Go to the main menu
Skip to content
Go to bottom
REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
Journal of Microbiology and Biotechnology
Journal Basic Information
Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
Editor in Chief :
Volume & Issues
Volume 14, Issue 6 - Dec 2004
Volume 14, Issue 5 - Oct 2004
Volume 14, Issue 4 - Aug 2004
Volume 14, Issue 3 - Jun 2004
Volume 14, Issue 2 - Apr 2004
Volume 14, Issue 1 - Feb 2004
Selecting the target year
Glycoantigen Biosyntheses of Human Hepatoma and Colon Cancer Cells are Dependent on Different N-Acetylglucosaminyltransferase-III and -V Activities
Kim, Cheorl-Ho ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 891~900
-l ,4N-acetylglucosaminyltransferase-III (GnT-III) and UDP-N-GlcNAc:
-1,6N-acetylglucosaminyltransferase-V(GnT - V) activities were determined in human hepatoma cell lines and metastatic colon cancer cells, and their activities were compared with those of normal liver cells and fetal hepatocytes. GnT-III activities were higher than those of GnT-V in hepatic carcinoma cells. When the two enzyme activities were assayed in highly metastatic colon cancer cells, GnT - V activities were much higher than those of GnT-III. When GlcN, GlcN-biant-PA and UDP-GlcNAc were used as substrates, the enzymes displayed different kinetic properties between hepatic and colon cancer cells, depending on their metastatic potentials. Normal cells of two origins had characteristically very low levels of GnT-III and -V activities, whereas hepatoma and colon cancer cells contained high levels of activities. These data were supported by RT-PCR and Northern blot analyses, showing that the expression of GnT-III and -V mRNAs were increased in proportion to the enzymatic activities. The increased GnT-III, md -V activities were also correlated with increased glycosylation of the cellular glycoproteins in hepatoma and colon cancer cells, as examined by lectin blotting analysis by using wheat germ glutinin (WGA), erythroagglutinating phytohemagglutinin (E-PHA), leukoagglutinating phytohemagglutinin (L-PHA), and concanavalin A (Con A). Treatment with retinoic acid, a differentiation agent, resulted in decreases of both GnT-III and -V activities of HepG2 and HepG3 cells. In colon carcinoma cells, however, treatment with retinoic acid resulted in a reduction of GnT-V activity, but not with GnT-III activity. Although the mechanism underlying the induction of these mzymes is unclear, oligosaccharides in many glycoproteins have been observed of cancer cells.
Effects of Soil Types on the Biodegradation of Crude Oil by Nocardia sp. H17-1
Yoon, Byung-Dae ; Baek, Kyung-Hwa ; Kim, Hee-Sik ; Moon, Seong-Hoon ; Lee, In-Sook ; Oh, Hee-Mock ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 901~905
The degradation and mineralization of crude oil were investigated over 50-days in three soils, loamy sand, sand, and combusted loamy, which were artificially contaminated with crude oil (50 g
) and inoculated with Nocardia sp. H17-1. The degradation efficiency of total petroleum hydrocarbon (TPH) in sand was the highest at 76% among the three soils. The TPH degradation rate constants
in loamy sand, sand, and combusted loamy sand were 0.027
, and 0.016
, respectively. In contrast, the total amount of
evolved was the highest at 146.1 mmol in loamy sand. The
evolution rate constants
in loamy sand, sand, and combusted loamy sand were 0.057
, and 0.037
, respectively. Therefore, it seems that the degradation of crude oil in soils can be proportional to the soil pore space and that mineralization can be accelerated with the increase of organic substance.
Cold-Seep Sediment Harbors Phylogenetically Diverse Uncultured Bacteria
Cho, Jae-Chang ; Lee, Sang-Hoon ; Oh, Hae-Ryun ; Lee, Jung-Hyun ; Kim, Sang-Jin ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 906~913
A culture-independent molecular phylogenetic survey was carried out on the bacterial community in cold-seep sediment at Edison Seamount, south of Lihir Island, Papua New Guinea. Small-subunit rRNA genes were amplified directly from the sediment DNA by PCR and cloned. The majority of the cloned 16S rRNA gene sequences were most closely related to as-yet-uncultivated microorganisms found in deep-sea sediments, and were primarily affiliated with one of four groups: the
-subdivisions of Proteobacteria, and Cytophaga-Flavobacterium-Bacteroides. We did not recover any sequences related to cyanobacteria, prochlorophytes, and
-Proteobacteria, which are known to occur in great abundance within the surface mixed layer of the Atlantic and Pacific Oceans. The majority of the cloned
-Proteobacterial sequences were closely related to chemoautotrophic sulfur-oxidizing symbionts of marine benthic fauna, and the
-Proteobacterial sequences to sulfate- and sulfur-reducing bacteria, indicating that they might play an important role in chemoautotrophic primary production and the sulfur cycle in the cold-seep area. There results demonstrate the high diversity of the bacterial community in the cold-seep sediment, and substantially expand knowledge of the extent of bacterial diversity in this formidable and unique habitat.
Biocatalytic Oxidation-Reduction of Pyruvate and Ethanol by Weissella kimchii sk10 Under Aerobic and Anaerobic Conditions
Kang, Hye-Sun ; Park, Sun-Mi ; Park, Doo-Hyun ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 914~918
This study was carried out to analyze the metabolic flux of W. kimchii sk10 on pyruvate and ethanol as a carbon source. The sk10 grown on ethanol produced acetate under aerobic conditions rather than under anaerobic conditions. The lactate and acetate were produced on ethanol plus pyruvate by the sk10 grown under aerobic and anaerobic conditions, respectively. The resting cell of sk10 produced 99.1 mM acetate and 17.3 mM lactate under aerobic conditions and 51.1 mM acetate and 62.4 mM lactate under anaerobic conditions from ethanol plus pyruvate, respectively. This result is thought to be due to the difference in the
ratio depending on the growth conditions. The 11-fold overproduction of NADH peroxidase results in a low
ratio under aerobic growth conditions. At the low
ratio, the metabolic flux of pyruvate toward lactate has to be shifted to a flux toward acetate without NADH oxidation to
, and ethanol oxidation to acetate coupled to
reduction to NADH has to be activated.
