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Transfer of RP4:Mu cts from E. coli to Rhizovium leguminosarum (RP4:Mu cts의 E.coli로 부터 Rhizovium leguminosarum으로의 전달)

  • 이인렬;허연주;이영록
    • Korean Journal of Microbiology
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    • v.23 no.2
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    • pp.107-114
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    • 1985
  • In order to use for recipient strains of RP4:Mu cts, 5 strainsof Rhizobium were selected among 32 strains, which were isolated and identified in this study. Hybrid plasmin RP4::Mu cts, which, is temperature sensitive and confers resistance to ampicillin, kanamycin and tetracycline was transfered by conjugation from E. coli to other atrains of C. coli and the symbiotic nitrogen fixer, Rhizobium leguminosarum. Transfer frequencies of RP4::Mu cts plasmid from E. coli to Rhizobium were about $10^{-8}-10^{-7}$ in LB agar and YMA media. The transconjugants were confirmed by demonstrating that the drug-resistant and temperature-sensitive clones isolated were drug-resistant and temperature-sensitive clones isolated were capable of releasing phage and forming plaques. The plaque-forming units of transconjugants were about $10^2\;to\;10^3$. Stability test of RP4::Mucts in Rhizobium represented that most of the transconjugants had drug resistance and produce phage Mu cts.

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Effect of Chaff Vinegar on the Growth of Food-Borne Pathogenic Bacteria

  • Jo, Seong-Chun;Rim, A-Ram;Park, Hee-Jin;Park, Sun-Min;Lee, Seung-Cheol
    • Preventive Nutrition and Food Science
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    • v.10 no.2
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    • pp.130-133
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    • 2005
  • The growth inhibitiory effect of chaff vinegar against various food-borne pathogenic bacteria was evaluated. Bacterial growth was evaluated in chaff vinegar at concentrations of 15, 30, 50, 65, 80, and $100\%$ using the paper disc diffusion method and 0.5, 1.0, 1.5, 1.8, 2.0, 2.2 and $2.5\%$ in broth. In the paper disc diffusion assay, chaff vinegar showed a clear zone on both the Gram-positive bacteria; Listeria monocytogenes, Staphylococcus aureus and Gram-negative bacteria; Escherichia coli O157:H7, Salmonella Typhimurium, and Vibrio parahaemolyticus. Chaff vinegar exhibited the greatest growth inhibition for V. parahaemolyticus. The bactericidal effect of chaff vinegar on the E. coli O157:H7 was tested at concentrations ranging from 0.5 to $2.5\%$ (v/v) in the LB broth media. Chaff vinegar retarded the lag phase time of the growth curve in proportion in a concentration-dependent manner. Chaff vinegar at $2.5\%$ completely inhibited the growth of E. coli O157:H7.

Effect of Glycine Supplement on Extracellular Secretion of Levansucrase form Pseudomonas aurantiaca S-4380 in Recombinant Escherichia coli (Glycine 첨가에 의한 Pseudomonas aurantiaca S-4380 유래 재조합 levansucrase 효소의 세포 외 분비촉진 효과)

  • 김승환;장은경;김인환;장기효;강순아;장병일
    • KSBB Journal
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    • v.18 no.4
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    • pp.312-317
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    • 2003
  • The addition of glycine up to 0.5% (w/v) to Luria broth (LB) media on the secretion of levansucrase in a recombinant strain Escherichia coli JM109/pUPLK1 was observed to enhance the release of periplasmic proteins from the cell to the broth, without significantly affecting the cell growth rate and protein productivity. However, the glycine concentration at 1 % (w/v), the cell density attainable at the stationary phase fell to about 50% and the extracellular activity of levansucrase corresponded to about 80% of the total (extracellular plus intracellular) activity and increased by 2.6-fold, comparing to the cells grown in the absence of glycine. The increased pH at stationary phase accelerated the degradation of levansucrase. Maximal extracellular activity was attained when 1 % glycine was supplemented at the onset of strain growth.

