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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Microbiology and Biotechnology
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 10, Issue 6 - Dec 2000
Volume 10, Issue 5 - Oct 2000
Volume 10, Issue 4 - Aug 2000
Volume 10, Issue 3 - Jun 2000
Volume 10, Issue 2 - Apr 2000
Volume 10, Issue 1 - Feb 2000
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Mechanism Used by White-Rot Fungus to Degrade Lignin and Toxic Chemicals
Chung, Nam-Hyun ; Lee, Il-Seok ; Song, Hee-Sang ; Bang, Won-Gi ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 737~752
Wood-rotting basidiomycetous fungi are the most efficient degraders of lignin on earth. The white-rot fungus Phanerochaete chrysosporium has been used as a model microorganism in the study of enzymology and its application. Because of the ability of the white-rot fungus to degrade lignin, which has an irregular structure and large molecular mass, this fungus has also been studied in relation to degrading and mineralizing many environmental pollutants. The fungus includes an array of enzymes, such as lignin peroxidase (LiP), manganese-dependent peroxidase (MnP), cellobiose:quinone oxidoreductase, and
-producing enzymes and also produces many other components of the ligninolytic system, such as veratryl alcohol (VA) and oxalate. In addition, the fungus has mechanisms for the reduction of degradation intermediates. The ligninolytic systems have been proved to provide reductive reactions as well as oxidative reactions, both of which are essential for the degradation of lignin and organopollutants. Further study on the white-rot fungus may provide many tools to both utilize lignin, the most abundant aromatic polymer, and bioremediate many recalcitrant organopollutants.
Secretory Production of Human Leptin in Bacillus subtilis
Jeong, Ki-Jun ; Lee, Sang-Yup ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 753~758
Human leptin is identified as a 16kDa (146 amino acids) protein secreted from adipocytes which influences body weight homeostasis. In order to produce active leptin, the human obese gene coding for leptin was expressed in Bacillus subtilis WB600 strain which is deficient in six extracellular proteases. The recombinant leptin was produced in a culture supernatant, and in a culture supernatant, it was contained as high as 48% for total proteins. After simple purification steps, which consisted of ammonium sulfate precipitation and anion-exchange column chromatography, 2.3 mg of leptin with a purity greater than 95% was obtained from the 0.51 culture with the recovery yield of 38.3%. The purified leptin showed the correct folding structure with one disulfied bond.
Biodegradation of 2,4,5-Trichlorophenol Using Cell-Free Culture Broths of Phanerochaete chrysosporium
Choi, Sueh-Yung ; Moon, Seung-Hyeon ; Lee, Jae-Suk ; Gu, Man-Bock ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 759~763
Cell-free culture broth of Phanerochaete chrysosporium has been adopted to biologically degrade 2,4,5-trichlorophenol. Two different medium compositions of nitrogen-sufficient and nitrogen-limited were compared for their distribution of isozymes, activity of lignin peroxidase, and production of oxalate. The two different culture broths were tested for their ability to degrade 2,4,5-trichlorophenol, and the biodegradation efficiency was estimated in terms of the disappearance of 2,4,5-trichlorophenol. The degradation efficiency for the nitrogen-limited culture broth was higher than that of the nitrogen-sufficient culture broth, since the nitrogen-limited culture broth induced lignin peroxidases (LiPs) and manganese peroxidases (MnPs), and contained sufficient oxalate for producing necessary radicals. Finally, the possible mechanism of 2,4,5-CP degradation using the nitrogen-limited culture broth was proposed.