Metabolic Flux Shift of Weissella kimchii sk10 Grown Under Aerobic Conditions
Park, Sun-Mi ; Kang, Hye-Sun ; Park, Doo-Hyun ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 919~923
The sk10 isolated from kimchi was identified as W. kimchii on the basis of l6s-rDNA sequencing. Studies were made to analyze the metabolic flux shift of the sk10 on glucose under aerobic growth conditions. The sk10 produced 38.2 mM acetate, 16.3 mM ethanol, and 33.2 mM lactate under aerobic conditions, but 2.4 mM acetate, 48.0 mM ethanol, and 44.1 mM lactate under anaerobic conditions. The NADH peroxidase (NADH-dependent hydrogen peroxidase) activity of sk10 grown under aerobic conditions was 11 times higher than that under anaerobic conditions. Under the low ratio of
, the metabolic flux toward lactate and ethanol was shifted to the flux through acetate kinase without NADH oxidation. The kinds of enzymes and metabolites of sk10 were close to those in the pathway of Leuconostoc sp., but the metabolites produced under aerobic growth conditions were different from those of Leuconostoc sp. The stoichiometric balance calculated using the concentrations of metabolites and substrate was about 97%, coincident with the theoretical values under both aerobic and anaerobic conditions. From these results, it was concluded that the metabolic flux of W. kimchii sk10 was partially shifted from lactate and ethanol to acetate under aerobic conditions only.
Isolation, Identification, and Characterization of a Bacteriocin-Producing Enterococcus sp. from Kimchi and Its Application to Kimchi Fermentation
Moon, Gi-Seong ; Kang, Chang-Hoon ; Pyun, Yu-Ryang ; Kim, Wang-June ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 924~931
A bacteriocin-producing lactic acid bacterium, which strongly inhibited the Lactobacillus plantarum recognized as an important acid spoilage microorganism in kimchi fermentation, was isolated from kimchi. From morphological, physiological, sugar fermentation, biochemical tests, and l6S rDNA sequencing results, the isolate was identified as an Enterococcus sp. and designated as Enterococcus sp. K25. The bacteriocin produced by Enterococcus sp. K25 inhibited several Gram-positive bacteria, including Lb. plantarum, whereas it did not inhibit Gram-negative bacteria and yeasts. Optimal temperature and pH for the bacteriocin production were
and 5.5, respectively. Enterococcus sp. K25 was applied to kimchi manufacturing alone and together with other preservatives (i.e., chitosan and fumaric acid). In addition, growth of lactic acid bacteria, pH, and titratable acidity (TA) were measured during aging at
. Inoculation of Enterococcus sp. K25 together with fumaric acid showed the most synergistic effect on extension of kimchi shelf-life. Compared to control (no addition), the treatment prolonged the kimchi shelf-life up to 6 days, whereupon the eight-point TA value recognized as the edible limit was reached.
Improvement of Hydrocarbon Recovery by Two-Stage Cell-Recycle Extraction in the Cultivation of Botryococcus braunii
An, Jin-Young ; Sim, Sang-Jun ; Kim, Byung-Woo ; Lee, Jin-Suk ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 932~937
In situ extraction by organic solvent was studied in order to improve the recovery yield of hydrocarbon from the culture of Botryococcus braunii, a green colonial microalga. When the solvent mixture of octanol as an extractive solvent and n-octane as a biocompatible solvent was added to a two-phase column, the algal growth was seriously inhibited, even at a low concentration of polar octanol. Therefore, a two-stage cell-recycle extraction process was proposed to improve the contact area between the organic phase and the aqueous phase. The hydrocarbon recovery with in situ cell-recycle extraction showed a three-fold increase (57% of cell) in yield over that with two-phase extraction. In addition, over 60% of the hydrocarbon could be recovered without serious cell damage by downstream separation when this process was applied to the culture broth after batch fermentation.
Biotransformation of the Fungicide Chlorothalonil by Bacterial Glutathione S-Transferase
Kim, Young-Mog ; Park, Kun-Bawui ; Choi, Jun-Ho ; Kim, Jang-Eok ; Rhee, In-Koo ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 938~943
A gene responsible for the chlorothalonil-biotransformation was cloned from the chromosomal DNA of Ochrobactrum anthropi SH35B, an isolated bacterium strain from soil. We determined the nucleotide sequences and found an open reading frame for glutathione S-transferase (GST). The drug-hypersensitive Escherichia coli KAM3 cells transformed with a plasmid carrying the GST gene can grow in the presence of chlorothalonil. The GST of O. anthropi SH35B was expressed in E. coli and purified by affinity chromatography. The fungicide chlorothalonil was rapidly transformed by the purified GST in the presence of glutathione. No significant difference in the chlorothalonil-biotransformation effect was observed among the thiol compounds (cysteine, reduced glutathione, and
-mercaptoethanol). Thus, the result reported here is the first evidence on the chlorothalonil-biotransformation by conjugation with the cellular free thiol groups, especially glutathione, catalyzed by the bacterial GST.