Use of Vegetable Waste as a Culture Medium Ingredient Improves the Antimicrobial and Immunomodulatory Activities of Lactiplantibacillus plantarum WiKim0125 Isolated from Kimchi

  • Seul-Gi Jeong;Ho Myeong Kim;Moeun Lee ;Jung Eun Yang;Hae Woong Park
    • Journal of Microbiology and Biotechnology
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    • v.33 no.1
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    • pp.75-82
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    • 2023
  • Lactic acid bacteria (LAB) isolated from kimchi (a traditional Korean dish typically made of fermented cabbage) can provide various health benefits, including anti-obesity, antioxidant, antiinflammatory, anticancer, and antimicrobial effects. In this study, we examined the antimicrobial and immunomodulatory effects of Lactiplantibacillus plantarum WiKim0125 cultured in de Man, Rogosa, and Sharpe (MRS) medium containing vegetable waste. Live bacterial cells were eliminated via supernatant filtration or heat treatment. The cell-free supernatant (CFS) obtained from culture broth containing kimchi cabbage waste (KCW), cabbage waste (CW), or onion waste (OW) showed significantly higher antimicrobial activity against skin pathogens (Propionibacterium acnes and Staphylococcus aureus) and foodborne pathogens (Escherichia coli and Salmonella typhimurium), with inhibition zones ranging between 4.4 and 8.5 mm, compared to that in conventional MRS medium (4.0-7.3 mm). In lipopolysaccharide-stimulated RAW264.7 cells, both supernatant and heat-inactivated Lb. plantarum WiKim0125 from culture media containing KCW and CW suppressed the production of inflammatory cytokines (72.8% and 49.6%, respectively) and nitric oxide (62.2% and 66.7%, respectively) without affecting cell viability. These results indicate that vegetable waste can potentially increase the antimicrobial and immunoregulatory potency of LAB while presenting a molecular basis for applying postbiotics to health products.

Production of Vanillin from Ferulic Acid Using Recombinant Strains of Escherichia coli

  • Yoon Sang-Hwal;Li Cui;Lee Young-Mi;Lee Sook-Hee;Kim Sung-Hee;Choi Myung-Suk;Seo Weon-Taek;Yang Jae-Kyung;Kim Jae-Yeon;Kim Seon-Won
    • Biotechnology and Bioprocess Engineering:BBE
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    • v.10 no.4
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    • pp.378-384
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    • 2005
  • Vanillin is one of the world's principal flavoring compounds, and is used extensively in the food industry. The potential vanillin production of the bacteria was compared to select and clone genes which were appropriate for highly productive vanillin production by E. coli. The fcs (feruloyl-CoA synthetase) and ech (enoyl-CoA hydratase/aldolase) genes cloned from Amycolatopsis sp. strain HR104 and Delftia acidovorans were introduced to pBAD24 vector with $P_{BAD}$ promoter and were named pDAHEF and pDDAEF, respectively. We observed 160 mg/L vanillin production with E. coli harboring pDAHEF, whereas 10 mg/L of vanillin was observed with pDAHEF. Vanillin production was optimized with E. coli harboring pDAHEF. Induction of the fcs and ech genes from pDAHEF was optimized with the addition of 13.3 mM arabinose at 18 h of culture, from which 450 mg/L of vanillin was produced. The feeding time and concentration of ferulic acid were also optimized by the supplementation of $0.2\%$ ferulic acid at 18 h of culture, from which 500 mg/L of vanillin was obtained. Under the above optimized condition of arabinose induction and ferulic acid supplementation, vanillin production was carried out with four different types of media, M9, LB, 2YT, and TB. The highest vanillin production, 580 mg/L, was obtained with LB medium, a 3.6 fold increase in comparison to the 160 mg/L obtained before the optimization of vanillin production.

