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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
Journal Basic Information
Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 31, Issue 4 - Aug 1993
Volume 31, Issue 3 - Jun 1993
Volume 31, Issue 2 - Apr 1993
Volume 31, Issue 1 - Feb 1993
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Isolation and Characterization of a Restricted Facultatively Methylotrophic Bacterium Methylovorus sp. Strain SS1
Seo, Sung A. ; Kim, Young M. ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 179~183
A restricted facultatively methanol-oxidizing bacterium, Methylovorus sp. strain SS1, was isolate dfrom soil samples from Kuala Lumpur, Malaysia, through methanol-enrichment culture technique. The isolate was nonmotile Gram-negative rod and did not have complex internal membrane system. The colonies were small, pale-yellow, and raised convex with entire margin. The cell did not produce any spores and capsular materials. The cell was obligately aerobic and exhibited catalase, but no oxidase, activity. Plasmid, carotenoid pigment, and poly-.betha.-hydroxybutyric acid were not found. The guanine plus cytosine content of the DNA was 55%. The isolate was found to grow only on methanol methylamine, or glucose. Growth factors were not required. Cells growing on methanol was found to produce extracellular polysaccharides containing glucose, lactose, and fructose. Growth was optimal (t
= 1.7) with 0.5%(v/v) methanol at 40.deg.C and pH 6.5. No Growth was observed at over 60.deg.C. Cell-free extracts of the methanol grown cells exhibited the phenazine methosulfate-linked methanol dehydrogenase activity Methanol was found to be assimilate dthrough the ribulose monophosphate pathway.y.
Genomic Variation and Toxin Specificity of Ustilago maydis Virus Isolated in Korea
Hee, Hwang-Seon ; Yie, Se won ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 184~188
Novel Ustilagomaydis strains, designated as SH1 to 14 containing new types of ds RNA segments, are identified from corn smut in Korea. Among 14 isolates, 7 isolates appear to posses virus particles and the other isolates may contain dsRNA as a plasmid form. The pattern of dsRNA is highly diverse form a typical P-type containing one or more of H, M, and L dsRNAs to the one containing one or move M dsRNAs. It is likely that the strains containing H dsRNA posses virus particles which were confirmed by sucrose density gradient followed with different range of specificity and the activity of the strain (SH14) is stronger than A4 toxin. The sensitivity of 14 isolates is also very diverse and two strains (SH10, SH11) appear tobe universal sensitve strains against 5 tested toxin samples.
Nucleotide Sequences of nodD and nodA from Bradyrhizobium sp. SNU001
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 189~196
Nucleotide sequences of nodD and nodA from Bradyrhizobium sp. SNUOOI were determined. The open reading frame (ORF) of nodD was 942 bp in length and encoded 314 amino acids. while ORF of nodA, sequence of which is the first one among legume symbionts Bradyrhizobium, was 630 bp and encoded 210 amino acids. The nucleotide sequence of nodD showed 99.4% homology with nodDI of B. japonicum USDAllO. while that of nodA showed 81.5% with B. sp. (Parasponial. At the 5' of nodYAB operon and nodD, consensus nod box sequences composed of 9 bp unit repeated four times and two times respectively were found. Also an A.T-rich sequence was found at 5' of nodD.
Molecular Cloning of the Gene in Schizosaccharomyces pombe Related to the CDC3 Gene in Saccharomyces cerevisiae
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 197~202
The budding yeast S. cerevisiae contains 10-nm filament ring that lies just inside the plasma memhrane in the region of the mother-bud neck. It is possihle that CDC3. CDCIO, CDCII. CDCI2 genes encode the filaments. Recently it has been shown that the CDC3 and CDCI2 gene products arc localized to [he vicinity of the neck lilaments by immunolluorescence. However. the role of the lilament ring is not clear. In order to find out the role of filament ring. I have tried to clone the similar gene in S. pomhe to the CDC3 in S. cerevisiae. Genomic library was constructed by use of
gtll expression vector and screened with CDC3 antibodies. From sequencing data, there were more than two introns in the newly cloned gene. There was 62% homology between the part of the predicted amino acid sequence of cloned gene and CDC3 amino acid sequence.
The Glucoamylase Signal Sequence Directs the Efficient Secretion of Human
1-Antitrypsin in Yeast Cells
Song, Moo-Young ; Kwon, Ki-Sun ; Kang, Dae-Ook ; Yu, Myeong-Hee ; Park, Hee-Moon ; Kim, Jinmi ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 203~207
Five different secretion vectors were constructed by varying the signal sequences and .alpha.-antitrypsin (.alpha.1-AT) a numan secretory protein, was produced from yeast cells. The signal sequences used are those of acid phosphatase (PH05) and .alpha.-factor (M f.alphal1) of Saccharomyces cerevisiae, glucoamylase (STA1) of Saccharomyces diastaticus, and human .alpha.1-AT. Four vectors directed the efficient secretion of .alpha.1-AT ito the culture media. The secretion vector carrying the glucoamylase signal sequence (pGAT11) showed the highest efficiency of secretion. About 70% of .alpha.1-AT produce dwere secreted into the media. The endo H treatment of partially purified .alpha.1-AT indicates that the secreted .alpha.1-AT appeared to be glycosylated. This glycosylation pattern was altered when amino acid substitution mutations were introduced at the three glycosylation sites of .alpha.-AT.
