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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 11, Issue 6 - Dec 2001
Volume 11, Issue 5 - Oct 2001
Volume 11, Issue 4 - Aug 2001
Volume 11, Issue 3 - Jun 2001
Volume 11, Issue 2 - Apr 2001
Volume 11, Issue 1 - Feb 2001
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Inducible Periplasmic Chromate Reducing Activity in Pseudomonas aeruginosa Isolated from a Leather Tannery Effluent
GANGULI, A. ; TRIPATHI, A.K. ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 355~361
A Chromate tolerant strain of Pseudomonas aeruginosa isolated from the effluent of a tannery showed significant enzymatic activity of chromate reduction. Cells grown in chromate-supplemented medium reduced 8
chromate/mg protein/h in the presence of NADH/NADPH. The chromate reducing activity was inducible as cells pregrown in chromate showed higher chromate reduction. In contrast, the periplasmic fraction of cells gown in chromate reduce
chromate in 4 h and the spheroplast fraction failed to do so, indicating that chromate reductase may be located in the periplasm. The presence of a 30 kDa protein in the periplasmic extracts of cells grown in the presence of chromate, but its absence of the protein in cells grown without chromate, points out a possible role of this protein in chromate reduction.
Hydrolysis of Paper Mill Sludge Using an Improved Enzyme System
Lin Jianqiang ; Lee, Sang-Mok ; Koo, Yoon-Mo ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 362~368
The effects of water soluble materials in paper mill sludge on cellulase and
-glucosidase activities were studied while the optimization of enzyme system for hydrolysis of the paper mill sludge for production of glucose was made. Water soluble materials in the paper mill sludge showed stimulatory effect on carboxymethyl cellulose (CMC) activity, inhibitory effect on filter paper (FP) activity, and no effect on avicelase and
-glucosidase activities. CMC and
-glucosidase activities at 5 and 10, 5 or 10 and 10, and 10 and 10 U/ml were optimal for hydrolysis of 5, 10, and 20% of the paper mill sludge, respectively.
A New Detergentless Micro-Emulsion System Using Urushiol as an Enzyme Reaction System
Kim, John-Woo-Shik ; Yoo, Young-Je ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 369~375
Urushiol, a natural monomeric oil, was used to prepare a detergentless micro-emulsion with water and 2-propanol The formation of micro-emulsion was verified by conductivity measurements and dynamic light scattering. The conductivity data showed phase change dynamics, a characteristics of micro-emulsions, and subsequent dynamic light scattering study further confirmed the phenomenon. Average water droplet diameter was 10 nm to 500 nm when the molar ratio of 2-propanol ranged from 0.40 to 0.44 . Earlier studies were performed on toluene and hexane, in which the insoluble substrate in water phase was added to the solvents to be reacted on by enzymes. However, in the present urushiol system, urushiol was used as both solvent and substrate in the laccase polymerization of urushiol. The laccase activity in the system was examined using polymerization of urushiol. The laccase activity in the system was examined using syringaldezine as a substrate, and the activity increased rapidly near the molar ratio of 2-propanol at 0.4, where micro-emulsion started. The activity rose until 0.46 and fell dramatically thereafter. The study of laccase activity in differing mole fractions of 2-propanol showed the existence of an ‘optimal zone’, where the activity of laccase was significantly higher. In order to analyze urushiol polymerization by laccase, a bubble column reactor using a detergentless micro-emulsion system was constructed. Comparative study using other organic solvents systems were conducted and the 2-propanol system was shown to yield the highest polymerization level. The study of laccase activity at a differing mole fraction of 2-propanol showed the existence of an ‘optimal zone’ where the activity was significantly higher. Also, 3,000 cP viscosity was achieved in actual urushi processing, using only 1/100 level of laccase present in urushi.
Improvement of the Stability of Lactobacillus casei YIT 9018 by Microencapsulation Using Alginate and Chitosan
Koo, Sun-Mo ; Cho, Young-Hee ; Huh, Chul-Sung ; Baek, Young-Jin ; Park, Ji-Yong ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 376~383
Lactobacillus casei YIT 9018 was microencapsulated within alginate or alginate/chitosan double membrane using an air atomizer. Microbiological analysis revealed that the viability of encapsulated L. casei in gastric juice, hydrogen peroxide, and pepsin was 2-3 log cycle higher than that of the nonencapsulated cell. However, the encapsulated cells did not show a signifciant increase in survival when subjected to in virto high acid and 0.6% bile salt condition. Alginate-encapsulated, alginate/chitosan-encapsulated, and nonencapsulated cells were stored at different temperatures eencapsulated cells showed similar stability at
. However, at
, the alginate/chitosan-encapsulted cell was the most stable.
