• Microbiology and Biotechnology Letters
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• v.26 no.2
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• pp.151-160
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• 1998

The Leuconostoc paramesenteroides dominated at refrigeration temperature range was isolated from kimchi, and improved its growth properties by mutation for competitive growth against Lactobacillus plantarum at lower pH. It was found that the minimal pH for the wild type Leuconostoc paramesenteroides Pw growth was pH 4.5 adjusted with HCI and pH 5.0 adjusted with organic-mixture (lactic acid:acetic acid=1:2), respectively. The mutant P-100 could grow in pH 4.0, 4.5, respectively, in MRS broth. Two strains Pw and P-100 were added into kimchi as starter and compared the quality characteristics of kimchi. The total acceptability of Pw and P-100 inoculated kimchi were evaluated better than that of control kimchi (no starter added) by sensory test and extended the optimal pH range of kimchi up to about 2.2, 2.5 times, respectively. In kimchi added P-100, the succinic acid was more abundant than others and the total number of Lactobacillus plantarum was down about 2.5 times in contrast to control kimchi.

• Journal of Microbiology and Biotechnology
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• v.10 no.6
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• pp.789-796
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• 2000

In order to analyze the effects of tktA, $aroF^{FBR}$, and aroL expression in a tryptophan-producing Escherichia coli, a series of plasmids carrying the genes were constructed. Introduction of tktA, $aroF^{FBR}$, and aroL into the E. coli strain resulted in approximately 10-20 fold increase in the activities of transketolase, the feedback inhibition-resistant 3-deoxy-D-arabinoheptulsonate-7-phosphate synthase, and shikimate kinase. Expression of $aroF^{FBR}$ in the aroB mutant strain of E. coli resulted in the accumulation of 10 mM of 3-deoxy-D-arabinoheptulsonate-7-phosphate (DAHP) in the medium. Simultaneous expression of tktA and $aroF^{FBR}$ in the strain further increased the amount of excreted DAHP to 20 mM. In contrast, the mutant strain which has no gene introduced accumulated 0.5 mM of DAHP. However, the expression of tktA and $aroF^{FBR}$ in a tryptophan-producing E. coli strain did not lead to the increased production of tryptophan, but instead, a significant amount of shikimate, which is an intermediate in the tryptophan biosynthetic pathway, was excreted to the growth medium. Despite the fact that additional expression of shikimate kinase in the strain could possibly remove 90% of excreted shikimate to 0.1 mM, the amount of tryptophan produced was still unchanged. Removing shikimate using a cloned aroL gene caused the excretion of glutamate, which suggests disturbed central carbon metabolism. However, when cultivated in a complex medium, the strain expressing tktA, $aroF^{FBR}$, and aroL produced more tryptophan than the parental strain. These data indicate that additional rate-limiting steps are present in the tryptophan biosynthetic pathway, and the carbon flow to the terminal pathway is strictly regulated. Expressing tktA in E. coli cells appeared to impose a great metabolic burden to the cells as evidenced by retarded cell growth in the defined medium. Recombinant E. coli strains harboring plasmids which carry the tktA gene showed a tendency to segregate their plasmids almost completely within 24h.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04a
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• pp.69-75
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• 2002

Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd/dwf3 were shown to be blocked in $D^4$ reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bri1/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus It is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRI1 could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

• Korean Journal of Microbiology
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• v.41 no.3
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• pp.177-182
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• 2005

