• Title/Summary/Keyword: Strain specificity

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Synchronization of Cell Cycle in Korean Hydrogen Producing Cyanobacterial Strains (한국산 수소생산 남세균 종주들의 세포주기 동조화)

  • Park, Jong-Woo;Ahn, Se-Hee;Kim, Hyung-Seop;Yih, Won-Ho
    • Journal of Hydrogen and New Energy
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    • v.22 no.5
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    • pp.663-670
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    • 2011
  • Under a daily photoperiod of 14h light and 10h dark synchronization of cell cycle in Korean Cyanothece spp. strains and $Synechococcus$ sp. strain Miami BG043511 was analyzed as to be applicable to enhanced hydrogen production. For all strains peaks of double cell were observed during the light period of a daily cycle. Peaks of maximal cell size measured by a coulter counter appeared at the peak of double cells observed under light microscope reconfirming the synchronization of daily cell cycle. The cell cycle synchronization became weakened within two days when treated with continuous illumination. Rapid detection of the peak time of double cell percentage by coulter counters may contribute to quasi-realtime feedback control for efficient production of photobiological hydrogen by unicellular cyanobacterial strains.

Plant Terpene-Induced Expression of Multiple Aromatic Ring Hydroxylation Oxygenase Genes in Rhodococcus sp. Strain T104

  • Kim, Byung-Hyuk;Oh, Eun-Taex;Ahn, Yeong-Hee;Koh, Sung-Cheol
    • Journal of Microbiology
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    • v.41 no.4
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    • pp.349-352
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    • 2003
  • Recent studies have shown that some of the PCB (polychlorinated biphenyl)-degraders are able to effectively degrade PCB in the presence of monoterpenes, which act as inducers for the degradation pathway. Rhodococcus sp. T104, an effective PCB degrader, has been shown to induce the degradation pathway by utilizing limonenes, cymenes, carvones, and pinenes as sole carbon sources which can be found in the natural environment. Moreover, the strain T104 proved to possess three separate oxidation pathways of limonene, biphenyl, and phenol. Of these three, the limonene can also induce the biphenyl degradation pathway. In this work, we report the presence of three distinct genes for aromatic oxygenase, which are putatively involved in the degradation of aromatic substrates including biphenyl, limonene, and phenol, through PCR amplification and denaturing gradient gel electrophoresis (DGGE). The genes were differentially expressed and well induced by limonene, cymene, and plant extract A compared to biphenyl and/or glucose. This indicates that substrate specificity must be taken into account when biodegradation of the target compounds are facilitated by the plant natural substrates.

A pathogen of New Zealand Pyropia plicata (Bangiales, Rhodophyta), Pythium porphyrae (Oomycota)

  • Diehl, Nora;Kim, Gwang Hoon;Zuccarello, Giuseppe C.
    • ALGAE
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    • v.32 no.1
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    • pp.29-39
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    • 2017
  • Geographic distributions of pathogens are affected by dynamic processes involving host susceptibility, availability and abundance. An oomycete, Pythium porphyrae, is the causative agent of red rot disease, which plagues Pyropia farms in Korea and Japan almost every year and causes serious economic damage. We isolated an oomycete pathogen infecting Pyropia plicata from a natural population in Wellington, New Zealand. The pathogen was identified as Pythium porphyrae using cytochrome oxidase subunit 1 and internal transcribed spacer of the rDNA cistron molecular markers. Susceptibility test showed that this Pythium from New Zealand was able to infect several different species and genera of Bangiales including Pyropia but is not able to infect their sporophytic (conchocelis) phases. The sequences of the isolated New Zealand strain were also identical to Pythium chondricola from Korea and the type strain from the Netherlands. Genetic species delimitation analyses found no support for separating P. porphyrae from P. chondricola, nor do we find morphological characters to distinguish them. We propose that Pythium chondricola be placed in synonymy with P. porphyrae. It appears that the pathogen of Pyropia, both in aquaculture in the northern hemisphere and in natural populations in the southern hemisphere is one species.