A Biological Activity of Serratia marcescens Strains Isolated from Dead Larva of the Diamondback Moth, Plutella xylostella (Plutellidae, Lepidoptera) (배추좀나방(Plutella xylostella)의 죽은 유충에서 분리한 Serratia marcescens 균주의 생물활성)

  • Jun, Jun Hack;Jin, Na Young;Lee, You Kyoung;Lee, Bo Ram;Youn, Young Nam;Yu, Yong Man
    • The Korean Journal of Pesticide Science
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    • v.20 no.2
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    • pp.152-158
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    • 2016
  • The cause of death was investigated with several dead cabbage moth larvae in breeding box. Bacterial strains were isolated and selected from the dead larvae by bioassay. One of them was identified as Serratia marcescens used by morphological characteristics and gene sequencing. S. marcescens were cultured by Luria Bertani (LB) media broth for bioassay. When 100-fold dilution of culture broth to third larvae of diamondback moth, Plutella xylostella, it was showed a 100% mortality at 2 days after treatment, and only 10-fold dilution of supernatant liquid was showed 86.6% mortality. When the culture broth of S. marcescens was applied to the larvae of beet armyworm, Spodoptera exigua, contact and feeding toxicity were 20 and 8% of mortality, respectively. Otherwise, when the culture broth of S. marcescens was applied to 5 major plant pathogens, antibacterial activities against Fusarium oxysporum, Rhizoctonia solani, Colletotrichum gloeosporioides, Phytophthora capsici and Sclerotinias clerotiorum were 4.7, 11.3, 20, 15.7 and 42.6%, respectively. Also, degradation ability of S. marcescens against protein and chitin were examined.

Characteristics of Biosurfactant Producing Pseudomonas sp. HN37 (생물계면활성제 생성 세균 Pseudomonas sp. HN37의 특성)

  • Jung, Da Hee;Chang, Dong Ho;Kim, Yeong Eun;Jeong, Mi Rang;Hahn, Kyu Woong;Kim, Hyong Bai;Park, Kyeong Ryang
    • Korean Journal of Microbiology
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    • v.50 no.1
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    • pp.33-39
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    • 2014
  • One hundred forty four bacterial colonies which were able to degrade crude oil were isolated from soil samples that were contaminated with oil in Daejeon area. Among them, one bacterial strain was selected for this study based on its emulsification activity, growth rate and surface tension activity, and this selected bacterial strain was identified as Pseudomonas sp. HN37 through physiological- biochemical tests and analysis of its 16S rRNA sequence. Pseudomonas sp. HN37 utilize the several aliphatic hydrocarbons, 3,5-dinitrosalicylic acid and 2,4-dichlorophooxyacetic acid as a sole carbon source. And this bacterial strain showed a high resistance to antibiotics such as ampicillin and chloramphenicol, as well as heavy metals such as Ba, Cr, Li and Mn. Also, it was found that the optimal pH and temperature for the cell growth, surface tension, and emulsification activity of Pseudomonas sp. HN37 were pH 6.0-9.0 and $30^{\circ}C$, respectively. The emulsification and surface tension activity was reached the maximum to 1% (V/V) crude oil and 1% (W/V) NaCl concentration. The surface tension of the culture broth was decreased from 62 to 27 dyne/cm after fifteen hours of inoculation in LB media.

Optimum Conditions of Freezing Lyophilization and Bioluminescence Activity Recovery for Environmental Applications Using a Recombinant Strain (유전자 재조합 균주를 환경에 적용하기 위한 (동결) 건조 및 활성회복 조건 최적화)

  • Ko Kyung-Seok;Kim Myung-Hee;Kong In-Chul
    • Journal of Soil and Groundwater Environment
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    • v.11 no.5
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    • pp.43-50
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    • 2006
  • Bioreporter bacteria, such as recombinant bioluminescent bacteria, have been used for the detection of specific compounds in complex environmental media. In this study, optimum conditions for the preparation and application of deep-freezed and Iyophilized recombinant bioluminescent strain KG1206 were investigated for the future application on contaminated environmental sites. Genetically engineered microorganism, Pseudomonas putida mt-2 KG1206, contains TOL plasmid and the plasmid inserted $P_{m}$, promoter on the upper part of lux gone in vector pUCD615, and m-toluate and benzoate are considered direct inducers for bioluminescence. Optimum conditions determined for the preparation and application of the deep-freezed and lyophilized strain were followings: cryoprotective agent (24% sucrose), lyophilization time (12 hrs), strain concentration ($OD_{600}=0.6$), reconstitution for freezed strain (quick reconstitution at $35^{\circ}C$), reconstitution for lyophilized strain ($3{\sim}6$ hrs exposure on LB medium), carrying conditions (keep at $20^{\circ}C$ after reconstitution). These results demonstrate the feasibility of deep-freezed or lyophilized state of genetically engineered bioluminescent strain for environmental usage.

