• Journal of Microbiology and Biotechnology
• /
• v.1 no.2
• /
• pp.116-120
• /
• 1991

The transformant Bacillus subtilis ED213 carrying the pSO100 which cloned the cdd gene encoding cytidine deaminase (cytidine /2'-deoxycytidine aminohydrolase, EC 3.5.4.5, CDase) originated from wild type B. subtilis was cultivated in Spizizen minimal medium (SMM). To overcome poor expression of the cdd gene in SMM medium, the medium compositions and growth conditions were optimized. The optimized medium compositions and growth conditions were cytidine concentration of 80 mg/l, glycerol of 25 g/l, and $(NH_4)_2SO_4$ of 10 g/l, along with $37^{\circ}C$ and pH 7.0. The intracellular CDase production was increased 3 times from 1,000 unit/ml to 3,200 unit/ml, and extracellular CDase also increased from nearly undetectable amounts to 1,500 unit/ml. The cytidine concentration was signified as the most critical compositional factor for overproduction of CDase by increasing the cell density mainly in culture broth. The plasmids were more stable in cells that were grown in original SMM medium with stability of 90% compared to those grown in optimized SMM medium with stability of 80% after 48 hours cultivation. The most active amplification of plasmid was occurred in the logarithmic phase, which showed a value around four times higher than the initial copy number. In the exponential phase, the CDase production was closely related to the plasmid copy number along with the cell density. However, it was not accorded with cell density at the stationary phase.

• Journal of Mushroom
• /
• v.9 no.2
• /
• pp.87-89
• /
• 2011

This study was performed to investigate the antibacterial activities and the minimal inhibitory concentration (MIC) of 6 strains of Beauveria bassiana against mulberry pathogenic bacteria. The antibacterial activities and the MIC were measured using paper disc method and broth dilution method, respectively. The antibacterial activities were found out just B. bassiana J200, and shown at 13 mm from Erwinia rhapontici KACC 10407 and at 17 mm from Pseudomonas syringae KACC 10390 and Xanthomonas campestris KACC 12134. The MIC were all observed at 4.0% from E. rhapontici KACC 10407, P. syringae KACC 10390 and X. campestris KACC 12134. The results suggest that B. bassiana could play a good role for biological control against mulberry pathogenic bacteria.

• Food Science of Animal Resources
• /
• v.28 no.1
• /
• pp.99-104
• /
• 2008

A lactic acid bacterium showing growth inhibitory activity against Enterobacter sakazakii was isolated from bovine intestinal tracts. From biochemical and molecular biological studies, the isolate was identified and named as Enterococcus faecium JH95. This strain was resistant to kanamycin and streptomycin at a concentration of $100{\mu}g/mL$. E. faecium JH95 had high antimicrobial activity against food-borne pathogens such as Escherichia coli O157:H7, Listeria monocytogenes, Salmonella typhimurium, Staphylococcus aureus, and Clostridium perfrigens. The culture supernatant of this strain did not have antimicrobial activity. The culture broth of this strain failed to show the antimicrobial activity by heat treatment at $100^{\circ}C$ for 5 min or by pretense treatments for 2 hr. This result suggested that the putative antimicrobial substance produced by E. faecium JH95 is likely a protein which is not secreted into culture medium.

• Korean Journal of Microbiology
• /
• v.38 no.1
• /
• pp.50-53
• /
• 2002

The optimum temperature and pH for mycelial growth of B. bassiana DGUM 34001 were $24^{\circ}C$ and pH 7.0, respectively. Among the complex media used, mushroom complex medium (MCM) was the most favorable for mycelial growth. When Czapek-Dox medium was used as a minimal medium, glucose was an excellent source for carbon and energy. Soytone and sodium phosphate were favorable constituent for culture medium as a source of organic nitrogen and phosphorus, respectively. When the fungus was grown in MCM broth, the specific activity of extracellular enzyme of ${\alpha}$-amylase, lipase, chitinase, CMCase and pretease were 297.0, 0.058, 0.33, 0.21 and 22.8 units/mg protein, respectively. When various sources of organic nitrogen and chitin were supplemented to determine the production of enzymes, casein and soluble chitosan enhanced the production of extracellular protease and chitinase.

