• Journal of Microbiology and Biotechnology
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• 제9권5호
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• pp.646-649
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• 1999

A thermophilic bacterium producing D-stereospecific dipeptidase was isolated from Korean soil samples. The enzyme hydrolyzed the peptide bond between D-alanyl-D-alanine (D-Ala-D-Ala). The isolated bacterial strain was rod shaped, gram-positive, motile, and formed an endospore. Morphological and physiological characteristics suggested this microorganism a thermophilic Bacillus species, and was named as Bacillus sp. BCS-l. The production of D-stereospecific dipeptidase was growth-associated and optimal at $55^{\circ}C$. The enzyme was applied for the synthesis of D-amino acid-containing peptide, N-benzyloxycarbonyl-L-aspartyl-D-alanine benzyl ester (Z-L-Asp-D-AlaOBzl), as a model reaction. A thermodynamically controlled synthesis of Z-L-Asp-D-AlaOBzl was achieved in an organic solvent.

• 대한화학회지
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• 제57권6호
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• pp.731-737
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• 2013

A convenient synthesis of bifunctional sulfonamide-amide derivatives was reported. Amide coupling of 4-methyl benzoic acid 1 followed by reaction with chlorosulfonic acid produce ethyl-4-(3-(chlorosulfonyl)-4-methylbenzoyl)piperazine-1-carboxylate 4. The resulted compound on further treatment with various anilines produces the title sulfonamide-amide derivatives 5a-n. The configurations of these compounds were established by elemental analysis, IR, $^1H$ NMR, mass spectra, and by their preparation from the corresponding 4-methyl benzoic acid 1 and chlorosulfonic acid. All these new compounds demonstrate significant in vitro antibacterial and antifungal activities against all bacterial and fungal strains.

• 약학회지
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• 제47권5호
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• pp.331-338
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• 2003

A butyrolactone ester of cefazolin (CFZ-BTL) was synthesized by the esterification of cefazolin (CFZ) with $\alpha$-bromo-${\gamma}$-butyrolactone. The synthesis was confirmed by the spectroscopic analysis. The CFZ-BTL was more lipophilic than the CFZ when assessed by n-octanol/water partition coefficients at various pH. The CFZ-BTL itself did not show any antimicrobial activity in vitro, but after oral administration of CFZ-BTL to rabbits, exerted significant anti-microbial activity in serum samples when measured by the inhibion zone method in nutrient agar plates, due to conversion of CFZ-BTL to an active metabolite, probably CFZ, in the body. The CFZ-BTL was also converted into CFZ as confirmed by in vitro incubation study, with tissue homogenates (liver, blood and intestine) of rabbits. The liver showed the fastest conversion rate, probably via the hydrolysis mechanism. In vivo metabolism of CFZ-BTL to CFZ was also confirmed in vivo serum samples by HPLC. The oral bioavailability of CFZ-BTL in rabbits was 1.6-fold increased when compared to CFZ, resulting from followed by enhanced lipophilicity increased passive absorption in the intestine.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.163.4-164
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• 2003

Peptide deformylase (PDF) is essential and unique to bacteria for cytoplasmic protein synthesis, but not required in eukaryotes, thus making it an attractive target for the discovery of novel antibacterial drugs. Protein synthesis in eubacteria, under normal conditions, is initiated by formyl-methionyl-tRNA. PDF removes the formyl-group of N- formylmethionine of newly synthesized polypeptides to produce a mature protein. In this study, a pdf gene from Staphylococcus aureus 6538p was cloned in pET-14b vector and transformed in Escherichia coli BL21 (DE3). (omitted)

• Asian-Australasian Journal of Animal Sciences
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• 제14권6호
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• pp.885-893
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• 2001