Estimation of Theoretical Yield for Ethanol Production from D-Xylose by Recombinant Saccharomyces cerevisiae Using Metabolic Pathway Synthesis Algorithm
Lee, Tae-Hee ; Kim, Min-Young ; Ryu, Yeon-Woo ; Seo, Jin-Ho ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 384~388
The metabolic pathway synthesis algorithm was applied to estimate the maximum ethanol yield from xylose in a model recombinant Saccharomyces cerevisiae strain containing the genes involved in xylose metabolism. The stoichiometrically independent pathways were identified by constructing a biochemical reaction network for conversion of xylose to ethanol in the recombinant S. cerevisiae. Two independent pathways were obtained in xylose-assimilating recombinant S. cerevisiae as opposed to six independent pathways for conversion of glucose to ethanol. The maximum ethanol yield from xylose was estimated to be 0.46 g/g, which was lower than the known value of 0.51 g/g for glucose-fermenting and wild-type xylose-fermenting yeasts.
Characterization of an
-Glucosidase Inhibitor Produced by Streptomyces sp. CK-4416
Chun, Hyoung-Sik ; Chang, Heung-Bae ; Kwon, Young-In ; Yang, Han-Chul ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 389~393
-glucosidase inhibitor, CK-4416, was identified from the culture broth of Streptomyces sp. CK-4416. CK-4416, which had some specificity against intestinal disaccharidases, especially sucrase and matlase activities, was purified by adsorption and cationic ion exchange chromatographies. The molecular formula was determined to be
(MW 1001.31) by FAB-MS and NMR analyses. In vitro studies found CK-4416 to show a potent inhibitory activity against sucrase and maltase, but it had low inhibition against
White Spot Syndrome Virus in Penaeid Shrimp Cultured in Korea
Shin, En-Joo ; Park, Jae-Hak ; Lee, Yeon-Hee ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 394~398
Because of the great concern over the possibility of contamination from the rod-shaped nuclear virus (PRDV) from Japan and white spot virus (WSSV) from Taiwan, most eggs used in Korean shrimp farms are currently obtained from local broodstock. In addition, the screening of imported broodstock for any viral presence at the National Fisheries Research and Development Institute is also mandatory. Nonetheless, massive mortality from white spot syndrome continues in Korea. In the present study, we present an improved PCR method to use tissue-extracted DNA instead of viral DNA extracted from a purified virus based on a sucrose density gradient, and produced results within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected within 8 h. In 1998, this modified PCR method was able to detect that diseased Penaeus japonicus were infected only with PRDV, while Fenneropenaeus chinensis were infected with both PRDV and WSSV. In 1999, PRDV and WSSV were detected in F. chinensis with signs of infection, but not with WSSV alone.
Cytotoxic Effect of Urushiol on Human Ovarian Cancer Cells
Choi, Ju-Youn ; Park, Chang-Soo ; Choi, Jong-Oh ; Rhim, Hyang-Shuk ; Chun, Heung-Jae ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 399~405
Urushiol, a natural pro-electrophilic quinone compound, has potential structural characteristics as antitumor chemotherapeutic agents. However, urushiol's use as an antitumor drug has some problems, because it is hardly miscible with an aqueous solution. Purified urushiol is highly viscous and soluble only in strong solvents. for this study, we prepared an urushiol-ethanol micro-emulsion with a unimodal size distribution by high-speed homogenization. This generated effective delivery of urushiol to its action wites, so that we could investigate its cytotoxic activity against cancer cells. Using a colony-forming assay, we were able to show that urushiol selectively inhibited the growth of the ovarian cancer cells PA-1 and 2774 at a concentration of
, whereas it had only a negligible effect on normal CHO cells at the same concentration. The data suggest that urushiol may have potential as an effective antitumor agent in the treatment of ovarian cancer. In addition, we addressed the question of whether the specific cytotoxic effect of urushiol is linked to apoptosis, by DNA fragmentation and DAPI staining assays. The inhibitory effects of urushiol on the growth of ovarian cancer cells was found to be associated with DNA fragmentation and the fragmented nuclei formation, both of which represent markers for the induction of apoptosis. Therefore, the results suggested that urushiol affected its profound cytotoxicity by triggering apoptosis in ovarian cancer cells.
Bioelectrochemical Denitrification by Pseudomonas sp. or Anaerobic Bacterial Consortium
Park, Doo-Hyun ; Park, Yong-Keun ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 406~411
In a bacterial denitrification test with Pseudomonas sp. and anaerobic consortium, more nitrates and less substrate were consumed but less metabolic nitrite was produced under an anaerobic
condition rather than under
condition. In a bioelectrochemical denitrification test with the same organisms, the electrochemically reduced neutral red was confirmed to be a substitute electron donor and a reducing power like
. The biocatalytic activity of membrane-free bacterial extract, membrane fraction, and intact cell for bioelectrochemical denitrification was measured using cyclic voltammetry. When neutral red was used as an electron mediator, the electron transfer from electrode to electron acceptor (nitrate) via neutral red was not observed in the cyclic voltammogram with the membrane-free bacterial extract, but it was confirmed to gradually increase in proportion to the concentration of nitrate in that of the membrane fraction and the intact cell of Pseudomonas sp.