Substitution of Glycine 275 by Glutamate (G275E) in Lipase of Bacillus stearothermophilus Affects Its Catalytic Activity and Enantio- and Chain Length Specificity
Kim, Myung-Hee ; Kim, Hyung-Kwoun ; Oh, Byung-Chul ; Oh, Tae-Kwang ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 764~769
The lipase gene(lip) from Bacillus stearothermophilus was recombined in vitro by utilizing the DNA shuffling technique. After four rounds of shuffling, transformation, and screening based on the initial rate of clear zone formation on a tricaprylin plate, a clone (M10) was isolated, the cell extract of which showed about 2.8-fold increased lipase activity. The DNA sequence of the mutant lipase gene (m10) showed 3 base changes, resulting in two cryptic mutations and one amino acid substitution: S113(
), L252 (
), and G275E (
). SDS-PAGE analysis revealed that the increased enzyme activity observed in M10 was partly caused by high expression of the m10 lipase gene. The amount of the expressed G275E lipase was estimated to comprise as much as 41% of the total soluble proteins of the cell. The maximum velocity (
) of the purified mutant enzyme for the hydrolysis of olive oil was measured to be 3,200 U/mg, which was 10% higher than that of the parental (WT) lipase (2,900 U/mg). Its optimum temperature for the hydrolysis of olive oil was
and it showed a typical
-dependent thermostability, properties fo which were the same as those of the WT lipase. However, the mutant enzyme exhibited a high enantiospecificity towards (S)-naproxen compared with the WT lipase. In addition, it showed increased hydrolytic activity towards triolein, tricaprin, tricaprylin, and tricaproin.
Solid Substrate and Submerged Culture Fermentation of Sugar Cane Bagasse for the Production of cellulase and Reducing Sugars by a Local Isolate, Aspergillus terreus SUK-1
Wan Mohtar, Yusoff ; Massadeh, Muhannad Illayan ; Kader, Jalil ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 770~775
Several process parameters were studied to ascertain the effect on degradation of sugar cane bagasse in relation to the production of cellulase enzyme and reducing sugars by Solid Substrate Fermentation (SSF) and Submerged Culture Fermentation (SCF) of Aspergillus terreus SUK-1. The effect of air-flow rate (0-1.3 v/v/m), of different ratios of substrate weight to liquid volume (1:6, 1:10, 1:20, and 1:30 w/v, g/ml), scale-up effect (10, 20, and 100 times of 1:10 ration, w/v) and the effect of temperature (30, 40, 50, and
) in SSF were studied. Air-flow rate of 1.0 v/v/m gave the highest enzyme activity (FPase 0.25 IU/ml, CMCase 1.24 IU/ml) and reducing sugars concentration (0.72 mg/ml). Experiment using 1:10 ratio (w/v) was found to support maximum cellulase activity (FPase 0.58 IU/ml, CMCase 1.97 IU/ml) and reducing sugar concentration (1.23 mg/ml). Scaling-up the ratio of 1:10(w/v) by a factor of 20 gave the highest cellulase activity (FPase 0.71 IU/ml, CMCase 2.25 IU/ml) and reducing sugar concentration (3.67 mg/ml). The optimum temperature for cellulase activity and reducing sugar production was
(FPase 0.792 IU/ml, CMCase 2.25 IU/ml and 3.85 mg/ml for reducing sugar concentration). For SCF, the activity of cellulase enzyme and reducing sugar concentration was found to be lower than that obtained for SSF. The highest cellulase activity obtained in SCF was 50% lower than the highest cellulase activity in SSF, while for reducing sugar concentration, the highest concentration obtained in SCF was 90% lower than that obtained in SSF.
Fermentation and Sporulation Characteristics of Saccharomyces cerevisiae SHY111 Isolated from Korean Traditional Rice Wine
Kim, Seung-Hwan ; Chung, Oon-Chan ; Woo, Im-Sun ; Shin, Jae-Ho ; Rho, Dong-Hyun ; Rhee, In-Koo ; Park, Heui-Dong ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 776~783
Various alcohol yeast strains have been isolated from main mashes of Korean traditional liquors, and their genetic diversities were previously reported . In this study, the strain SHY111, showing the highest alcohol production, was tested for its fermentation and sporulation characteristics. Additionally, its haploid cells were isolated and tested for their growth and fermentation patterns. The strain was identified as Saccharomyces cerevisiae based on its morphological and physiological characteristics. The sequences of the ITS(internal transcribed spacer) and 5.8S rDNA regions of S. cerevisiae SHY111 were found to be identical to those of S. cerevisiae that was obtained from through the yeast genome project. The maximum fermentation ratio obtained by the strain SHY111 (96.7%) was almost the same as that by S. cerevisiae Balyun No. 1 (96.5%) that was a little higher than that by S. cerevisiae KCCM11215(95.8%). The strain was induced for sporulation in a sporulation liquid medium using log phase cells grown in different types of pre-sporulation media, and its haploid cells were obtained by spore dissection using a micromanipulator. The majority of the spores formed a small colony on a YPD agar plate, and the haploid yeast cells derived from the strain SHY111 showed a variety of growth and alcohol fermentation patterns. It was proposed that the fermentation patterns were related to their growth phenotypes in the most haploid strains, but possible not in some strains.