This review summarizes some recent research into ways of improving the productivity of ruminal fermentation by increasing protein flow from the rumen and decreasing the breakdown of protein that results from the action of ruminal microorganisms. Proteinases derived from the plant seem to be of importance to the overall process of proteolysis in grazing animals. Thus, altering the expression of proteinases in grasses may be a way of improving their nutritive value for ruminants. Inhibiting rumen microbial activity in ammonia formation remains an important objective: new ways of inhibiting peptide and amino acid breakdown are described. Rumen protozoa cause much of the bacterial protein turnover which occurs in the rumen. The major impact of defaunation on N recycling in the sheep rumen is described. Alternatively, if the efficiency of microbial protein synthesis can be increased by judicious addition of certain individual amino acids, protein flow from ruminal fermentation may be increased. Proline may be a key amino acid for non-cellulolytic bacteria, while phenylalanine is important for cellulolytic species. Inhibiting rumen wall tissue breakdown appears to be an important mechanism by which the antibiotic, flavomycin, improves N retention in ruminants. A role for Fusobacterium necrophorum seems likely, and alternative methods for its regulation are required, since growth-promoting antibiotics will soon be banned in many countries.

• Journal of Microbiology and Biotechnology
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• 제17권2호
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• pp.226-233
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• 2007

Members of Methylobacterium, referred as pink-pigmented facultative methylotrophic bacteria, are frequently associated with terrestrial and aquatic plants, tending to form aggregates on the phyllosphere. We report here that the production of autoinducer molecules involved in the cell-to-cell signaling process, which is known as quorum sensing, is common among Methylobacterium species. Several strains of Methylobacterium were tested for their ability to produce N-acyl-homoserine lactone (AHL) signal molecules using different indicators. Most strains of Methylobacterium tested could elicit a positive response in Agrobacterium tumefaciens harboring lacZ fused to a gene that is regulated by autoinduction. The synthesis of these compounds was cell-density dependent, and the maximal activity was reached during the late exponential to stationary phases. The bacterial extracts were separated by thin-layer chromatography and bioassayed with A. tumefaciens NTI (traR, tra::lacZ749). They revealed the production of various patterns of the signal molecules, which are strain dependent. At least two signal molecules could be detected in most of the strains tested, and comparison of their relative mobilities suggested that they are homologs of N-octanoyl-$_{DL}$-homoserine lactone ($C_8-HSL$) and N-decanoyl-$_{DL}$-homoserine lactone ($C_{10}-HSL$).

• Bulletin of the Korean Chemical Society
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• 제32권5호
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• pp.1645-1649
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• 2011

[ ${\beta}$ ]Ketoacyl acyl carrier protein synthase I (KAS I) is involved in the elongation of unsaturated fatty acids in bacterial fatty acid synthesis and a therapeutic target of designing novel antibiotics. In this study, we performed receptor-oriented pharmacophore-based in silico screening of E. coli KAS I (ecKAS I) with the aim of identifying novel inhibitors. We determined one pharmacophore map and selected 8 compounds as candidates ecKAS I inhibitors. We discovered one antimicrobial compound, YKAe1008, N-(3-pyridinyl) hexanamide, displaying minimal inhibitory concentration (MIC) values in the range of 128-256 ${\mu}g/mL$ against MRSA and VREF. YKAe1008 was subsequently assessed for binding to ecKAS I using saturation-transfer difference NMR spectroscopy. Further optimization of this compound will be carried out to improve its antimicrobial activity and membrane permeability against bacterial cell membrane.

• Asian-Australasian Journal of Animal Sciences
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• 제11권6호
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• pp.661-672
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• 1998

Two experiments were conducted to examine the effects of a range of concentrations of ruminal fluid ammonia ($NH_3$-N) on forage digestibility, microbial growth efficiency and the mix of microbial species. Urea was either continuously infused directly into the rumen of sheep fed 33.3 glh of oaten chaff (Exp. I) or sprayed onto the oaten chaff (750 g/d) given once daily (Exp. 2). Concentrations of $NH_3$-N increased with incremental addition of urea (p < 0.01). Volatile fatty acids (VFA) concentrations and 24 h in sacco organic matter digestibility in the rumen were higher when supplemental urea was given (p < 0.01). The (C2 + C4) : C3 VFA ratio was lower (p < 0.05) when $NH_3$-N was above 200 mgN/I. The fungal sporangia appearing on oat leaf blades were significantly higher when urea was supplemented, indicating that $NH_3$-N was a growthlimiting nutrient for fungi at levels of $NH_3$-N below 30 mgN/l. The density of protozoa was highest when $NH_3$-N concentrations were adjusted to 30 mgN/I for continuously fed ($4.4{\times}10^5/ml$) and to 168 mgN/1 for once daily feeding ($2.9{\times}10^5/ml$). Thereafter increasing concentrations of $NH_3$-N, were associated with a concomitant decline in protozoal densities. At the concentration of $NH_3$-N above 200 mgN/l, the density of protozoa was similar to the density of protozoa in ruminal fluid of the control sheep ($1.8{\times}10^5/ml$). The efficiency of net microbial protein synthesis in the rumen calculated from purine excretion was 17-47% higher when the level of $NH_3$-N was above 200 mgN/1. The possibilities are that 1) there is less bacterial cell lysis in the rumen because of the concomitant decrease in the protozoal pool and/or 2) microbial growth per se in the rumen is more efficient with increasing $NH_3$-N concentrations.

