• Title/Summary/Keyword: gel electrophoresis

Search Result 2,290, Processing Time 0.022 seconds

Analysis of the Bacterial Composition During Kochujang, a Korean Traditional Fermented Hot Pepper-soybean Paste, Fermentation

  • Park, Sun-Jung;Chang, Jin-Hee;Cha, Seong-Kwan;Moon, Gi-Seong
    • Food Science and Biotechnology
    • /
    • v.18 no.4
    • /
    • pp.1035-1037
    • /
    • 2009
  • In this study we analyzed the dynamic changes in microbiota composition during kochujang fermentation at $30^{\circ}C$. During fermentation, the viable cell counts slowly increased and reached $3.2{\times}10^7$ for aerobic bacteria, $8.3{\times}10^3$ for yeast, and $1.4{\times}10^3$ CFU/mL for fungi after 60 days. Bacilli were found to be the most dominant microorganisms throughout the fermentation process. Using the culture dependent method Bacillus subtilis, Bacillus licheniformis, and Bacillus amyloquefaciens were found to be the main species during the early stages of fermentation; however, Bacillus pumilus and Bacillus stearothermophilus became the most dominant species during the late stage of fermentation. In contrast, when the polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) method was used Bacillus ehimensis was found to be the dominant species during the early stage of fermentation and Bacillus megaterium, B. pumilus, B. subtilis, and B. licheniformis were dominant in the ate stages. These results indicate various other Bacillus species rather than just B. subtilis and B. licheniformis might be involved in the fermentation of kochujang.

Molecular Association of Glucose Transporter in the Plasma Membrane of Rat Adipocyte

  • Hah, Jong-Sik
    • The Korean Journal of Physiology
    • /
    • v.25 no.2
    • /
    • pp.115-123
    • /
    • 1991
  • Molecular association of glucose transporters with the other proteins in the plasma membrane was assessed by gel electrophoresis and immunoblot techniques. Approximately $31.5{\pm}5.1%$ of GLUT-4, $64.8{\pm}2.7%$ of clathrin, 48.7% of total protein in the plasma membrane (PM) were found insoluble upon extraction with 1% Tx-100. Sodium dodecyl sulfate polyacrylamide gel electrophoresis revealed that the Tx-100 insoluble PM fraction contained about 4 major polypeptides with apparent molecular weight of above 200, 100-120, 80 and 30-35 KDa that were readily removed upon wash with a high pH buffer which is known to remove clathrin and 0.5 M Tris-buffer which is known to remove assembly proteins (AP). Immunoblotting of GLUT4 and clathrin against specific antibodies showed that GLUT-4 and clathrin were co-solubilized up to 84.6% and 82.7% respectively by wash with a high pH buffer and 1% Tx-100. When the membrane was pre-washed with a high pH buffer and 0.5 M Tris solution, GLUT4 and clathrin were not solubilized further suggesting that GLUT4 molecules are in molecular association with clathrin, AP and/or other extrinsic membrane proteins in plasma membrane and the formation of clathrin-coated structures might be involved in insulin stimulated glucose transporter translocation mechanism.

  • PDF

Purification and Characteristics of Pullulanase from Bacillus cereus subsp. mycoides (Bacillus cereus subsp. mycoides가 생산하는 Pullulanase의 정제와 특성)

  • Chung, Man-Jae;Woo, Jeong-Suk;Cho, Dae-Sun;Lee, Myong-Yur;Park, Nam-Kyu
    • Microbiology and Biotechnology Letters
    • /
    • v.22 no.1
    • /
    • pp.73-79
    • /
    • 1994
  • The optimum cultural temperature and time for the pullulanase production by Bacillus cereus subsp. mycoides were 35$\circ $C and 48 hrs, respectively. The addition of egg albumin and casein to the basal medium increased the enzyme production. The enzyme was purified by ammonium sulfate fractionation and DEAE-cellulose column chromatography. specific activity of the purified enzyme was 82.37 U/mg protein and yield of theenzyume activity was 62.1%. The purified enzuyme showed a single band on ployacrylamide disc gel electrophoresis and its molecular weight was estimated to be 66.,000 by SDS-polyacrylamide disc gel electrophoresis. The isoelcular point for the purified enzyme was pH 5.0. The optimum temperature and pH were 50$\circ $C and pH 6.5, respectively. The purified enzyme was stable below 40$\circ $C and in the pH range of 6.5~10.0 The pullulanase activity was greatly inhited by Ag$^{+}$, Hg$^{2+}$ and EDTA, and its heat stability was increased by the addition of Ca$^{2+}$. The tydrolysis product with the enzyme on pullulan was maltotriose.

