• Title/Summary/Keyword: resistant strains

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Effects of tktA, $aroF^{FBR}$and aroL Expression in the Tryptophan-Producing Escherichia coli

  • Kim, Tae-Hyun;Namgoong, Suk;Kwak, Joon-Hyeok;Lee, Se-Yong;Lee, Heung-Shick
    • 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.

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Relationship between Eradication of Helicobacter pylori Infection and Clarithromycin Resistance (Helicobacter pylori 감염의 치료와 Clarithromycin 내성간의 연관성)

  • Sohn Seung Ghyu;Lee Jong Hwa;Lee Jung Hun;Lee Sang Hee
    • 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.

Dual Coating Improves the Survival of Probiotic Bifidobacterium Strains during Exposure to Simulated Gastro-Intestinal Conditions (위장관내 조건에서 이중코팅 처리 된 프로바이오틱 비피도박테리움의 생존력 향상)

  • Kang, Joo Yeon;Lee, Do Kyung;Park, Jae Eun;Kim, Min Ji;Lee, Joong-Su;Seo, Jae-Gu;Chung, Myung Jun;Shin, Hea Soon;Ha, Nam Joo
    • Korean Journal of Microbiology
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    • v.49 no.3
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    • pp.275-281
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    • 2013
  • Probiotics have been reported to benefit human health by modulating immunity, lowering cholesterol, improving lactose tolerance, and preventing some cancer. Once ingested, probiotic microorganisms have to survive harsh conditions such as low pH, protease-rich condition, and bile salts during their passage through the gastro-intestinal (GI) tract colonize and proliferate to exert their probiotic effects. The dual coating technology, by which the bacteria are doubly coated with peptides and polysaccharides in consecutive order, was developed to protect the ingested bacteria from the harsh conditions. The aim of the study was to evaluate the viable stability of a doubly coated blend of four species of Bifidobacterium by comparing its bile/acid resistance and heat viability in vitro with that of the non-coated blend. After challenges with acid, bile salts, heat, and viable cell counts (VVCs) of the dual coated and non-coated blend were determined by cultivation on agar plates or flow cytometric measurement after being stain with the BacLigtht kit$^{TM}$. The results showed that the dual coated blend was much higher resistant to the acidic or bile salt condition than the non-coated blend and heat viability was also higher, indicating that the dual coating can improve the survival of probiotic bacteria during their transit through the GI tract after consumption.

Profiles of Toxin Genes and Antibiotic Susceptibility of Bacillus cereus Isolated from Perilla Leaf and Cultivation Areas (들깻잎과 생산환경에서 분리한 Bacillus cereus의 독소 유전자와 항생제 감수성 분석)

  • Kim, Se-Ri;Lee, Ji-Young;Lee, Seo-Hyun;Ryu, Kyoung-Yul;Park, Kyeong-Hun;Kim, Byung-Seok;Yoon, Yo-Han;Shim, Won-Bo;Kim, Kyoung-Yul;Ha, Sang-Do;Yun, Jong-Chul;Chung, Duck-Hwa
    • Korean Journal of Food Science and Technology
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    • v.43 no.2
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    • pp.134-141
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    • 2011
  • Two-hundred Bacillus cereus isolated from perilla leaf cultivation areas in Miryang, Korea were investigated for toxin genes and antibiotic susceptibility. Toxigenic patterns of isolates were identified to be 11 groups through toxin gene profiles. 21% of strains isolated from the perilla leaves had both enterotoxin and emetic toxin. Toxin genes entFM (100%), nheA (100%) and hblA, C, D (65.5%) were frequently found in the perilla leaves, whereas EM (21.0%) was less common. Most isolates were susceptible to 10 antibiotics, but they were highly resistant to penicillin (100%), ampicillin (100%), oxacillin (94.9%), amoxicillin-clavulanic acid (95.6%), cefazolin (78.2%), and rifampicin (58.0%). These results indicate that food-borne outbreak caused by B. cereus might lead to diarrhea and emetic syndromes.

