• Microbiology and Biotechnology Letters
• /
• v.28 no.2
• /
• pp.63-70
• /
• 2000

To determine an increased acid tolerance of an adipic acid-resistant mutant Leuconostoc mesenteroides(ANaM100) developed for use as a Kimchi starter, proton permeability of cytoplasm, activities of H+-ATPase, Mg++ release and fatty acid composition of cytoplasmic membranes of strain ANaM100 were studied and compared with those of its wild type (LMw). The value of protons permeability of LMw after an acid shock at pH 5.0 was 5.4 min., while the value of ANaM100 cells was 8.4 min. at the same pH. The pH of maximal specific activ-ities of ATPase originated from the LMw and ANaM100 were 0.87 unit/mg protein at pH 6.0 and 0.92 unit/mg pro-tein at pH 5.5, respectively. The release of magnesium ion from ANaM100 was observed about 12.8% at pH 4 after 2 hours, while the wild strains of LMw released Mg++ about 27.6% under the same conditions. The content of C19:0,cyclo and C18:1 in a membrane fatty acid of ANaM100 was higher and lower, respectively than that of LMw. These results indicated that acid tolerance of adipic acid-resistant strain, ANaM100 was significantly improved in comparison with that of its wild type, LMw. In addition, the strain ANaM100 was adipic resistance based on the result of growth of the strain in comparison with that of strain LMw in a broth containing adipic acid.

• Journal of Microbiology
• /
• v.34 no.3
• /
• pp.289-293
• /
• 1996

A cadimium-resistant strain isolated from Anyang stream, Azomonas agilis PY101 exhibited strong resistance to 1000 ppm of cadmium ion $(Cd^{2+})$. A agilis PY101 also exhibited resistance to various antibiotics, such as amoxicillin, amplicillin, bacitracin, cefazolin, erythromycin, penicillin, tetracycline, and vancomycin. In the presence of $Cd^{2+}$, the growth of A. agilis PY101 started after an extended lag phase and produced a green-fluorescent pigment induced by cadmium. The dramatic decrease (approximately 400ppm) of concentration of $cd^{2+}$ in the culture medium during the growth phase of A. agilis PY101 was confirmed by the inductively coupled plasma-atomic emission spectrophotometer. Transmission electron microscopic analysis revealed that A. agilis PY 101 actively accumulated $Cd^{2+}$ in the cytoplasm.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.12
• /
• pp.2043-2050
• /
• 2016

Exploring novel antibiotics is necessary for multidrug-resistant pathogenic bacteria. Because the probiotics in soybean food have antimicrobial activities, we investigated their effects on multidrug-resistant Acinetobacter baumannii. Nineteen multidrug-resistant A. baumannii strains were clinically isolated as an experimental group and 11 multidrug-sensitive strains as controls. The growth rates of all bacteria were determined by using the analysis for xCELLigence Real-Time Cell. The combination of antibiotics showed synergistic effects on the strains in the control group but no effect on the strains in the experimental group. Efflux pump gene adeS was absent in all the strains from the control group, whereas it exists in all the strains from the experimental group. Furthermore, all the strains lost multidrug resistance when an adeS inhibitor was used. One strain of probiotics isolated from soybean food showed high antimicrobial activity for multidrug-resistant A. baumannii. The isolated strain belongs to Bacillus subtilis according to 16S RNA analysis. Furthermore, E. coli showed multidrug resistance when it was transformed with the adeS gene from A. baumannii whereas the resistant bacteria could be inhibited completely by isolated Bacillus subtilis. Thus, probiotics from soybean food provide potential antibiotics against multidrug-resistant pathogenic bacteria.

