• The Korean Journal of Food And Nutrition
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• v.10 no.2
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• pp.174-179
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• 1997

The antimicrobial and GTase(Glucosyltransferase) inhibition activity were investigated for solvent fractions of Gall-nut, variety of fork drugs and Red-grape husk water extracts. Among them, Gall-nut and Red-grape husk water extracts were selected for the powerful antimicrobial and GTase inhibition activity. The methanol fractions of Gall-nut and Red-grape husk were showed very powerful antimicrobial activity on both B. subtilis and E. coli. The MIC(Minimum Inhibitory Concentration) of gall-nut methanol fraction were 1.0mg/ml for B. subtilis and 3.0mg/ml for E. coli. Red-grape husk were 2.0mg/ml for B. subtilis and 3.0mg/ml for E. coli. The methanol fractions of Gall-nut and Red-grape husk were showed very powerful Gtase inhibition activity. The concentrations of these fractions for 80% inhibition of GTase activity were 1.08$\times$10-3mg/ml and 1.08$\times$10-2mg/ml, respectively. The principal compound for the antimicrobial and GTase inhibition activity in tese extracts seems to be polyphenol derivatives.

• Food Science and Biotechnology
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• v.14 no.1
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• pp.164-167
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• 2005

The growth responses of materials extracted from Juniperus virginiana leaves against Bifidobacterium bifidum, B. longum, Clostridium perfringens, Escherichia coli, Lactobacillus acidophilus, L. casei, and Streptococcus mutans were examined using impregnated paper disk agar diffusion. The biologically active constituent isolated from the J. virginiana extracts was characterized as ${\alpha}$-cedrene using various spectroscopic analyses including IR, EI-MS, and NMR. The responses varied according to the dose, chemicals, and bacterial strain tested. Methanol extracts of J. virginiana leaves exhibited a strong and moderate inhibitory activity against C. perfringens and E. coli at 5 mg/disk, respectively. However, in tests conducted with B. bifidum, B. longum, L. acidophilus, L. casei, and S. mutans, the methanol extracts showed no or weak inhibitory response. At 2 mg/disk, a-cedrene strongly inhibited the growth of C. perfringens and moderately inhibited the growth of E. coli and S. mutans, without any adverse effects on the growth of four lactic acid-bacteria. Of the commercially available compounds originating from J. virginiana leaves, cedrol and ${\alpha}$-pinene exhibited strong and moderate growth inhibition against C. perfringens, and ${\alpha}$-copaene revealed moderate growth inhibition against E. coli at 1 mg/disk. Furthermore, cedrol exhibited moderate and weak growth inhibition against S. mutans at 2 and 1 mg/disk, respectively. However, little or no activity was observed for camphene, (+)-2-carene, p-cymene, limonene, linalool, and a-phellandrene against B. bifidum, B. longum, C. perfringens, L. acidophilus, L. casei, and S. mutans at 2 mg/disk. The observed inhibitory activity of the J. virginiana leaf-extracted materials against C. perfringens, E. coli, and S. mutans may be an indication of at least one of the pharmacological actions of the J. virginiana leaf.

• Microbiology and Biotechnology Letters
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• v.20 no.3
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• pp.271-279
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• 1992

Genomic DNA of Bacillus stearothemzophilus, which expressed alkalophilic and thermophilic xylanases, was partially digested with HindIII, cloned into pBR322, and subsequently transferred into the Escherichra coli HB101 cells. Three among 5, 000 transformants screened formed clear zones around their colonies. From the functional clones, three recombinant plasmids (pMG11, pMG12 and pMG13) had been isolated, and they were identified to carry the same 4 kb HindIII fragment originated from B. stearothemzophilus which was responsible for the xylanase activity. pMGl3, however, had the foreign DNA of opposite orientation compared to the other two recombinant plasmids. This recombinant plasmid gave much lower xylanase activity. B. stearothermophilus was observed to produce at least three xylanase activities as evidenced by the PAGE-xylan zymogram. The xylanase from E. coli HB101/pMG12 was judged to correspond to the largest among the three B. stearothermophilus xylanases observed in the zymogrom. The enzyme hydrolyzed xylooligosaccharides larger than xylotriose and degraded xylan to produce xylobiose and xylotriose as major products. The xylanase was considered to have trans-xylosidase activity, too.