H. pylori strains were isolated from antral biopsies taken during upper endoscopy in 114 dyspeptic patients with no previous therapy against H. pylori. Rapid urease test, PCR amplification of SSA and cagA gene for H. pylori detection, and Western blot for CagA expression detection were performed. H. pylori infected patients were treated with omeprazole, clarithromycin (a macrolide), and amoxicillin. At 6 weeks after the discontinuation of therapy, the bacterial eradication rate was determined by endoscopy. The resistance rate to clarithromycin and amoxicillin was $20.2\%$ and $0.0\%$, respectively. The clarithromycin resistance was mainly caused by the A2142G mutation in the 23S rRNA gene of H. pylori. MICs of clarithromycin for the A2142G mutant isolates were significantly higher than MICs for the A2143G mutant isolates. H. pylori eradication was obtained in all patients with clarithromycin-susceptible isolates but not in patients with clarithromycin-resistant isolates (P = 0.0001). These results did not appear to be biased by any differences in CagA expression. The resistance of H. pylori to clarithromycin included in the therapeutic regimens is the most important reason for treatment failure. H. pylori antimicrobial susceptibility testing of the gastric biopsy culture should be performed before choosing the first triple therapy in infected patients and the increase in prevalence of clarithromycin resistance in Korea was problematic.

• Microbiology and Biotechnology Letters
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• v.35 no.1
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• pp.45-51
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• 2007

Arthrospira (Spirulina) platensis is one of the commercially important filamentous cyanobacteria. The heterotrophic cultivation of Arthrospira can be an alternative strategy for commercial mass production. In heterotrophic culture, the specific growth rate of A. platensis M9108, a glucose-resistant mutant of A. platensis PCC 9108, was $0.014h^{-1}$ which was 1.8 higher than that oi the previous report. The mutant possessed the facility to assimilate and to metabolize glucose efficiently under heterotrophic condition. However, the r-linolenic acid content of 6 Arthrospira strains was not increased in heterotrophic culture. Four Arthrospira strains out of 6 tested strains were able to utilize maltose as a carbon source under heterotrophic condition. The biomass production of these strains on maltose was similar to that on glucose. The specific growth rate of A. platensis M9108 increased with glucose concentration up to 5.0 g/L and then decreased at a glucose concentration of 10.0 g/L. Additionally, A. platensis M9108 under heterotrophic condition showed no aggregation during the cultivation in contrast to A. platensis PCC 9108.

• Korean Journal of Fisheries and Aquatic Sciences
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• v.20 no.1
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• pp.57-62
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• 1987

The transformational conditions of plasmids $pPL-\lambda$ and pAS 1 are as follows in E. coli, Ts-mutant M 5248 strain at $30^{\circ}C$. When the culture time was 2.5 hours of mid logarithemic phase, the cell concentration was $4.5\times10^7\;cells/ml$, the optical density was equal to 0.45 at 590 nm wave length, the transformational frequencies of plasmid$pPL-\lambda$ and pAS 1 had the highest values as $2\times10^{-6}\;and\;1.5\times10^{-6}\;and\;1.5\times10^{-6}$ and respectively. For $9\times10^6$ competent cells in $200{\mu}l$, the transformational frequency was as high as $4.4\times10^{-6}$ at 510 ng plasmid concentration. The competent cells treated with the mixture of calcium chloride and thymidine twice rates of transformation than those treated with calcium chloride. The ampicillin resistance of transformants was expressed in LB broth after 2 hours at $30^{\circ}C$.

• Proceedings of the Korean Society of Plant Biotechnology Conference
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• 2002.04b
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• pp.69-75
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• 2002

Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd/dwf3 were Shown to be blocked in $D^4$ reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bri1/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRI1 could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

• Microbiology and Biotechnology Letters
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• v.11 no.3
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• pp.233-239
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• 1983

Bacillus thuringiensis was mutagenized with UV light irradiation and nitrosoguanidine. Twenty-four tem perature-sensitive ts mutants were isolated at 42$^{\circ}C$ and classified into two groups by growth on nutrient agar at 42$^{\circ}C$. First is the lethal group, which did not grow at the nonpermissive temperature, the second is the reduced group whose growth was restricted from one-half to one-fourth, Thirteen ts mutants belong to the lethal group and eleven ts mutants belong to the reduced group. Auxotrophic mutant, A-N28 required five amino acids as growth factors, A-N65 also five amino acids, A-N92 seven, A-N115 four and A-N156 three. Bacillus thuringiensis wild type is resistant to penicillin, ampicillin, and cephalothin. The ts-Ul7l, A-N92 and A-Nl15 are sensitive to the three antibiotics. The ts -U601, -U603, -U604 and -Ul71 did not grow at the permissive temperature after temperature-shifting from 42$^{\circ}C$. Four auxotrophic mutants (A-N38, A-N65, A-N92 and A-Nl15) did not form spores in their cells.