Survival of Bifidobacterium breve in Acidic Solutions and Yogurt, Following Immobilization in Calcium Alginate Beads
Lee, Ki-Yong ; Kim, Ji-Youn ; Yu, Won-Kyu ; Lee, Yoon-Jong ; Yoon, Sung-Sik ; Heo, Tae-Ryeon ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 412~417
Sodium alginate was used to immobilize Bifidobacterium breve ATCC 15700 cells. The ability of the Ca-alginate beads to protect the B. breve ATCC 15700 was evaluated under different conditions including alginate concentration, bead size, pH, hydrogen peroxide, and storage period. The survival of the B. Breve ATCC 15700 was estimated in pasteurized yogurt, containing either the immobilized or free cells, throughout the storage period. The survival cells in bead after exposure to acidic solution (pH 3.0) increased with increase of both the alginate gel concentration and bead size. Also, immobilized cells in alginate bead were more resistant than the free cells to hydrogen peroxide, storage period, and the environment inside yogur. When retreated beads with skim milk and nonretreated beads were tested in acidified pH 3.0 TPY media including acetic and lactic acid, the number of viable cells in the retreated bead was approximately 10-fold higher than that of nonretreated beads. This suggests that the skim milk operated as a material decreasing the diffusion of acid and hydrogen perosicde into alginate gels. From this research, it was found that yogurt itself supported immobilized cells with an improved protection from the extreme acidity in yogurt.
Inhibition of Compylobacter jejuni in Chicken by Ethanol, Hydrogen Peroxide, and Organic Acids
Shin, Soon-Young ; Hwang, Han-Joon ; Kim, Wang-June ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 418~422
Growth inhibition of Compylobacter jejuni ATCC 33291 was observed in the presence of various preservatives at various temperatures. The addition of ethanol (0.5% to 5%), hydrogen peroxide (0.05%), acetic acid (1%), propionic acid, benzoic acid, and sorbic acid showed strong antibacterial activities against C. jejuni at pH 5.5 or 6.5. The addition of 1% acetic acid and lactic acid were most effective at
, followed by
. This indicated that the inhibitory effect was temperature dependent. In the chicken model system, the practical death rate of C. jejuni in the FBP-media with 1% acetic temperatures (
). Therefore, precaution has to be taken in the use of organic acids as a disinfectant in the chicken slaughterhouse.
Effect of Collagen Concentration on the Viability and Metabolic Function of Encapsulated Hepatocytes
Kim, Sung-Koo ; Yu, Sun-Hee ; Lee, Ji-Hyun ; Axel Racemacher ; Lee, Doo-Hoon ; Park, Jung-Keug ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 423~427
Chitosan/alginate capsules were formed by electrostatic interactions and had appropriated mechanical strength, permeability to albumin, and stability to hepatocytes. Rat hepatocytes were isolated and immobilized in chitosan/alginate capsules. During the encapsulation process with hepatocyte, 10% of viability was decreased mainly due to the low pH of the chitosan solution. Among various capsule fabrication methods, the chitosan-alginate capsule showed the highest mechanical strength. Addition of collagen in the capsule with hepatocytes enhanced hepatic metabolism as well as the cell viability for 2 weeks of culture. The hepatocyte in the capsule without collagen decreased the viability to 10% for 2-week cultures.
Levan-Producing Bacillus subtilis BS 62 and Its Phylogeny Based on Its 16S rDNA Sequence
Choi, Seong-Hyun ; Chang, Sung ; Choi, Woo-Young ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 428~434
A viscous substance producer strain BS62, which was isolated from conventional Chungkookjang, was examined for its productivity of levansucrase and levan during soybean fermentation at
. After one day of cultivation, the enzyme activity reached the highest level, 8 units
. Extracts of fermented soybeans were precipitated by ethanol and hydrolyzed by either 0.1 N HCl or invertase, and the hydrolyzates were analyzed using thin layer and ion chromatographies. Fructose was the only sugar detected. This suggest that fructose was derived from the levan produced by the strain BS62 during soybean fermentation. The aerobic, endospore-forming bacterium BS62 was identified as a Bacillus subtilis sp., based on the composition of its cellular fatty acids and phylogeny, which was determined by its 16S rDNA sequence.