Optimization of Submerged Culture Conditions for Mycelial Growth and Exopolysaccharides Production by Agaricus blazei
Kim, Hyun-Han ; Na, Jeong-Geol ; Chang, Yong-Keun ; Chun, Gie-Taek ; Lee, Sang-Jong ; Jeong, Yeon-Ho ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 944~951
The influences of inoculum size, pH, and medium composition on mycelial growth and exopolysaccharides (EPS) production were investigated in shake flasks and in a bioreactor. The optimum inoculum size for both mycelial growth and EPS production was identified to be 10% (v/v) in shake flask cultures. The optimal initial pH for mycelial growth and EPS production in shake flask cultures were found to be 5.0 and 7.0, respectively. However, the optimal pH was 5.0 for both mycelial growth and EPS production in bioreactor cultures where the pH was regulated. The optimal mass ratio of the two major carbon sources, glucose to dextrin, was 1:4. The optimal mass ratio of the two major nitrogen sources, yeast extract to soy tone peptone, was 2:1. When 500 mg
was added to the bioreactor culture, both mycelial growth and EPS production were enhanced by approximately 10%. Under the optimized conditions, a mycelial biomass of 9.85 g
and an EPS concentration of 4.92 g
were obtained in 4 days.
Screening and Characterization of Psychrotrophic, Lipolytic Bacteria from Deep-Sea Sediments
Zeng, Xiang ; Xiao, Xiang ; Wang, Peng ; Wang, Rengping ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 952~958
Of 23 psychrotrophic bacteria isolated from the west Pacific deep-sea sediments, 19 were assigned to the
-Proteobacteria, 3 to the <
-Proteobacteria, and 1 to the Gram-positive bacteria, as determined by their 16S rDNA sequences. Ten psychrotrophs, affiliated to the Psychrobacter, Pseudoalteromonas, and Pseudomonas genera in the
-Proteobacteria group, were screened for lipolytic bacteria. The majority of the lipolytic isolates had growth temperatures between 4-
, and all of them were neutrophilic, aerobic, or facultatively anaerobic, and some were able to produce multiple kinds of ectohydrolytic enzymes. The deep-sea strains Psychrobacter sp. wp37 and Pseudoalteromonas sp. wp27 were chosen for further lipase production analysis. Both strains had the highest lipase production when grown at 10 to
; their highest lipase production occurred at the late-exponential growth stage; and the majority of the enzymes were excreted to the outside of the cells. Lipases from both strains had the same optimal reaction temperature and pH (20-
, pH 7-8) and could retain about 60% of their highest activity at
. Furthermore, SDS-PAGE and an in-gel activity test showed that they had the same high molecular mass of about 85 kDa.
Phylogenetic Analysis of Harmful Algal Bloom (HAB)-Causing Dinoflagellates Along the Korean Coasts, Based on SSU rRNA Gene
Kim, Se-Hee ; Kim, Keun-Yong ; Kim, Chang-Hoon ; Lee, Woo-Sung ; Chang, Man ; Lee, Jung-Hyun ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 959~966
Twenty-three cultures of harmful algal bloom (HAB)-(causing dinoflagellates were isolated from the coastal waters of Korea. For each of the 14 morphospecies, the nuclearencoded small subunit (SSU) rDNA was analyzed to determine the phylogenetic relatedness of the species. Despite temporal and spatial isolation, 3-4 clonal cultures of Alexandrium catenella, Cochlodinium polykrikoides, and Gymnodinium catenatum had 100% identical SSU rDNA sequences. In contrast, heterogeneities in the SSU rDNA sequences were observed in Akashiwo sanguinea and Lingulodinium polyedrum strains. Extreme sequence polymorphism was shown within the SSU rRNA genes of an Al. tamarense clonal culture. A homology search in GenBank revealed that 11 dinoflagellate species were located in clusters corresponding to their morphological classification. The SSU rDNA sequences of C. polykrikoides, Gyrodinium instriatum, and Pheopolykrikos hartmannii, which were determined for the first time in this study, showed the following phylogenetic relationships: C. polykrikoides formed an independent branch separated from other dinoflagellates; Gyr. instriatum was placed in a monophyletic group with Gyr. dorsum and Gyr. uncatenum; and Ph. hartmanii, which forms a distinct two-celled pseudocolony, belonged to Gymnodinium sensu Hansen and Moestrup.
Evidence for Sulfite Proton Symport in Saccharomyces cerevisiae
Park, Hoon ; Alan T. Bakalinsky ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 967~971
The kinetics of sulfite uptake were examined in a wild-type laboratory strain of Saccharomyces cerevisiae to determine if carrier-mediated sulfite uptake involved a proton symport, as previous studies on sulfite uptake have suggested both an active process and facilitated diffusion. Accumulation of intracellular sulfite was initially rapid and linear up to 50 sec. Uptake was saturable at final concentrations equal to or greater than 3 mM sulfite, and increased 2-fold in the presence of 2% glucose. Uptake was significantly reduced in cells pretreated with 100-500
M carbonyl cyanide mchlorophenylhydrazone (CCCP) or 2,4-dinitrophenol (DNP), both of which dissipate proton gradients. Uptake was also significantly inhibited in the presence of 1 mM arsenate, an inhibitor of ATP synthesis. Extracellular alkalization was observed in cells incubated with 1-2 mM sulfite in a weak tartrate buffer at pH 3.5 and 4.5. These findings suggest that the bisulfite ion,
, an anionic form of sulfite, is taken up by a carrier-mediated proton symport. A met16 sull sul2 mutant, impaired in both sulfite formation and sulfate uptake, was found able to grow on a medium with sulfite as the sole Sulfur source, indicating that the sulfate transporters Sul1p and Sul2p are not required for sulfite uptake.
