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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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The Korean Journal of Microbiology
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Journal DOI :
The Microbiological Society of Korea
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Volume & Issues
Volume 36, Issue 4 - Dec 2000
Volume 36, Issue 3 - Sep 2000
Volume 36, Issue 2 - Jun 2000
Volume 36, Issue 1 - Mar 2000
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Expression of the Genes Involved in the Synthesis of Riboflavin from Photobacterium species of Bioluminescent Marine Bacteria
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 1~7
The genes involved in riboflavin synthesis (ribI, II, III, and IV) were found immediately downstream of luxG in the lux operon from Photobacterium species. The single stranded DNA containing the intergenic region of lux genes and rib genes from Photobacterium phosphoreum was fully protected by P. phosphoreum mRNA from the S1 nuclease mapping assay suggesting that a transcriptional terminator was not present in the region. In addition, the levels of riboflavin synthase activity in P. phosphoreum was increased during the development of bacterial bioluminescence in the same fashion as the luciferase and fatty acid reductase activities. Insertion of the Photobacterium leiognathi DNA extending from luxB to ribII, between a strong lux promoter and a reporter gene (chloramphenicol acetyltransferase, CAT) and transferred by conjugation into P. leiognathi, did not affect expression of reporter gene. Moreover the CAT gene was not expressed in an analogous construct missing the lux promoter indicating that a promoter was not present in this region. Based on the data here, it can be concluded that the lux genes and rib genes in Photobacterium species are under common regulation.
The Action Mode of
-glucosidase Purified from Trichoderma koningii
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 8~13
We have examined the mode of transglycosylation, catalyzed by an extracellular
-glucosidase purified from Trichoderma koningii ATCC 26113, using cellobiose, sophorose, laminaribiose and gentiobiose as substrates. The dimers separated from the reaction mixture by HPLC were analyzed by
-NMR spectroscopy. When cellobiose was subjected to the action of the
-glucosidase, the products included laminaribiose, sophorose and gentiobiose. When laminaribiose, sophorose or gentiobiose was used as a substrate, the
-glucosidase accumulated transglycosylation products possessing different types of
-glycosidic linkages from the original one. The amount of dimers accumulated as reaction proceeded seemed to be dependent on the velocity of hydrolysis but not on that of formation.
Isolation, Purification, and Characterization of the Lytic Enzyme of Anabaena cylindrica by Penicillium oxalicum (HCLF-34)
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 14~19
Algal lytic enzyme, an extracellular enzyme, was purified from the culture filtrate of Penicillium oxalicum(HCLF-34) by ultrafiltration, gel filtration chromatography, and anion exchange chromatography. The enzyme has a molecular mass of approximately 22 kDa, an it is a monomer by renaturation SDS-PAGE. The amino acid sequences of the enzyme was revealed to be NH2-Glu-Ser-Tyr-Ser-Ser-Asn-Ala-Ala-Gly-Ala-Val-Leu-Ile---, had about 84% identity with the mature light chain of aspergillopepsin II precursor and 81% identity with the mature protein of the acid proteinase EapC precursor.
Characterization of Endoglucanase (F-II-II) Purified from Trichoderma sp. C-4
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 20~25
One of endoglucanases(F-II-II) was purified from the culture filtrate of Trichoderma sp. C-4 through two step procedures including chromatography on Sephacryl S-200 and Sephacryl S-100. The molecular weight of the enzyme was determined to be about 26,000 by SDS-PAGE and the isoelectric point as 8.0 by analytical isoelectric focusing. The optimum temperature of the enzyme was
and the optimum pH was 5.0. No loss of activity was observed when the enzyme was preincubated at
for 24 hours. The specific activity of the enzyme toward carboxymethylcellulose (CMC) was estimated to be 776.2 U/mg. The internal amino acid sequence was analysed.
Purification and Characterization of Catechol 2,3-Dioxygenase from Recombinant Strain E. coli CNU312.
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 26~32
Catechol 2,3-dioxygenase was purified from recombinant strain E. coli CNU312 carrying the tomB gene which was cloned from toluene-degrading Burkholderia cepacia G4. The purification of this enzyme was performed by acetone precipitation, Sephadex G-75 chromatography, electrophoresis and electro-elution. The molecular weight of native enzyme was about 140.4 kDa and its subunit was estimated to be 35 kDa by SDS-PAGE. It means that this enzyme consists of four identical subunits. This enzyme was specifically active to catechol, and
value of this enzyme were 372.6
M and 39.27 U/mg. This enzyme was weakly active to 3-methylcatechol, 4-methylcatechol, and 4-chlorocatechol, but rarely active to 2,3-DHBP. The optimal pH and temperature of the enzyme were pH 8.0 and
. The enzyme was inhibited by
ions, and was inactivated by adding the reagents such as N-bromosuccinimide, and
-diazobenzene sulfonic acid. The activity of catechol 2,3-dioxygenase was not stabilized by 10% concentration of organic solvents such as acetone, ethanol, isopropyl alcohol, ethyl acetate, and acetic acid, and by reducing agents such as 2-mercaptoethanol, dithiothreitol, and ascorbic acid. The enzyme was inactivated by the oxidizing agent
, and by chelators such as EDTA, and ο-phenanthroline.