Flavonoids Biotransformation by Human Gut Bacterium Dorea sp. MRG-IFC3 Cell-Free Extract

  • Huynh Thi Ngoc Mi;Heji Kim;Jong Suk Lee;Bekir Engin Eser;Jaehong Han
    • Journal of Microbiology and Biotechnology
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    • v.34 no.6
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    • pp.1270-1275
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    • 2024
  • Human gut bacterium Dorea sp. MRG-IFC3 is unique in that it is capable of metabolizing puerarin, an isoflavone C-glycoside, whereas it shows broad substrate glycosidase activity for the various flavonoid O-glycosides. To address the question on the substrate specificity, as well as biochemical characteristics, cell-free biotransformation of flavonoid glycosides was performed under various conditions. The results showed that there are two different enzyme systems responsible for the metabolism of flavonoid C-glycosides and O-glycosides in the MRG-IFC3 strain. The system responsible for the conversion of puerarin was inducible and comprised of two enzymes. One enzyme oxidizes puerarin to 3"-oxo-puerarin and the other enzyme converts 3"-oxo-puearin to daidzein. The second enzyme was only active toward 3"-oxo-puerarin. The activity of puerarin conversion to daidzein was enhanced in the presence of Mn2+ and NAD+. It was concluded that the puerarin C-deglycosylation by Dorea sp. MRG-IFC3 possibly adopts the same biochemical mechanism as the strain PUE, a species of Dorea longicatena.

Avirulence Gene Diversity of Xanthomonas axonopodis pv. glycines Isolated in Korea

  • Park, Hyoung-Joon;Han, Sang-Wook;Oh, Chang-Sik;Lee, Seung-Don;Ra, Dong-Soo;Lee, Suk-Ha;Heu, Sung-Gi
    • Journal of Microbiology and Biotechnology
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    • v.18 no.9
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    • pp.1500-1509
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    • 2008
  • The hybridization patterns with the avrBs3 gene that is known to determine the recognition of host specificity were used to study the diversity of Xanthomonas axonopodis pv. glycines causing bacterial leaf pustule in soybean. A total of 155 strains were isolated from diverse tissues of soybean cultivars collected in Korea and were classified into six different type strains of OcsF, SL1017, SL1018, SL1045, SL1157, and SL2098 according to the patterns of avrBs3-homologous bands. When these type strains were inoculated on various cultivars, most of the Korean strains mildly induced disease symptoms on the resistant CNS1 cultivars. Unlike other type strains, strain SL2098, which appeared not to contain any avrBs3 homolog, induced only a few pustules on even highly susceptible cultivars. When a plasmid carrying the 3.7-kb avrBs3-homologous gene from strain SL1045 was introduced into SL2098, the transformant could not recover the pathogenicity in susceptible host plants. However, when avrBs3-homologous genes of strain SL1018 were mutated by transposon mutagenesis, one of the mutants in which a 5.2-kb chromosomal band homologous to avrBs3 was disrupted could not induce the hypersensitive response on resistant cultivars such as William82 or CNS2. Our results suggest that the avrBs3 homologs may play important roles in the pathogenicity of Xanthomonas axonopodis pv. glycines and the recognition of soybean cultivars.

Studies on avian infectious bronchitis: II. Standardization of an indirect enzyme-linked immunosorbent assay (ELISA) for antibody measurement (닭 전염성 기관지염에 관한 연구: II. 간접 enzyme-linked immunosorbent assay(ELISA)에 의한 항체가 측정)