Hypoglycemic Effect of Exo- and Endo-biopolymers Produced by Submerged Mycelial Culture of Ganoderma lucidum in Streptozotocin-Induced Diabetic Rats
Yang, Byung-Keun ; Michael A Wilson ; Cho, Kai-Yip ; Song, Chi-Hyun ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 972~977
The hypoglycemic effect of an exo-biopolymer (EXO) and endo-biopolymer (ENDO) produced from submerged mycelial culture of Ganoderma lucidum was investigated in streptozotocin (STZ)-induced diabetic rats. Both the EXO and ENDO showed hypoglycemic potential, however, the former proved to be more potent than the latter. The administration of the EXO at the dose of 100 mg/kg body weight (BW) significantly reduced the plasma glucose level (23.5%) and increased the plasma insulin level (2.2 fold) in the diabetic animals. The EXO also lowered the plasma total cholesterol, triglyceride, low-density lipoprotein (LDL) cholesterol, and athrogenic index by 14.7, 31.4, 24.1, and 45.4%, respectively, and reduced the liver total cholesterol and triglyceride levels by 6.7 and 25.8%, respectively. It increased the plasma high-density lipoprotein (HDL) cholesterol (37.7%), compared to the control group. Furthermore, the alanine transaminase (ALT) and aspartate transaminase (AST) showed lower activities in the EXO administered groups than the other experimental groups. Taken together, these results suggest that the exo-biopolymer may alleviate the blood glucose level by increased insulin secretion.
Inhibition of Cell Cycle Progression and Induction of Apoptosis in HeLa Cells by HY558-1, a Novel CDK Inhibitor Isolated from Penicillium minioluteum F558
Lim, Hae-Young ; Kim, Min-Kyoung ; Cho, Youl-Hee ; Kim, Jung-Mogg ; Lim, Yoong-Ho ; Lee, Chul-Hoon ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 978~984
In the course of screening for a novel inhibitor of CDC2, HY558-1 was isolated from a culture broth of Penicillium minioluteum F558. Moreover, it was found that HY558-1 had an effect on both the cell cycle regulation and apoptosis of human cervical adenocarcinoma HeLa cells. A flow cytometric analysis of HeLa cells revealed appreciable cell cycle arrest at the G1 and G2/M phases following treatment with HY558-1. Furthermore, DNA fragmentation due to apoptosis was observed in HeLa cells treated with HY558-1. To obtain further information on the cell cycle arrest and apoptotic induction induced by HY558-1, the expression of certain cell cycle and apoptosis-associated proteins was examined using a Western blot analysis. The results revealed that HY558-1 inhibited the phosphorylation of pRb and decreased the expression levels of CDK2, CDC2, and cyclin A in the cell cycle progression. It was also shown that the level of
was increased in HeLa cells treated with 0.52 mM of HY558-1. Accordingly, HY558-1 was found to inhibit the proliferation of HeLa cells through the induction of G1 phase arrest by inhibiting pRb phosphorylation via an upregulation of
, and G2/M phase arrest by directly inhibiting CDC2 and cyclin A. Moreover, HeLa cells treated with 0.52 mM of HY558-1 exhibited apoptotic induction associated with the cleavage of Bid and release of cytochrome c from mitochondria into the cytosol. Subsequent investigation of the activation of caspase-3 and cleavage of poly (ADP-ribose) polymerase (PARP) suggested that the mitochondrial pathway was primarily involved in the HY558-1-induced apoptosis in HeLa cells.
Effects of Microbacterium laevaniformans Levans Molecular Weight on Cytotoxicity
Oh, Im-Kyung ; Yoo, Sang-Ho ; Bae, In-Young ; Cha, Jae-Ho ; Lee, Hyeon-Gyu ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 985~990
Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages.
-NMR analysis proved that the polysaccharide was mainly a
-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of
. The M2 fraction
, 80,000) at 400
had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1
, 50,000) at
showed the greatest (46.32%) on HepG2. To obtain more uniform
fractions of levan, the levan was further fractionated from S1
1,000,000) to S5
10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4
80,000) on SNU-1 and S5
10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller
, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.
Quantification of Oxygen Transfer in Test Tubes by Integrated Optical Sensing
Wittmann, Christoph ; Schutz, Verena ; John, Gernot ; Heinzle, Elmar ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 991~995
Immobilized sensor spots were applied for online measurement of dissolved
, in test tubes. Oxygen transport was quantified at varied shaking frequency and filling volumes. The k
a increased with increasing shaking frequency and decreasing filling volume. In non-baffled tubes the maximum
, equivalent to a maximum
transfer capacity of
. Monitoring of the hydrodynamic profile revealed that the liquid bulk rotated inside the tube with an inclined liquid surface, whereby the angle between the surface and tube wall increased with increasing shaking frequency. The
clearly correlated to the surface area. Placement of four baffles into the tubes improved the oxygen transfer up to 3-fold. The highest increase in
was observed at high filling volume and high shaking frequency. The maximum
in baffled tubes was
Protective Effect of Astaxanthin Produced by Xanthophyllomyces dendrorhous Mutant on Indomethacin-Induced Gastric Mucosal Injury in Rats
Kim, Jeong-Hwan ; Choi, Seok-Keun ; Lim, Wang-Jin ; Chang, Hyo-Ihl ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 996~1003
Nonsteroidal anti-inflammatory drugs such as indomethacin induce severe gastric mucosal damage in humans and rodents. In the present study, the in vivo protective effect of astaxanthin on indomethacin-induced gastric lesions in rats was investigated. The test groups were injected with indomethacin (25 mg/kg) after the oral administration of astaxanthin (25 mg/kg) for 1, 2, and 3 days, while the control group was treated only with indomethacin. Thiobarbituric acid reactive substances in the gastric mucosa, as an index of lipid peroxidation, increased significantly after indomethacin administration and this increase was inhibited by oral administration of astaxanthin. In addition, pretreatment with astaxanthin resulted in a significant increase of the activities of superoxide dismutase (SOD), catalase, and glutathione peroxidase (GSH-px). Histologic examination clearly revealed acute gastric mucosal lesions induced by indomethacin in the stomach of the control group, but were not observed in that of the test group. These results indicate that astaxanthin activates SOD, catalase, and GSH-px, and removes the lipid peroxides and free radicals induced by indomethacin. It is evident that astaxanthin acts as a free radical quencher and antioxidant, and is an effective molecule in the remedy of gastric mucosal lesions.
