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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Microbiology and Biotechnology
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 3, Issue 4 - Dec 1993
Volume 3, Issue 3 - Sep 1993
Volume 3, Issue 2 - Jun 1993
Volume 3, Issue 1 - Mar 1993
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Nucleotide Sequence and Analysis of a Xylanase gene (xynS) from Alkali-tolerant Bacillus sp. YA-14 and Comparison with Other Xylanases
Yu, Ju-Hyun ; Park, Young-Seo ; Yum, Do-Young ; Kim, Jin-Man ; Kong, In-Soo ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 139~145
The nucleotide sequence of the xylanase gene (xynS) from alkali-tolerant Bacillus sp. YA.14 was determined and analyzed. A 639 base pairs open reading frame for xynS gene was observed and encoded for a protein of 213 amino acids with a molecular weight of 23, 339. S1 nuclease mapping showed that the transcription initiation site of the xynS gene did not exist in the cloned DNA. Ribosome binding site sequence with the free energy of -18.8 Kcal/mol was observed 8 base pairs upstream from the initiation codon, ATG. The proposed signal sequence consisted of 28 amino acids, of which 3 were basic amino acid residues and 21 were hydrophobic amino acid residues. When the amino acid sequences of xylanases were compared, Bacillus sp. YA-14 xylanase showed 48% homology with Bacillus sp. YC-335 xylanase and 96% homology with xylanases from B. subtilis and B. circulans.
Structure and Regulation of a Complex Promoter Region from an Alkali-tolerent Bacillus sp.
Kim, Jin-Man ; Park, Hee-Kyung ; Park, Young-Seo ; Yum, Do-Young ; Bai, Dong-Hoon ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 146~155
A DNA fragment from an alkali-tolerent Bacillus sp., conferring strong promoter activity, was subcloned into the promoter probe plasmid pPL703 and the nucleotide sequence of this promoter region was determined. The sequence analysis suggested that this highly efficient promoter region containing the complex clustered promoters comprised three kinds of promoters (P1, P2 and P3), which are transcribed by
RNA polymerase holoenzymes which play major rules at the onset of endospore formation, during sporulation and at the vegetative phase of growth, respectively. S1 nuclease mapping experiments showed that all three promoters had staggered transcription initiation points. The results of chloramphenicol acetyltransferase assay after the subcloning experiments also indicated that the expression of these clustered promoters was correlated with the programs of growth and endospore development. Promoter P1, P2 and P3 were preceded by 75% AT, 79% AT and 81% AT regions, respectively, and a partial deletion of AT-rich region prevented transcription from promoter P1 in vivo. Two sets of 5 -AGTGTT-3 sequences and inverted repeat sequences located around the promoter P1 were speculated as the possible cis acting sites for the catabolite repression in B. subtilis. In vivo transcripts from these sequence regions may be able to form a secondary structure, however, the possibility that a regulatory protein induced by the excess amount of glucose could be bound to such a domain for crucial action remains to be determined.
Expression of Chimeric Chicken-Yeast-Chicken H2B Histone Gene
Son, Seung-Yeol ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 156~160
A chicken H2B histone gene was cloned and expressed in Rat 3 cell line. Its messenger RNA level was about 10 times higher during S phase than during
phase. A chimeric chicken-yeast-chicken H2B histone gene was made to change some of wobble sequences of chicken H2B gene. When the chimeric H2B gene was transfected into the Rat 3 cell line, it showed a pattern of expression similar to that of the original chicken H2B gene. At least in this gene, it was concluded that the wobble sequences were not required for the cell-cycle regulated pattern of expression.
Immobilized Metal Ion Affinity Chromatography of Genetically Engineered Hirudin Variants
Chung, Bong-Hyun ; Chu, Chang-Woong ; Chang, Yong-Keun ; Sohn, Jung-Hoon ; Rhee, Sang-Ki ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 161~167
Immobilized metal ion affinity chromatography (IMAC) was used to separate various types of recombinant hirudins from the culture broth. The wild type hirudin exhibited a retention in Cu(II)-chelated affinity chromatgoraphy since it contained a single exposed histidine at position 51. To obtain a stronger retention on an IDA-Cu(II) column, the hirudin variants were genetically engineered to contain one or two histidine (s) more than the wild type. While the affinity of the variants for IDA-Cu(II) ligand increased in comparison to that of the wild type, the antithrombin activities reduced to a certain degree. Cu(II), Ni(II) and Zn(II) ions were applied separately to the metal chelate column to investigate ligand specificity with respect to protein retention. As a result, the Cu(II) chelated chromatography gave the best resolution for all the hirudins tested and appeared to be the only IMAC that could be used generally for the purification of hirudins with a decreasing pH gradient.
