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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Microbiology and Biotechnology Letters
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 31, Issue 4 - Dec 2003
Volume 31, Issue 3 - Sep 2003
Volume 31, Issue 2 - Jun 2003
Volume 31, Issue 1 - Mar 2003
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High-Value Materials from Microalgae
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 95~102
Microalgae are a diverse group of photosynthetic organisms and abundant in every ecosystem in the biosphere. They are common in aqueous environments including marine, brackish and fresh waters and in some habitats that lack eukaryotic life such as some hot springs and highly alkaline lakes. Microalgal biotechnology that is focused on the microalgae-based production of a variety of useful materials such as pharmaceutical comfounds, health foods, natural pigments, and biofuels is considered as an important discipline with the development of biotechnology. In addition, the mass cultivation of microalgae can also contribute to improving the environmental quality by reducing the concentration of
which is one of major gases lead to global warming. Consequently, it seems that the microalgae can be used as an efficient, renewable, environmentally friendly source of high-value biomaterials such as chemicals, pigments, energy, etc. and the microalgal biotechnology will most likely represent a larger portion of modern biotechnology.
Characterization and Functional Study of PyrR Orthologues from Genome Sequences of Bacteria
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 103~110
The regulation of pyrimidine nucleotide synthesis has been proved to be controlled by a regulatory protein PyrR-mediated attenuation in the Gram-positive bacteria. After several bacterial genome sequencing projects, we have discovered the PyrR orthologues in the databases for Haemophilus influenzae and Synechocystis and sp. PCC6803 genome sequences. To investigate whether these PyrR orthologue proteins regulate pyrimidine nucleotide synthesis as well as the cases of Bacillus, the PyrR regions of each strains were amplified by PCR and cloned with pUC19 or T-vector in Escherichia coli and with a shuttle vector pHPS9 for E. coli and B. subtilis. For the regulation test of the PyrR orthologues, the aspartate-transcarbamylase (ATCase) assay was carried out. From the results of the ATCase assay, it was confirmed that Synechocystis sp. PCC6803 could not restore by pyrimidines to a B. subtilis, PyrR but H. influenzae PyrR could. For Purification of PyrR orthologue proteins, PyrR orthologue genes were cloned into the expression vector (pET14b). Over-expressed product of PyrR orthologue genes was purified and analyzed by the SDS-PACE. The purified PyrR orthologue proteins from H. influenzae and Synechocystis sp. PCC6803 turned out to be molecular mass of 18 kDa and 21 kDa, respectively. The result of uracil phosphoribosyl transferase (UPRTase) assay with purified PyrR orthologue proteins showed that H. influenzae PyrR protein only has UPRTase activity. In addition, we could predict several regulatory mechanisms that PyrR orthologue proteins regulate pyrimidine de novo synthesis in bacteria, through phylogenetic analysis for PyrR orthologue protein sequences.
Strength of the Mutant Promoters for the \beta-xylosidase gene of Bacillus stearothermophilus No. 236
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 111~116
The xylA gene of Bacillus stearothermophilus No. 236 encoding
-xylosidase was cloned and sequenced previously. The transcriptional start site of the xylA gene cloned in E. coli was identified to be the guanine (G) by primer extension analysis. This supports that the expression of xylA gene is also directed in the E. coli cells by the previously determined transcription initiation signals, -10 sequence (CATAAT) and -35 sequence (TTGTTA) separated by 12 bp. To increase the expression of
-xylosidase, firstly the spacer region of xylA promoter was extended from 12 to 17 bp, and then the -10 and -35 elements were converted into their respective consensus sequences. The mutant promoters thus obtained were tested for their activities in both the E. coli and B. subtilis host cells. The change of the length of the spacer region from 12 to 17 bp resulted in a 1.6- and 2.5-fold increase in promoter strength in comparison with the wild type promoter in E. coli and B. subtilis cells, respectively. Also, strength of the promoter with the fourth T to A transversion on its -35 element increased in the transcription level by about 35 times compared with that of wild-type promoter. However, surprisingly the 5' end C-to-T transition of the -10 hexamer showed a 5- to 15-fold reduction in
-xylosidase activity in both E. coli and B. subtilis. Together, the present data demonstrated that the 5' end nucleotide C of the -10 sequence CATAAT and the fourth nucleotide A of the -35 hexamer are two most critical nucleotides for the promoter activity in the context of the xylA promoter.