  • PDF

Microbial Community Structure in Hexadecane- and Naphthalene-Enriched Gas Station Soil

  • Baek, Kyung-Hwa;Kim, Hee-Sik
    • Journal of Microbiology and Biotechnology
    • /
    • v.19 no.7
    • /
    • pp.651-657
    • /
    • 2009
  • Shifts in the activity and diversity of microbes involved in aliphatic and aromatic hydrocarbon degradation in contaminated soil were investigated. Subsurface soil was collected from a gas station that had been abandoned since 1995 owing to ground subsidence. The total petroleum hydrocarbon content of the sample was approximately 2,100 mg/kg, and that of the soil below a gas pump was over 23,000 mg/kg. Enrichment cultures were grown in mineral medium that contained hexadecane (H) or naphthalene (N) at a concentration of 200 mg/l. In the Henrichment culture, a real-time PCR assay revealed that the 16S rRNA gene copy number increased from $1.2{\times}10^5$to $8.6{\times}10^6$with no lag phase, representing an approximately 70-fold increase. In the N-enrichment culture, the 16S rRNA copy number increased about 13-fold after 48 h, from $6.3{\times}10^4$to $8.3{\times}10^5$. Microbial communities in the enrichment cultures were studied by denaturing gradient gel electrophoresis and by analysis of 16S rRNA gene libraries. Before the addition of hydrocarbons, the gas station soil contained primarily Alpha- and Gammaproteobacteria. During growth in the H-enrichment culture, the contribution of Bacteriodetes to the microbial community increased significantly. On the other hand, during N-enrichment, the Betaproteobacteria population increased conspicuously. These results suggest that specific phylotypes of bacteria were associated with the degradation of each hydrocarbon.

Comparative Study of Rhizobacterial Community Structure of Plant Species in Oil-Contaminated Soil

  • Lee, Eun-Hee;Cho, Kyong-Suk;Kim, Jai-Soo
    • Journal of Microbiology and Biotechnology
    • /
    • v.20 no.9
    • /
    • pp.1339-1347
    • /
    • 2010
  • In this study, the identity and distribution of plants and the structure of their associated rhizobacterial communities were examined in an oil-contaminated site. The number of plant species that formed a community or were scattered was 24. The species living in soil highly contaminated with total petroleum hydrocarbon (TPH) (9,000-4,5000 mg/g-soil) were Cynodon dactylon, Persicaria lapathifolia, and Calystegia soldanella (a halophytic species). Among the 24 plant species, the following have been known to be effective for oil removal: C. dactylon, Digitaria sanguinalis, and Cyperus orthostachyus. Denaturing gradient gel electrophoresis (DGGE) profile analysis showed that the following pairs of plant species had highly similar (above 70%) rhizobacterial community structures: Artemisia princeps and Hemistepta lyrata; C. dactylon and P. lapathifolia; Carex kobomugi and Cardamine flexuosa; and Equisetum arvense and D. sanguinalis. The major groups of rhizobacteria were Beta-proteobacteria, Gamma-proteobacteria, Chloroflexi, Actinobacteria, and unknown. Based on DGGE analysis, P. lapathifolia, found for the first time in this study growing in the presence of high TPH, may be a good species for phytoremediation of oil-contaminated soils and in particular, C. soldanella may be useful for soils with high TPH and salt concentrations. Overall, this study suggests that the plant roots, regardless of plant species, may have a similar influence on the bacterial community structure in oil-contaminated soil.

Phage typing and pulsed-field gel electrophoresis of Salmonella typhimurium and S enteritidis isolated from domestic animals in Gyeongbuk province (경북지역 가축에서 분리된 Salmonella typhimurium과 S enteritidis의 phage typing 및 pulsed-field gel electrophoresis)

  • Kim, Sang-Yun;Lee, Hee-Moo;Kim, Sin;Hong, Hyon-Pyo;Kwon, Heon-Il
    • Korean Journal of Veterinary Service
    • /
    • v.24 no.3
    • /
    • pp.243-253
    • /
    • 2001
  • Forty-five Salmonella typhimurium isolates were encountered 8 phage types in which DT197 and U302 were the predominant types. The DT104 type which was first found from pig in Korea, and was resistant to chloramphenicol, streptomycin, sulfamethoxazole/trimethoprim, tetracycline, gentamicin and nalidixic acid. Twenty-two S enteritidis isolates were encountered 5 phage types in which PT4 were the representative (predominant). S enteritidis isolates were susceptible to all antimicrobial agents. As a result of PFGE analysis for S typhimurium and S enteritidis, PFGE patterns was better than phage typing in discriminating of strains. PFGE patterns were not in accord with phage type even though some strain had the same phage types.