• The Microorganisms and Industry
• /
• v.19 no.2
• /
• pp.34-41
• /
• 1993

Industrial strain improvement is concerned with developing or modifying microorganisms used in production of commercially important fermentation products. The aim is to reduce the production cost by improving productivity of a strain and manipulating specific characteristics such as the ability to utilize cheaper raw materials or resist bacteriophages. The traditional empirical approach to strain improvement is mutation combined with selection and breeding techniques. It is still used by us to improve the productivity of organisms in amino acids, organic acids and enzymes production. The breeding of high L-lysine-producing strain Au112 is one of the outstanding examples of this approach. It is a homoserine auxotroph with AEC, TA double metabolic analogue resistant markers. The yield reaches 100 g/l. Besides, the citric acid-producing organism Aspergillus niger, Co827, its productivity reaches the advanced level in the world, is also the result of a series mutations especially with $^60Co{\gamma}$-radiation. The thermostable .alpha.-amylase producing strain A 4041 is the third example. By combining physical and chemical mutations, the strain A 4041 becomes an asporogenous, catabolite derepressed mutant with rifamycin resistant and methionine, arginine auxotroph markers. The .alpha.-amylase activity reaches 200 units/ml. The fourth successful example of mutation in strain improvement is the glucoamylase-producing strain Aspergillus niger SP56, its enzyme activity is 20,000 units/ml, 4 times of that of the parental strain UV-11. Recently, recombinant DNA approach provides a worthwhile alternative strategy to industrial strain improvement. This technique had been used by us to increase the thermostable .alpha.-amylase production and on some genetic researches.

• Journal of Microbiology and Biotechnology
• /
• v.26 no.3
• /
• pp.452-460
• /
• 2016

Acid stress can affect the viability of probiotics, especially Bifidobacterium. This study aimed to improve the acid tolerance of Bifidobacterium longum BBMN68 using adaptive evolution. The stress response, and genomic differences of the parental strain and the variant strain were compared by acid stress. The highest acid-resistant mutant strain (BBMN68m) was isolated from more than 100 asexual lines, which were adaptive to the acid stress for 10^th, 20^th, 30^th, 40^th, and 50^th repeats, respectively. The variant strain showed a significant increase in acid tolerance under conditions of pH 2.5 for 2 h (from 7.92 to 4.44 log CFU/ml) compared with the wild-type strain (WT, from 7.87 to 0 log CFU/ml). The surface of the variant strain was also smoother. Comparative whole-genome analysis showed that the galactosyl transferase D gene (cpsD, bbmn68_1012), a key gene involved in exopolysaccharide (EPS) synthesis, was altered by two nucleotides in the mutant, causing alteration in amino acids, pI (from 8.94 to 9.19), and predicted protein structure. Meanwhile, cpsD expression and EPS production were also reduced in the variant strain (p < 0.05) compared with WT, and the exogenous WT-EPS in the variant strain reduced its acid-resistant ability. These results suggested EPS was related to acid responses of BBMN68.

• Korean Journal of Microbiology
• /
• v.38 no.4
• /
• pp.306-311
• /
• 2002

PCR of the mecA gene for the rapid detection of methicillin-resistant staphylococci was perfomed and compared with the antibiotic sensitivity test. A total of 43 strains of staphylococi from clinical specimens were used in this study. An antibiotic sensitivity test by the agar dilution method of NCCLS (The National Commitee for Clinical Laboratory Standard) was performed for the strains. Among them, 39 isolates were methicillin-resistant (MRS), and 4 isolates were methicillin-susceptible (MSS). With the exception for one strain (Staphylococcus cohnii, HRC2-4), all MRS strains amplified the expected 533 bp fragments of the mecA gene by PCR, However, one strain (Staphylococcus aureus, HSA1-10) that was classified as a sensitive strain by the antibiotic sensitivity test was mecA positive by PCR. All 35 methicillin-resistant Staphylococcus aureus (MRSA) strains were mecA positive, but overall, concordance between the results of the mecA PCR and antibiotic sensitivity test was 95.6%.