Isolation and Characterization of Growth Stimulating Thermophilic Fungi on Oyster Mushroom from Oyster Mushroom (Pleurotus ostreatus) Compost
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 33~39
Some of thermophilic fungi which has growth-promoting effect on Pleurotus ostreatus were isolated from compost during high temperature fermentation process. The temperature optima of 7 isolated thermophilic fungi were
on PDA media. Isolated strains S-1 and S-2 have the best mycelial growing rate, so these isolates were expected as excellent thermophilic fungi for high temperature composting and mycelial growing of oyster mushroom. In liquid culture, the optimal pH of thermophilic fungi observed variously, pH 7.0-10.0 but most of thermophilic fungi grow well in pH 8.0-pH 9.0 and the final pH of media after cultured was done pH 5.5-6.0. In liquid culture of thermophilic fungi on the optimal condition, S-2 have the best mycelial growing rate. The growing rate of thermophilic fungi S-1, S-2, S-5, and S-10 on lignocellulosic substrates was good but Humicola grisea var. thermoidea, well know thermophilic fungi which has growth-promoting effect on Agaricus bisporus, was poor and which was well grown on PDA at
, pH 7.0. Isolated strain S-1 was identified as Trichophyton sp. and other 6 strains were identified as Sepedonium sp. by morphological characteristics.
Molecular Systematics of Rhizoctonia solani Isolates from Various Crops with AFLP
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 40~45
Rhizoctonia solani Kuhn[Thanatephorus cucumeris (Frank) Donk]is a destrutive soil-borne plant pathogen affecting many agricultural crops worldwide. R. solani is divided into anastomosis groups based on the ability of the hyphae to fuse, and into subgroups based on morphological, physiological characteristics. AG classifications are convenient and useful in identifying primary causal agents of Rhizoctonia diseases, although the mechanism of anastomosis is not fully understood. Beacause of the difficulties, we sought to develop a more direct method for genetic identification and charaterization of R. solani. Twenty nine isolates of R. solani were used for the analysis of genetic relationships among themselves and for rapid anastomosis grouping with AFLP method. All isolates studied were divided into five groups. Isolate 6 was included in AG-3 with 67% genetic similarity. When isolates 3 was compared with 13 and 10 each, they showed more than 84% and 83% similarity, respectively. Isolates 3, 4, 5, 13, and 16 were included in AG-1 with 83% genetic similarity. Isolates 1, 7, and 8 were included in AG-1(IB).
Cloning and Nucleotide Sequence Analysis of the aroA Gene from Salmonella typhi KNIH100
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 46~51
Salmonella typhi is one of important causes of human enteric infections. S. typhi KNIH100 was isolated from a patient of typhoid fever in Korea. We cloned a 5.0 kb SalⅠ fragment containing the aroA gene encoding a 5-enolpyruvylshikimate-3-phosphate synthetase from chromosomal DNA of this strain. This recombinant plasmid was named pSAL80. E. coli CGSC2829, an aroA- mutant, was not grown on the M9 minimal medium but E. coli CGSC2829 (pSAL80) was grown on the M9 minimal medium. The aroA gene was composed of 1,284 base pairs with ATG initiation codon and TAA termination codon. Sequence comparison of the aroA gene exhibited 99%, 98%, and 77% identity with those of S. typhi Ty2, S. typhimurium, and E. coli respectively. As in the cases of Shigella sonnei and E. coli, the serC and aroA genes lie in a single operonic structure.
Effect of Selected Environmental Factors on the Production of Geosmin in Phormidium Sp.
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 52~57
A method for quantitative and qualitative analysis of geosmin, odorant produced by several actinomycetes and cyanobacteria, was established and optimized. The effects of environmental conditions on the growth of Phormidium sp. NIVA-CYA7 were examined and the production and release of geosmin by the species was analyzed by using the purge and trap-gas chromatographic technique. One of the major advantages of the technique established in the present study is that the preparation of sample is simpler and purge time is shorter. Under the culture conditions (pH 7.9,
/s and Z8 medium), Phormidium showed growth characteristics with a lag phase for 8 days and an exponential phase for 14 days followed by a stationary phase. Reduction of inorganic nitrogen concentrations in the culture medium from 250 to 100 or 25
M brought no significant effect on the cell growth. However, the cell growth was significantly inhibited with decreasing concentrations of inorganic phosphorus from 25 to 10 or 2.5
M. When the inorganic phosphorus concentration in the medium was lowered from 25 to 10
M, the levels of geosmin in the organism expressed as percentages per unit TOC and chlorophyII-
increased by 35% and 68%, respectively. When the initial pH of the medium was 9.4, geosmin content was 0.0824
g/mg C, which was 2-fold higher than that at pH 7.9 Consequently, the level of geosmin in Phormidium was found to vary with growth phases of the culture, external inorganic phosphorus concentration and external pH, while the release of geosmin was not significantly affected by the factors.