• BMB Reports
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• v.29 no.3
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• pp.221-224
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• 1996

To examine the growth-phase dependent control of Escherichia coli rnpB gene we used a combination of Northern analysis for RNA determination and Southern analysis for plasmid DNA determination. The relative amounts of metabolically unstable transcript derived from the internally deleted rnpB gene harbored on a multicopy plasmid as well as the relative plasmid contents were measured by Northern analysis and Southern analysis, respectively, of total nucleic acids from E coli cells containing the plasmid. The relative transcription activity of the rnB was represented by a ratio of the relative amount of the transcript to that of the plasmid DNA during bacterial growth. The rnpB transcription increased rapidly with time during exponential growth, but started to decrease before the transition period of an exponential growing cell culture into the stationary phase. Although the expression pattern was similar to the changes of ${\beta}-galactosidase$ activity expressed from the lysogenic strain carrying the chromosomal rnpB-lacZ fusion which were shown in a previous work, the present data appears to represent a more actual growth-phase control of the rnpB transcription than the previous data by the ${\beta}-galactosidase$ assay. In addition the present method described for a direct analysis of both RNA and plasmid DNA provides a rapid and efficient method that can applied to an examination of transcription control by using a multicopy plasmid.

• Journal of Microbiology and Biotechnology
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• v.11 no.4
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• pp.720-723
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• 2001

The effect of long-term storage on garlic antibacterial activity was investigated. A concentration of 5% or more garlic was found to be necessary to completely inhibit Eschrichia coli growth in tryptic soy broth. This value is substantially higher than the minimum inhibitory concentration of 1% for E. coli reported previously. pH-modified garlic extract was stored at different temperatures to investigate the impact of storage conditions (i.e., temperature, pH, period of storage) on the stability of the antibacterial activity of the garlic extract used against E. coli B34. The antibacterial effectiveness of the garlic extract against E. coli remained stable when both the storage temperature and the pH of the extract were kept low. When the garlic extract was stored at $40^{\circ}C and above, most or all of the garlic antibacterial activity disappeared after a 24-h storage period, regardless of the storage pH. The antibacterial activity was weakened when the pH of the garlic extract was adjusted to 8, and at low temperatures.

• Journal of Microbiology and Biotechnology
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• v.16 no.6
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• pp.901-910
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• 2006

Poly(3-hydroxybutyrate) [P(3HB)] is a microbial polyester intracellularly accumulated as distinct granules in numerous microorganisms as an energy and carbon storage material. Recombinant Escherichia coli harboring the heterologous P(3HB) biosynthesis genes accumulates large amounts of P(3HB) granules, yet the granule-associated proteins have not been identified. Therefore, this study reports on an analysis of the P(3HB) granule-associated proteome in recombinant E. coli. Fiye proteins out of 7 spots identified were found to be involved in functions of translation, heat-stress responses, and P(3HB) biosynthesis. Two of the major granule-associated proteins, IbpA/B, which are already known to bind to recombinant proteins forming inclusion bodies in E. coli, were further analyzed. Immunoblotting and immunoelectron microscopic studies with IbpA/B antibodies clearly demonstrated the binding and localization of IbpA/B to P(3HB) granules. IbpA/B seemed to play an important role in recombinant E. coli producing P(3HB) by stabilizing the interface between the hydrophobic P(3HB) granules and the hydrophilic cytoplasm. Thus, IbpA/B were found to act like phasins in recombinant E. coli, as they are the major proteins bound to the P(3HB) granules, affect the morphology of the granules, and reduce the amount of cytosolic proteins bound to the P(3HB) granules.