Purification and Properties of an Extracellular Acid Phytase from Pseudomonas fragi Y9451
In, Man-Jin ; Jang, Eun-Seok ; Kim, Young-Jin ; Oh, Nam-Soon ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1004~1008
An extracellular acid phytase from Pseudomonas fragi Y9451 was purified to homogeneity from the culture supernatant by salting-out, DEAE-Sepharose column chromatography, CM-Sepharose column chromatography, and Sephacryl S-300 gel filtration. The molecular weight of the purified enzyme was estimated to be 74 kDa on gel filtration and 54 kDa and 25 kDa on SDS-PAGE, suggesting that the native enzyme was a heterodimeric protein. The purified enzyme was most active at pH 4.5 and
and fairly stable from pH 4.0- 6.0. It was specific for phytate and exhibited a
value of 27 mM (sodium phytate, pH 4.5,
). The phytase activity was strongly inhibited (at maximum by 87%) by
at 5 mM concentration, and greatly inhibited by
at 10 mM concentration. However, EDTA notably stimulated the phytase activity at 10 mM concentration. With optimum pH and stability, Pseudomonas fragi phytase could be a potential candidate for animal feed applications.
Polyvinyl Alcohol Degradation by Microbacterium barkeri KCCM 10507 and Paenibacillus amylolyticus KCCM 10508 in Dyeing Wastewater
Choi, Kwang-Keun ; Park, Chul-Hwan ; Kim, Sang-Yong ; Lyoo, Won-Seok ; Lee, Sang-Hun ; Lee, Jin-Won ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1009~1013
The purpose of this study was to investigate the degradation of PVA (polyvinyl alcohol) contained in dyeing wastewater by a mixed culture of Microbacterium barkeri KCCM 10507 and Paenibacillus amylolyticus KCCM 10508. Firstly, synthetic wastewater which contained different initial concentrations of PVA varying from 50 to 3,500 mg/l were tested to obtain optimal PVA biodegradation activity of isolated strains, and the above two strains were found to degrade PVA up to 90%, when the initial concentration of PVA was 750 mg/l and below. Next, dyeing wastewater was tested by a nixed culture of the two isolated strains, and 42% and 55% of the initial concentrations of PVA and COD, respectively, was removed after five days. MLSS was gradually increased from an initial 1,400 to 2,500 mg/l, and the pH was also increased from 5.1 to 7.8. Sterilized dyeing wastewater was tested to find the effect of strains only on the biodegradation of PVA, and PVA degradation ratio and COD removal ratio were 50% and 72.8%, respectively. Thus, the results indicated that these two strains have good ability to degrade PVA and remove COD in dyeing wastewater, Finally, it is expected that if these two strains were used in the dyeing wastewater treatment, good efficiency for PVA degradation and COD removal could be achieved.
Purification and Characterization of Two Thermostable Xylanases from Paenibacillus sp. DG-22
Lee, Yong-Eok ; Lim, Pyung-Ok ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1014~1021
Two thermostable xylanases, designated XynA and XynB, were purified to homogeneity from the culture supernatant of Paenibacillus sp. DG-22 by ion-exchange and gel-filtration chromatography. The molecular masses of xylanases A and B were 20 and 30 kDa, respectively, as determined by SDS-PAGE, and their isoelectric points were 9.1 and 8.9, respectively. Both enzymes had similar pH and temperature optima (pH 5.0-6.5 and
), but their stability at various temperatures differed. Xylanase B was comparatively more stable than xylanase A at higher temperatures. Xylanases A and B differed in their
values. XynA had a
of 2.0 mg/ml and a
of 2,553 U/mg, whereas XynB had a
of 1.2 mg/ml and a
, of 754 U/mg. Both enzymes were endo-acting, as revealed by their hydrolysis product profiles on birchwood xylan, but showed different modes of action. Xylotriose was the major product of XynA activity, whereas XynB produced mainly xylobiose. These enzymes utilized small oligosaccharides such as xylotriose and xylotetraose as substrates, but did not hydrolyzed xylobiose. The amino terminal sequences of XynA and XynB were determined. Xylanase A showed high similarity with low molecular mass xylanases of family 11.
Cloning, Expression, and Characterization of DNA Polymerase from Hyperthermophilic Bacterium Aquifex pyrophilus
Choi, Jeong-Jin ; Kwon, Suk-Tae ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1022~1030
The gene encoding Aquifex pyrophilus (Apy) DNA polymerase was cloned and sequenced. The Apy DNA polymerase gene consists of 1,725 bp coding for a protein with 574 amino acid residues. The deduced amino acid sequence of Apy DNA. polymerase showed a high sequence homology to Escherichia coli DNA polymerase I-like DNA polymerases. It was deduced by amino acid sequence alignment that Apy DNA polymerase, like the Klenow fragment, has only the two domains, the
exonuclease domain and the
polymerase domain, containing the characteristic motifs. The Apy DNA polymerase gene was expressed under the control of T7lac promoter on the expression vector pET-22b(+) in E. coli. The expressed enzyme was purified by heat treatment, and Cibacron blue 3GA and
Q column chromatographies. The optimum pH of the purified enzyme was 7.5, and the optimal concentrations of KCl and
were 20 mM and 3 mM, respectively. Apy DNA polymerase contained a double strand-dependent
proofreading exonuclease activity, but lacked any detectable
exonuclease activity, which is consistent with its amino acid sequence. The somewhat lower thermostability of Apy DNA polymerase than the growth temperature of A. pyrophilus was analyzed by the comparison of amino acid composition and pressure effect.