Characteristics of Vibrio mimicus K-1 Isolated from Coastal Sea Water (연안 해수에서 분리된 Vibrio mimicus K-1의 특성)

  • KOH Byeong-Ho;LEE Won-Jae;LEE Myung-Suk
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.27 no.3
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    • pp.292-298
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    • 1994
  • An environmental study was done to examine the distribution of Vibrio mimicus in aquatic environments of Kwangan and Minrak beach, Pusan, Korea. Moreover, both bacteriological characteristics and lethal effects of isolated V. mimicus were observed. Sea water samples were collected monthly from January to September, 1993, and quantitatively analyzed for V. mimicus. This organism was isolated from April(water temperature was $16.3^{\circ}C$), whereas it was not isolated when the water temperature fell below $15^{\circ}C$. V. mimicus counts were not remarkably high, however this study at least describes the distribution and occurrence of the possible highest density in aquatic environments of this region. Among the confirmed V. mimicus strains, the author chose the strongest antibiotic resistant bacterium and named it V. mimicus K-1. This strain has antibiotic resistance to colistin, erythromycin, tetracycline, and penicillin, and most isolates had a higher level of antibiotic resistance than V. mimicus ATCC 33653. The optimum growth for V. mimicus K-1 was observed at $37^{\circ}C$, pH 7.5, and $1\%$ NaCl, respectively. This organism was mostly inactivated by Ultra Violet irradiation (30W, $50^{\circ}C$) for 70 seconds and death lethality increased in proportion to treatment temperature ($D_{50}=5.7min,\;D_{60}=\;2.1min,\;and\;D_{70}=0.7min$).

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Transfer of foreign Genes into the Bradyrhizobium japonicum and their Inoculation Effects on Soybean Plants (Bradyrhizobium japonicum에 외부유전자(外部遺傳子)의 도입(導入)과 대두(大豆)에 대한 접종효과)

  • Kim, Yong-Woong;Kim, Kil-Yong;Rhee, Young-Hwan;Kim, Kwang-Sik
    • Korean Journal of Soil Science and Fertilizer
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    • v.25 no.4
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    • pp.387-393
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    • 1992
  • The fate of inoculum strain of Bradyrhizobium japonicum was studied by using genetically marked strain. RJB6 $str^rnal^rneo^r$. A spontaneous mutant of B. japonicum isolated from nodules was made to have antibiotic resistance against streptomycin and nalidixic acid. In order to make genetically marked strain, neomycine resistant gene(Tn5) was introduced into this spontaneous mutant by conjugation with E. coli containing pSUP2021. The southern hybridization was carried out to confirm the plasmid insertion. Hybridization of chromosome DNA using pSUP2021(Tn5) as a probe showed that Tn5 was located on the 4.9kb fragment of chromosome. Soybean seeds were planted into a soil previously cultivated with soybean and inoculated with different cell densities of marked strain. Fourty days after planting, the inoculation effects on nodule number, nodule fresh weight, plant height and nitrogen content in the plot inoculated with heavy cell suspension was a little better than those in the plot with low inoculation. The recovery percentage of the marked strains was about 12% in the plot inoculated with heavy density cell suspension, while 5% in the plot inoculated with low cell suspension.

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Transferable R plasmid of Streptococci Ioslation from Diseased Olive Flounder (Paralichthys olivaceus) in Jeju (제주도 양식넙치병어에서 분리된 연쇄상구균의 약제내성 전이성 plasmid)

  • Kim, Jong-Hun;Lee, Chang-Hoon;Kim, Eun-Heui
    • Journal of fish pathology
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    • v.19 no.3
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    • pp.267-276
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    • 2006
  • Seventy-five streptococci were isolated from diseased olive flounder, Paralichthys olivaceus in Jeju. Their drug susceptibility and transferable multiple drug resistance were characterized. All isolates were resistant to flumequine (AR) and oxolinic acid (OA) and 26 isolates (34.7%) showed 4~6 multiple resistance of ampicillin (ABPC), AR, doxycycline (DOXY), erythromycin (EM), norfloxacin(NOR), OA and oxytetracycline (OTC) in various combinations. pST9 of a transferable R plasmid was detected from a multiple drug resistance strain, Streptococcus sp., ST9 originated from diseased flounder in Jeju, previously. We performed DNA hybridization to know the distribution of plasmid with the same DNA structure as pST9 in streptococci. Thirteen out of 60 isolates analyzed were positive in colony DNA hybridization and the part of bacteria isolated from raw meal was also hybridized with pST9. It suggested that raw meal is one of the origin of the resistance plasmid and R plasmid with DNA structure differing from pST9 is also involving in multiple drug resistance of the streptococci. In conjugation experiment, we found transferable R plasmid carrying OTC, DOXY and/or EM resistance determinant in the 13 resistance strains. all of the streptococci carrying the transferable R plasmid were similar in RAPD patterns. However, pST -type R plasmid was rare in S. iniae most frequently appearing in flounder farm.