  • Chang, Chong-ho;Kim, Sun-joong
    • Korean Journal of Veterinary Research
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    • v.29 no.4
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    • pp.503-515
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    • 1989
  • Critical parameters affecting sensitivity and specificity of an enzyme-linked immunosorbent assay(ELISA) for detection of antibodies to avian infections bronchitis virus(IBV) were standardized. By adopting the optimized conditions an equation calculating ELISA antibody titers from the observations at single serum dilution was formulated. The purified antigen of IBV Mass-41 strain was dispensed into polystyrene microplate wells at a concentration of 300ng per well($100{\mu}l$) and the plates were coated by completey drying at $37^{\circ}C$. Diluted chicken serum and horseradish peroxidase conjugated goat anti-chicken IgG were added in order in $100{\mu}l$ volumes per well and allowed to react for 30 minutes each at room temperature. Just before use and after each reaction the plates were washed three times with distilled water. Finally o-phenylenediamine solution was added as an enzyme substrate. After incubation for another 15 minutes at room temperature absorbances were read at 492nm. Hyperimmune serum against Mass-41 strain was used as internal reference positive(IRP) serum. After repeated titration of IRP and negative serum, a constant titer of IRP was determined. Serum titrations were carried out for various sample sera together with IRP and negative sera and the observed titers of sample sera were corrected by reflecting the ratio between observed and constant titers of IRP serum. These corrected titers of the sample sera were plotted against sample/positive(S/P) OD ratios. All the OD's measured in the serum titrations were also corrected by substracting negative serum OD. The following equation was formulated from the above data; $Log_{10}$ ELISA titer=$5.568({\log}_{10}S/P)+4.161$ Thus it was possible to calculate ELISA titer by measuring absorbance at 1/400 single serum dilution. Titer measured by cross ELISA tests employing Mass-41 strain and three local IBV isolates were similar. These results suggest that the ELISA tests standardized in this study can be used for evaluating not only vaccinal immunity but also for infection status against fields IBV's.

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Evaluation and cloning of a (R)-stereospecific esterase from Bacillus stearothermophilus JY144

  • Kim, Ji-Yeon;Kim, Yun-Jeong;Choe, Gi-Seop;Kim, Geun-Jung;Yu, Yeon-U
    • 한국생물공학회:학술대회논문집
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    • 2002.04a
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    • pp.457-460
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    • 2002
  • In an effort to isolate novel strains expressing a thermostable esterase that hydrolyzed the rac-ketoprofen ethyl ester to ketoprofen in the stereospecific manner, we screened various soils and composts from broad ecological niches in which the activity was expected to be found. Three hundreds of microbial strains were tested to determine their ester-hydrolyzing activity by using an agar plate containing insoluble tributyrin as an indicative substrate, and then further screened by activity on the (R,S)-ketoprofen ethyl ester. Twenty-six strains were screened primarily at high growth and incubation temperature and further compared the ability to ethyl ester-hydrolyzing activity in terms of conversion yield and chiral specificity. Consequently, a strain JYl44 was isolated as a novel strain that produced a (R)-stereospecific esterase with high stability and systematically identified as a Bacillus stearothermophilus JY144. The enzyme indeed stables at a broad range of temperature, upto 65 $^{\circ}C$, and pH ranging from 6.0 to 10.0. The optimal temperature and pH for enzymatic conversion were 50 $^{\circ}C$ and 9.0, respectively. Based on the observations that resulted a poor cell growth, and enzyme expression in wild type strain, we further attempted the gene cloning into a general host Escherichia coli and determined its primary structure, concomitantly resulting a high level expression of the enzyme. The cloned gene had an open reading frame (250 amino acids) with a calculated molecular mass of 27.4 kDa, and its primary structure showed a relative high homology (45-52 %) to the esterases from Streptomyces and Bacillus strains. The recombinant whole cell enzyme could efficiently convert the rac-ketoprofen ethyl ester to (R)-ketoprofen, with optical purity of 99 % and yield of 49 %.