Production of Exo-Polymers by Submerged Mycelial Culture of Cordyceps militaris and Its Hypolipidemic Effect
Yang, Byung-Keun ; Ha, Ji-Young ; Jeong, Sang-Chul ; Das, Surajit ; Yun, Jong-Won ; Lee, Young-Se ; Choi, Jang-Won ; Song, Chi-Hyun ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 784~788
Hypolipidemic effect of exo-polymers(EPs) form submerged mycelial culture of Cordyceps militaris was investigated in male Sprague-Dawley rats. For a dose-dependent study, EPs were administered at the level of 50-100 mg/kg body weight (BW) and the data was compared with the saline administered control group. A significant reduction of both the plasma total cholesterol (TC) and triglyceride (TG) was registered under the influence of EPs. It reduced the lowe density lipoprotein (LDL) cholesterol level as much as 40.5%. Levels of high density lipoprotein (HDL) choloesterol did not vary significantly within the different experimental groups, but the HDL: TC ratio showed consistently a high value with the increasing dose. The effects of cultural conditions (pH and temperature) in mycelial growth and EPs production were studied. Both the biomass and EPs were produced in a wide range of pH and temperature.
Effects of tktA,
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, volume 10, issue 6, 2000, Pages 789~796
In order to analyze the effects of tktA,
, and aroL expression in a tryptophan-producing Escherichia coli, a series of plasmids carrying the genes were constructed. Introduction of tktA,
, 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
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
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
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,
, 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.
Generation and Characterization of Cell-Permeable Greem Fluorescent Protein Mediated by the Basic Domain of Human Immunodeficiency Virus Type 1 Tat
Park, Jin-Seu ; Kim, Kyeong-Ae ; Ryu, Ji-Yoon ; Choi, Eui-Yul ; Lee, Kil-Soo ; Choi, Soo-Young ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 797~804
The human immunodeficiency virus type 1 (HIV-1) Tat is one of the viral gene products essential for HIV replication. The exogenous Tat protein is transduced through the plasma membrane and then accumulated in a cell. The basic domain of the Tat protein, which is rich in arginine and lysine residues and called the protein transduction domain (PTD), has been identified to be responsible for this transduction activity. To better understand the nature of the transduction mediated by this highly basic domain of HIV-1 Tat, the Green Fluorescent Protein (GFP) was expressed and purified as a fusion protein with a peptide derived from the HIV-1 Tat basic domain in Escherichia coli. The transduction of Tat-GFP into mammalian cells was then determined by a Western blot analysis and fluorescence microscopy. The cells treated with Tat-GFP exhibited dose- and time-dependent increases in their intracellular level of the protein. the effective transduction of denatured Tat-GFP into both the nucleus and the cytoplasm of mammalian cells was also demonstrated, thereby indicating that the unfolding of the transduced protein is required for efficient transduction. Accordingly, the availability of recombinant Tat-GFP can facilitate the simple and specific identification of the protein transduction mediated by the HIV-1 Tat basic domain in living cells either by fluorescence microscopy or by a fluorescence-activated cell sorter analysis.