-Phenylalkanoic Acids with Butyric Acid for Efficient Production of Aromatic Polyesters in Pseudomonas putida BM01
Song, Jae-Jun ; Choi, Mun-Hwan ; Yoon, Sung-Chul ; Huh, Nam-Eung ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 435~442
Poly(3-hydroxy-5-phenylvalerate) [P(3HPV)] was efficiently accumulated from 5-phenylvalerate (5PV) in Pseudomonas putida BM01 in a mineral salts medium containing butyric acid (BA) as the cosubstrate. A nove aromatic copolyester, poly(5 mol% 3-hydroxy-4-phenylbutyrate-co- 95 mol% 3-hydroxy-6-phenylhexanoate) [P(3HPB-co-3HPC)] was also synthesized from 6-phenylhexanoate (6PC) plus Ba. The two aromatic polymers, P(3HPV) and P(3HPB-co-3HPC), were found to be amorphous and showed different glass-transition temperatures at
, respectively. When the bacterium was grown ina medium containing 20 mM 5PV as the sole carbon source for 140 h, 0.4 g/l of dry cells was obtained in a flask cultivation and 20 wt% of P(3HPV) homopolymer was accumulated in the cells. However, when it was grown with a mixture of 2 mM 5PV and 50 mM BA for 40 h, the yield of dry biomass was increased up to 2.5 g/l and the content of P(3HPV) in the dry cells was optimally 56 wt%. This efficient production of P(3HPV) homopolymer from the mixed substrate was feasible because BA only supported cell growth and did not induce any aliphatic PHA accumulation. The metabolites released into the PHA synthesis medium were analyzed using GC or GC/MS. Two
-oxidation derivatives, 3-phenylpropionic acid and trans-cinnamic acid, were found in the 5V-grown cell medium and these comprised 55-88 mol% of the 5PV consumed. In the 6PC-grown medium containing Ba, seven
-oxidation and related intermediates were found, which included phenylacetic acid, 4-phenylbutyric acid, cis-4-phenyl-2-butenoic acid, trans-4-phenyl-3-butenoic acid, trans-4-phenyl-2-butenoic acid, 3-hydroxy-4-phenylbutyric acid, and 3-hydroxy-6-phenylhexanoic acid. Accordingly, based on the metabolite analysis, PHA synthesis pathways from the two aromatic carbon sources are suggested.
Physiological Responses of Oxygen-Tolerant Anaerobic Bifidobacterium longum under Oxygen
Ahn, Jun-Bae ; Hwang, Han-Joon ; Park, Jong-Hyun ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 443~451
In order to investigate what kind of response anaerobic bifidobacteria has on oxygen stress, five oxygen-tolerant bifidobacteria were isolated from human fecal samples. All were temporarily identified as Bifidobacterium longum through an analysis of carbohydrate utilization patterns and cellular fatty acid profiles. In the presence of oxygen, the lag phase became extended and the cell growth was suppressed. Bifidobacterial cell was able to remove dissolved oxygen in an early stage of growth and to overcome oxygen stress to a certain extent. The cell became long n size and showed a rough surface containing many nodes which were derived from abnormal or incomplete cell division. Cellular fatty acid profiled changed remarkably under a partially aerobic condition, so that the carbon chain of cellular fatty acid became short. All the dimethyl acetals originated from plasmalogen were reduced, any cyclopropane fatty acid, 9, 10-methyleneoctadecanoic acid (
), was increased remarkably. Oxygen stress induced a 5.5 kD protein in B. longum JI 1 of the oxygen-teolerant bifidobacteria, that was named Osp protein, and its N-terminal amino acid sequence was as follows: unknown amino acid-Thr-Gly-Val-Arg-Phe-Ser-Asp-Asp-Glu. Therefore, the oxygen-tolerant bifidobacteria seemed to defend against oxygen stress byincreasing the content of short fatty acid and cyclopropane fatty acid, and induction of an oxygen stress protein, but not the plasmalogen.
Characterization of Glutaryl 7-ACA Acylase from Pseudomonas diminuta KAC-1
Kim, Dae-Weon ; Kang, Sang-Mo ; Yoon, Ki-Hong ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 452~457
The glutaryl 7-aminocephalosporanic acid (glutaryl 7-ACA) acylase was purified from Pseudomonas diminuta KAC-1 cells isolated from soil, and characterized. The acylase was purified by procedures including ammonium sulfate fractionation and column chromatographies on DEAE-Sepharose, Phenyl-Sepharose, Q-Sepharose, and Superose 12H/R. The negative acylase was found to be composed of two subunits with molecular masses of approximately 55 kDa and 17 kDa, respectively. The isoelectric point of the enzyme was 4.0. The specific activities of the purified acylase were 8.0 and 7.0 U/mg on glutaryl 7-ACA and glutaryl 7-aminodesacetoxy cephalosporanic acid (glutaryl 7-ADCA), respectively, and
values were 0.45 mM for glutaryl 7-ADCA and 0.67 mM for glutaryl 7-ADCA. The enzyme had a pH optimum at 8.0 and a tmperature optimum at
. The acylase catalyzed the synthesis of glutaryl 7-ACA from glutaric acid and 7-ACA as well as the hydrolysis of glutaryl 7-ADCA, although the reaction rate of the synthesis was slower than that of the hydrolysis. In addition, it was found that the enzyme had a glutaryl transferase activity, thereby transferring the glutaryl group from one cephalosporin nucleus to another.
