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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Microbiology and Biotechnology Letters
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The Korean Society for Applied Microbiology and Biotechnology
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Volume 34, Issue 4 - Dec 2006
Volume 34, Issue 3 - Sep 2006
Volume 34, Issue 2 - Jun 2006
Volume 34, Issue 1 - Mar 2006
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Lessons from the Sea : Genome Sequence of an Algicidal Marine Bacterium Hahella chehuensis
Jeong Hae-Young ; Yoon Sung-Ho ; Lee Hong-Kum ; Oh Tae-Kwang ; Kim Ji-Hyun ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 1~6
Harmful algal blooms (HABs or red tides), caused by uncontrolled proliferation of marine phytoplankton, impose a severe environmental problem and occasionally threaten even public health. We sequenced the genome of an EPS-producing marine bacterium Hahella chejuensis that produces a red pigment with the lytic activity against red-tide dinoflagellates at parts per billion level. H. chejuensis is the first sequenced species among algicidal bacteria as well as in the order Oceanospirillales. Sequence analysis indicated a distant relationship to the Pseudomonas group. Its 7.2-megabase genome encodes basic metabolic functions and a large number of proteins involved in regulation or transport. One of the prominent features of the H. chejuensis genome is a multitude of genes of functional equivalence or of possible foreign origin. A significant proportion (
) of the genome appears to be of foreign origin, i.e. genomic islands, which encode genes for biosynthesis of exopolysaccharides, toxins, polyketides or non-ribosomal peptides, iron utilization, motility, type III protein secretion and pigment production. Molecular structure of the algicidal pigment was determined to be prodigiosin by LC-ESI-MS/MS and NMR analyses. The genomics-based research on H. chejuensis opens a new possibility for controlling algal blooms by exploiting biotic interactions in the natural environment and provides a model in marine bioprospecting through genome research.
Selection and Cultural Characteristics of Whole Chicken Feather-Degrading Bacterium, Bacillus sp. SMMJ-2
Park Sung-Min ; Jung Hyuck-Jun ; Yu Tae-Shick ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 7~14
Feather, generated in large quantities as a byproduct of commercial poultry processing, is almost pure keratin, which is not easily degradable by common professes. Four strains, SMMJ-2, FL-3, NO-4 and RM-12 were isolated from soil for production of extracellular keratinolytic protease. They were identified as Bacillus sp. based on their morphological and physiological characteristics. They shown high protease activity on 5.0% skim milk agar medium and produced a substrate like mucoid on keratin agar medium. Bacillus sp. SMMJ-2 had a faster production time for producing keratinolytic protease than other strains. This strain did not completely degrade whole chicken feather for five days in basal medium but completely degraded whole chicken feather when supplied with nitrogen source for 40hours in keratinolytic producing medium (
fructose, 1.2% whole chicken feather,
, pH 7.0). When supplied with chicken feather as nitrogen source, keratinolytic protease activity was 89 units/ml/min. When soybean meal was used as nitrogen source, the keratinolytic protease production reached a maximum of 106 units/ml/min after 48 hours under
, 180 agitation. To isolate the keratinolytic protease, the culture filtrate was precipitated with
and acetone. The recovery rate of keratinolytic protease was about 96% after treatment with 50% acetone. The enzyme was stable in the range of
Cloning and Characterization of the Paraquat Resistance-Related Genes from Ochrobactrum anthropi JW-2
Bae Eun-Kyung ; Lee Hyo-Shin ; Won Sung-Hye ; Lee Byung-Hyun ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 15~22
A 4,971 bp chromosomal DNA fragment containing the pqrA, paraquat resistance gene, was cloned from Ochrobactrum anthropi JW-2, and the complete nucleotide sequence was determined. Nucleotide and deduced amino acid sequences of the fragment revealed the presence of 4 complete ORFs (orf2, pqrA, orf3, orf4) and two incomplete ORFs(orf1, orf5). Orf1, pqrA, orf4 and orf5 exists at the direct strand but orf2 and orf3 exists at the reverse complementary strand. Orf1 which of incomplete sequences without start codon shares homology with ATP binding region of the response regulator receiver. Orf2 shares high homology with members of the tetR family of transcriptional repressor which have a helix-turn-helix (H-T-H) motif. Therefore, the orf2 is predicted as a transcriptional repressor of pqrA and is designated as pqrR2. Orf3 shares high homology with the members of the lysR family acting as a transcriptional activator which have both of a H-T-H motif at the N-terminal region and substrate binding domain at the C-terminal region. Therefore, the orf3 is predicted as a transcriptional activator of pqrA and is designated as pqrR1. Orf4 shows homology with the periplasmic substrate-binding protein of amino acid ABC transporter. Orf5 which of incomplete sequences without stop codon revealed the homology with the permeases protein of amino acid ABC transporter.
