• pp.1110-1117
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• 2014

Tidal flats are continuously contaminated by human activities. This study assessed the bioremediation efficiency of tidal flat soil using microcosm reactors and microorganisms originating from the tidal area. We screened 135 bacterial strains that produce extracellular enzymes from the tidal area located in the North port of Incheon bay. Two bacterial strains (Pseudoalteromonas sp. and IC35 Halothiobacillus neapolitanus IC_S22) were selected and used in the microcosm reactors, which were specially designed to functionally mimic the ecological conditions of the tidal flats. Pseudoalteromonas sp. IC35 was selected based on its relatively high activity of the enzymes amylase, cellulose, lipase, and protease. Halothiobacillus neapolitanus IC_S22 was selected for oxidation of sulfur. The M1 and M2 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M2 was first inoculated with Pseudoalteromonas sp. IC35 before the seawater feeding. The initial COD in both the M1 and M2 microcosm reactors was 320 mg/l. The final COD was 21 mg/l (M1) and 7 mg/l (M2). The M3 and M4 microcosm reactors were operated by continuous feeding of seawater under the same conditions, but M4 was first inoculated with H. neapolitanus IC_S22. The initial sulfate concentration in both the M3 and M4 microcosm reactors was 660 mg/l, and the maximum sulfate concentration was 1,360 mg/l (M3) and 1,600 mg/l (M4).

• The Korean Journal of Food And Nutrition
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• v.25 no.4
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• pp.807-819
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• 2012

To evaluate the functional characteristic and availability for drinking of the fermented Smilax china leaf tea by using different microbial species, various fermented leaf tea was prepared by non-fermentation (C), or the fermentation of Saccharomyces cerevisiae (S), Bacillus sp. (B), Bifidobacterium bifidus (L), Monascus pilosus (M) and Aspergilus oryzae (A), and sensory and antioxidant parameter of each brewed tea was observed. The color of the A tea was red, but the other teas were yellow in color. Furthermore, the aesthetic quality of the A and M tea was 3.95 and 3.30 point, respectively, and other teas (2.55~2.28) were similar to that of the C tea. TP of fermented tea water extract was lower than that of the C, although TF was not significantly different between the fermented and non-fermented tea. Especially, TF of the A tea was significantly lower than those of the other teas. The range of EDA ($1mg/m{\ell}$) of water and ethanol extracts of tea C and the fermented teas was 19.25~22.48%; however, tea A was only 8.04~12.49%. In addition, FRAP, FICA and LPOIA of teas were not significantly different between the fermented and non-fermented teas. On the other hand, XOIA and AOIA of tea ethanol extracts were slightly higher than those of water extracts. XOIA of water extract derived from the teas was 4.83~9.20%, while ethanol extract of these was 9.00~19.00%. However, XOIA of B and L teas water extract was not detected. Furthermore, AOIA of fermented tea water extract (30.17~48.52%) were lower than those of ethanol extract (44.09~66.93%). In this study, interestingly, antioxidant parameters, such as FRAP, FICA, LPOIA and AOIA, of the A tea water extract (0.1%) was higher than that of the other tea in spite of high decreasing rate in the contents of TP and TF. Therefore, above results imply the possibility of fermented Smilax china leaf tea as a functional food.

• Childhood Kidney Diseases
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• v.8 no.2
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• pp.214-222
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• 2004

Purpose: Appropriate antibiotic therapy is important in childhood urinary tract infection and the selection of anibiotics is based on antimicrobial sensitivity of Escherichia coli. Extended-Spectrum ${\beta}-Lactamase(ESBL)$ is an enzyme produced by gram-negative bacilli that has the ability to hydrolyse penicillins, broad-spectrum cephalosporin and monobactam. There have been many reports of outbreaks of hospital infection by ESBL-producing organism. However, community-acquired infection with ESBL-producing organism are rare. This study was performed to retrospectively identify the incidence, characteristics and risk factors of ESBL (+) E. coli in community-acquired childhood UTI. Methods: In 288 children admitted in Ewha Womans University Hospital with E. coli UTI from Mar 2001 to February 2003, ESBL was isolated. ESBL was confirmed by the utilization of an automatized machine(Vitek GNS 433 card) using liquid medium dilution method according to National Committee for Clinical Laboratory Standard. The clinical characteristics, risk factors, antimicrobial resistance and treatment effectiveness were compared with ESBL(-) E. coli UTI. Results: Of 288 E. coli isolates, 31(10.8%) produced ESBL and 93.5%(29/31) occurred in infants younger than 6 month of age(P<0.01). No significant differences were noted in prior antibiotic use, prior admission history and underlying urogenital anomaly. Antimicrobial resistance was significantly higher in ESBL(+) E. coli compared with control patients (P<0.05). Although ceftriaxone showed 100% resistance in ESBL(+) E. coli, bacteriologic sterilization rate after ceftriaxone therapy was higher(96.8%). However, the recurrence rate of febrile UTI within 6 months was higher(25.8%) than control patients(6.6%). Conclusion: Epidemiologic study is required to find out any new risk factors of community-acquired ESBL(+) E. coli UTI and changes in selection of empirical antibiotics should be considered.