• The Microorganisms and Industry
• /
• v.15 no.1
• /
• pp.38-42
• /
• 1989

Industrial strain Improvement is concerned with developing or modifying microorga-nisms used In production of commercially important fermentation products. The aim is to reduce the production cost by improving productivity of a strain and manipulating specific cilarafteristic such as the ability to utilize cheaper raw materials or resist bacteriophages. The traditional empiri-cal approach to strain improvement is mutation combined with selection and breeding techniques. It is still used by us to improve the productivity of organisms in amino acids. organic acids andenzymes production. The breeding of high L-lysine-producing strain Au112 is one of the outstanding examples of this approach. It is it homoserine auxotroph with AEC, TA double metabolicanalogue resistant markers. The yield reaches 100g/1. Resides, the citric acid-producing organism Aspergillus nuger, Co827, its productivity reches the advanced level in the world, is also the result of a series mutations expecially with Co Y-radiation. The thermostable a-amylaseroducing strain A 4041 is the third example. By combining physical and chemical multations. the strain ,A 4041becomes an asporogenous, catabolite derepressed mutant with rifamycin resistant and methionine, arginine auxotroph markers. The a-amylase activity reaches 200 units/ml. The fourth successful example of mutation in strain improvement is the glucoamylase-producing strain Aspergillus nigerSP56 its enzyme activity is 20,000 units/ml, 4 times of that of the parental strain UV_11. Recently recombinant DNA approach Provides a worth while alternative strategy to Industrial strain improve-ment. This technique had been used by us to increase the thermostable a-amylase production and on some genetic researches.

• Journal of Microbiology and Biotechnology
• /
• v.16 no.10
• /
• pp.1513-1517
• /
• 2006

One single lactic acid producing bacterium, isolated from kimchi, inhibited the growth and adherence of Helicobacter pylori to the human gastric epithelial cell line MKN-45. This isolate was identified as Lactobacillus plantarum and termed L. plantarum strain PL9011. The adherence of H pylori, in the presence of live or nonviable L. plantarum strain PL9011 (10-fold CFU), decreased to 14-20%. The spent culture supernatant of L. plantarum strain PL9011 resulted in the eradication of H pylori. This activity remained stable following neutralization and heat treatment, but not following pepsin treatment, thereby suggesting small peptides as the inhibitory factor. L. plantarum strain PL9011 did not produce any harmful metabolites or enzymes. The results obtained in this study suggest that the L. plantarum strain PL9011 may be a potential novel probiotic for the stomach.

• Archives of Pharmacal Research
• /
• v.18 no.3
• /
• pp.173-178
• /
• 1995

Ciprofloxacin resistance mechanisms were studied by investigating the inhibitory effect of ciprofloxacin on the gyrase-mediated DNA supercoiling and the intracellular accumulation of ciprofloxacin in clinical isolates of Pseudomonas aeruginosa. A higher amount of ciprofloxacin was required to inhibit the gyrases purified from the ciprofloxacin-resistant strains than that from the sensitive strain. Reconstitution of heterologous gyrase subunits from different strains revealed alterations in the A and/or the B subunits of gyrase in these strains. In addition, the resistant strains accumulated approximately a half amount of ciprofloxacin inside the cells, compared to the sensitive strain. However, when the active efflux was blocked by carbonyl cyanide m-chlorophenyl hydrazone treatment, intracellular concentration of ciprofloxacin was elevated about 4-7 fold in these strains, while the sensitive strain was not significantly affected by this treatment, indicating that the ciprofloxacin-resistant strains developed a drug efflux system. Interestingly, these resistant strains expressed an envelope protein of approximately 51 kD. These studies suggest that alterations in the gyrase as well as the active drug-efflux system conferred dual ciprofloxacin resistance mechanisms to these clinical isolates of P. aeruginosa.

• Korean Journal of Microbiology
• /
• v.17 no.2
• /
• pp.49-57
• /
• 1979

Cadimium ion-resistant microorganism was isolated from the sludge of wastewater. The physiological, morphological and other cultural data showed that this strain belonged to Citrobacter freudii. A clearcut distinction of growth among nutrient broth, typtic soy broth and synthetic medium was demonstrated. The resistant cells showed only slight mutagenic action. During the growth of bacterial population in resting state, the organisms reduced the initial level of resistance to cadmium ions when they were not kept in contact with cadmium ions in bacteral multiplication. And cadmium ion-resistant and cadmium ion-sensitive strain were found to show equal, lower or higher sensitivity to other heave metals.