Localization of Paclitaxel in Suspension Culture of Taxus chinensis
Choi, Hyung-Kyoon ; Kim, Sang-Ic ; Song, Jai-Young ; Son ; Hong, Seung-Suh ; Durzan, Don-J. ; Lee, Hyong-Joo ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 458~462
The localization of paclitaxel was investigated in suspension culture cells of Taxus chinensis. Over 93% of the cell-associated paclitaxel were detected throughout the entire culture period. Intracellular localization of paclitaxel over the culture time was analyzed further by cell fractionation for days 21 and 42. Paclitaxel contents in intracellular organelles were decreased at day 42, while the content in the cell wall fraction was increased at day 42 compared to the value for day 21. The localization of paclitaxel in the cell wall was confirmed by using the immunocytochemical method with the aid of a confocal laser scanning microscope.
Inclusion Complexation of a Family of Cyclsohoraoses with Indomethacin
Lee, Sang-Hoo ; Kwon, Chan-Ho ; Choi, Young-Jin ; Seo, Dong-Hyuk ; Kim, Hyun-Won ; Jung, Seun-Ho ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 463~468
Cyclosophoraoses are a class of unbranced cyclic-(1longrightarrow2)-
-D-glucans found in the Rhizobium species. Their unique cyclic structures and high solubility make them potent for inclusion complexation as a host for an insoluble guest molecule. A family of neutral cyclosophoraoses (DP 17-27) isolated from Rhizobium meliloti 2011 was used as a host for inclusion complexation with an insoluble guest drug, indomethacin. A high performance liquid chromatographic analysis indicated that the inclusion complexation of cyclosophoraoses greatly ehanced the solubility of indomethacin compared with
-cyclodextrin. The estimated value of the association constant of the complex in water for
-cyclodextrin and cyclosophoraoses was
, respectively. NMR spectroscopy showed that the inclusion complex was characterized by the interaction of the indole ring moiety of indomethacin with the cavity of cyclosophoraoses.
Identification of Streptomyces sp. AMLK-335 Producing Antibiotic Substance Inhibitory to Vancomycin-Resistant Enterococci
Rhee, Ki-Hyeong ; Choi, Kyung-Hee ; Kim, Chang-Jin ; Kim, Chang-Han ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 469~474
The actinomycete strain AMLK-335 was antagonistic to vancomycin-resistant enterococci (VRE). Based on the diaminopimelic acid (DAP) type, and morphological and physiological characteristics revealed by scanning electron microscopy (SEM), AMLK-335 was confirmed to belong to the genus Streptomyces. Analysis of the 16S rDNA nucleotide sequences found AMLK-335 to have a relationship with Streptomyces platensis. The production of antibiotic from this strain was most favorable when cultured on glucose, polypeptone, yeast extract (PY) medium for 6 days at
. The antibiotic was identified as cyclo(L-phenylalanyl-L-prolyl) by comparing ti with the reported MS and NMR spectral data. Cyclo(phe-pro) from the PY cultures of AMLK-335 was most effective (K-98-258). Futhermore, cyclo(phe-pro) had antimicrobial activity against Bacillus subtilis, Microcuccs luteus, Staphylococcus aureus, and Saccharomyces cerevisiae, but it wa ineffective against Candida albicans, Streptomyces murinus, and Aspergillus niger.
Phylogenetic Classification of Antrodia and Related Genera Based on Ribosomal RNA Internal Transcribed Spacer Sequences
Kim, Seon-Young ; Park, So-Yeon ; Jung, Hack-Sung ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 475~481
Sequences of ribosomal internal transcribed spaces (ITS) obtained from two Antrobia species and two related species were compared to investigate intrageneric and intergeneric phylogenetic relationships of Antrodia. The results showed that Antrodia species causing a brown rot in wood did not form a monophyletic clade and were separated into three distinct groups. Antrodia gossypina and A. vaillantii formed a clade having rhizomorphs as a homologous character. Antrodia serialis, A. sinuosa, and A. malicola formed a group together with Daedalea, Fomitopsis, and Postia species with brown rot habit. Antrodia xantha with a trimitic hyphal system and amyloid skeletal hyphae formed another distinct clade form other Antrodia species. The Antrodia species were separated from white rot genera such as Antrodiella, Diplomitoporus, Junghuhnia, and Steccherinum, indicating the phylogenetic importance of the rot type in the classification of the Polyporaceae.
