• Journal of Microbiology and Biotechnology
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• v.29 no.3
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• pp.401-409
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• 2019

Heat-resistant microbial hosts are required for bioprocess development using high cell density cultivations at the industrial scale. We report that the thermotolerance of Escherichia coli can be enhanced by overexpressing ybeD, which was known to encode a hypothetical protein of unknown function. In the wild-type E. coli BL21(DE3), ybeD transcription level increased over five-fold when temperature was increased from $37^{\circ}C$ to either $42^{\circ}C$ or $46^{\circ}C$. To study the function of ybeD, a deletion strain and an overexpression strain were constructed. At $46^{\circ}C$, in comparison to the wild type, the ybeD-deletion reduced cell growth half-fold, and the ybeD-overexpression promoted cell growth over two-fold. The growth enhancement by ybeD-overexpression was much more pronounced at $46^{\circ}C$ than $37^{\circ}C$. The ybeD-overexpression was also effective in other E. coli strains of MG1655, W3110, DH10B, and BW25113. These findings reveal that ybeD gene plays an important role in enduring high-temperature stress, and that ybeD-overexpression can be a prospective strategy to develop thermotolerant microbial hosts.

• Microbiology and Biotechnology Letters
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• v.19 no.3
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• pp.259-264
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• 1991

Glutamine synthetase (GS) of Kkbsiellu pmumonzae was purified and identified it's properties. It was determined to be composed of 12 identical subunits and it's molecular weight was about 600,000. It's optimum pH and temperature were identified as pH 7.0 and $37^{\circ}C$ respectively, and also there was no considerable variation of activity between pH 5 and 8. When GS was incubated at $57^{\circ}C$ for 10 min, it's activity was decreased to half of maximum activity. It was observed that K. pneumoniae has adenylylation-deadenylylation system which regulates activity of GS according to the quality and quantity of nitrogen source like GS of E. coli Also it's GS was very similar to that of E. coli. in structure deduced from the immunodiffuslon experiment using anti-E. coli GS antibody.

• Biomedical Science Letters
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• v.2 no.2
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• pp.231-240
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• 1996

Human blood clotting (coagulation) factor 9 cDNA which codes for 461 amino acid has been cloned by screening human fetal liver cDNA library using PCR. This 1.4 kb cDNA spanning from the ATG initiation codon to the TAA termination codon was cloned into bacterial .expression vector pGEX-2T, generating pGEX-F9 plasmid. The plasmid pGEX-F9 expresses about 73 kDa GST (Glutathione S-transferase)-Factor 9 fusion protein when introduced into E. coli. Western blot analysis using polyclonal antibody raised against human factor 9 confirmed this fusion protein contains factor 9 protein. The level of GST-factor 9 expression was about 20% of total protein and the purification of fusion protein was efficiently achieved by using GST agarose bead based on one step purification protocol.

• Korean Journal of Microbiology
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• v.26 no.1
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• pp.6-12
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• 1988

Cellobiase ($\beta$-glucosidase) is an enzyme of the cellulase system in cellulolytic microor-ganisms. The chromosomal DNA fragment which include cellobiase gene of Cellulomonas biazotea was cloned in Eschericia coli via plasmid pBR 322 vector. Restriction enzyme Sal I was used to obtain adequate size of fragments from C. biazotea. chromosomal DNA. The transformant of E. coli HB101 with recombinant plasmid pBG101 showed cellobiase activity, which is not ordinary in E. coli HB101. The enzyme activity of the transformant was as of 20% lower than that of C. biazotea.

• BMB Reports
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• v.30 no.5
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• pp.320-325
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• 1997

Thioredoxin is a multi-functional protein which is ubiquitous in microorganisms, animals and plants. Previously, expression of the E. coli thioredoxin gene was found to be negatively regulated by cAMP. In the present study, the effect of cAMP on two separate promoters of the E. coli thioredoxin gene was investigated. Cyclic AMP had a repressible effect on P1 and P1P2 promoter activity of the constructs. This effect was also observed in the cya strain. The P2 promoter construct gave very high -galactosidase activity, and its expression was not affected by exogenous cAMP. It was assumed that a cis-acting negative element, probably the cAMP-CRP binding site, might have been deleted in the P1 promoter construct. Repression of the thioredoxin gene expression by cAMP appeared to be independent of ppGpp.

• Microbiology and Biotechnology Letters
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• v.27 no.3
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• pp.260-265
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• 1999

The structural $\beta$-galactosidase gene (lacZ) from Lactococcus lactis ssp. lactis 7962 was cloned into plamid vector pKF18, which was designated as pKF-gal. Expression of the lacZ from L. lactis 7962 was found to be higher when cells were grown at 3$0^{\circ}C$ than 37$^{\circ}C$. Maximum $\beta$-galactosidase activity was obtained when E. coli/pKF-gal was cultivated for 6hr at 3$0^{\circ}C$ and for 3hr at 37$^{\circ}C$, and L. lactis 7962 was grown for 8hr at 3$0^{\circ}C$. Enzyme induction was achieved by the addition of lactose, galactose, or lactose+IPTG to growing culture. The addition of glucose had no effect on enzyme induction.