• Asian-Australasian Journal of Animal Sciences
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• 제20권1호
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• pp.75-81
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• 2007

The object of this study was to determine the influence of supplementation of concentrate containing high levels of cassava chip on rumen ecology, microbial protein and digestibility of nutrients. Four, rumen fistulated crossbred beef steers with initial body weight of 400${\pm}$10 kg were randomly assigned according to a 4${\times}$4 Latin square design. The dietary treatments were concentrate cassava chip based offering at 0, 1, 2 and 3% BW with urea-treated rice straw fed ad libitum. It was found that ruminal pH was significantly decreased with increase of concentrate. Volatile fatty acids (VFA) concentration in the rumen was significantly different among treatments. In addition, a molar proportion of propionate was higher in supplemented groups at 2 and 3% BW (p<0.05), leading to significantly decreased acetate:propionate ratio. Furthermore, microbial N supply was significantly improved and was highest at 2% BW supplementation. The efficiency of rumen microbial-N synthesis based on organic matter (OM) truly digested in the rumen was highest in level of concentrate supplementation at 2% BW (80% of cassava chip in diets). Moreover, bacterial populations such as amylolytic bacteria was linearly increased, while cellulolytic bacteria was linearly decreased (p<0.01) when cattle received concentrate supplementation in all levels. The total protozoal counts were significantly increased, while fungal zoospores were dramatically decreased in cattle receiving increased levels of concentrate. In conclusion, cassava chip can be use as energy source at 80% in concentrate and supplementation of concentrate at 2% BW with urea-treated rice straw as roughage could improve rumen fermentation efficiency in beef cattle.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 1998년도 Proceedings of UNESCO-internetwork Cooperative Regional Seminar and Workshop on Bioassay Guided Isolation of Bioactive Substances from Natural Products and Microbial Products
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• pp.158-158
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• 1998

The non-mevalonate pathway is a newly discovered isoprenoid biosynthetic pathway in some bacteria, cyanobacteria, algae and plants. Because isoprenoid metabolites (ubiquinone, menaquinone, undecaprenol) are essential for bacterial growth, this pathway may represent a novel target for antibacterial agents. Antibiotics with a unique mechanism of action are needed to combat the risk of antibiotic resistance that is a current worldwide problem. In order to study this pathway as viable target, it was necessary to verify use of the pathway in our model system, the bacterium Bacillus subtilis. Incubation experiments with [6,6-$^2$H$_2$]-D-glucose and [l-$^2$H$_3$]-deoxy-D-xylulose were conducted to provide labeled menaquinone-7 (MK -7), the most abundant isoprenoid in B. subtilis. $^2$H-NMR analysis of the MK-7 revealed labeling patterns that strongly support utilization of the non-mevalonate pathway. Another approach to study the pathway is by structure activity relationships of proposed inhibitors of the pathway. Fosmidomycin is a phosphonic acid with antibacterial activity known to inhibit isoprenoid biosynthesis in susceptible bacteria and may act by inhibiting the non-mevalonate pathway. Fosmidomycin and an N-methyl analog were synthesized and tested for antibacterial activity. Fosmidomycin was active against Escherichia coli and B. subtilis, while N-formyl-N-methyl-3-amino-propylphosphonic acid was inactive.