• Korean Journal of Microbiology
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• v.41 no.2
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• pp.140-145
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• 2005

Streptomyces natalensis ATCC27448 produces natamycin, a commercially important macrolide antifungal antibiotic. For molecular genetic study of S. natalensis, we have developed a system for introducing DNA into S. natalensis via conjugal transfer from Escherichia coli. An effective transformation procedure for S. natalensis was established based on transconjugation from E, coli ET12567/pUZ8002 using a ${\Phi}C31$-derived integration vector, pSET152, containing oriT and attP fragments. The high frequency was obtained on MS medium containing 10 mM $MgCl_2$ using $6.25\times10^8$ of E.coli donor cells without heat treatment of spores. In addition, southern blot analysis of exconjugants and the sequence of plasmids containing DNA flanking the insertion sites from the chromosome revealed that S. natalensis contains a single ${\Phi}C31$ attB site and at least a secondary or pseudo attB site. Similar to the case of various Streptomyces species, a single ${\Phi}C31$ attB site of S. natalensis is present within an ORF encoding a pirin-homolog, but a pseudo-attB site is present within a distinct site (GenBank accession no. $YP\_117731$) and also its sequence deviates from the consensus sequences of attB sequence.

• Korean Journal of Clinical Laboratory Science
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• v.53 no.3
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• pp.217-224
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• 2021

Fluoroquinolone (FQ) resistant gram-negative pathogens have emerged worldwide, and the recent increase in FQ resistant Escherichia coli is of great concern in Korea. This study investigated FQ resistance determinants and the epidemiological relationship of 56 ciprofloxacin-resistant E. coli isolated from a tertiary hospital in Daejeon, South Korea from June to December 2018. Molecular epidemiology was investigated by multilocus sequence typing (MLST). Polymerase chain reaction (PCR) and sequence analysis were performed to identify chromosomal mutations in the quinolone resistance determining regions (QRDR) of gyrA, gyrB, parC, and parE and to describe the occurrence of the following plasmid-mediated quinolone resistance (PMQR) genes: aac(6)-Ib-cr, qepA, qnrA, qnrB, qnrC, qnrD, and qnrS. MLST analysis showed 12 sequence types (STs) and the most prevalent ST was ST131 (31/56, 55.4%), followed by ST1193 (13/56, 23.2%), and ST405 (3/56, 5.4%). In 56 ciprofloxacin-resistant E. coli isolates, Ser83→Leu and Asp87→Asn in gyrA and Ser80→Ile and Glu84→Val in parC (51.8%, 29/56) were the most frequent amino acid substitutions and aac(6)-Ib-cr (33.9%, 19/56) was the most common PMQR gene. These results of FQ resistance determinants were more frequently observed in ST131 compared with other clones. Continuous monitoring of the epidemiological characteristics of ciprofloxacin-resistant E. coli isolates and further investigation of FQ resistance determinants are necessary.

• Journal of Microbiology and Biotechnology
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• v.16 no.11
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• pp.1684-1689
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• 2006

Airborne microorganisms, which are currently termed bioaerosols, have received attention owing to the harmful effects they have on human health. As the concern over airborne microorganisms grows, there also grows an urgent need to study and develop efficient methods for controlling them. In this study, thermal energy using a thermal tube was tested as a control method, mainly against airborne E. coli. For a comparison, B. subtilis var. niger spores were utilized in the experimentation. It was found that the widely known inactivation conditions for microorganisms were not adequate against airborne microorganisms. The experimental results demonstrated the need for extensive studies that should investigate adequate and economic conditions to control against airborne bacteria. In this study, thermal energy exposed by the thermal tube demonstrated an inactivation performance for controlling E. coli bioaerosols.

• Journal of Microbiology and Biotechnology
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• v.10 no.2
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• pp.275-280
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• 2000

The inulin fructotransferse (depolymerizing) (IFTase, EC 2.4.1.93) gene of Arthrobacter sp. A-6 was cloned and expressed in Escherichia coli and Bacillus subtilis. The IFTase gene consisted of an ORF of 1.311 nucleotides encoding a polypeptide of 436 amino acids containing a signal peptide of 31 amino acids in the N-terminus. The molecular mass of the IFTase based on the nucleotide sequence was calculated to be 46.116 Da. The recombinant E. coli $DH5{\alpha}$ cells expressing the Arthrobacter sp. A-6 IFTase gene produced most of the IFTase intracelularly. In contrast, the recombinant B. subtilis DB 104 carrying the IFTas gene on a B. subtilis-E. Coli expression vector secreted the IFTase into the culture fluid efficiently.