Cloning of Autoregulator Receptor Gene form Saccharopolyspora erythraea IFO 13426
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 117~123
For screening of autoregulator receptor gene from Saccharopolyspora erythraea, PCR was performed with primers of receptor gene designed on the basis of amino acid sequences of autoregulator receptor proteins with known function. PCR products were subcloned into the BamHI site of pUC19 and transformed into the E. coli DH5
. The isolated plasmid from transformant contained the fragment of 120 bp, which was detected on 2% gel after BamHI treatment. The insert, 120 bp PCR product, was confirmed as the expected internal segment of gene encoding autoregulator receptor protein by sequencing. Southern and colony hybridization using Saccha. erythraea chromosomal DNA were performed with the insert as probe. The plasmid (pEsg) having 3.2 kbp SacI DNA fragment from Saccha. erythraea is obtained. The 3.2 kbp SacI DNA fragment was sequenced by the dye terminator sequencing. The nucleotide sequence data was analyzed with GENETYX-WIN (ver 3.2) computer program and DNA database. frame analyses of the nucleotide sequence revealed a gene encoding autoregulator receptor protein which is a region including KpnI and SalI sites on 3.2 kbp SacI DNA fragment. The autoregulator receptor protein consisting of 205 amino acid was named EsgR by author. In comparison with known autoregulator receptor proteins, homology of EsgR showed above 30%.
Isolation and Characterization of a Novel Aspergillus tubingensis with a Hydrolyzing Activity of Cellulose-pectin Complex
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 124~128
In order to isolate and characterize a novel fungal strain capable of producing cellulase, each samples of the old rice straw, soil, and the old tree were screened by congo red test. One of the fungi screened has been identified as Aspergillus tubingensis strain from the results of the phylogenic analysis based on partial DNA sequence and the basis of its biochemical properties. A carboxymethyl cellulase activity of the strain was higher than that of A. oryzae KCTC 6291. In CMCase activity measurement, it wasn't sensitive about pH 2.0, 3.0, 4.0, but the enzyme was more stable than A. oryzae under the various pH and temperature conditions and the enzyme activity was more similar to neutrality and alkali. Therefore, it could be suggested that the isolated strain has a potential possibility for the developing of the probiotics.
Screening and Characterization of Probiotic Strains for Prevention of Bacterial Fish Diseases
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 129~134
The purpose of the present study was to screen the effective of lactic acid bacteria (LAB) as probiotics, which are able to protect aquacultural fish pathogenic bacteria, and investigate their characterization. Twenty strains of lactic acid bacteria were isolated from fish intestine, fermented fish foods and kimchis. These bacteria were screened for antagonistic activity against fish pathogenic bacteria. Seven tested LAB strains were able to inhibit the fish pathogenic bacteria, including Vibrio anguillarum, Edwardsiella tarda, and Streptococcus sp.. Of the probiotic candidates, BK19 strain isolated from fermented pollack viscera indicated the largest inhibition activity. Moreover, this strain showed a resistance over low pH and antibiotic agents. Therefore this probiotic candidate BK19 was finally selected and identified as a probiotic strain. This particular probiotic bacteria was identified as Lactobacillus sakei BK19 by biochemical characteristics and 165 rRNA PCR amplification.
Melanin Biosynthesis Inhibitory Activities of Coumarins Isolated from Angelica polymorpha MAXIM
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 135~139
During the screening for inhibitors of melanin biosynthesis from plant extract, Angelica polymorpha MAXIM which showed a high level of inhibition was selected. The inhibiting substances were purified form methanol extract of Angelica polymorpha MAXIM followed by silica gel column chromatography and HPLC. The inhibitors were identified as heraclenin, isosaxalin and heraclenol 3'-Me ether, by spectrescopic methods of ESI-MS, H-NMR, C-NMR, DEPT, HMQC and HMBC. These compounds did not have mushroom tyrosinase inhibitory activity, but showed a highly potent melanin biosynthesis inhibition zone in the plate culture of Streptomyces bikiniensis, a bacterium used as an indicator organism in this work. These compounds did not show any growth inhibition against S. bikiniensis at the same concentration of melanin biosynthesis test.