  • PDF

Antimicrobial resistance and pulsed -field gel electrophoresis patterns of Salmonella gallinarum isolated from broiler (육계에서 분리한 Salmonella gallinarum 의 약제내생 및 PFGE 양상)

  • Kim Seong-Guk;Kim Yeong-Hwan;Eom Hyun-Jung;Jang Seong-Jun;Jo Gwang-Hyeon;Lee Yang-Soo
    • Korean Journal of Veterinary Service
    • /
    • v.29 no.3
    • /
    • pp.297-308
    • /
    • 2006
  • Fowl typhoid (FT) is a septicemic disease caused by Salmonella gallinarum. The purpose of this study was to investigate the antimicrobial resistance and pulsed -field gel electrophoresis (PFGE) patterns of S gallinarum isolated from broiler. During 1999 to 2004, there was isolated a total of 26 strains in liver and spleen. The biochemical characteristics of S gallinarum isolates was nonmotile, no production of $H_2S$, glucose gas, non-fermented rhamnose, indole-negative, fermentation of dulcitol, mannitol, maltose, and ornithine decarboxylase. At antimicrobial susceptibility, all of isolates were susceptible to amoxicillin/clavulanic acid, amikacin, neomycin, kanamycin, and cephalothin. Twenty-six isolates were divided into 19 resistant patterns and 6 strains was 8-multi-drug resistance. PFGE of Xba I restriction fragments of S gallinarum isolates was 22 patterns.

Genotyping of Brucella abortus isolated in Gyeongbuk province by PFGE (PFGE를 이용한 경북지역에서 분리된 Brucella abortus의 유전형별)

  • Jo, Min-Hee;Kim, Seong-Guk;Kim, Young-Hoan;Kim, Soon-Tae;Eom, Hyun-Jung;Jang, Young-Sui;Ko, Young-Hwal
    • Korean Journal of Veterinary Service
    • /
    • v.32 no.3
    • /
    • pp.257-264
    • /
    • 2009
  • Subtyping of Brucella abortus isolates is epidemiologically important for monitoring of bovine brucellosis outbreaks. Pulsed-field gel electrophoresis (PFGE) is considered as a gold standard of molecular typing methods to study the DNA polymorphisms of bacteria. In this study, we analyzed using PFGE the DNA fragment profiles of B. abortus isolated in Gyeongbuk province from 1998 to 2006. The genomic DNA was digested with the restriction endonuclease Xba I, Xho I and Smi I followed gel electrophoresis. No distinguishable patterns of the genomic DNA digested with Xba I and Xho I were observed among the field isolates of B. abortus tested in this study. But Smi I restriction enzyme resulted in two PFGE patterns consisting of 13-15 bands that ranged in size from 33 to 668bp by standard marker. The cluster analysis by DNA fingerprinting software showed 93.75% similarity between two PFGE patterns. No different PFGE patterns were recognized among the isolates originated from various years, regions and cow breeds.

Studies on the Purification and Biochemical Properties of Vitellin in the Antheraea yamamai Guerin-Meneville I. Isolation and Purification of Vitellin and its Change to Embryonic Development (천잠(Antheraea yamamai) Vitellin의 분리와 생화학적 특성에 관한 연구 I. Vitellin의 분리와 동정 및 배자발생에 따른 변동)

  • 김철명;문재유
    • Journal of Sericultural and Entomological Science
    • /
    • v.31 no.2
    • /
    • pp.72-81
    • /
    • 1989
  • Antheraea yamamai vitellin was purified from matured eggs by polyacryamide gel electrophoresis, also stage dependent appearance, immunological comparison and relative content of the protein were investigated. 1. Vitellogenin, the precursor of vitellin, was first detected in the larval hemolymph at the late spinning stage by polyacrylamide gel electrophoresis and immunoelectrophoresis. 2. The electrophoretic mobility of the vitellin was identical with that of Bombyx mori and of Bombyx mandarina. However, the specific antiserum against A. uamamai vitellin did not react with either that of Bombyx mori or Bombyx mandarina in immumo-diffusion test. 3. Relative content of A. yamamai vitellin to the total soluble egg protein was 46.0 percent and did not change till eight days after oviposition. But the content started to decline from ten days after oviposition and was negligible in the five or serventeen month old eggs.

  • PDF

Smoking Related DNA Damage in Human Lymphocytes Assessed by the Comet Assay (단세포전기영동법으로 평가한 흡연자의 백혈구 DNA손상)

  • 선수진;정해원;한정호
    • Environmental Mutagens and Carcinogens
    • /
    • v.22 no.2
    • /
    • pp.83-89
    • /
    • 2002
  • The single cell gel electrophoresis (comet) assay is one of the useful tools for the study of genetic damage in humans exposed to environmental mutagens and carcinogens. This study was undertaken to evaluate the status of DNA damage in peripheral lymphocytes depending on their sex, age, smoking habits, and other factors in normal healthy Korean population. The 99 volunteers included in the study and out of these, 36 volunteers were smoker and 63 volunteers were non-smoker aged between 20-59 years. All individual answered a questionnaire that assessed their general information including smoking habits and the extent of the environmental tobacco smoke (ETS) exposure, and blood samples were obtained. There was a statistically significant difference in the extent of DNA damage between smoker and non-smoker (p<0.001). A significant difference was also observed between male and female (p<0.001) and amongst the different group of age (p<0.005), however, correlation analysis showed that only smoking habit was a significant factor for DNA damage. No significant effect of smoking duration, number of cigarettes smoking a day, SPY (smoke pack years) in smokers and environmental tobacco smoke exposure in non-smokers on the status of DNA damage was observed.

  • PDF