Effect of Galactose and Dextrose on Human Lipocortin I Expression in Recombinant Saccharomyces cerevisiae Carrying Galactose-Regulated Expression System
Nam, Soo-Wan ; Seo, Dong-Jin ; Rhee, Sang-Ki ; Park, Young-Hoon ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 168~173
The expression kinetics of human lipocortin I (LCI), a potential anti-inflammatory agent, was studied in the shake-flask and fermenter cultures of Saccharomyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LCI, and the plasmid stability were investigted under various galactose induction conditions. The expression of LCI was repressed by the presence of a very small amount of dextrose in the culture medium, but it was induced by galactose after dextrose became completely depleted. The optimal ratio of dextrose to galactose for lipocortin I production was found to be 1.0 (10 g/l dextrose and 10 g/l galactose). With optimal D/G ratio of 1.0 and the addition of galactose prior to dextrose depletion, LCI of about 100~130 mg/l was produced. LCI at a concentration of 174 mg/l was porduced in the fed-batch culture, which was nearly a twice as much of that produced in the batch culture. The plasmid stability was very high in all culture cases, and thus was considered to be not an important parameter in the expression of LCI.
Production and Characterization of Extracellular
-Lactamase from Streptomyces aureofaciens SMF14
Kim, Myung-Kuk ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 174~180
A strain SMF14 producing an extracellular
-lactamase was isolated from soil and identified to be a strain of Streptomyces aureofaciens.
-Lactamase was purified from the cell free culture broth through batchwise hydroxyapatite adsorption, anion exchange chromatography on DEAE Sephadex A-50, gel filtration on Sephadex G-75, and adsorption chromatography on hydroxyapatite. The molecular mass was estimated to be about 43 kDa by SDS-PAGE. The
-lactamase had substrate specificity to penicillins and it was inhibited by clavulanic acid, being classified to the group 2a of penicillinase.. The optimal reaction pH and temperature were pH 6.0~7.5 and
-lactamase for penicillin G were calculated to be 1.72 mM and 5.4
Purification of a Steroid
-dehydrogenase from Arthrobacter simplex
BAE. MOO ; MI-KYUNG LEE ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 181~187
-dehydrogenase which introduces a double bond into the 1, 2 positions of steroid ring A was purified from Arthrobacter simplex, an excellent biotransformer of hydrocortisone into prednisolone. Hydrocortisone-induced cells were disrupted by vigorous agitation with glass beads, and a solubilized enzyme was obtained after centrifugation at 100, 000
g for 90 minutes. The enzyme was purified 123-fold in three steps of chromatographic procedures with 13% yield. The last step of testosterone-agarose affinity column decisively contributed to the successful purification. The molecular weight of the enzyme was estimated to be 98, 000 by SDS-PAGE and 100, 000 by gel filtration, indicating that this enzyme behaves as a monomer. The enzyme showed demands for artificial electron acceptor, and among the several reagents tested, phenazine methosulfate acted as the most effective electron acceptor. Subcellular distribution of this enzyme was studied by centrifugation experiment. Comparison of the enzyme activities in pelleted membrane and cytosol fractions suggests that the enzyme may be a weakly attached peripheral membrane protein in vivo. But considerable amounts of enzyme was solubilized without any additional treatments for membrane protein.
Purification and Characterization of Superoxide Dismutase from Pseudomonas polycolor
LEE SANG-OK ; IL-CHUN SEO ; SOOK-HYUN CHUNG ; TAE-HO LEE ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 188~193
Superoxide dismutase (SOD) was purified from Pseudomonas polycolor to an electrophoretically homogeneous state and partially characterized. SOD was purified by ammonium sulfate fractionation, column chromatography on DEAE-Sephadex A-50, phenyl-Toyopearl 650 M, and gel filtration on Sephadex G-100. The molecular weight and subunit molecular weight of the purified enzyme were estimated to be 40, 000 and 20, 000, respectively. The purified enzyme remained stable at pH 9.0~11.0,
for 40 hr, but rapidly became inactive below 9.0. SOD was stable up to
at pH 9.0 with about 80% relative activity, but rapidly became inactive at temperature above that. The enzyme was insensitive to cyanide and fluoride, and sensitive to hydrogen peroxide and azide. The results suggest that the enzyme be an iron-containing SOD.