D99 Type I Signal Peptidase Implicated Stabilizing the Protein Structure
Sung, Meesook ; Eunyoung Han ; Lee, Hoyoung ;
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 140~144
Type Ⅰ signal peptidase is an integral membrane protein that functions to cleave signal peptides from secreted and membrane proteins. The enzyme serves as a potential target for the development of novel antibacterial agents due to its unique physiological properties. Despite being one of the best characterized enzymes, the catalysis of Type Ⅰ signal peptidase still remains controversy over the catalytic serine/lysine dyad mechanism. It appears that the dyad proteases are generally less efficient than the prototypical serine/histidine/aspartic acid triad found in most enzymes, although Type Ⅰ signal peptidase is an exception to this rule. In this paper, we have proposed that Type Ⅰ signal peptidase may act as the serine/lysine/aspartic acid triad cataltytic mechanism. Therefore, the aspartic acid 99 residue in the E. coli signal peptidase was chosen and mutated to an alanine to see if there is any possible role of the aspartic acid in the catalytic function. Type Ⅰ signal peptidase D99A protein was inactive in vitro assay using the procoat synthesized by in vitro transcription translation. However, the mutant was active using a highly sensitive in vivo assay. Pulse-chase experiments show that the replacement of aspartic acid 99 with alanine results in a very unstable signal peptidase molecule. Therefore, we conclude that it is unlikely that the residue is directly involved in catalysis, but rather plays an important role in stabilizing the protein structure.
Production of Enantioselective Lipase from Acinetobacter sp. SY-01
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 145~150
Lipase from Acinetobacter sp. SY-01 plays an important role enzyme that products chiral drug. We investigated optimum condition for mass production of Acinetobacter sp. SY-01 lipase. Addition of among the different oils to medium. olive oil was optimal for enzyme production. When 0.2% olive oil was added as a carbon source, the production of lipase was increased to a maximum. The optimum pH and temperature were pH 7 and
. In the presence of
, the lipase activity was dramatically enhanced by 280% and 160%, respectively. SY-01 lipase was stable in the most of the DMSO among organic solvents. The addition of triton-X 100 increased the SY-01 lipase by 100-fold. The optimum composition of medium for production of the enzyme was 0.8% yeast extract, 0.2% olive oil, 0.4% triton X-100＋40% DMSO. 0.1%
Characterization of the \beta-Galactosidase Produced by Streptomyces sp. YB-10
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 151~156
A strain YB-10 was isolated from soil as a producer of the extracellular
-D-galactosidase, which catalyzes the hydrolysis of lactose. The strain YB-10 was identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. After treating culture supernatant of the isolate with ammonium sulfate, the precipitated protein was used as a crude
-galactosidase for analyzing its reaction properties with para-nitrophenyl-
Gal) as a substrate. The
-galactosidase showed its maximal activity at pH 6.0 and 6
. The enzyme was also active on lactose. The hydrolyzing activity of
-galactosldase for pNP-
Gal and lactose was decreased by galactose. Its hydrolyzing activity far lactose was also decreased by glucose, but the activity for pNP-
Gal was increased to 1.8-folds by glucose.
Isolation of Bacillus alcalophilus AX2000 Producing Alkaling Xylanase and Its Enzyme Production
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 157~164
An alkali-tolerant bacterium producing the xylanase was isolated from soil and identified as Bacillus alcaiophilus. This strain, named B. alcalophilus AX2000, was able to grow and produce xylanase optimally at pH 10.5 and
. The maximum xylanase production was obtained when 0.5%(w/v) birchwood xylan and 0.5%(w/v) polypeptone and yeast extract were used as carbon source and nitrogen source, respectively. The biosynthesis of xylanase was under the catabolite repression by glucose in the culture medium, and inhibited in the presence of high concentration of xylose. The maximum activity of xylanase was observed at pH 10.0 and
and the enzyme activity remained was over 80% at
and from pH 5.0 to 11.0.
Effect of Neuronal Differentiation Activity of Hot Water Extracts of Marine Alga, Chlorella capsulata
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 165~170
Hot water extracts of Chlorella capsulata(CCE) is a biological response modifier (BRM) which exhibits neuronal differentiation activity. The effect of CCE on the growth of nerve cells, PC12 was observed as follows: The viable cell density in adding CCE was increased up to 2.5 times, compare to that in no addition. The neurite of the cells was also lengthened up to 40
longer than 5
in no addition. The number of neurite-bearing cells were about four times higher than that in no addition.