• KSBB Journal
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• v.13 no.2
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• pp.168-173
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• 1998

Gamma irradiated and non-irradiated Agrimonia pilosa Ledebour were extracted by 70% ethanol. The combined effects of the Agrimonia pilosa Ledebour extract and NaCl on survival of Escherichia coli O157:H7 in tryptic soy broth were investigated. E. coli O157:H7 decreased ca 1 log cycle by the addition of 2% sample extract, and the anthbacterial activity was increased as the concentration of sample extract was increased. The irradiation effect of the sample on antibacterial activity was not observed. On the treatment of NaCl alone, E. coli O157:H7 was inactivated (ca 3~4 log cycle reduction within 48 hr) in more than 7% NaCl. The higher inactivation(ca 5 log cycle reduction within 48 hr) occurred in the presence of 2% sample extract and 5% NaCl than in the addition of each alone. The extracted antibacterial substance was stable in the pH range of 4.0 to 7.0, heat treatment at 121$^\circ C$ for 15 min, and freezing at -18$^\circ C$ and thawing at 37$^\circ C$. There fore, the sample extract, would substantially increase the food-safety in terms of E. coli O157:H7.

• Korean Journal of Microbiology
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• v.29 no.2
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• pp.80-84
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• 1991

We cloned and expressed hepatitis B viral core antigen (HBcAg) gene in E. coli using $P_{L}$ promoter system. For optimal expression of the gene, we undertook the studies on the effects of the distance between Shine-Dalgarno (SD) sequence and start codon, copy number of repressor gene, induction temperature, and the stability of the core antigen. The results demonstrated that the induction at 37.deg.C was more efficient than at 42.deg.C, and the 11 base pairs (bp) distance between SD sequence and start codon of HBcAg gene was more efficient than the 15 bp distance in E. coli. The copy number of cI857 repressor gene did not influence on the expression of HBcAg, and the expression level of HBcAg in mutant type (low protease activity) and wild type strains was almost the same. The produced core antigen appeared to be HBcAg not HBeAg judged by two different radioimmunoassat (RIA) kits. This result suggested that the antigen was stable in E. coli.i.

• Bulletin of the Korean Chemical Society
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• v.35 no.8
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• pp.2460-2464
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• 2014

Recently studies indicate that the microcalorimetry is a suitable measurement for metabolic activities of organisms by recording the rate of heat outputs. In this work, we investigated the growth thermogenic curves of Escherichia coli (E. coli) affected by three kinds of Glycyrrhiza extracts by microcalorimetry. The power-time and exponential phase power-time curves of the E. coli growth at various concentrations of extracts were generated. The kinetic parameters such as the growth rate constants (C), maximum power outputs (Pm), peak times (Tm), and inhibition ratios were calculated and the relationships between Pm or Tm and C were established. Also, the clear correlations among the antimicrobial effects, Pm and C were obtained. The results revealed the Glycyrrhiza extracts had inhibitory activities towards E. coli while the Glycyrrhiza polysaccharides showed the most potent effects.

• Food Science and Preservation
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• v.9 no.4
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• pp.434-440
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• 2002

The growth characteristics of 5. coli O157:H7 CDFI, A. sobria CDF3 and S. aureus CDF2 isolated from surface of carcass were investigated to improve hygienic quality of beef. The total count of carcass surface before washing was higher than that of after washing. Total count of after cooling decreased about 10$^1$∼ 10$^2$/㎠ compare with before cooling. Total count of carcass surface after transfer increased regardless seasons. The growth E. coli O157:H7 CDF1 occurred at pH 4 and 6% NaCl but A. sobria CDF3 and S. aureus CDF2 did not grow at the same conditions. Although the growth of E. coli O157:H7 CDF1 and S. aureus CDF2 was inhibited by 0.3％ lactic acid, but A sobria CDF3 did not grow in TSB containing 0.3％ lactic acid. E. coli O157:H7 CDF1 grew rapidly after 3 days incubation at 10$\^{C}$ but did not grow at 4$\^{C}$. But A. sobria CDF3 grew rapidly after 3 days incubation at 4$\^{C}$. E. coli O157:H7 CDF1 and A. sobria CDF3 were destroyed by heat treatment for 3 min at 60$\^{C}$. S. aureus CDF2 did not detect after heat treatment for 2 min at 70$\^{C}$.