Antimicrobial activities of Burkholderia sp. strains and optimization of culture conditions (Burkholderia sp. OS17의 항균활성 증진을 위한 배양최적화)

  • Nam, Young Ho;Choi, Ahyoung;Hwang, Buyng Su;Chung, Eu Jin
    • Korean Journal of Microbiology
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    • v.54 no.4
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    • pp.428-435
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    • 2018
  • In this study, we isolated and identified bacteria from freshwater and soil collected from Osang reservoir, to screen antimicrobial bacteria against various pathogenic bacteria. 38 strains were isolated and assigned to the class Proteobacteria (22 strains), Actinobacteria (7 strains), Bacteroidets (6 strains), and Firmicutes (3 strains) based on 16S rRNA gene sequence analysis. Among them, strain OS17 showed a good growth inhibition against 5 methicillin-resistant Staphylococcus aureus subsp. aureus strains and Bacillus cereus, Bacillus subtilis, Filobasidium neoformans. As a result of the 16S rRNA gene sequence analysis, strain OS17 show the high similarity with Burkholderia ambifaria $AMMD^T$, B. diffusa $AM747629^T$, B. tettitorii $LK023503^T$ 99.8%, 99.7%, 99.6%, respectively. We investigated cell growth and antimicrobial activity according to commercial culture medium, temperature, pH for culture optimization of strain OS17. Optimal conditions for growth and antimicrobial activity in strain OS17 were found to be: YPD medium, $35^{\circ}C$ and pH 6.5. When the strain was cultured in LB, NB, TSB, R2A media at $20^{\circ}C$ and $25^{\circ}C$, the antimicrobial activity did not show. Culture filtrate of strain OS17 showed antimicrobial activity against 5 MRSA strains, Bacillus cereus, Bacillus subtilis, and Filobasidium neoformans with inhibition zones from 2 to 8 mm. Optimal reaction time was 48 h in YPD medium, 100 rpm and 0.3 vvm in 2 L-scale fed-batch fermentation process for antimicrobial activity. Culture optimization of strain OS17 can be improved on antimicrobial activity. Therefore, the antimicrobial activity of Burkholderia sp. OS17 had potential as antibiotics for pathogens including MRSA.

Antifungal Activity of Pseudomonas sp. DGUM 5051 Against Apple Bitter-rot Causing Fungus, Glomerella cingulata (사과탄저병균(Glomerella cingulata)에 대한 Pseudomonas sp. DGUM 5051의 항진균 활성)

  • Kim, Jung-Mi;Lee, Min-Woong;Han, Yeong-Hwan
    • The Korean Journal of Mycology
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    • v.26 no.4 s.87
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    • pp.458-465
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    • 1998
  • The strain DGUM 5051, an antagonistic bacterium against apple-bitter rot causing Glomerella cingulata, was isolated from soil in Kyongju. Based on the morphological and physiological characteristics, the bacterium was identified as Pseudomonas sp. and named as Pseudomonas sp. DGUM 5051. The optimal pH and temperature for cell growth were pH 6.0 and $30^{\circ}C$, whereas those for antifungal activity were pH 7.0 and $24^{\circ}C$, respectively. Among the complex media tested, brucella medium, brain heart infusion medium and Luria-Bertani medium were good for both cell growth and antifungal activity. The high antifungal activity was found in the mineral salts medium, in which sucrose, $KNO_3$ and $K_2HPO_4$ were used as sources of carbon, nitrogen and phosphorus, respectively.

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