Sequencing and Baculovirus-Based Expression of the Glycoprotein B2 Gene of HSV-2 (G)
Uh, Hong-Sun ; Park, Jong-Kuk ; Kang, Hyun ; Kim, Soo-Young ; Lee, Hyung-Hoan ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 482~490
The gene for glycoprotein B (gB2) of HSV-2-strain G was subcloned, sequenced, recombinated into the lacZ-HcNPV, expressed in insect cells, and compared with the homologous gene of other HSV-2 strains. The ORF of the gB2 gene was 2,715 bp. The overall nucleotide sequence homology of te gB2 gene compared ith that of the two previously reported HSV-2 strains appeared to be over 98%. A recombinant virus named Baculo-gB2 protein in insect cells. The recombination was confirmed by a PCR and the expression was demonstrated by radio immunoprecipitation. Insect cells infected with the Baculo-gB2 virus synthesized and processed gB2 with approximately 120 kDa in the cells, and then secreted it into the culture media, where it reacted with a nomoclonal antibody to gB2. The gB2 polypeptide contained two main hydrophobic regions (a signal sequence from 1 to 23 amino acid residues, and a membrane anchor sequence from aa 745 to 798), eight N-glycosylation sites evenly distributed, and was rich in alanine (11.2%). Antibodies to this recombinant protein that were raised in mice recognized the viral gB2 and neutralized the infectivity of the HSV-2 in vitro. There results show that the gB2 protein was successfully porduced in insect cells and could be used to raise a protective neutralizing antibody. Accordingly, this particular recombinant protein may be useful in the development of a subunit vaccine.
Improvement of Decarboxylating Agar Medium for Screening Biogenic Amine-Producting Bacteria in Kimchi
Mah, Jae-Hyung ; Shin, Soon-Young ; Lee, Heung-Shick ; Cho, Hong-Yon ; Hwang, Han-Joon ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 491~496
A modification of decarboxylating agar medium as described by niven was performed to improve the detection method of biogenic amine-producing bacteria and to eliminate the false-positive. A total of 120 bacterial strains isolated from kimchi were used to evaluate different dicarboxylating agar media and for screening biogenic amines. Potential false-positives ranged from approximately 66 to 79% of the strains tested in the already well-known media. In our improved medium, none of the 120 strains showed the potential false-positives. There was a good agreement (81.7%-87.5%) between the results obtained by the improved medium and by HPLC analysis. Consequently, this medium was greatly improved in screening biogenic amine-producing bacteria and discarding false-positives. Of the 120 kimchi isolates, 14.2, 18.3, 37.5, and 0.8% were found by HPLC to be the producers of histamine, tyramine, putrescine (as a form of spermine), and cadaverine, respectively. The proportion of biogenic amine producer during kimchi fermentation increased to a maximum at an immature period and decreased thereafter.
Removal and Inactivation of Viruses during Manufacture of a High Purity Antihemophilic Factor VII Concentration from Human Plasma
Kim, In-Seop ; Choi, Yong-Woon ; Lee, Sung-Rae ; Woo, Hang-Sang ; Lee, Soung-Min ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 497~503
The purpose of this study was to examine the efficacy and mechanism of the cryo-precipitation, solvent/detergent (S/D) treatment, monoclonal anti-FVIIIc antibody (mAb) column chromatography, Q-Sepharose column chromatography, and lyophilization involved in the manufacture of antithemophilic factor VII(GreenMono) from human plasma, in the removal and/or inactivation of blood-borne 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 all selected for this study. BHV and EMCV were effectively partitioned from a factor VII during the cryo-precipitation with a log reduction factor of 2.83 and 3.24, respectively. S/D treatment using the organic solvent, tri(n-butyl) phosphate (TNBP), and the detergent, Triton X-100, was a robust and effective step in inactivating enveloped viruses. The titers of BHV and BVDV were reduced from the initial titer of 8.85 and
, respectively, reaching undetectable levels within 1 min of the S/D treatment. The mAb chromatography was the most effective step for removing nonenveloped viruses, EMCV and PPV, with the log reduction factors of 4.86 and 3.72, respectively. Q-Sepharose chromatography showed a significant efficacy for partitioning BHV, BVDV, EMCV, and PPV with the log reduction the log reduction factors of 2.32, 2.49, 2.60, and 1.33 respectively. Lyophilization was an effective step in inactivating g nonenveloped viruses rather than enveloped viruses, where the log reduction factors of BHV, BVDV, DMCV, and PPV were 1.41, 1.79, 4.76, and 2.05, respectively. The cumulative log reduction factors of BHV, BVDV, EMCV, and PPV were
7.88, 15.46, and 7.10, respectively. These results indicate that the production process for GreenMono has a sufficient virus-reducing capacity to achieve a high margin of the virus safety.