Cultural Characteristics of a Biosurfactant-Producing Microorganism Pseudomonas aeruginosa F722
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 171~176
Productivity of biosurfactant (rhamnolipid) by Pseudomonas aeuginosa F722 was investigated in the several culture conditions and culture composition. Biosurfactant production by P. aeuginosa F722 was amounted to 0.78 g/l as the result of the nitrogen sources and carbon sources without investing of optimum conditions. As for that one was investigated, biosurfactant production by P. aeruginosa F722 was amounted to 1.66 g/l. Biosurfactant production increased twofold because the composition of a modified C-medium was investigated efficiently.
inorganic nitrogens and yeast extract or trypton organic nitrogens were effective, but others inorganic nitrogens and organic nitrogens tested were not efficient far biosurfactant production by P. aeruginosa F722. The optimum concentration of
Cl; inorganic nitrogen and yeast extract; organic nitrogen were 0.05% and 0.1%, respectively. In various carbon sources, others with the exception of hydrophobic property substrate (n-alkane) and hydrophilic property substrate (glucose, glycol) were not found to be effective fur biosurfactant production, and 3.0% was better in yield than other concentration of glucose. This yielded C-to-N ratios between 17 and 20. In our experiment, the highest biosurfactant production by P. aeruginosa F722 were observed in 5 days cultivation, containing glucose 3.0%,
Cl 0.05%, and yeast extract 0.1% and C-to-N ratio was 20. Optimal pH and temperature for biosurfactant production were 7.0 and
, respectively. Under the optimal culture conditions with glucose, biosurfactant production was amounted to 1.66 g/l. Velocity of biosurfactant production and strain growth increased after nitrogen depletion. The average surface tension of 30 mN/m after the 3 days of incubation under optimal culture condition was measured by ring tensionmeter.
PCR Detection of Terephthalic Acid Degrading Comamonas testosteroni in Soil
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 177~181
Eleven bacterial strains which are able to utilize terephthalic acid as a carbon and an energy source for growth were isolated from the soil of 7 water quality evaluation points in Kyonggi area of Korea. Phthalic acid isomer degrading activity of the isolates from the 4 contaminated points was higher than those from the 3 clean points. Among 11 isolates, 4 isolates which have high terephthalic acid degrading activity and degrade two phthalic acid isomers were identified by partal 16S rDNA sequence determination. One of them was identified as Pseudomonas putida, and the others as Comamonas testosteroni. Thus a large number of phthalic acid isomer degrading bacteria in domestic soil were inferred as C. testosteroni. On the basis of these results, the PCR detection of C. testosteroni in soil was applied to monitor soil contamination by phthalic acid isomers. The DNA of C. test-osteroni extracted from 4 g soil was directly detected by PCR with C. testosteroni specific primer pair. The amount of PCR products was different according to sampling sites and more PCR products were obtained from contaminated sites than those from clean sites (Gulpo-chun＞Anyang-chun＞Hwangguji-chun>Shin-chun＞Huk-chun＞Pukhan-river>Kapyeong-chun). This result was coincided with that of the viable cell counts for terephthalic acid degrading bacteria.
Design and Implementation of Integrated System for Microarray Data
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 182~190
As DNA microarrays are widely used recently, the amount of microarray data is exponentially increasing. Until now, however, no domestic system is available for the efficient management of such data. Because the number of experimental data in a specific laboratory is limited, it is necessary to avoid redundant experiments and to accumulate the results using a shared data management system for microarrays. In this paper, a system named WEMA (WEb management of Micro Arrays) was designed and implemented to manage and process the microarray data. WEMA system was designed to include the basic feature of MIAME (Minimal Information About a Microarray Experiment), and general data units were also defined in the system in order to systematically manage the data. The WEMA system has three main features: efficient management of microarray data, integration of input/ouput data, and metafile processing. The system was tested with actual microarray data produced by a molecular biology laboratory, and we found that the biologists could systematically manage and easily analyze the microarray data. As a consequence, the researchers could reduce the cost of data exchange and communication.