Classification of Listeria monocytogenes Isolates from Korean Domestic Foods Using Random Amplification of Polymorphic DNA and Serotyping Analysis
Kim Hyun-Joong ; Park Si-Hong ; Kim Hae-Yeong ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 23~27
Molecular subtyping of Listeria monocytogenes, including type strains and isolates from Korean foods, were performed using random amplification of polymorphic DNA (RAPD). Each Listeria species showed specific RAPD band patterns, and L. monocytogenes serotypes and isolates were divided into two clusters. RAPD results showed that L. monocytogenes isolates from Korean foods were divided into two groups. Group I contained L. monocytogenes serotypes 1/2b and 4b, whereas Group II contained serotypes 1/2a and 1/2c. These results suggested RAPD as possible subtyping methods for Listeria species. Also, RAPD Results showed significant correlation between molecular subtyping and serotyping of L. monocytogenes, and classified two different groups of L. monocytogenes isolated from Korean foods.
Characterization of Phytase from Bacillus coagulans IDCC 1201
Lee Seung-Hun ; Kwon Hyuk-Sang ; Koo Kyo-Tan ; Kang Byung-Hwa ; Kim Tae-Yong ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 28~34
A native extracellular acid phosphatase, phytase (EC 22.214.171.124), from Bacillus coagulans IDCC 1201 (commercially known as Lactobacillus sporogenes) used as probiotics, was characterized. Though some strains of B. coagulans have been evaluated with regard to several health-promoting effects, it has not been reported to produce phytase. Partially purified phytase front the strain IDCC 1201 had a pH optimum of 4.0 and a temperature optimum of
, respectively. The requirement for divalent cations was studied and cobalt ion remarkably increased the enzyme activity. The removal of metal ions from the enzyme by EDTA decreased activity below 50%. The enzyme activity depleted restored when the assay was performed in the presence of
is the most active stimulator and has unique activation effect at high temperature. The phytase was specific for sodium phytate and p-nitrophenylphosphate, which is different from other known Bacilli phytases. The putative amino acid sequences of the phytase from B. coagulans IDCC 1201 were very similar to that of the phytase from B. subtilis strain 168. Based on these data, we concluded that the phytase from B. coagulans IDCC 1201 is a
-dependent acid phosphatase. Therefore, the strain B. coagulans IDCC 1201 is thought to be a valuable addititive for livestocks as well as a beneficial probiotics for human.
Imitation of Phosphoenolpyruvate to Oxaloacetate Pathway Regulation of Rumen Bacteria in Enteric Escherichia coli and Effect on C4 Metabolism
Kwon Yeong-Deok ; Kwon Oh-Hee ; Lee Heung-Shick ; Kim Pil ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 35~39
One of the fermentative metabolism of enteric Escherichia coli was imitated after rumen bacteria, which have high C4 metabolism. E. coli expresses phosphenolpyruvate carboxylase (PPC) for the pathway between phosphoenolpyruvate (PEP) and oxaloacetate (OAA) during glycolytic condition while expresses phosphoenolpyruvate carboxykinase (PCK) during gluconeogenic condition. In contrast to enteric E. coli, rumen bacteria express the PEP-OAA pathway only by PCK. To verify the effect of the regulation imitation on the C4 metabolism of E. coli, PPC-deficient E. coli strain with PCK expression in glycolytic condition was constructed. The PEP-OAA regulation modified E. coli strain increased 2.5-folds higher C4 metabolite than the wild type strain. The potential use of C4 metabolism by regulation control is discussed.
