• Journal of Microbiology and Biotechnology
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• v.23 no.11
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• pp.1529-1535
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• 2013

Tuberculosis is a worldwide epidemic disease caused by Mycobacterium tuberculosis, with an estimated one-third of the human population currently affected. Treatment of this disease with aminoglycoside antibiotics has become less effective owing to antibiotic resistance. Recent determination of the crystal structure of the M. tuberculosis Rv3168 protein suggests a structure similar to that of Enterococcus faecalis APH(3')-IIIa, and that this protein may be an aminoglycoside phosphotransferase. To determine whether Rv3168 confers antibiotic resistance against kanamycin, we performed dose-response antibiotic resistance experiments using kanamycin. Expression of the Rv3168 protein in Escherichia coli conferred antibiotic resistance against $100{\mu}M$ kanamycin, a concentration that effected cell growth arrest in the parental E. coli strain and an E. coli strain expressing the $Rv3168^{D249A}$ mutant, in which the catalytic Asp249 residue was mutated to alanine. Furthermore, we detected phosphotransferase activity of Rv3168 against kanamycin as a substrate. Moreover, docking simulation of kanamycin into the Rv3168 structure suggests that kanamycin fits well into the substrate binding pocket of the protein, and that the phosphorylation-hydroxyl-group of kanamycin was located at a position similar to that in E. faecalis APH(3')-IIIa. On the basis of these results, we suggest that the Rv3168 mediates kanamycin resistance in M. tuberculosis, likely through phosphotransferase targeting of kanamycin.

• Korean Journal of Microbiology
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• v.50 no.3
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• pp.239-244
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• 2014

ErmSF is one of the four antibiotic resistance factor proteins expressed by Streptomyces fradiae, antibiotic tylosin producer, which renders $MLS_B$ (macrolide-lincosamide-streptogramin B) antibiotic resistance through dimethylating A2058 of 23S rRNA, thereby reducing the affinity of antibiotic to ribosome. Unlike other Erm proteins, ErmSF harbors long N-terminal end region. To investigate its role in enzyme activity, mutant ErmSF deleted of 1-38 amino acids was overexpressed and activity in vivo and in vitro was observed. In vitro enzymatic assay showed that mutant protein exhibited reduced activity by 20% compared to the wild type enzyme. Due to the reduced activity of the mutant protein, cells expressing mutant protein showed weaker resistance to erythromycin than cells with wild type enzyme. Presumably, the decrease in enzyme activity was caused by the hindrance in substrate binding and (or) product release, not by defect in the methyl group transfer occurred in active site.

• The Korean Journal of Pain
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• v.36 no.1
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• pp.11-50
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• 2023

As the field of interventional pain management (IPM) grows, the risk of surgical site infections (SSIs) is increasing. SSI is defined as an infection of the incision or organ/space that occurs within one month after operation or three months after implantation. It is also common to find patients with suspected infection in an outpatient clinic. The most frequent IPM procedures are performed in the spine. Even though primary pyogenic spondylodiscitis via hematogenous spread is the most common type among spinal infections, secondary spinal infections from direct inoculation should be monitored after IPM procedures. Various preventive guidelines for SSI have been published. Cefazolin, followed by vancomycin, is the most commonly used surgical antibiotic prophylaxis in IPM. Diagnosis of SSI is confirmed by purulent discharge, isolation of causative organisms, pain/tenderness, swelling, redness, or heat, or diagnosis by a surgeon or attending physician. Inflammatory markers include traditional (C-reactive protein, erythrocyte sedimentation rate, and white blood cell count) and novel (procalcitonin, serum amyloid A, and presepsin) markers. Empirical antibiotic therapy is defined as the initial administration of antibiotics within at least 24 hours prior to the results of blood culture and antibiotic susceptibility testing. Definitive antibiotic therapy is initiated based on the above culture and testing. Combination antibiotic therapy for multidrug-resistant Gram-negative bacteria infections appears to be superior to monotherapy in mortality with the risk of increasing antibiotic resistance rates. The never-ending war between bacterial resistance and new antibiotics is continuing. This article reviews prevention, diagnosis, and treatment of infection in pain medicine.

• Korean Journal of Microbiology
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• v.42 no.3
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• pp.165-171
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• 2006

ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA to confer the resistance to MLS (macrolide-lincosamide-streptogramin B) antibiotics on microorganism ranging from antibiotic producers to pathogens. To define the functional role of peptide segment encompassing amino acid residues 1 to 25 in NTER (N-terminal end region) of ErmSF, one of the ERM proteins, DNA fragment encoding mutant protein deprived of that peptide was cloned and overexpressed in E. coli to obtain a purified soluble form protein to the apparent homogeneity in the yield of 12.65 mg per liter of culture. The in vitro activity of mutant protein was found to be 85% compared to wild type ErmSF, suggesting that this peptide interact with substrate to affect the enzyme activity. This diminished activity of mutant protein caused the delayed expression of antibiotic resistance in vivo, that at fIrst cells expressing mutant protein showed the retarded growth due to the antibiotic action but with time cells inhibited by antibiotic gradually recovered the viability to exert the resistance to the same extent as those with wild type protein.