Diversity and Antibacterial Activity of Lactic Acid Bacteria Isolated from Kimchi
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 191~196
This study was carried out to investigate the isolation, identification, and antibacterial activity of lactic acid bacteria related to kimchi fermentation. Diluted kimchi soup was plated on the MRS agar media with CaCO
and incubated at
for 2 days. A total of 27 strains of lactic acid bacteria from various indigenous, spontaneously fermented vegetables (kimchi) were isolated. Combined methods of Bergey's manual of systematic bacteriology, BPB media analysis and 16S rDNA sequence analysis were applied for identification, however, their results did not coincide in several cases. Isolated lactic acid bacteria could be classified by the 16S rDNA sequence analysis as Leuconostoc mesenteriodes, Leu. carnosum, Lactobacillus curvatus, Lac. pentosus, Weisselia kimchi, W. cibaria, and Pediococcus pentosaceus. Leu. carnosum has not been reported in kimchi lactic acid bacteria. In addition, antibacterial activities of the isolates were tested with Bacillus subtilis, Escherichia coli, Salmonella enteritidis, S. paratyphica, S. typhi, Staphylococcus aureus, Shigella boydii, and S. sonnei. Some of isolates showed significant antibacterial activities to those pathogens.
Effect of Electron Acceptor on Anaerobic Toluene Biodegradation in Rice Field and Tidal Mud Flat
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 197~200
In oil-contaminated environments, anaerobic biodegradation of toluene depended on the concentration and distribution of terminal electron acceptor as well as the physicochemical properties such as DO concentration, redox potential and pH. This study showed the anaerobic biodegradation of toluene in two different soils by using nitrate reduction, ferric iron reduction, sulfate reduction and methanogensis. Toluene degradation rates in the soil samples taken from rice filed and tidal mud flat by nitrate reduction were higher than those by other processes. Tho soil samples from the two fields were enriched for 130 days by providing toluene as a sole carbon source and nitrate or sulfate as a terminal electron acceptor. The toluene degradation rates in the enriched denitrifying consortia obtained from the rice field and tidal mud flat soil were 310.7 and 200.6
, respectively. The toluene (legradation rates in the enriched sulfate-reducing consortia from the fields ranged fi-om 149.1 to 86.1
Separation and Purification of Teicoplanin by Diaion HP-20 and Conacnavalin A Chromatography
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 201~204
Glycopeptide antibiotics, teicoplanin was purified from a mutant strain of Actinoplanes teichomyceticus ATCC31121, A. teichomyceticus MSL2211. We developed a simple procedure to separate and purify the teicoplanin from the fermentation broth. Teicoplanin was purified by two-step purification system, hydrophobic adsorption and sugar affinity chromatography in combination with HPLC analysis based on the properties of hydrophobic acyl chain and sugar moiety in teicoplanin. Teicoplanin was separated from the culture broth by Diaion HP-20 and further purified by concanavalin A affinity column chromatography. As an adsorbent resin, Diaion HP-20 in broth eliminated toxic effects on growth, reduced feedback repression of teicoplanin production, and assisted In rapid recovery of teicoplanin. The teicoplanin displayed the final yield of 80% and 95% of purity.
Identification of Prevotella intermedia ATCC 25611 Using Pi29-L DNA Probe.
Microbiology and Biotechnology Letters, volume 31, issue 2, 2003, Pages 205~209
Recently, we introduced a new method for rapid screening of bacterial species- or subspecies-specific DNA probes, named “inverted dot blot hybridization screening method”. We then applied this method to develop species- or strain- specific DNA probes for Prevotella intermedia and Prevotella nigrescens. In those studies, among 96 candidate DNA probes which were screened by the new method, 5 probes were confirmed as being putatively strain-specific : 3 probes for P. nigrescens 9336 (ATCC 33563), one for each p. intermedia ATCC 25611 and one for P. nigrescens G8-9K-3 (ATCC 49046). In the present study, we evaluated by Southern blot analysis a DNA probe Pi29-L, one of the 96 candidate probes described above, whether it is specific for the strain ATCC 25611 off. intermedia. Our data show that the probe Pi29-L is potentially P. intermedia ATCC 25611-specific, which can be useful for the detection and identification of the strain, particularly in maintenance of the strain.