Purification and Characterization of Phytase from Bacillus subtilis
Koh Hyun-Jung ; Chu In-Ho ; Chung Kun-Sub ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 40~46
A bacterial strain producing high level of a phytase was isolated from cattle feces and identified as Bacillus subtilis, and designated as Bacillus sp. CF 5-26. The production of the phytase from Bacillus sp. CF 5-26 reached the highest level after 72 hours at
. The optimum condition of the media for the production of phytase was 10% rice bran extract, 0.1% whey protein powder,
. The phytase was purified 20.3 folds with ethanol precipitation, Sephadex G-100, CM Sepharose CL-6B and Sephacryl S-100-HR column chromatography. The molecular weight of the purified enzyme was estimated to be 66 kDa on SDS-polyacrylamide gel electrophoresis. The purified phytase activity was stable up pH 5.0, 7.0, 11.0 and the remaining activity was 50% when it was treated at
for 1 hour. The substrate specificity of phytase was most active against sodium phytate and inositol polyphosphate compound. And the phytase hydrolysed tripolyphosphate and pyrophosphate a little. The Km value for the sodium phytate was 0.64 mM and the Vmax value was
Production of Humanised Anti-hepatitis B Antibody in Butyrate-Treated Chinese Hamster Ovary Cells
Park Se-Cheol ; Lee Jae-Sun ; Lee Byung-Kyu ; Kang Heui-Il ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 47~51
Sodium butyrate (NaBu) is used as an enhancer for the production of recombinant proteins in Chinese hamster ovary (CHO) cells. However, NaBu is well-known for its cytotoxic effect, thereby inducing apoptosis. CHO cells which had been engineered to express a humanised anti-HBV antibody were cultured using serum-free medium, Ex-cell 301. From a seeding density of
cells/ml, CHO cells grown with serum-free medium reached a maximum cell density of
cells/ml after 9 days in culture and produced a maximal antibody concentration of 130 mg/l after 13 days in culture. In the perfusion culture system, CHO cells producing anti-HBV antibody grown in an 7.5 1 bioreactor seeded with
cells/ml reached a maximal antibody concentration of 85 mg/1 after 720 h in culture. The addition of 0.3 mM NaBu and lowering culture temperature to
elongated the culture period to 60 days and increased the production yield by 2-fold, compared to control culture.
Production of an Antihyperlipemial HMG-CoA Reductase Inhibitor from Bacillus cereus D-3
Lee Dae-Hyoung ; Lee Jae-Won ; Jeong Jae-Hong ; Lee Jong-Soo ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 52~57
For the purpose of production of a novel antihyperlipemial HMG-CoA reductase inhibitor from bacteria, a bacterium which showed the highest HMG-CoA reductase inhibitory activity was isolated from traditional Doenjang. This strain was identified as Bacillus cereus (D-3) based on its microbiological characteristics and 165 rRNA sequence analysis. The maximal HMG-CoA reductase inhibitor production from Bacillus cereus D-3 was obtained by cultivation in a Glucose-CSL broth containing 2% glucose, 0.6% corn steep liquor,
for 36 h. The final HMG-CoA reductase inhibitory activity under the above conditions was 39.4%.
High Throughput Screening and Directed Evolution of Tyrosine Phenol-Lyase
Choi Su-Lim ; Rha Eu-Gene ; Kim Do-Young ; Song Jae-Jun ; Hong Seung-Pyo ; Sung Moon-Hee ; Lee Seung-Goo ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 58~62
Rapid assay of enzyme is a primary requirement for successful application of directed evolution technology. Halo generation on a turbid plate would be a method of choice for high throughput screening of enzymes in this context. Here we report a new approach to prepare turbid plates, by controlling the crystallization of tyrosine to form needle-like particles. In the presence of tyrosine phenol-lyase (TPL), the needle-like tyrosine crystals were converted to soluble phenol rapidly than the usual rectangular tyrosine crystals. When an error-prone PCR library of Citrobacter freundii TPL was spread on the turbid plate, approximately 10% of the colonies displayed recognizable halos after 24 hours of incubation at
. Representative positives from the turbid plates were transferred to LB-medium in 96-wellplates, cultivated overnight, and assayed for the enzyme activity with L-tyrosine as the substrate. The assay results were approximated to be proportional to the halo size on turbid plates, suggesting the screening system is directly applicable to the directed evolution of TPL. Actually, two best mutants on the turbid plates were identified to be
and 1.5-fold improved in the activity.