Purification and Characterization of a Novel Antifungal Protein from Paenibacillus macerans PM1 Antagonistic to Rice Blast Fungus, Pyricularia oryzae
Bae, Dong-Won ; Kawk, Weon-Sik ; Lee, Joon-Taek ; Son, Dae-Young ; Chun, Sung-Sik ; Kim, Hee-Kyu ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 805~810
An antifungal protein antagonistic to the rice blast fungus, Pyricularia oryzae was purified from Paenibacillus macerans PM-1 by ammonium sulfate fractionation, Q Sepharose Fast Flow column chromatography, Phenyl Sepharose CL-4B column chromatography and Superose 12 gen filtration. An apparent molecular mass of the purified antifungal protein was determined as 8 kDa by SDS-PAGE and 9 kDa by analytical gel filtration, respectively, suggesting that the purified protein is a monomer. The antifungal protein was stable at pH range from 7-12 and up to
. The protein was also stable at 0.1-1% Tween 20 and Triton X-100. The N-terminal amino acid sequence of the antifungal protein was Thr-Glu-Leu-Pro-Leu-Gly-Ile-Val-Met-Asp-Lys-Tyr-Thr-Asp-Ala-Phe-Lys-Phe-Asp-Met-Phe. Comparison of the determined sequence with other peptide and DNA sequences did not reveal homology at all. Therefore, the purified antifungal protein was speculated to be a novel protein. The condidial germination in vitro of P. oryzae KJ301:93-39 by the purified protein (
) was limited to
only, compared with
of the control. Ungerminated conidia were swollen at basa and mid cell by the purified protein. In vivo bioassay for inhibition of conidial germination of P. oryzae KJ 301, one of the most predominating racesin Korea. the purified protein (
)strongly inhibited the conidial germination. The conidia, even though germinated, could not develop any further to produce appressoria efficiently.
Isolation of Bacterial Strain Antagonistic to Pyricularia oryzae and Its Mode of Antifungal Action
Bae, Dong-Won ; Kawk, Weon-Sik ; Lee, Joon-Taek ; Son, Dae-Young ; Chun, Sung-Sik ; Kim, Hee-Kyu ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 811~816
An antagonistic bacterium PM-1 which strongly inhyibits the growth of Pyricularia oryzae was isolated and identified as paenibacillus macerans. The antifungal substances of the strain PM-1 showed the broad antifungal spectra against P.oryzae races. Relating to the localization test, it was found that the antifungal substances existed not only in the cytoplasm but also in the culture supernatant, and importantly the antifungal activity of the latter was stronger than that of the former. The extracellular antifungal substances were extremely heat-stable up to
for 15 min. The substances were optimally produced at
and pH 10.0 in a potato dextrose broth. The culture filtrate of the strain PM-1 caused a partial swelling of the mycelia of P.oryzae, and it prevents the normal growth of the fungus as well. This result suggested that the antifungal substances secreted by the strain PM-1 potentially inhibited the germination of P.oryzae.
Effect of Flashing Light on Oxygen Production Rates in High-Density Algal Cultures
Park, Kyong-Hee ; Kim, Dong-Il ; Lee, Choul-Gyun ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 817~822
A proper flashing light is expected to enhance microalgal biomass productivity and photosynthetic efficiency. The effect of flashing light on high-density Chlorella kessleri (UTEX 398) cultures was studied using light-emitting diodes. A frequency modulator was designed to flash LEDs, and the device successfully provided wide range of frequencies and various duty cycles of flashing. A relatively high frequencies of 10, 20 and 50 kHz were used in this study. These frequencies have very short flashing time (
), which corresponded to the time constant of the light reaction of photosynthesis. The specific oxygen production rates of photosynthesis under flashing light were compared with those under an equivalent continuous light in specially designed illumination cuvette. The specific oxygen production rates under flashing light were 5-25% higher than those under the continuous light. A range of cell concentration was discovered, where the benefit of flashing light was maximized. The photosynthetic efficiency was also higher under flashing light with frequencies of over 1 kHz, which was a clear indication of flashing light effect and the degree of mutual shading could by overcome by flashing lights, particularly at high-density algal cultures.