Isolation and Identification of Klebsiella oxytoca C302 and Its Degradation of Aromatic Hydrocarbons
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 58~63
A bacterial isolate capable of degrading benzoate was selected from wastewater of Yocheon industrial complex and examined its biochemical characteristics and fatty acid composition. The isolate was identified as Klebsiella oxytoca strain C302. The strain C3O2 degraded catechol, protocatechuate, and 4-hydroxybenzoate as well as benzoate. The strain grew on and degraded 0.5 to 1.0 mM catechol most actively in MM2 medium at pH 7.0 and
Immunopotentiating Effect of Polysaccharides Extracted from Agrocybe cylindracea
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 64~68
The immunopotentiating effects of the polysaccharides, both intracellular and extracellular, was examined by an animal feeding test. The results are summarized as follows. The oral administration of intracellular and extracellular polysaccharides of Agrocybe cylindracea for 10 days resulted in the enhanced phagocytic activity of peritoneal exudate cells(PEC), spleen cells(SC), and monolymphocytes(ML). In the experiment of PFC(plaque forming cell) and RFC(rosette forming cell), the results showed that all the polysaccharide fractions enhanced the immune related cells. The EAC II group(the extracellular polysaccharide of Agrocybe cylindracea 10 mg/0.2 ml distilled water/day/ mouse) increased the PFC and RFC by 46~50% and 43%, respectively, compared to the control group. On the other hand, the IAC I group(the intracellular polysaccharide of Agrocybe cylindracea 1 mg/0.2 ml distilled water/day/mouse) increased the PFC and RFC by 49~70% and 91%, respectively. In terms of the mitogenic activity, the extracellular polysaccharides of A. cylindracea showed a higher activity than the intracellular polysaccharides.
Cultivation of Arthrobactor sp. A-6 and Production of DFA III(Di-Fructofuranose Dianhydride) from Chicory Root Extract
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 69~73
Arthrobacter sp. A-6 was cultivated and DFA III(di-fructofuranose dianhydride) was produced with inulin fructotransferase from the chicory root. The specific growth rate, yield of cell mass and yield of enzyme from the culture in variable chicory root extracts were studied and the results compared. Standard inulin solution(10%) was treated with the crude enzyme solution of inulin fructotransferase from the cell culture, 1.14mg/ml of DFA III was produced. The enzyme reactions were processed with various preparations of chicory root extracts in the same conditions. The highest yield of DFA III production(2.29 mg/ml) was obtained from the chicory roots without washing or extraction. The yield of DFA III from the washed chicory roots without extraction was at lowest(0.44 mg/ml). The production process of inulin fructotransferase and DFA III from the chicory root without prewashing or extraction steps were more efficient.
Studies on the Culture Media and the Optimal Storage Conditions of Bioluminescent Bacteria Photobacterium phosphoreum
The Korean Journal of Microbiology, volume 36, issue 1, 2000, Pages 74~78
Vibrio, Photobacterium, Alteromonas and Xenorhabdus species are capable of emitting light, called bioluminescence. They exist in marine, freshwater and terrestrial environments. Bacterial bioluminescent reaction is that reduced riboflavin phosphates and a long-chain aldehyde are oxidized in the presence of molecular oxygen and enzyme luciferase. This experiment aims to develop the proper culture media and to optimize the storage condition for the recovery of bioluminescent activity in Photobacterium phosphoreum. The Luria broth (LB) medium was modified for cultivation of Photobacterium phophoreum, called as modified LB(mLB) medium. The mLB medium is LB fortified with 3% glycerol and 1.5% NaCl. In mLB medium. bacterial growth and bioluminescent activity are 25% higher than those in a Nutrient broth medium. When the cell stocks were stored at
and LN2 for 3 months, cell growth and bioluminescent activity of culture after stored at
were better than those of other treatments. The highest bioluminescent activity obtained at the late exponential phase in all treatments. When the cell stock was freeze-dried with 5% adonitol as a cryoprotectant, the recovery of cell was better than those of control and freeze-dried cell stock without addition of cryoprotectant.