Antifungal Activity of Bacillus sp. KMU-1011 Against Gray Mold Causing Botrytis cinerea
Park Sung-Min ; Kim Hyun-Soo ; Yu Tae-Shick ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 63~69
We isolated a bacterium which produces antifungal substances from the Lake of Saimaa soils in Fin-land. The isolated strain was identified as Bacillus sp. and shown a strong antifungal activity on plant pathogenic fungi. Bacillus sp. KMU-1011 produced maximum level of antifungal substances under incubation aerobically at
for 48 hours in nutrient broth containing 1.0% glucose and 1.0% polypeptone at 180 rpm and initiated pH adjusted to 6.0. Precipitate of culture broth by
ammonium sulfate precipitation exhibited strong antifungal activity against Botrytis cinerea KACC 40573 by dry cell weight. Chloroform extract of cultured broth also shown fungal growth inhibitory activity against C. gloeosporioides KACC 40804, D. bryoniae KACC 40669, F. oxysporum KACC 40037, F. oxysporum KACC 40052, F. oxysporum f. sp. radicis-lycopersici KACC 40537, F. oxysporum KACC 40902, M. cannonballus KACC 40940, P. cambivora KACC 40160, R. solani AG-1 KACC 40101, R. solani AG-4 KACC 40142, and S. scleotiorum KACC by agar diffusion method.
Antibacterial Effects of Oriental Herb Extract Against Gardnerella vaginalis
Kim Youn-Hee ; Lee Heung-Shick ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 70~73
To investigate the potential of treatment, antimicrobial activity of various oriental herb extracts were tested for Gardnerella vaginalis, which is the predominant organism in bacterial vaginosis. Among the tested 14 oriental herbs, water-extracts of Kalkeun, Kosam, Nuro, Pakjakyak, Sukchangpo, Shiyup, Junghyang and Hwangryun represented antibacterial activities against G. vaginalis. The minimal inhibition concentration (MIC) of Shiyup against G. vaginais was 0.63 mg/mL, and those of Pakjakyak and Hwangryun, Kalkeun and Nuro, Kosam, Sukchangpo and Junghyang were 1.25 mg/mL, 2.5 mg/mL, and 5 mg/mL, respectively. There-fore, the water-extracts of Kalkeun, Kosam, Nuro, Pakjakyak, Sukchangpo, Shiyup, Junghyang and Hwangryun were considered to be potential treatment of bacterial vaginosis caused by G. vaginalis.
Characterization of Pediococcus pentosaceus Isolated from Porcine Intestine
Lee Mi-Sung ; Yoon Ki-Hong ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 74~77
A lactic acid bacterial strain resistant to several antibiotics such as oxytetracycline, tylosin, neomycin and sulfathiazole, which have been often used as a therapeutic agent in livestock, was isolated from the porcine gastrointestinal tract. The isolate YB-55 was identified as belonging to the genus Pediococcus with the highest similarity to P. pentosaceus on the basis of its 16S rRNA sequence and biochemical properties. The isolated strain showed viability of over 85% at pH 3.0 and was resistant to bile salt. The strain produced lactic acid of 12.3 g/L by jar fermentation and maintained its viability in the presence of antibiotics at dosage related therapeutic effect, suggesting P. pentosaceus YB-55 may of ffr potential as a probiotics for livestock.
Plant Growth-Promoting Capabilities of Diazotrophs from Wild Gramineous Crops
Lee Su-Jin ; Lee Sang-Eun ; Seul Keyung-Jo ; Park Seung-Hwan ; Ghim Sa-Youl ;
Microbiology and Biotechnology Letters, volume 34, issue 1, 2006, Pages 78~82
Since there could be more and rather various diazotrophs in rhizosphere of wild crops than those in rhizosphere of cultivars, some wild gramineous crops grown in Korea were collected for isolating nitrogen-fixing bacteria. Six diazotrophs were purified from their roots using nitrogen-free media. The isolated bacteria were partially identified as 4 genera by 16S rDNA sequence analysis: Stenotrophomonas sp., Bosea sp., Klebsiella sp., and Azorhizobium sp. By PCR amplification and sequence analysis, DNA fragments extracted from all isolates turned out to have an individual nifH homologous gene. Five isolates (KNUC163, KNUC165, KNUC169, KNUC170, and KNUC171) showed auxin activity and four isolates (KNUC163, KNUC166, KNUC170, and KNUC171) produced siderophores. Especially,3 strains of S. maltophilia showed both auxin and siderophore activities. In conclusion, the isolated nitrogen-fixing bacteria might have capabilities for plant growth promotion.