The Phylogenetic Relationship of Several Oscillatorian Cyanobacteria, Forming Blooms at Daecheong Reservoirs, Based on Partial 16S rRNA Gene Sequences
Lee, Wook-Jae ; Bae, Kyung-Sook ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 504~507
The partial 16S gene sequences of six filamentous cyanobacterial strains, Oscillatoria lmosa KCTC AG10168, Oscillatoria princeps KCTC AG10153, Oscillatoria sp. KCTC AG 10184, Phormidium tenue KCTC AG10158, Phormidium parchydematicum KCTC AG10164, and Lyngbya hieronymusii KCTC AG10199, which were isolated in the late summer at Daecheong Reservoirs, were determined and assigned their phylogenetic and taxonomic position among taxa of order Ocillatoriales whose partial 16S rRNA gene sequences aligned in this suty, were very heterogeneously clustered with other taxa. The two strains, Oscillatoria limosa KCTC AG10168 and O. princeps KCTC AG10153, formed a cluster with O. sancta PCC7515, which supported 64% of the bootstrap trees with high similarity (19-96.15%). Strain Oscillatoria sp. KCTC AG10184, that was known to produce a nasty substance, was closely related to the toxic Oscillatoria group. The study on morphological variation in various environments and toxin production will confirm the taxonomic status of these species. Phormidium tenus KCTC AG10158 and Phormidium parchydematicum KCTC AG10164 made a cluster with other oscillatorian species of Phormidium, Oscillatoria, and Leptolynbya, which supproted 100% of the bootstrap trees with a very high sequence smilarity (96.8-99.8%) in thsi study. The sequence analysis in this study also supported that taxa of oscillatoriales are not monophyletic. Some of the fractures, such as the presence or absence of sheath and cell shape, which were used to define them, would be inadequate and should be reconfirmed. We suggest that sequences of partial 16S rRNA gene fragments aligned in this study should be more useful than morphological features in the identification and reconfirmation of the taxonomic status of these oscillactorian cyanobacteria.
Cloning and Expression of a Yeast Cell Wall Hydrolase Gene (ycl) from Alkalophilic Bacillus alcalophilus subsp. YB380
Ohk, Seung-Ho ; Yeo, Ik-Hyun ; Yu, Yun-Jung ; Kim, Byong-Ki ; Bai, Dong-Hoon ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 508~514
A stuructural gene (ycl) encoding novel yeast cell wall hydrolase, YCL, was cloned from alkalophilic Bacillus alcalophilus subsp. YB380 by PCR, and transformed into E. coli JM83. Based on the N-terminal and internal amino acid sequences of the enzyme, primers were designed for PCr. The positive clone that harbors 1.8 kb of the yeast cell wall hydrolase gene was selected by the colony hybridization method with a PCR fragment as a probe. According to the computer analysis, this gene contained a 400-base-paired N-terminal domain of the enzyme. Based on nucletide homology of the cloned gene, a 850 bp fragment was amplified and the C-terminal domain of the enzyme was sequenced. With a combination of the two sequences, a full nucleotide sequence for YCL was obtained. This gene, ycl, consisted of 1,297 nucleotides with 27 nucleotides with 27 amino acids of signal sequence, 83 redundant amino acids of prosequence, and 265 amino acids of the mature protein. This gene was then cloned into the pJH27 shuttle vector and transformed into the Bacillus subtilis DB104 to express the enzyme. It was confirmed that the expressed cell wall hydrolase that was produced by Bacillus subtilis DB104 was the same as that of the donor strain, by Western blot using polyclonal antibody (IgY) prepared from White Leghorn hen. Purified yeast cell wall hydrolase and expressed recombinant protein showed a single band at the same position in the Western blot analysis.
Cloning and Characerization of the Ribosomal RNA Gene from Gonyaulax polyedra
Lee, Hee-Gyun ; Lee, Ji-Yeon ; Lee, Dong-Hee ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 515~523
The dinoflagellates have some primitive nuclear features and are evolutionarily intermediate between prokaryotes and eukaryotes. The small subunit ribosomal RAN gene, the 5.8S ribosomal RNA gene, and the internal transcribed spacer (ITS) of Gonyaulax polyedra were cloned, and their sequences were analyzed to better understand their evolutionary position. The small subunit ribosomal RNA gene was 1,794 nt long, the large subunit ribosomal RNA gene was approximately 3,500 nt long, and the 5.8S ribosomal RNA gene was 159 nt long. The first internal transcribed spacer (ITS1) was 191 nt long, and the second internal transcribed spacer (ITS2) was 185 nt long. The intergenic spacer of the ribosomal RNA gene (IGS) was about 2,200 nt long, indicating that 5,800 nt of transcribed sequences were separated by roughly 2,200 nt of intergenic spacer. The ribosomal RNA genes were repeated many times and arranged in a head-to-tail, tandemly repeated manner. The repeating unit of ribosomal RNA gene of G. polyedra was proposed to be 8,000 nt long. Based on the lengths of ribosomal RNA, sequence alignments with representative organisms, and phylogenetic analysis on ribosomal RNA, G. polyedra appears to be one of the alveolates branched from the eukaryotic crown and, among dinoflagellates, it seems to not have emerged early.