The Preparation of Crystalline Mannobiose from Brown Copra Meal Using the Enzyme System and Yeast Fermentation
Park, Gwi-Gun ; Chang, Hak-Gil ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 194~198
, 4-Mannobiose was prepared by the enzymatic hydrolysis of brown copra meal and the subsequent elimination of mono-saccharides from the resultant hydrolysate with a yeast. The enzyme system hydrolyzed brown copra meal and produced monosaccharides and
-1, 4-mannobiose without other oligomers at the final stage of the reaction. Brown copra meal (30 g) was hydrolyzed at
and pH 5 for 48 hr with the crude enzyme solution (300 ml) from Penicillium purpurogenum. By the elimination of monosaccharides from the hydrolysis products with a yeast (Candida parapsilosis var. komabaensis k-75), 5.2 g of crystalline mannobiose was obtained without the use of chromatographic techniques. After 50 hours of yeast cultivation, the total sugar content fell from 3.5% to 2.4%, and the average degree of polymerization rose from 1.8 to 2.2.
Use of Bacteriocinogenic Pediococcus acidilactici in Sausage Fermentation
Kim, Wang-June ; Hong, Seok-San ; Cha, Seong-Kwan ; Koo, Young-Jo ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 199~203
The bacteriocin produced by Pediococcus acidilactici KFRI 168 exhibited a wide antimicrobial spectrum including many strains of lactic acid bacteria, Listeria monocytogenes, Staphylococcus aureus, and Enterococcus faecium by both disk and deferred assay methods. Inhibition of Lis. monocytogenes and Stph. aureus were observed only from deferred assay. Gram-negative bacteria were not inhibited. Bacteriocin production was observed at 10 h, and was maximized at 16 h in MRS broth incubated at
. In a beaker sausage fermented with P. acidilactici KFRI 168, viable counts of Stph. aureus, Salmonella, Escherichia coli, Clostridium perfringens, and Lis. monocytogenes were reduced by 2.8, 2.3, 2.4, 0.7, and 0.5 log CFU/g, respectively. Inoculated P. acidilactici KFRI 168 maintained its viable count of more than
CFU/g during the whole fermentation period, and it took less than 8 h to reduce sausage pH below 5.
Morphological Measurements of Submerged Culture of Aspergillus niger by Fully Automatic Image Analysis
OH, SUNG-HOON ; JONG-IL KIM ; PYONG-SU O ; CHERL-HO LEE ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 204~208
A fully automatic image analysis method was applied to obtain detailed data on morphological parameters of a glucoamylase fermentation broth with Aspergillus niger No. PFST-38. a mutant strain for glucoamylase hyperproducer. In the initial stage of fermentation. there was an increase in hyphal length. whereas at the end of the fermentation a decrease in hyphal length and increase in hyphal thickness were observed. The percentage of clumps declined with dilution and the influence of shear stress upon hyphal length was negligible. It was found that the slower the decrease in the main hyphal length and the number of tips with the fermentation time. the higher the glucoamylase production rate was recorded. The production rate of glucoamylase was closely related to the increase in the hyphal thickness.
Rheological Properties of Mycelial Broth in Submerged Culture of Aspergillus niger No. PFST-38
Oh, Sung-Hoon ; O, Pyong-Su ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 209~213
The flow behavior of the mycelial broth of glucoamylase hyperproducer Asp. niger No. PFST-38 for the production of glucoamylase were studied. The mycelial broth followed Bingham-pseudoplastic flow model described by Herschel-Bulkley equation. The yield stress increased with the increase in mycelial concentration. The dependency of the consistency index and the flow behavior index on the mycelial concentration could be expressed by a linear relationship. The consistency index increased proportionally with the mycelial concentration while the flow behavior index decreased with the increase in mycelial concentration. The flow property of the broth was related to the morphological data obtain in the previous study. The changes in apparent viscosity of the broth could be expressed as a function of the hyphal thickness as shown below.
Removal of Inorganic Nitrogen and Phosphorus from Cow s Liquid Manure by Batch Algal Culture
KIM, MAM-SOO ; MOO-YOUNG PACK ;
Journal of Microbiology and Biotechnology, volume 3, issue 3, 1993, Pages 214~216
Cow's liquid manure (CLM), an animal waste, was treated by a batch algal culture to remove inorganic nutrients. CLM used in this study was especially high in concentrations of inorganic nitrogen and phosphorus. The optimum dilution ratio of the CLM for maximum algal growth was 1:25. Ninety five percent of inorganic nitrogen and 100％ of inorganic phosphorus were removed from the CLM with a dilution ratio of 1:25.