Functional Role of the Internal Guide Sequence in Splicing Activity of T4 Thymidylate Synthase Gene in vivo
Shin, Sook ; Park, In-Kook ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 208~213
The structural and functional roles of IGS element of T4 td intron in thymidylate synthase activity in vivo were investigated Site-directed mutagenesis was employed to crete mutations of IGS element of T4 td intron, When a U-G pari was changed to a U-C pari in the 5' splice site of P1 stem of td intron, the activity of thymidylate synthase was completely abolished whereas the wild type retained the normal activity of enzyme. When U at 12 position within IGS element was changed to C, the activity of thymidylate synthase was approximately 32% of that of the wild type. Comparison of enzyme activities suggests that IGS element within P1 structure is an essential requirement for splicing of td gene in vivo.
Constitutive Expression of Carbon Monoxide Dehydrogenase in Acinetobacter sp. Strain JC1 DSM 3803
Ro, Young T. ; Kim, Young M. ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 214~217
Carbon monoxide dehydrogenase (CO-DH) was found to be present in Acinetobacter sp. strain JC1 grown on CO and also on methylotrophic and heterotrophic substrates, except for pyruvate and nutrient broth. The amounts of CO-DH in cells grown on methylamine, glucose, galactose, and succinate were comparable to that of the CO-grown cells. CO-DH activity, however, was onot deteted by the dye-linked assay method in cell extracts prepared from cells grown on organic substrates, except on ethanol and succinate. THe activity was detected when the CO-DH was stained by activity using CO as a substrate. CO-DHs in cells grown on different substrates were found to be identical in immunological properties.
Intergeneric Protoplast Fusion between Rhizopus oryzae and Aspergillus oryzae
Lee, Soo-Youn ; Jung, Sung-Won ; Kim, Seong-Han ; Lee, Yung-Nok ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 218~223
Conditions for the release and regeneration of protoplasts form Rhizopus oryzae and intergeneric protoplast fusion between Rhizopus oryzae and Aspergillus oryzae were studied. High yields of protoplast fusion between Rhizopus oryzae and Aspergillus oxyzae were studied. High yield of protoplasts from young germilings of R. oryzae were obtained by using lytic enzymes containing chitosanase (3 mg/ml), chitinase (3 mg/ml) and Novozym 234 (5 mg/ml). 0.5M glucose was used as the osmotic stabilizer and optimum pH of buffer was determined to be pH 7.5-8.0. Under these conditions, protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts were formed after about 3-4 hrs incubation. Approximately, 1.0%-4.9% of these protoplasts regenerated on solid medium with a soft agar overlay. We have also carried out protoplasts fusion between R. oryzae and A. oryzae and have succeeded in obtaining three types of intergeneric fusants. In these experiments, 35% PEG-4000 and 10 mM CaCl
were used as fsogenic agents, and auxotrophic properties were used as a genetic marker to select fusants. Complementation frequency be protoplasts fusion of A. oxyzae and R. oryzae was 4.4% * 10
. The fusant strains of the first type were prototrophs showing an Aspergillus type morphology with dark-yellow sporulation, those of the second type were also Apergillus type morphology but showed no sporulation. And the strains of the third type stopped growing when fusion products grown on regeneration minimal medium were transferred to fresh minimal medium. The formation of fusion products was observed by fluorescent vital stains for complementary labelling of protoplats from R. oryzae and A. oryzae. Rhodamine 6G and fluorescein diacetate wer useful complementary vital stains of Rhizopus and Aspergillus protoplasts for visualization of requency and type (dicell, multicell) of fusion.
Inhibitory Actions of Mycotoxins on Brain
Lee, Su-Jin ; Lee, Kil-Soo ; Choi, Soo-Young ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 224~229
GABA transminase (4-aminobutyrate aminotransferase), which catalyzes the breakdown of the major inhibitory neurotransmitter, GABA, in mammalian brain, was inactivated by preincubation with the mycotoxin patulin. The time course of the reaction was significantly affected by the substrate .alpha.-ketoglutarate, which aforded complete protection against the loss of catalytic activity. The recovery from the inhibition of patulin by the addition of dithiothreitol (DTT) supports that patulin reacts with the sulfhydryl residue in the catalytic domain of the enzyme. The reconstitution of the reduced enzyme and apoenzyme with pyridoxal-5-P(PLP) was inhibited by another mycotoxin, penicilic acid. This mycotoxin may interact with lysyl residue of the enzyme. Therefore, it is postulated that the critical sulfhydryl and lysyl residues in the catalytic domain of the enzyme react with mycotoxin patulin and penicillic acid, respectively.