Novel Dosimeter for Low-Dose Radiation Using Escherichia coli PQ37
Park, Seo-Hyoung ; Kim, Tae-Hwan ; Cho, Chul-Koo ; Lee, Yeon-Hee ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 524~528
The measurement of radiation response using simple and informative techniques would be of great value in studying the genetic risk following occupational, therapeutic, or accidental exposure to radiation. When patients receive radiation therapy, many suffer from side effects. Since each patient receives a different dose due to different physical conditions, it is important to measure the exact dose of radiation received by each patient to lessen the side effects. Even though several biological dosimetric systems have already been developed, there is no ideal system that can satisfy all the criteria for an idean dosimetric system, especially for low-dose radiation as used in radiation therapy. In this study, an SOS Chromotest of E. coli PQ37 was evaluated as a novel dosimeter for low-dose gamma-rays. E. coli PQ37 was originally developed to screen chemical mutagens using the SOS Chromotest-a colorimtric assay, based on the induction of
-galactosidase ue to DNA damage. The survival fraction of E. coli PQ37 decreased dose-dependently with an increasing dose of cobalt-60 gamma-rays. Also, a good linear correlation was found between the biological damage revealed by the
-galactosidase expression and the doses of gamma-rays. The expression of
-galactosidase activity that responded to low-dose radiation under 1 Gy was
(Y, absorbance at 420 nm; D, Dose of irradiation) as calculated using Graph Pad In Plot and Excel. When a rabbit was fed with capsules containing an agar block embdded with E. coli PQ37 showed a linear response to the radiation doses. Accordingly, the results confirm that E. coli PQ37 can be used as a sensitive biological dosimeter fro cobalt-60 gamma-rays. To the best of our knowledge, this is the first time that a bacterium has been used as a biological dosimeter, especially for low-dose radiation.
A Gene Encoding
-amylase from Saprolegnia parasitica and Its Expression in Saccharomyces cerevisiae
Kim, Hee-Ok ; Park, Jeong-Nam ; Shin, Dong-Jun ; Blaise Lee, Hwang-Hee ; Chun, Soon-Bai ; Bai, Suk ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 529~533
-Amylase cDNA fragment from the oomcete Saprolegnia parasitica was cloned by reverse transcription-polymerase chain reaction (RT-PCR) using degenerate oligonucleotide primers derived from conserved
-amylase sequences. The 5'and 3'regions of the
-amylase gene were amplified using the rapid amplification of cDNA ends (rACE) system. It consisted of an open reading frame of 1,350 bp for a protein of 450 amino acids. Comparison between the genomic and cDNA sequences revealed that the intron was not present in the coding region. The deduced amino acid sequence of the
-amylase gene had a 97% similarity to the
-amylase of Saprolegnia ferax, followed by 41% similarity to those of Arabidopsis thaliana, Hordeum vulgare, and Zea mays. The
-amylase gene was also expressed in Saccharomyces cerevisiae by placing it under the control of the alcohol dehydrogenase gene (ADC1) promoter.
Asparagine Residue at Position 71 is Responsible for Alkali-Tolerance of the Xylanase from Bacillus Pumilus A-30
Liu, Xiang-Mei ; Qi, Meng ; Lin, Jian-Aiang ; Wu, Zhi-Hong ; Qu, Yin-Bo ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 534~538
The xynA gene encoding an alikali-tolerant endo-1,4-
-xylanase (XYN) was cloned from the alkalophilic Bacillus pumilus A-30. The nucleotide sequence of a 974-bp DNA fragment containing the xynA was determined. An ORF of 684 nucleotides that encoded a protein of 228 amino aicds was detected. Asparagine-71 of XYN from B. Pumilus A-30 showed to be highly conservative in alkaline xylanases of family G/11, upon comparing the amino acid sequences of 17 family G/11 xylanases. Site-directed mutation of N71D of the xynA gene resulted in a decrease of 12.4% in the specific acitivity and a significant decline in the enzyme activity in the alkaline pH range.
In Vitro Detection of Apoptosis in Human Promyleoytic Leukemia HL-60 Cells by
Lee, Chul-Hoon ; Lee, Min-A. ; Cho, Youl-Hee ; Lim, Hae-Young ; Jung, Ji-Hyun ; Kim, Kyung H. ; Lim, Yoong-Ho ;
Journal of Microbiology and Biotechnology, volume 11, issue 3, 2001, Pages 539~542
-NMR spectroscopy was used to detect apoptosis in HL-60 cells in vitro. The relationship between cell apoptosis and NMR data was validated by the flow cytometry assay. To evaluate the NMR apoptosis results, the ratio of methylene and methyl groups caused by lipids was used. In addition, an identical analysis was applied to HepG2 cells. Detection of apoptotic cell death by NMR spectroscopy was oserved.