Phylogenetic Analysis of Hepatitis B Virus Genome Isolated from Korean Patient Serum
Kim, Seon-Young ; Kang, Hyen-Sam ; Kim, Yeon-Soo ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 823~828
The complete nucleotide sequence of hepatitis B virus DNA isolated from Korean patient serum was determined and characterized, and its phylogenetic relation was then investigated. The viral genome was 3,215 base pairs long and included four well known open reading frames (i.e. surface antigens, core antigens, X protein and DNA polymerase). The sequence of the surface antigen showed that the HBV genome under investigation, designated HBV 315, was characteristic of subtype adr. A phylogenetic analysis using the total genome sequence revealed that HBV315 was grouped into genomic group C together with isolates from Japan, China, Thailand, Polynesia, and New Caledonia. The mean percent similarity between HBV315 and other HBV isolates in genomic group C was 97.25%, and that with other genomic groups ranged from 86.16% to 91.25%. The predicted amino acid sequences of HBV315 were compared with two closely related subtype adr isolates, M38636 and D12980. The results showed that the X gene product was identical in the three strains, while there were significant amino acid sequence differences between HBV315 and M38636 in the Pre-S1 and Pre-S2 regions.
Cloning and Characterization of the Lactococcus lactis subsp. lactis ATCC 7962 pts HI Operon
Kim, Tea-Youn ; Park, Rae-Jun ; Chang, Hae-Choon ; Chung, Dae-Kyun ; Lee, Jong-Hoon ; Lee, Hyong-Joo ; Kim, Jeong-Hwan ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 829~835
The ptsH and ptsI genes of Lactococus lactis subsp. lactis ATCC 7962 (L. lactis 7962), encoding the general proteins of phosphotransferase system (PTS) components, HPr and enzyme I, respectively, were cloned and characterized. A 1.3 kb PCR product was obtained using a primer set that was hybridized to the internal region of the L. lactis 7962 pts HI genes and then subcloned into a low-copy number vector, pACYC184. The 5' upstream and 3' downstream region from the 1.3 kb fragment were subsequently clone using the chromosome walking method. The complete ptsHI operon was constructed and the nucleotide sequences determined. Two ORFs corresponding to HPr (88 amino acids) and enzyme I (575 amino acids) were located. The ptsHI genes of L. lactis 7962 showed a very high homology (84-90%) with those genes from other Gram-positive bacteria. A primer extension analysis showed that the transcription started at either one of two adjacent bases upstream of the start codon. Using a Northern analysis, two transcripts were detected; the first, a 0.3 kb transcript corresponding to ptsH and the second, a 2 kb transcript corresponding to ptsH and ptsI. The transcription level of ptsH was higher than that of ptsI. The concentration of the ptsH transcript in cells grown on glucose was similar to that in cells grown on lactose, yet higher than that in cells grown on galactose. The ptsI transcript was scarcely detected in cell grown on lactose or galactose. The ptsI transcript was scarcely detected in cells grown on lactose or galactose. The results of a sequence analysis and Northern blot confirmed that the ptsH and ptsI genes of L. lactis 7962 were arranged in an operon like other known ptsHI genes and the expression of the ptsHI genes was regulated at the transcriptional level in response to the carbon source.
Beef Tallow Hydrolysis by Immobilized Lipase
Kim, Dong-Joon ; Shin, Dong-Hoon ; Hur, Byung-Ki ; Kim, Eun-Ki ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 836~839
Beef tallow, which is an industrial lipid substrate, was hydrolyzed by lipase immobilized on a high-density polyethylene (HDPE) powder. Ethanol pre-washing process affected the immobilization efficiency. Half-life of storage of the HDPE at
was 150 days. And after 10 times of repeated use, more than 50% of initial activity remained. An apparent Michaelis constant (
) and maximum velocity (
) were 2.7M, and 1.4 mmol/min/l for immobilized lipase, and 0.5 M, and 1.9 mmol/min/l for soluble lipase, respectively.