Characterization of Segments of
Subunit Required for Efficient Coupling with Chemoattractant C5a, IL-8, and fMLP Receptors
Eia, Ji-Hee ; Lee, Chul-Hoon ; Lee, Chang-Ho ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1031~1037
The interaction of chemoattractant receptors and
was studied to provide the molecular basis to elucidate the interaction of chemoattractant receptors with
subunit, thereby possibly contributing to finding novel targets for designing new type of G protein antagonists with anti-inflammatory effects. Experiments were performed to characterize the
subunit domains responsible for efficient coupling to chemoattractant receptors. Thus, a series of chimeric
cDNA constructs were expressed, and the ability of chimeric proteins to mediate C5a, IL-8, and fMLP-induced release of inositol phosphate in transfected Cos-7 cells was tested. The results showed that short stretches of residues 154 to residue 167 and from residue 174 to residue 195 of
contribute to efficient coupling to the C5a receptor. On the other hand, a stretch of amino acid residues 220-240 of
that is necessary for interacting with C5a receptor did not play any role in the interaction with IL-8 receptor. However, a stretch from residue 155 to residue 195 of
was found to be crucial for efficient coupling to IL-8 receptor in concert with C-terminal 30 amino acid residues of this
subunit. Coupling profiles of a variety of chimeras, composed of
to fMLP receptor indicate that the C-terminal 30 amino acids are most critical for the coupling of
to fMLP receptor. Taken together,
subunit recruits multiple and distinctive coupling regions, depending on the type of receptors, to interact.
Photoswitching Characteristics of Biodevice Consisting of Chlorophyll
Nam, Yun-Suk ; Choi, Jeong-Woo ; Lee, Won-Hong ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1038~1042
The photoelectric responses of a biodevice consisting of chlorophyll
Langmuir-Blodgett film were investigated. Chlorophyll
Langmuir-Blodgett films were deposited onto ITO and Au coated glass. To confirm film formation, surface analysis of chlorophyll
Langmuir-Blodgett film was carried out by measurement using atomic force microscopy. The metal/insulator/metal structured biodevice was constructed by depositing aluminum onto the chlorophyll
Langmuir-Blodgett film surface. To investigate the photoelectric response, the current-voltage characteristic was measured by the conducting metal tip. The photoswitching function and transient photovoltage characteristics of the proposed device were measured by irradiation with Ar ion laser and
pulse laser, respectively. This research suggested that the proposed biodevice consisting of chlorophyll
could be applied to the molecular scale biosensor and/or bioelectronic device.
Electrochemical Property of Immobilized Spinach Ferredoxin on HOPG Electrode
Nam Yun-Suk ; Kim, You-Sung ; Shin, Woon-Sup ; Lee, Won-Hong ; Choi, Jeong-Woo ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1043~1046
The stability and electrochemical properties of a self-assembled layer of spinach ferredoxin on a quartz substrate and on a highly oriented pyrolytic graphite electrode were investigated. To fabricate the ferredoxin self-assembly layer, dimyristoylphosphatidylcholine was first deposited onto a substrate for ferredoxin immobilization. Surface analysis of the ferredoxin layer was carried out by atomic force microscopy to verify the ferredoxin immobilization. To verify ferredoxin immobilization on the lipid layer and to confirm the maintenance of redox activity, absorption spectrum measurement was carried out. Finally, cyclic-voltammetry measurements were performed on the ferredoxin layers and the redox potentials were obtained. The redox potential of immobilized ferredoxin had a formal potential value of -540 mV. It is suggested that the redox-potential measurement of self-assembled ferredoxin molecules could be used to construct a biosensor and biodevice.
Analysis of cel and pel Genes from Pectobacterium chrysanthemi PY35 for Relatedness to Pathogenicity
Park, Sang-Ryeol ; Lim, Woo-Jin ; Kim, Min-Keun ; Hong, Su-Young ; Shin, Eun-Chule ; Kim, Eun-Ju ; Lee, Jong-Yeoul ; Woo, Jong-Gyu ; Kim, Hoon ; Yun, Han-Dae ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1047~1051
The phytopathogenic bacterium Pectobacterium chrysanthemi secretes multiple isozymes of plant cell wall disrupting enzyme such as pectate lyase and cellulase. The cel gene, existing in tandem with the pel gene, was isolated previously . The role of Cel5Z and PelL1 in P. chrysanthemi PY35 pathogenicity on potato tissues was assessed by mutagenizing cloned cel gene and pel gene in tandem and recombining them with the chromosomal alleles. Strains with the Km cassette interposon in pelL1 or a double mutant showed a delay in the appearance of symptoms, suggesting that P. chrysanthemi PY35 pectate lyase PelL1 may playa minor role in soft-rot pathogenesis.
Screening and Characterization of Secretion Signals from Lactococcus lactis ssp. cremoris LM0230
Jeong, Do-Won ; Choi, Youn-Chul ; Lee, Jung-Min ; Seo, Jung-Min ; Kim, Jeong-Hwan ; Lee, Jong-Hoon ; Kim, Kyoung-Heon ; Lee, Hyong-Joo ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1052~1056
A secretion signal sequence-selection vector (pGS40) was constructed based on an
-amylase gene lacking a secretion signal and employed for selecting secretion signals from Lactococcus lactis ssp. cremoris LM0230 chromosomal DNA. Six fragments were identified based on their ability to restore
-amylase secretion in E. coli, and among these, a fragment, S405, conferred the highest secretion activity (84%) in E. coli. Meanwhile, S407, which conferred poor secretion activity in E. coli, was quite active in L. lactis. The results suggested that the efficiency of a secretion signal depended on the host. All six fragments had an open reading frame (ORF) fused to the reporter gene, and the potential Shine-Dalgamo (SD) sequence and putative promoter sequences were located upstream of the ORF. Deduced amino acid sequences from the six fragments did not show any homology with known secretion signals. However, they contained three distinguished structural features and cleavage sites, commonly found among typical secretion signals. The characterized secretion signals could be useful for the construction of food-grade secretion vectors and gene expression in LAB.