Characterization of Isocitrate Lyase from Micrococcus luteus
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 230~236
The isocitrate lyase extracted from Micrococcus luteus was purified 38.8 folds with the overall yield of 10.2%, by the ammonium sulfate fractionation, DEAE-cellulose, 1st Sephadex G-200 and 2nd Sephadex G-200 column chromatography. The purified enzyme showed to be a single protein band by polyacrylamide gel electrophoresis. The molecular weight of the purified enzyme was estimated 60,000 by the SDS-polyacry]amide gel electrophoresis. The apparent Michaelis constant, Km value for isocitrate was 0.95 mM. The optimum pH and temperature of the purified enzyme were pH 7.5 and
, respectively. The enzyme was activated by
and inhibited by
. In addition, the activity of isocitrate lyase was increased by glutathione and 2-mercaptocthanol at 5 mM and cysteine at I mM.
Characterization of Aspartate Aminotransferase Purified from Streptomyces fradiae
Lee, Sang-Hee ; Lee, Kye-Joon ;
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 237~244
Aspartate aminotransferase (ASAT) (L-aspartate : 2-oxyoglutarate, EC 2.6. 1. 1.) from Streptomyces fradiae NRRL 2702 has been purified by acetone precipitation, DEAE-cellulose, hydroxyapatite, and preparative electrophoresis (Prep cell), of which the last was the most effective step in the purification of ASAT. The molecular mass was estimated to be 54,000 dalton by SDS-PAGE and 120,000 dalton by gel filtration chromatography. Preparative isoelectric focusing of purified ASAT resulted in one polypeptide band with a pI of 4.2, showing homogeneity and indicating that the enzyme is composed of two identical subunits. The enzyme was specific for L-aspartate as an amino donor ; the
values were determined to be 2.7 mM for L-aspartate, 0.7 mM for 2-oxoglutarate, 12.8 mM for L-glutamate, and 0.15 mM for oxaloacetate. The enzyme was relatively heat-stable, having maximum activity at 55.deg.C, and it had a broad pH optimum ranging from 5.5 to 8.0. The activity of the purified enzyme was not inhibited by ammonium ions. This paper reports the first purification and characterization of the aspartate aminotransferase from a species of Streptomyces.s.
Bacterial Distribution and Variation in Water Supply Systems
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 245~254
Distribution and variation of bacterial densities of heterotrophic plate count (HPC) and Enterobacteriaceae in the water supply systems comprising raw, treated, and three tap water samples of a water treatment plant in Seoul were studied 23 times from 1991 to 1992. HPC bacteria of raw. treated, and tap waters on
agar media were at a density of
and 2 to
cfu/ml, respectively. Densities of Enterobacteriaceae in raw, treated, and tap waters on mENDO-LES agar media ranged from 0.] to 8200 cfu/ml, 0 to 17.5 cfu/JOO mI. and 0 to 47.5 cfu/IOO ml, respectively. Injured Enterobacteriaceae of treated and tap waters on m-T7 agar media were at a density of o to 27 and 0 to 35 cfu/100 mI. These results showed that the density of bacteria in the treated water outflowing from the water plant significantly increased as the water flowed along the distribution sytems, which is so-called bacterial regrowth. The predominant bacteria] types in the water supply system were Pw'udomonas and Acinerobacter. In raw water, the ratio of Pseudomonas was higher than that of Acinetobaeter, but in treated and tap waters. both ratios were reversed. The most predominant species of Enterobacteriaceae was Enterobacter agglomerans. Some species such as Citrobacter freundii. Escherichia coli. Klebsiella pneumoniae. and Shigella dysenteriae which are opportunistic pathogens or pathogens were not found in the treated water but additionally detected in tap waters.
Resistance of Biofilm Bacteria to Chlorination
The Korean Journal of Microbiology, volume 31, issue 3, 1993, Pages 255~260
The Enterobacter cloacae biofilms developed on slide glasses and galvanized-iron coupons were applied to test the attached bacterial resistance to chlorination. The chlorine resistances of biofilm bacteria grown on the slide glasses and galvanized-iron coupons were 14 and 480 times that of the suspended bacteria, respectively. The chlorine resistance of particleattached bacterial populations was 48 times that of suspended bacterial populations. The biofilm bacterial densities developed on the slide glasses and galvanized-iron coupons which were immersed in the flowing tap water for 75 days were
It is concluded that main mechanisms of enteric or HPC bacterial resistance to chlorination in tap waters are bacterial attachment or . adsorption to particles or bacterial aggregations and formation of biofilms on the inner wall of distribution systems by escaped bacteria from chlorination in water treatment processes, which results in bacterial regrowth in water distribution systems.