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Polyclonal Antibody against Paenibacillus larvae and its Application (Paenibacillus larvae에 대한 다클론 항체 및 그 응용)

  • 백경찬;양옥순;정규회;윤병수
    • Korean journal of applied entomology
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    • v.41 no.1
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    • pp.49-53
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    • 2002
  • Paenibacillus larvae is a gram-positive, spore-forming bacterium that is etiological agent for american foulbrood disease (AFB), which is the most severe disease in honey bee. To detect P. larvae from infected honeybee-comb or larvae, polyclonal antibody against whole bacterium was produced from guineapig and its specificity was evaluated. After optimization of ELISA-based detection system using these antibodies, a number of different P. larvae strains were analysed. Polyclonal antibody against P. larvae ATCC 25747 showed high affinity to most strains of P. larvae including P. larvae. strain ATCC 9545 (type strain), ATCC 25747 and other korean strain, SJl5 but exhibited no cross-reaction with other bacterial species. Additionally, this type of ELISA system was used for the detection of AFB in field-application The results have shown that this antibody could be useful for the rapid identification and monitoring of P. larvae in honeybee-comb.

Optimal Temperature for H2 Production and Population Growth of the N2-fixing Unicellular Cyanobacterial Strains from Korean Coasts (한국 연안산 질소고정 단세포 남세균 종주의 최적 성장 및 수소생산 온도)

  • Park, Jongwoo;Kim, Hyungseop;Yih, Wonho
    • Journal of Hydrogen and New Energy
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    • v.24 no.1
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    • pp.20-28
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    • 2013
  • Photobiological hydrogen production by nitrogen-fixing unicellular cyanobacteria has long been considered to be an environmentally sound and very promising method for the future supply of renewable clean energy. Using six Korean nitrogen-fixing unicellular cyanobacterial strains and the Synechococcus sp. strain Miami BG043511 we performed cultivation experiments to find out the strain-specific optimal temperature for population growth and $H_2$ production. Under $20^{\circ}C$ the population growth of all the tested strains was significantly retarded in contrasts to the faster and higher growth under 25, 30 or $35^{\circ}C$. The highest growth rates in all the 7 strains were measured under $30^{\circ}C$ while the maximal biomass yields were under $30^{\circ}C$ (strains CB-MAL 026, 054, and 055) or $35^{\circ}C$ (strains 002, 031, 058, and Miami BG043511). The difference between the maximal biomass yields at $30^{\circ}C$ and $35^{\circ}C$ was not greater than 10%. The quantity of photobiologically produced $H_2$ was only slight larger under $35^{\circ}C$ than that under $20^{\circ}C$. Our result may suggest a two-step process of $H_2$ production which includes rapid and sizable production of biomass at $30^{\circ}C$ and the following high $H_2$ production at $20^{\circ}C$ by the test strains of marine nitrogen-fixing unicellular cyanobacteria.

Studies on the mechanisms of suppression in Aspergillus nidulans (Aspergillus nidulans의 역제의 유전자의 작용기작에 관한 연구 II)

  • 문은표;강현삼
    • Korean Journal of Microbiology
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    • v.20 no.1
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    • pp.1-10
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    • 1982
  • From Aspergillus nidulans, six suppressor mutants, MSI-MS6, were isolated, and their characteristics were analysed. These were the suppressor mutants for acriflavin resistant and nicotinamide auxotrophic mutant phenotypes. MS1, MS2, MS5 and MS6, were linked to the chromosome IV, I, II respectively, and MS2 was linked to one of the rest chromosomes, MS3 and MS6 mutants had both suppressors for acriflavin resistant marker and for nicotinamide auxotrophic marker. In order to know the stability and efficiency of the suppressors, their reversion frequencies, that is, frequencies of losing the suppressibility, were analysed. Only MS3 and MS5 were reversed with high frequency. The four mutants didn't lose their suppressibilities, and this meant that the suppressors of these four were very stable and highly effcient. The suppressor specificities of these mutants were tested for other mutant's phenotype marker. One of the six suppressors, MS1, had the suppressor specificity for acriflavin resistant marker of 163 strain.

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