Expression of the Galactose Mutarotase Gene from Lactococcus lactis ssp. lactis ATCC7962 in Escherichia coli
Lee, Jong-Hoon ; Choi, Jae-Yeon ; Lee, Jung-Min ; Kim, Jeong-Hwan ; Chang, Hae-Choon ; Chung, Dae-Kyun ; Lee, Hyong-Joo ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 840~843
The structure of gal/lac operon of Lactococcus lactis ssp. lactis ATCC7962 was partially characterized and the gene (galM) encoding galactose mutarotase was cloned together with the order; galA-galM-galK-galT. The galM was found to be 1,027 bp in length and encoded the protein of 37,609 Da calculated molecular mass. The deduced amino acid sequence showed a homology with GalM proteins from several other microorganisms. Thus, the galM gene was expressed in Escherichia coli and the product was identified as a 38 kDa protein which corresponded to the size estimated from DNA sequence. mutarotase activity of the IPTG inducedrecombinant was 2.7 times increased against that of the host strain.
Receptor-Mediated Endocytosis of Hepatitis B Virus PreS1d Protein in EBV-Transformed B-Cell line
Park, Jung-Hyun ; Cho, Eun-Wie ; Lee, Dong-Gun ; Park, Jung-Min ; Lee, Yun-Jung ; Choi, Eun-A ; Kim, Kill-Lyong ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 844~850
The specific binding and internalization of viral particles is an essential step for the successful infection of viral pathogens. In the case of the hepatitis B virus (HBV), virions bind to the host cell via the preS domain of the viral surface antigen and are subsequently internalized by endocytosis. HBV-preS specific receptors are primarily expressed on hepatocytes, however, viral DNA and proteins have also been detected in extrahepatic sites, suggsting that celluar recepators for HBV may also exist on extrahepatic cells. Recently, an EBV-transformed B-cell line was identified onto which the preS region binds in a receptor-ligand specific manner. In this study, this specific interaction was further characterized, and the binding region within the preS protein was locaized. Also the internalization after host cell attachment was visualized and analyzed by fluorescence-labeled HBV-preS1 proteins using confocal microscopy. Energy depletion by sodium azide treatment effectively inhibited the internalization of the membrane-bound preS1 ligands, thereby indicating an energy-dependent receptor-mediated endocytotic pathway. Accordingly, the interaction of HBV-pres! with this specific B-cell line may serve as an effective model for an infection pathway in extrahepatic cells.
Enhancement of Phase II and Antioxidant Enzymes in Mice by Soybeans Fermentation with Basidiomycetes
Shon, Yun-Hee ; Kim, So-Yeun ; Lee, Jae-Sung ; Nam, Kyung-Soo ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 851~857
The activities of phase II and antioxidant enzymes in the liver, lung, kidney, stomach, and colon of mice were examined following intragastric application of polysaccharides extracted from soybeans fermented with either Agrocybe Cylindracea (AC) or Phellinus ignarius (PI). The intragastric application of the extracts to mice for 14 days significantly increased the activities of quinone reductase (QP) and glutathione S-transferase (GST) in the liver and kidney, glutathione (GSH) and superoxide dismutase (SOD) in the liver, kidney, lung, and stomach, and glutathione peroxidase (GSH-Px) in the liver, lung, and kidney. In general, the elevation of the phase II and antioxidant enzymes activities was more pronounced in the liver and kidney as compared to the lung, stomach, and colon. Accordingly, these finding suggest that polysaccharides extracted from soybeans fermented with A. cylindracea or P. igniarius have a cancer chemopreventive potential in various target organs.