Characterization of Bacillus thuringiensis Having Insecticidal Effects Against Larvae of Musca domestica
Oh, Se-Teak ; Kim, Jin-Kyu ; Yang, Si-Yong ; Song, Min-Dong ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1057~1062
The entomopathogenic bacterium Bacillus thuringiensis is the most widely used biopesticide. Insecticidal proteins, coded by genes located in plasmids, form typical parasporal, crystalline inclusions during sporulation. We isolated a Bacillus thuringiensis strain having insecticidal activity against larvae of the house fly (M. domestica) from the soils at a pig farm in Korea, and named it Bacillus thuringiensis SM. The culture filtrate from Bacillus thuringiensis SM showed strong lethality (83.3%) against M. domestica larvae. The parasporal crystal is enclosed within the spores' outermost envelope, as determined by transmission electron microscopy, and exhibited a bipyramidal form. The crystal proteins of strain SM consisted of five proteins with molecular weights of approximately ~130, ~80, ~68, ~42, and ~27 kDa on a 10% SDS-PAGE (major band, a size characteristic of Cry protein). Examination of antibiotic resistance revealed that the strain SM showed multiple resistant. The strain SM had at least three different plasmids with sizes of 6.6, 9.3, and 54 kb. Polymerase chain reactions (PCRs) revealed the presence of cry1, cry4A2, and cry11A1 genes in the strain SM. The cry1 gene profile of the strain SM appeared in the three respective products of 487 bp [cry1A(c)], 414 bp [cry1D], and 238 bp [cry1A(b)]. However, the strain SM has not shown the cry4A2 md cry11A1 genes. In in vivo toxicity assays, the strain SM showed high toxicity on fly larvae (M. domestic) [with
of 4.2 mg/ml,
of 8.2 mg/ml].
Functional Complementation of Escherichia coli by the rpoS Gene of the Foodborne Pathogenic Vibrio vulnificus
Park, Kyung-Je ; Kim, Song-Hee ; Kim, Min-Gon ; Chung, Duck-Hwa ; Ha, Sang-Do ; Kim, Keun-Sung ; Jahng, Deok-jin ; Lee, Kyu-Ho ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1063~1066
The rpoS gene product is a global transcriptional factor, which is involved in bacterial survival under various stress conditions. An rpoS-homologous gene was cloned from a septicemia-causing pathogenic Vibrio vulnificus. Introduction of this gene as a multicopy plasmid into various E. coli strains displayed functional complementation, for examples, increased survivability of an rpoS-defective E. coli cell and induction of known
-dependent, stress-responding promoters of E. coli genes.
7-Oxostaurosporine Selectively Inhibits the Mycelial Form of Candida albicans
Hwang, Eui-Il ; Yun, Bong-Sik ; Lee, Sang-Han ; Kim, Soo-Kie ; Lim, Se-Jin ; Kim, Sung-Uk ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1067~1070
In the course of screening for specific inhibitors against the mycelial form of Candida albicans from natural resources, we have isolated and identified A6792-1 from Streptomyces sp. A6792 by using several chromatographies. By spectral analyses, this compound was determined as 7-oxostaurosporine, having a structure of staurosporine aglycon noiety. 7-Oxostaurosporine exhibited a selective growth inhibitory activity against the mycelial form of Candida spp. up to
in bioassay. It also exhibited a specific antifungal activity against the mycelial form of Candida spp. including C. krusei, C. albicans, C. tropicalis, and C. lusitaniae with MICs ranging from 3.1 to
7-Oxostaurosporine demonstrated no in vivo toxicity in SPF ICR mice. Therefore, this compound may have a considerable potential as an antifungal agent based on the preferential inhibition against growth of the mycelial form of Candida spp., dimorphic fungi.
Cold Adaptation of Lactobacillus paraplantarum C7 Isolated from Kimchi
Kim, Su-Jung ; Kim, Jong-Hwan ; Park, Jae-Yong ; Kim, Han-Taek ; Jeong, Seon-Ju ; Ha, Yeong-Lae ; Yun, Han-Dae ; Kim, Jeong-Hwan ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1071~1074
The effect of preadaptation at low temperature on cryoprotection was studied for Lactobacillus paraplantarum C7, a bacteriocin producer isolated from kimchi. L paraplantarum C7 cells in their log growth phase were incubated at
for 2, 4, and 6 h, respectively, before being frozen at
. After 24 h of freezing, viable cells were counted after brief thawing. The freezing-thawing cycles were repeated three more times. Cells preadapted at
before freezing survived better than control cells, but preadaptation at
did not confer cryoprotection. Chloramphenicol addition did not destroy the cryoprotection, indicating that protein synthesis was not required for the development of cryoprotection. SDS-PAGE showed induction of a 6.5-kDa protein, a major cold-shock protein, in preadapted cells.