• Journal of Plant Biotechnology
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• v.34 no.3
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• pp.243-251
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• 2007

It has been recently demonstrated the presence of not only $C_{28}-BRs$ biosysnthesis, but also $C_{27}-$ and $C_{29}-BRs$ biosynthesis in plants, suggesting that BRs biosynthesis are complicatedly connected to produce biologically active BR (s). This prompted us to investigation of metabolism of a $C_{29}-BR$, 28-homoCS in seedlings of rice from which $C_{29}-BRs$ such as 28-homoTE and 28-homoTY have been identified. In vitro enzyme conversion study using a crude enzyme solution prepared from rice seedlings revealed that 28-homoCS is converted into both CS and 26-nor-28-homoCS, but their reversed reaction did not occur. This indicated that 28-homoCS is biosynthetically converted into more biologically active $C_{28}-BR$, CS by C-28 demethylation and biodegraded into 26-nor-28-homoCS by C-26 demethylation. Next, bio-conversion of 28-homoCS to 28-homoBL was examined by the same enzyme solution. No 28-homoBL as a metabolite of 28-homoCS was detected, meaning that biosynthetic reaction for 28-homoCS to 28-homoBL is not contained, and main connection of $C_{28}-BRs$ and $C_{29}-BRs$ biosynthesis is between CS and 28-homoCS in the rice seedling. This study is the first demonstrated that $C_{29}-BRs$ and $C_{28}-BRs$ bionsynthetic pathways are connected, and that $C_{29}-BRs$ biosynthetic pathway is an alternative biosynthetic pathway to produce more biologically active $C_{28}-BR$, CS in plant.

• Korean Journal of Microbiology
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• v.49 no.1
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• pp.24-29
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• 2013

In the present study, the effects of prebiotics and prebiotics+probiotics on intestinal microflora and fermentation products were evaluated in a pig in vitro fermentation model. The substrates used in this study were iso-malto oligosaccharide (IMO), partially digested chicory-inulin (CI), raffinose (RA), and cyclodextrin (CD) as prebiotics and Lactobacillus reiteri as probiotics. For a pig in vitro fermentation, the experimental diet for growing pigs was predigested using digestive enzymes secreted by small intestine and this hydrolyzed diet was mixed with a buffer solution containing 5% fresh swine feces. The mixture was then incubated with either prebiotics or prebiotics+probiotics for 24 h. Samples were taken at 24 h, and viable counts of microflora, gas, pH, volatile organic compounds (VOCs) and short-chain fatty acid (SCFA) were analyzed. The viable count of Enterobacteriaceae was significantly decreased (p<0.001) in all treatments containing prebiotics and prebiotics+probiotics when compared to the control. However, the number of lactic acid bacteria increased in the prebiotics and prebiotics+probiotics treatment. The pH values in the fermentation fluid decreased in all treatments when compared to the control, and their effects were greater in the prebiotics+probiotics group than prebiotics group. Fermentation with prebiotics resulted in a reduction in malodorous compounds such as ammonia, hydrogen sulfide and skatole when compared to the prebiotics+probiotics group. Short-chain fatty acid production was also higher for treatment with prebiotics+probiotics than treatment with prebiotics. In conclusion, the results of this study demonstrated that fermentation with prebiotics was effective in reducing the formation of malodorous compounds and prebiotics+probiotics was effective in increasing lactic acid bacteria and SCFA and reducing the pH. Moreover, further studies will be needed to determine whether the results observed in the in vitro model would occur in pigs that ingest these prebiotics or probiotics.