Partitioning and Inactivation of Viruses by Cold Ethanol Fractionation and Pasteurization during Manufacture of Albumin from Human Plasma
Kim, In-Seop ; Eo, Ho-Gueon ; Chang, Chon-Geun ; Lee, Soung-Min ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 858~864
The purpose of the present study was to examine the efficacy and mechanism of the fraction IV cold ethanol fractionation and pasteurization (
heat treatment for 10h) steps, involved in the manufacture of albumin from human plasma, in the removal and/or inactivation of blood-born viruses. A variety of experimental model viruses for human pathogenic viruses, including the Bovine viral diarrhoea virus (BVDV), Bovine herpes virus (BHV), Murine encephalomyocarditis virus (EMCV), and Porcine parvovirus (PPV), were selected for this study. Samples from the relevant stages of the production process were spiked with the viruses, and the amount of virus in each fraction was then quantified using a 50% tissue culture infectious dose (
). The mechanism of reduction for the enveloped viruses (BHV and BVDV) during fraction IV fractionation was inactivation rather than partitioning, however, it was partitioning in the case of the non-enveloped viruses (EMCV and PPV). The log reduction factors achieved during fraction IV fractionation were
for BBDV, 4.9 for EMC, and 4.0 for PPV. Pasteurization was found to be a robust and effective step in inactivating the enveloped viruses as well as EMCV. The log reduction factors achieved during pasteurization were
for EMCV, and 1.7 for PPV. These results indicate that the production process for albumin has sufficient virus-reducing capacity to achieve a high margin for virus safety.
Evaluation of Factors that Can Affect Protective Immune Responses Following Oral Immunization of Recombinant Helicobacter pylori Urease Apoenzyme
Kim, Jang-Seoung ; Chang, Ji-Hoon ; Park, Eun-Jeong ; Chung, Soo-Il ; Yum, Jung-Sun ; Kim, Jang-Seoung ; Chang, Ji-Hoon ; Park, Eun-Jeong ; Chung, Soo-Il ; Yum, Jung-Sun ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 865~872
Helocobacter phylori is the major cause of gastritis, peptic ulcer, and a principal risk factor for gastric cancer. As the firs step towards a vaccine against H. pylori infection, Hy.pylori urease was expressed and purified as a recombinant apoenzyme (rUrease) in E. coli. In order to develop an effective immunization protocol using rUrease, the host immune responses were evaluated after the oral immunization of mice with rUrease preparations plus cholera toxin relative to various conditions, such as the physical nature of the antigen, the frequency of the booster immunization, the dose of the antigen, and the route of administration. The protective efficacy was assessed using a quantitative culture following an H. pylori SS1 challenge. It was demonstrated that rUrease, due to its particulated nature, was more superior than the UreB subunit as a vaccine antigen. The oral immunization of rUrease elicited significant systemic and secretory antibody responses, and activated predominantly Th2-type cellular responses. The bacterial colonization was significantly reduced (~100-fold) in those mice immunized with three or four weekly oran doses of rUrease plus cholera toxin (p<0.05), when compared to the non-immunized/challenged controls. The protection correlated well with the elicited secretory IgA level against rUrease, and these secretory antibody responses were highly dependent on the frequency of the booster immunization, yet unaffected by the dose of the antigen (25-200
). These results demonstrate the remarkable potential of rUrease as a vaccine antigen, thereby strengthening the possibility of developing an H. pylori vaccine for humans.
Optimization of a Multiplex DNA Amplification of Three Short Tandem Repeat Loci for Genetic Identification
Ryu, Jae-Song ; Noh, Jae-Sang ; Koo, Yoon-Mo ; Lee, Choul-Gyun ; So, Jae-Seong ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 873~876
Short tendem repeat (STR) loci have been used in the field of forensic science. There are literally hundreds of STR systems which have been mapped throughout the human genome. These STR loci are found in almost every chromosome in the genome. They may be amplified using a variety of PCR primers. In this study, a DNA genotyping system based on the multiplex amplification of highly polymorphic STR loci was developed. Three STR loci with nonoverlapping allele size ranges have been utilized in the multiplex amplification including the Neurotensin receptor gene, D21S11, and Human tyrosine hydroxylase gene. The optimal condition for triplex PCr was obtained in a solution with a total volume of
containing 2.0 U of Taq polymerase, 3 mM of
of dNTP, 10 pmole of each primer set, an annealing temperature of
, and 35 cycles. The optimized condition was successfully employed in a family paternity test.