Antimicrobial and Antioxidant activity of the Fruit of Prunus avium L (체리(Prunus avium L.)의 항균 및 항산화 활성)

  • Ahn, Seon-Mi;Ryu, Hee-Young;Kang, Dong-Kyoon;Jung, In-Chang;Sohn, Ho-Yong
    • Microbiology and Biotechnology Letters
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    • v.37 no.4
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    • pp.371-376
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    • 2009
  • The fruits of sweet cherry are highly appreciated by the consumer due to their precocity and quality, such as their sweetness, color and sourness. In this study, the hot-water extract and its sequential organic solvent fractions were prepared from domestic Napoleon sweet cherry (Prunus avium L.) to investigate antimicrobial and antioxidant activity. The hot-water extract contained about 40% sugars, and the solvent fraction yields for hexane, ethylacetate (EA), butanol, and water residue were 0.01%, 3.45%, 16.30%, and 80.24%, respectively. Contents of total polyphenol and total flavonoid of the fractions were 1.24~5.24%, and 0~3.76%, respectively. Among the fractions, EA fraction showed the highest total polyphenol and total flavonoid concentrations. Evaluation of antimicrobial activity of the extract and fractions revealed that EA fraction and butanol fraction contained strong antibacterial activity against Listeria monocytogenes, Staphylococcus aureus, and Salmonella typhimurium with minimal inhibitory concentration (MIC) of 0.5~1.0 mg/mL. But the extract and fractions tested were not active to Pseudomonas aeruginosa. In a while, only hexane fraction showed anti-Candida activity with 0.5~1.0 mg/mL of MIC. The fraction showed strong activity against different multi-antibiotics resistant strains such as C. albicans CCARM 14020. Antioxidative activity assay showed that EA fraction has a strong DPPH scavenging activity and a reducing power. The $IC_{50}s$ of vitamin E and EA fraction were 15.5 and $195.5\;{\mu}g/mL$, respectively. Our results suggest that the fruit of P. avium L. has high potentials with anti-Candida and antioxidative activity.

PILOT SCALE FISH PRODUCTION IN WATER RECYCLING SYSTEM (순환여과식 사육수정화 재사용방법에 의한 Pilot 규모의 어류양식시험)

  • KIM In-Bae
    • Korean Journal of Fisheries and Aquatic Sciences
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    • v.13 no.4
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    • pp.195-206
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    • 1980
  • Fish were grown in a water recycling rearing system consisting of a filter system, of which the area is about $30\;m^2$ and the volume of filter gravels is about $15\;m^3$, 6 rearing and 2 supplementary tans about $7\;m^2$ of surface area each. The results for 180 days from April 18 to October 14,1980 are summarized as following: One rearing tank was used for main experiment stocked with an equal number, and weight of each Israeli strain and $F_1$, hybrid between this strain and Korean local strain of common carp. Gross total production was 461 kg with a net production of 395 kg, and the Israeli strain grew to about 8 times initial weight and the hybrid about 6 times. Based on an analysis of results of the above mentioned main experiment and the other miscellaneous fish grown in the remaining tanks, it was considered to be feasible to implement a commercial production in this system by improving parts of the system and adding a number of the units. A special problem seems to be an adoption of more effective aeration device when fish loading increases. A solid waste removal system was deviced by use of by-pass flow ana this system worked very efficiently in view of supply water economy. A serious problem encountered in this experiment was heavy outbreaks of the gill-fluke, Dactylogyrus sp. which was relatively easily eradicated with Dipterex or DDVP treatment in the first trials, but later on strains which were resistant to any of these chemicals appeared.

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Insecticidal Activities against Major Lepidopteran Pests and Culture Condition of Bacillus thuringiensis sp. aizawai collected in Korea (국내에서 선발한 Bacillus thuringiensis sp. aizawai 균주의 주요 나방류 해충에 대한 살충 활성 및 배양특성)

  • Lee, Sang-Guei;Choi, Kee-Hyun;Lee, Young-Su;Oh, Kyeong-Seok;O, Jeung-Hun;Choi, Sung-Won
    • The Korean Journal of Pesticide Science
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    • v.10 no.2
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    • pp.131-137
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    • 2006
  • This experiment was conducted to select prominent microorganisms with a good insecticidal activity among the ten species, which isolated from soil at the near of Chung-buk, Chung-nam, and Gang-won provinces and made protein crystal endotoxin. As a result, GB-413 strain was finally selected, which showed the high insecticidal activity against susceptible diamondback moth (Plutella xylostella), beet army worm (Spodoptera exigua) and tobacco cutworm (Spodoptera litura) as well as resistant diamondback moth strains. By modifying the cultivation process f.g. lowing the glucose concentration at early cultivation stage and adding the carbon after inducing the spores, the percentage of making spore as well as the number of active spore were increased and the time for cultivation and spore forming was reduced without a reduction of insecticidal activity. These results were not only applied successfully for the optimized cultivation process for a fermentation tank containing five tons capacity, but also improved the possibility of mass cultivation for commercial production.