• Journal of the Korean Society of Food Science and Nutrition
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• v.33 no.8
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• pp.1279-1285
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• 2004

To evaluate the antimutagenic effect and hepato protective of bamboo trees and bamboo byproduct, hot-water extracts from four kinds of bamboo [wang-dae (Phyllostachys bambusoides S. et Z.), som-dae (Phyllostachys nigra var. henonis), maengjong-juk (Phyllostachys pubescens) and o-juk (Phyllostachys nigra Munro)] and maengjong-juk extract coated rice were evaluated for antimutagenicity by Ames test using Salmonella typhimurium TA100. Bamboo extracts showed strong antimutagenic activity in the Ames test which MNNG was used as mutagen in the absence and presence of S9 mix. Maengjong-juk extract coated rice diet suppressed the loss of body weight significantly. Food intake was increased in maengjong-juk extract coated rice supplemented group but showed no significant differences between control and maengjong-juk extract coated rice diet groups. Food efficiency of maengjong-juk extract coated rice supplemented group was significantly higher than that of the control group. Liver weight was significantly increased by maengjong-juk extract coated rice diet administration. Plasma GOT & GPT activities of rabbit were significantly suppressed in maengjong-juk extract coated rice supplemented group. These results suggest that bamboo trees extracts and maengjong-juk extract coated rice are bioavailable resource on treatment of cardiovascular disease, such as atherosclerosis and hypercholesterolemia.

• Korean Journal of Poultry Science
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• v.8 no.1
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• pp.1-5
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• 1981

In trying to predict the effect of genetics on the broiler in the year 2000, this is a relatively short period of time as far as broiler genetics in concerned. Modern broiler genetics started around 1945 and tremendous gains when made in past 35 years. Futher improvements on broiler will depend on the evolution and revolution: 1. Evolution: (1) Growth rate has been made 4-5% per year. (2) Feed conversion has improved approximately 1% per year. (3) Abdominal fat is becoming a major complaint in broiler. (4) Because of the changing life-style, broiler meat sales in the future will be more and more in cut-up form. (5) Breeding for stress resistance and selection for docile temperament can be important in order to funker improve fled efficiency. (6) In female parent stock, reproduction characteristics are in many can negatively correlated with the desired broiler traits. (7) Egg production and hatchability in moot commercial parent nod m at a fairly high level. (8) In male parent stock, the heavier and mon super-meat-type male lines are desired to Product better broilers. 2. Revolution: Trying to forecast revolutionary change in broiler genetics is highly speculative, as sudden change are aften unpredictable. (1) Species hybridization, such as a turkey-chicken cross (2) Biochemical tools, such as blood typing. (3) Mutation breeding by radiation or chemical mutagentia. (4) Broiler breeding would be to change the phenotypic appearance by single gene, such as naked, wingless. (5) Changes in production techniques. such as growing in cage or growing in filtered air positive pressure houses.

• The Korean Journal of Food And Nutrition
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• v.14 no.6
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• pp.512-520
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• 2001

Kimchi is one of the traditional Korean food and a very popular side dish in Korea. To obtain funda mental data on how to prevent over ripening in kimchi after acidity of 0.4% was reached during the lactate fermentation, the physicochemical characteristics such as pH. acidity. organic acids, enzyme activity were measured and the time dependent ecology of microorganism were observed. In the initial stages of fermentation, the pH of kimchi was markedly changed and slowly decreased in 0.5% acidity The acidity was slowly increased and markedly increased in pH 4 by growth of microorganism. HPLC analysis showed oxalic acid, lactic acid, acetic acid, malic acid and succinic acid and this results reconfirmed by GC-MSD. Lactic acid was changed a lot during fermentation period as the time of storage went on, where as malic was decreased. Kimchi A, having acidity of 0.75%, showed the highest acidic Protease and lipase activity. Also, the amylase activity was high in kimchi C, having 0.95% acidity. The total viable bacteria showed 8.1$\times$10$^{5}$ , 4.7$\times$10$^4$, 1.2$\times$10$^3$, 3.2$\times$10$^4$, 4.9$\times$10$^{5}$ cfu/ml in the kimchi A, B, C, D and E, respectively. The numbers of lactic acid bacteria counted 1.0$\times$10$^{5}$ , 1.3$\times$10s, 1.2$\times$10$^3$, 2.3$\times$ 10$^3$, 2.1$\times$10$^4$c1u/m1 in the kimchi A, B, C, D and E, respectively. The numbers of acetobactor were counted 1.8$\times$10$^{5}$ , 9.3$\times$10$^4$, 7.0$\times$10$^1$, 4.5$\times$10$^4$, 5.3$\times$10$^3$cfu/m1 in the kimchi A, B, C, D and E, respectively.

• Journal of Dairy Science and Biotechnology
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• v.32 no.1
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• pp.17-29
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• 2014

GRAS probiotics can be used to modulate intestinal microbiota and to alleviate various gastrointestinal disorders. In several recent studies, researchers have explored the potential expansion and usability of probiotics to reduce the risk factors associated with diseases, including obesity, hypercholesterolemia, arterial hypertension, hyperhomocysteinemia, and oxidative stress. In this review, our aim was to clarify the mechanism underlying interactions between hosts (animal or human) and probiotics and the beneficial effects of probiotics on human health.