Regulation of Cycloinulooligosaccharide Fructanotransferase Synthesis in Bacillus macerans and Bacillus subtilis
Kim, Hwa-Young ; Choi, Yong-Jin ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 877~880
Cycloinulooligosaccharide fructanotransferase (CFTase) converts inulin into cyclooligosaccharides consisting of six to eight molecules
)-linked cyclic D-fructofuranose through intramolecular transfructosylation. We have examined the regulation of CFTase synthesis in Bacillus macerans and Bacillus subtilis. Synthesis of the CFTase was induced by inulin and it was subject to carbon catabolite repression (CCR) by glucose in both microorganisms. The DNA sequence upstream of the promoter of the CFTase gene was not involved in the inulin induction and glucose repression of the CFTase gene expression in B. subtilis. This suggests that the DNA element(s) responsible for the inuline induction and glucose repression is located downstream of the promoter region. Unexpectedly, the CCR of the expression of CFTase gene was observed not to be dependent on CcpA protein in B. subtilis.
Rapid Identification of Lactobacillus plantarium in Kimchi Using Polymerase Chain Reaction
Kim, Tae-Woon ; Min, Sung-Gi ; Choi, Dong-Hun ; Jo, Jae-Sun ; Kim, Hae-Yeong ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 881~884
A polymerase chain reaction (PCR) was performed to rapidly identify Lactobacillus plantarum from type strains and kimchi samples. The PCR experiments were carried out using specific oligonucleotide primer sets based on the 16S rRNA gene sequences of L. plantarum. The expected DNA amplificate of 419 bp was obtained when either purified DNA or whole cells of L. plantarum strains reacted with LP primers, yet not with any of the other strains. The PCR product was confirmed by DNA sequencing. Accordingly, since the PCR method used is simple, specific, and rapid, it will be useful for monitoring and evaluation L. plantarum in the mixed microbial population found in kimchi.
Characteristics of Electroantennogram Parameters for the Detection of Odorants
Yun, Eung-Sik ; Ko, Hwi-Jin ; Sun, Jong-Hwan ; Park, Tai-Hyun ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 885~888
Electroantennogram(EAG) can be applied to detect odorants since insects have highly specialized olfactory receptors inside their antennae. The characteristics of EAG parameters were investigated for the quantitative measurement of a general odorant using ammonia as a model odorant. The antennae of male silkworm moth, Bombyx mori, were used for the EAG. The electical signal curves generated from a pair of antennae originating from the same silkworm moth were never identical; however, they exhibited a typical type I or type II characteristic curve pattern for every pair of antennae. The correlation between the EAG parameters and the ammonia concentration was analyzed for the type I and type II antennae. The stability of each parameter was also investigated for each type of antenna. The results show the possibility of the quantitative measurement of general odorants using the EAG technique.
Structural Study of Antisense Dimers, Modified Adenosine-Thymidine Phosphorothioate
Jung, Kyeong-Eun ; Yang, Mi-Rim ; Lee, Kwang-Jun ; Lim, Hong ; Jung, Ji-Hyun ; Lim, Yoon-Gho ; Cho, Youl-Hee ; Shin, Dong-Hoon ; Lee, Chul-Hoon ;
Journal of Microbiology and Biotechnology, volume 10, issue 6, 2000, Pages 889~892
Antisense molecules are structurally simple linear oligomers of nucleotides. They can recognize a complementary sequence by base pairing, therefore, antisense drugs composed of 15-16 bases are potentially useful, unlike drugs such as protein agonists, antagonists, and inhibitors. Since antisense oligomers are classified as nucleotides, they are subject to attack by nucleases. In order to be antisense drugs resistant to degradation by nucleases, the structural modifications in the linkages, bases, and sugars to satisfy this requirement are considerable. We attempted in this study, to synthesize 16-mer antisenses with a modified linkage and adenosine. When studying on the three-dimensional structure of the oligomer, however, the existence of isomers may complicate the interpretation of the NMR data. Therefore, an attempt was made to eliminate the above problem, thus, two dimers were synthesized and their structural studies were carried out.