• Journal of Korean Medicine Rehabilitation
• /
• v.30 no.1
• /
• pp.31-45
• /
• 2020

Objectives In this study, we investigated the antimicrobial activity of a 70% ethanol extract of Maneung-hwan (MEH), which is prescribed by practitioners of oriental medicine for use against methicillin-resistant Staphylococcus aureus (MRSA). Methods The antibacterial activity of MEH against MRSA strains was evaluated using the disc diffusion method, broth microdilution method (minimal inhibitory concentration, MIC), checkerboard dilution test, and time-kill test. The mechanism of action of MEH was investigated by bacteriolysis using detergents or ATPase inhibitors Additionally, mRNA and protein expression were investigated by quantitative reverse transcription-polymerase chain reaction and western blot assay, respectively. Results The MIC of MEH was 25~1,600 ㎍/mL against all the tested bacterial strains. We showed that MEH extract exerts strong antibacterial activity. In the checkerboard dilution test, the fractional inhibitory concentration index of MEH in combination with antibiotics indicated synergism or partial synergism against S. aureus. The time-kill study indicated that the growth of the tested bacteria was considerably inhibited after a 24-h treatment with MEH and selected antibiotics. To measure the cell membrane permeability, MEH (3.9 ㎍/mL) was combined with Triton X-100 (TX) at various concentrations N,N-dicyclohexylcarbodimide (DCCD) was also tested as an ATPase inhibitor. TX and DCCD cooperation against S. aureus exhibited synergistic action. Accordingly, the antimicrobial activity of MEH in the context of cell membrane rupture and ATPase inhibition was assessed. Additionally, the expression of genes and proteins associated with resistance was reduced after exposing MRSA to MEH. Conclusions These results suggest that MEH possesses antibacterial activity and acts as a potential natural antibiotic against MRSA.

• The Korean Journal of Mycology
• /
• v.24 no.3 s.78
• /
• pp.206-213
• /
• 1996

Talaromyces luteus 2004, a thermophilic mutant of T. luteus 6112 was obtained by mutagenesis with N-methyl-N'-nitro-N-nitrosoguanidine. T. luteus 2004 produced thermophilic carboxymethylcellulase (CMCase), and other polysaccharide enzymes: avicellase, xylanase, and ${\beta}-glucosidase$. Induction of CMCase production was shown at the highest level in 3% carboxymethylcellulose (CMC) minimal broth, indicating that CMC could work as an inducer. However, glucose and D-cellobiose showed catabolite repression for CMCase production which was under the control of CMC utilization. Optimal conditions for CMCase activity were at $70^{\circ}C$ and pH 4.0, suggesting that CMCase of T. luteus 2004 was a thermophilic enzyme.

• The Korean Journal of Mycology
• /
• v.26 no.1 s.84
• /
• pp.127-133
• /
• 1998

Two regulatory mutants of Trichoderma reesei QM 9414 were isolated by treatment with N-methyl-N'-nitro-N-nitrosoguanidine, and the effects of various inducers on the carboxymethyIcellulose (CMC) and filter paper (FP) production were investigated. Induction of CMCase and FPase production of mutants was shown higher level than wild type strain in 1% lactose minimal broth. When induced by glucose, wild type showed glucose-repression for CMCase and FPase production and mutants showed glucose-derepression. Mutant 1 showed 8.38 fold higher CMCase activity and 5.68 fold higher FPase activity than wild type stain. Mutant 2 showed about 8.42 fold higher CMCase activity and 5.41 fold higher FPase activity than wild type strain. Enzyme activities from the mutants and wild type had the same optimum pH of 4.8 and optimum temperature of $60^{\circ}C$.