Identification of Amino-Acids Residues for Key Role in Dextransucrase Activity of Leuconostoc mesenteroides B-742CB
Ryu, Hwa-Ja ; Kim, Do-Man ; Seo, Eun-Seong ; Kang, Hee-Kyung ; Lee, Jin-Ha ; Yoon, Seung-Heon ; Cho, Jae-Young ; Robyt, John-F. ; Kim, Do-Won ; Chang, Suk-Sang ; Kim, Seung-Heuk ; Kimura, Atsuo ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1075~1080
Dextransucrase (DSRB742) from Leuconostoc mesenteroides NRRL B-742CB is a glucosyltransferase that catalyzes the synthesis of dextran using sucrose, or the synthesis of oligosaccharides when acceptor molecules, like maltose, are present. The DSRB742 gene (dsrB742) was cloned and the properties were characterized. In order to identify critical amino acid residues, the DSRB742 amino acid sequence was aligned with glucosyltransferase sequences, and three amino acid residues reported as sucrose binding amino acids in Streptococcus glucosyltransferases were selected for site-directed mutagenesis experiments. Asp-533, Asp-536, and His-643 were independently replaced with Ala or Asn. D533A and D536A dextransucrases showed reduced dextran synthesis activities, 2.3% and 40.8% of DSRB742 dextransucrase, respectively, and D533N, D536N, H643A, end H643N dextransucrases showed complete suppression of dextran synthesis activities altogether. Additionally, D536N dextransucrase showed complete suppression of oligosaccharide synthesis activities. However, modifications at Asp-533 or at His-643 retained acceptor reaction activities in the range of 8.4% to 21.3% of DSRB742 acceptor reaction activity. Thus at least two carboxyl groups of Asp-533 and Asp-536, and His-643 as a proton donor, are essential for the catalysis process.
BioPlace: A Web-Based Collaborative Environment for Effective Genome Research
Ahn, Geon-Tae ; Kim, Jin-Hong ; Kang, Kyung-Mi ; Lee, Myung-Joon ; Han, In-Seob ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1081~1085
Genome research has become very popular in most nations. In order to enhance the efficiency of collaboration among genome research groups, ways to store and share data, communicate with each other, be guided through right research strategies, and to easily use well-established databases. In addition, since techniques and softwares for genome research groups are well established, a similar research road map could commonly be applied. In this study, we developed a web-based work place for effective genome research, named 'BioPlace.' From the beginning of writing a proposal, research members can work on the same environment with convenient aid to share files or data. BioPlace provides various ways of collaboration methods among genome researchers. The BioPlace system supports two types of workplaces, namely 'Personal Workspace' and 'Team Workspace.' For each BioPlace user, a Persona] Workspace is provided, while a Team Workspace is provided for each group with the same purpose. In addition, BioPlace provides a 'General Research Road Map' for genome research, and several Korean user interfaces for BLAST, PDB, and Primer3. We expect that BioPlace may facilitate collaboration of genome research among the experienced scientists and help beginners in many different ways as well.
Inhibition of Epstein-Barr Virus by the Triterpenoid Betulin Diphosphate and Uvaol
Muhammad, Amjad ; Carlson, Robert M. ; Krasutsky, Pavel ; Karim, M.Reza-Ul ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1086~1088
Betulin, a pentacyclic triterpenoid isolated from the bark of Betula papyrifera. Laboratory synthesized structural analogs were tested for antiviral activities against Epstein-Barr Virus (EBV) by immunofluorescent antiviral assay. Among the several analogs tested, betulin 3,28-diphosphate and uvaol exhibited significant antiviral activities against EBV. The
of betulin 3,28-diphosphate and uvaol was found to be
Stimulation of Actinorhodin Production by Streptomyces lividans with a Chromosomally-Integrated Antibiotic Regulatory Gene afsR2
Kim, Chang-Young ; Park, Hyun-Joo ; Yoon, Yeo-Joon ; Kang, Han-Young ; Kim, Eung-Soo ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1089~1092
An actinorhodin nonproducing Streptomyces lividans was converted to an actinorhodin overproducer through a single chromosomal integration of an antibiotic regulatory gene, afsR2. This strain exhibited early actinorhodin production and an average of 37.5% higher productivity than the S. lividans containing multiple copies of afsR2 plasmid in a glucose-containing liquid culture.
Identification of the Vibrio vulnificus cadC and Evaluation of Its Role in Acid Tolerance
Rhee, Jee-Eun ; Ju, Hyun-Mok ; Park, U-Ryung ; Park, Byoung-Chul ; Choi, Sang-Ho ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1093~1098
An open reading frame encoding CadC, consisting of 526 amino acids, was identified from the upstream region of the Vibrio vulnificus cadBA operon. The deduced amino acid sequences of the cadC were 22 to 78% similar to those reported from other Enterobacteriaceae. Functions of cadC gene on acid tolerance were assessed by comparing acid tolerances of V. vulnificus and its isogenic mutant, whose cadC gene was inactivated by allelic exchanges. The results demonstrated that the gene product of cadC contributes to acid tolerance of V. vulnificus, and that its contribution is dependent on prior exposure of cells to moderately acidic pH. The cellular level of cadB and cadA transcripts decreased in the cadC mutant, indicating that CadC exerts its effect on acid tolerance of V. vulnificus by enhancing the expression of cadBA in a pH-dependent manner.
Application of Flow Cytometry to Monitoring of Liposomal Restructuring Induced by Listeria monocytogenes
Kim, Hyung-Joo ; Bennetto, H.-Peter ; Halablab, Mahmoud-A. ;
Journal of Microbiology and Biotechnology, volume 14, issue 5, 2004, Pages 1099~1102
Liposomal restructuring induced by hemolytic Listeria monocytogenes was investigated by using flow cytometry. When added to calcein-entrapped liposomes, hemolytic, but not non-hemolytic, Listeria monocytogenes were able to induce reformation of vesicles. Such restructuring of liposomes was easily monitored by flow cytometry. Electron microscopy also indicated major changes in the challenged liposomal structures. The preliminary results described may offer a simple and fast method for monitoring liposomal restructuring and for differentiating between hemolytic and non-hemolytic bacteria.