• Applied Biological Chemistry
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• v.31 no.4
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• pp.371-375
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• 1988

Streptomycetes are attractive microorganisms for their production of various secondary metabolites such as antibiotics. Now, the development of gene manipulation in this microorganisms enables the cloning and analysis of the genes which coding for antibiotic biosynthesis and resistance to the drug. In this article, we reviewed the studies with respect to the mechanisms of self-protection and cloning of the genes cloning for antibiotic biosynthesis, particularly, in microorganisms which produce antibiotic inhibitors of protein synthesis.

• Proceedings of the Korea Society of Environmental Toocicology Conference
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• 2001.05a
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• pp.140-140
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• 2001

Porins are major outer membrane proteins which produce non-specific aqueous channels across the membrane that permit the diffusion into the bacterial cells of hydrophilic compounds including sugars, amino acids, and antibiotics. In some gram-negative organisms, antibiotic resistance can be induced by mutational loss of channel that causes a decrease in outer membrane permeability. (omitted)

• Journal of Korean Biological Nursing Science
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• v.8 no.1
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• pp.5-14
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• 2006

Staphyloccus aureus is one of the most important pathogens in clinical settings. It is also one of the leading causes of nosocomial infections and the dissemination of multiple drug-resistant strains, mainly methicillin resistant Staphyloccus aureus, and the recent emergence of a vancomycin resistant MRSA is the concern to hospital worldwide. MRSA strains have acquired multiple resistance to a wide range of antibiotics, including aminoglycosides and macrolides. $\beta$-Lactam resistance of methicillin-resistnat Staphyococcus aureus is determined by the function of penicillin binding protein 2'(PBP2') encoded by the methicillin resistance gene mec A. MRSA strains carry methicillin resistance gene mecA, encoded by a mobile genetic element designated staphylococoal cassette chromosome mec(SCCmec). MRSA clones are defined by the type of SCCmec element and the genotype of the methicilline-susceptible Staphyococcus aureus chromosome in which the SCCmec element is integrated.

• Korean Journal of Soil Science and Fertilizer
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• v.29 no.1
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• pp.60-66
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• 1996

Bradyrhizobium japonicum with different colony morphology populated in five Yeongnam soils of Korea was examined for intrinsic antibiotic resistance to eight antibiotics, serological property by immunoblot and immunodiffusion, and protein profile differentiation by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Colony morphological distribution of one hundred and twenty B. japonicum isolates was 47% for "dry". 41% for "wet", and 12% for "dry/wet" type. The total isolates showed such a strong correlation between the morphology and antibiotic resistance. Colony morphology, which though was dominantly consisted of the same type within a serogroup, wasn't absolutely linked to serological property of B. japonicum. Based on these data, colony morphology was too simple to identify variations with B. japonicum isolates : antibiotic resistance such complicated compared with serological analyses.

• Journal of Microbiology and Biotechnology
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• v.26 no.2
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• pp.263-269
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• 2016

Temperate phages have been suggested to carry virulence factors and other lysogenic conversion genes that play important roles in pathogenicity. In this study, phage TEM123 in wild-type Staphylococcus aureus from food sources was analyzed with respect to its morphology, genome sequence, and antibiotic resistance conversion ability. Phage TEM123 from a mitomycin C-induced lysate of S. aureus was isolated from foods. Morphological analysis under a transmission electron microscope revealed that it belonged to the family Siphoviridae. The genome of phage TEM123 consisted of a double-stranded DNA of 43,786 bp with a G+C content of 34.06%. A bioinformatics analysis of the phage genome identified 43 putative open reading frames (ORFs). ORF1 encoded a protein that was nearly identical to the metallo-β-lactamase enzymes that degrade β-lactam antibiotics. After transduction to S. aureus with phage TEM123, the metallo-β-lactamase gene was confirmed in the transductant by PCR and sequencing analyses. In a β-lactam antibiotic susceptibility test, the transductant was more highly resistant to β-lactam antibiotics than S. aureus S133. Phage TEM123 might play a role in the transfer of β-lactam antibiotic resistance determinants in S. aureus. Therefore, we suggest that the prophage of S. aureus with its exotoxin is a risk factor for food safety in the food chain through lateral gene transfer.

• Korean Journal of Microbiology
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• v.43 no.3
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• pp.166-172
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• 2007

ERM proteins transfer the methyl group to $A_{2058}$ in 23S rRNA, which reduces the affinity of MLS (macrolide-lincosamide-streptogramin B) antibiotics to 23S rRNA, thereby confer the antibiotic resistance on micro-organisms ranging from antibiotic producers to pathogens and are classified into monomethyltransferase and dimethyltransferase. To investigate the differences between mono- and dimethyltransferase, tirD, a representative monomethylase gene was cloned in Escherichia coli from Streptomyces fradiae which contains ermSF, dimethylase gene as well to overexpress the TlrD for the first time. T7 promoter driven expression system successfully overexpress tlrD as a insoluble aggregate at $37^{\circ}C$ accumulating to around 55% of the total cell protein but unlike ErmSF, culturing at temperature as low as $18^{\circ}C$ did not make insoluble aggregate of protein into soluble protein. Coexpression of Thioredoxin and GroESL, chaperone was not helpful in turning into soluble protein either as in case of ErmSF. These results might suggest that differences between mono- and dimethylase could be investigated on the basis of the characteristics of protein structure. However, a very small amount of soluble protein which could not be detected by SDS-PAGE conferred antibiotic resistance on E. coli as in ErmSF which was expected from the activity exerted by monmethylase in a cell.