• Journal of the Korean Society of Food Science and Nutrition
• /
• v.24 no.2
• /
• pp.293-298
• /
• 1995

Antimicrobial effect of tea extracts from green tea(steamed, roasted), oolong tea and black tea was investigated. Minimal inhibitory concentration(MIC) of tea extraxcts against 9 well known strains of foodborne pathogenic bacteria such as the Gram positive and Gram negative bacteria was determined at 37$^{\circ}C$. Significant antimicrobial activity was observed in the steamed green tea and the roasted green tea of the water-soluble fraciton, and the steamed green tea of the methanol-soluble fraction, and the steamed green tea, roasted green tea and the oolong tea of the crude catechin fraction. The MIC of these extracts against B. subtillis were 700$\mu\textrm{g}$/ml, 500$\mu\textrm{g}$/ml and 120 $\mu\textrm{g}$/ml, respectively. The crude catechin fraction possessed greater antimicrobial activity than did the other fractions. Among tea extracts, extracts of steamed green tea, roasted green tea and oolong tea showed higher antimicrobial activity than them of black tea. The MIC of the crude catechin fraction obtained from tea extracts against Gram-positive bacteria such as M. Iuteus, B. subtillis and S. mutans were 30~50$\mu\textrm{g}$/ml, 120~240$\mu\textrm{g}$/ml and 120~180$\mu\textrm{g}$/ml, and against Gram-negative bacteri such as e. aerogenes and V. parahaemolyticus were 50~60$\mu\textrm{g}$/ml and 60~70$\mu\textrm{g}$/ml in the broth medium, respectively. Especially, the MIC to Streptococcus mutans which has known as a causative bacteria of a decayed tooth were 120$\mu\textrm{g}$/ml, 140$\mu\textrm{g}$/ml, 180$\mu\textrm{g}$/ml and above 1,000$\mu\textrm{g}$/ml in steamed green tea, roasted green tea, oolong tea and black tea, respectively. Tea extracts had strong growth inhibition activity against foodborne pathogenic and dental bacteria.

• Journal of Korean Medicine Rehabilitation
• /
• v.28 no.2
• /
• pp.21-35
• /
• 2018

Objectives The objective of this study is to determine the antimicrobial effect of Daehwanggyeonu-san(Dahwangqianniu-san,DGE) and synergistic effects with antibiotics oxacillin, ampicillin, and gentamicin against Methicillin-Resistant Staphylococcus aureus(MRSA). Methods The antibacterial activity of DGE extract was evaluated againest MRSA strains by using the Disc diffusion method, broth microdilution method(minimal inhibitory concentration; MIC), checkerboard dilution test. The checkerboard dilution test was used to examined synergetic effect of oxacillin, ampicillin, gentamicin, ciprofloxacin with DGE extract. Results DGE showed antimicrobial activity against MRSA with an MIC value of $125{\sim}250{\mu}g/mL$. In the checkerboard test, the interation of DGE with all tested antibiotics produced almost synergy or partial synergy against MRSA. Conclusions This study shows that DGE reduced the MICs of several antibiotics tested, and a remarkable antibacterial effect of DGE, with membrane permeability enhancers and ATP synthase inhibitors. This study can be a valuable source for the development of a new drug with low MRSA resistance.

• Korean Journal of Microbiology
• /
• v.42 no.2
• /
• pp.131-134
• /
• 2006

The culture conditions for the enhanced mycelial of Clavicorona pyxidata DGUM 29005 were investigated. The optimal temperature and pH for the mycelial growth were $24^{\circ}C$ and 5.0, respectively. It was shown that trehalose was the best supplement of carbon sources in Czapek-Dox medium as a minimal medium for enhanced mycelial growth. In general, inorganic nitrogen sources were better than organic ones for mycelial growth. Calcium nitrate was the best out of the inorganic nitrogen test. The appropriate phosphorous and vitamin were $Na_2HPO_4$ and p-aminobenzoic acid, respectively. After the mycelial of C. pyxidata DGUM 29005 was cultivated at $24^{\circ}C$ for 20 days in MEM broth(pH 5.0), the specific activities of both exomycelial and endomycelial enzymes were determined. Among the exomycelial enzyme assayed, the specific activity of laccase was much higher than those of other enzymes. However, little or no enzyme activities of ${\alpha}$-amylase, chitinase, lipase and pretense were found.