• Journal of Plant Biotechnology
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• v.29 no.2
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• pp.139-144
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• 2002

Sterols play two major roles in plants: a bulk component in biological membranes and precursors of plant steroid hormones. Physiological effects of plant steroids, brassinosteroids (BRs), include cell elongation, cell division, stress tolerance, and senescence acceleration. Arabidopsis mutants that carry genetic defects in BR biosynthesis or its signaling display characteristic phenotypes, such as short robust inflorescences, dark-green round leaves, and sterility. Currently there are more than 100 dwarf mutants representing 7 genetic loci in Arabidopsis. Mutants of 6 loci, dwf1/dim1/cbb1, cpd/dwf3, dwf4, dwf5, det2/dwf6, dwf7 are rescued by exogenous application of BRs, whereas bri1/dwf2 shares phenotypes with the above 6 loci but are resistant to BRs. These suggest that the 6 loci are defective in BR biosynthesis, and the one locus is in BR signaling. Biochemical analyses, such as intermediate feeding tests, examining the levels of endogenous BR, and molecular cloning of the genes revealed that dwf7, dwf5, and dwf1 are defective in the three consecutive steps of sterol biosynthesis, from episterol to campesterol via 5-dehydroepisterol. Similarly, det2/dwf6, dwf4, and cpd /dwf3 were shown to be blocked in D$^4$reduction, 22a-hydroxylation, and 23 a-hydroxylation, respectively. A signaling mutant bril/dwf2 carries mutations in a Leucine-rich repeat receptor kinase. Interestingly, the bri1 mutant was shown to accumulate significant amount of BRs, suggesting that signaling and biosynthesis are dynamically coupled in Arabidopsis. Thus it is likely that transgenic plants over-expressing the rate-limiting step enzyme DWF4 as well as blocking its use by BRIl could dramatically increase the biosynthetic yield of BRs. When applied industrially, BRs will boost new sector of plant biotechnology because of its potential use as a precursor of human steroid hormones, a novel lead compound for cholesterol-lowering effects, and a various application in plant protection.

• Journal of Life Science
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• v.17 no.3 s.83
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• pp.356-361
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• 2007

To investigate the region of ErbB2 for the $ErbB2-{\beta}-catenin$ interaction, a proteasome $resistant-{\beta}-catenin$ and various ErbB2 constructs were transfected in COS7 cells. ErbB2 proteins were immunoprecipitated, and coimmunoprecipitated ${\beta}-catenin$ was examined by Western blotting. ${\beta}-catenin$ coimmunoprecipitated with full length ErbB2. Of the truncated ErbB2 proteins DT (1-1123), DHC (1-1031) and DK (1-750), the ErbB2 constructs containing the kinase domain, DT and DHC, precipitated together with ${\beta}-catenin$ but DK containing no kinase domain did not. To further test the requirement of the kinase domain for ${\beta}-catenin-ErbB2$ interaction, the presence of ${\beta}-catenin$ in the immunocomplex was examined following transfection with an ErbB2 mutant (${\triangle}750-971$) whose kinase domain is internally deleted and subsequent immunoprecipitation of the ErbB2 mutant. ${\beta}-catenin$ was not detected in the immunocomplex. These results suggest that the ErbB2 kinase domain comprises a potential site for ${\beta}-catenin$ binding to the receptor tyrosine kinase.