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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
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Volume & Issues
Volume 5, Issue 6 - Dec 1995
Volume 5, Issue 5 - Oct 1995
Volume 5, Issue 4 - Aug 1995
Volume 5, Issue 3 - Jun 1995
Volume 5, Issue 2 - Apr 1995
Volume 5, Issue 1 - Mar 1995
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Rapid and Direct Detection of Vibrio vulnificus in Small Octopus (Octopus variabilis) Using Polymerase Chain Reaction
Choi, Sang-Ho ; Lee, Jee-Yeon ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 181~187
The cells of Vibrio vulnificus can be induced to the viable but nonculturable (VBNC) state by natural environmental parameters. The V. vulnificus cells in the VBNC state can not be recovered by ordinary laboratory techniques. This nonculturability could often hamper development of effective processing strategies to minimize the number of V. vulnificus in seafoods. Even with V. vulnificus cells in a culturable state, the length of time required to identify the bacteria in contaminated food by phenotyphic characterization may prevent appropriate in-time responses by public health agencies to infections of the bacteria. In the present study, we used polymerase chain reaction (PCR) to develop a rapid and direct detection method for V. vulnificus in small octopus (Octopus variabilis) which is consumed as a raw food in Korea. The region targeted was a 704-base pair (bp) portion of the hemolysin gene, vvhA, of V. vulnificus. The primers designed for PCR amplification were specific for all V. vulnificus sp. tested. Several methods were examined to extract total DNA directly from V. vulnificus seeded into the octopus homogenate and the guanidine isothiocyanate (CITC) method appeared to be most effective. From the octopus homogenate seeded by V. vulnificus at an initial level of
CFU/ml of the homogenate and then incubated for 12 h, the targeted sequence was successfully amplified by PCR and the 704-bp DNA fragment was observed by gel electrophoresis. The total completion of this assay requires less than one day.
Cloning and Expression of the Gene Encoding Glucose Permease of the Phosphotransferase System from Brevibacterium flavum in Escherichia coli
Kwon, Il ; Lee, Kyu-Nam ; Lee, Jung-Kee ; Pan, Jae-Gu ; Oh, Tae-Kwang ; Lee, Hyung-Hoan ; Yoon, Ki-Hong ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 188~193
A Brevibacterium flavum gene coding for glucose permease of the phosphoenolpyruvate-dependent phosphotransferase system (PTS) was cloned by complementing the Escherichia coli ZSCl13 mutations affecting a ptsG gene with the B. flavum genomic library. From the E. coli clone grown as red colony on a MacConkey plate supplemented with glucose as an additional carbon source, a recombinant plasmid was isolated and named pBFT93. The plasmid pBFT93 was identified as carrying a 3.6-kb fragment of B. flavum chromosomal DNA which enables the E. coli transformant to use glucose or man nose as a sole carbon source in an M9 minimal medium. The non-metabolizable sugar analogues, 2-deoxy-D-glucose (2-DG) and methyl-
-D-glucopyranoside (MeGlc) affected the growth of ZSCl13 cells carrying the plasmid pBFT93 on minimal medium supplemented with non-PTS carbohydrate, glycerol, as a sole cabon source, while the analogues did not repress the growth of ZSCl13 cells without pBFT93. It was also found that both
could be effectively transported into ZSCl13 cells transformed with plasmid pBFT93. Several in vivo complementation studies suggested that the B. flavum DNA in pBFT93 encodes a glucose permease specific for glucose and mannose.
Cloning and Organization of the Ribosomal RNA Genes of the Mushroom Trichloma matsutake
Hwang, Seon-Kap ; Kim, Jong-Guk ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 194~199
A portion (7.4 kb) of ribosomal DNA tandem repeat unit from a genome of the mushroom T. matsutake has been cloned. A 1.75 kb EcoRI fragment was cloned first using S. cerevisiae 255 rRNA gene as a probe, and this was then used for further cloning. A chromosomal walking experiment was carried out and the upstream region of the 1.75 kb fragment was cloned using SmaI/BamHI enzyme, the size was estimated to be 5.2 kb in length. Part of the downstream region of the 1.75 kb fragment was also cloned using XbaI/BamHI enzymes. Restriction enzyme maps of three cloned DNA fragments were constructed. Northern hybridization, using total RNA of T. matsutake, and the restriction fragments of three cloned DNAs as probes, revealed that all four ribosomal RNA genes (large subunit[LSU], small subunit [SSU], 5.85 and 5S rRNA genes) are present in the cloned region. The gene organization of the rDNA are regarded as an intergenic spacer [IGS]2 (partial) - SSU rRNA - internal transcribed spacer [ITS]1 - 5.8S rRNA - ITS2 - LSU rRNA - IGS1 -5S rRNA - IG52 (partial).
Nutritional Regulation of Morphological and Physiological Differentiation on Surface Culture of Streptomyces exfoliatus SMF13
KYE JOON LEE ; KIM, IN SEOP ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 200~205
Nutritional factors regulating the morphological differentiation and physiological differentiation of Streptomyces exfoliatus SMF13 on surface cultures were evaluated. S. exfoliatus SMF13 produced leupeptin and chymotrypsin-like protease (CTP) at the stage of substrate mycelium growth, and leupeptin-inactivating enzyme (LIE) and trypsin-like protease (TLP) at the stage of aerial mycelium growth. The activity of leupeptin and CTP was high in the region of active growing substrate mycelium, whereas the activity of LIE and TLP was high in the region of aerial mycelium or spores. The differentiations were induced in glucose-limited conditions or by the addition of glucose anti-metabolite (methyl
-glucopyranoside), but repressed by high concentrations of glucose or casamino acids. Morphological differentiation (formation of aerial mycelia and spores) was closely related with physiological differentiation (formation of brown-pigment, LIE and TLP). The local distribution of leupeptin, CTP, LIE, and TLP in a developing colony showed that colony development correlated with the production and functions of the compounds: CTP is essential for providing a nitrogen source for mycelium growth: leupeptin regulates TLP activity: LIE inactivates leupeptin: TLP hydrolyzes nongrowing mycelium.
Purification and Characterization of A Cell Wall Hydrolyzing Enzyme Produced by An Alkalophilic Bacillus sp. BL-29
Hong, Soon-Duck ; Kim, Tae-Ho ; Hong, Soon-Duck ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 206~212
A strain BL-29, which produces a extracellular lytic enzyme on E. coli was isolated from the soil. The strain was identified as belonging to the genus Bacillus sp. The lytic enzyme was purified to homogeneity by ion exchange chromatography and gel filtration. Specific activity of the purified enzyme was 28, 850 U/mg protein and yield of the enzyme was 5
. The purified enzyme showed a single band on SDS-PAGE and its molecular weight was estimated to be 31, 000 by SDS-polyacrylamide gel electrophoresis and gel filtration column chromatography. The optimum temperature and pH were
and pH 10.0, respectively. The enzyme was stable at
but enzyme activity was reduced by up to 50
when the temperature was raised to
for 15 min. Stable range of pH was from 5.0 to 11.0. but Enzyme activity was inhibited by lead-acetate, mercuric chloride, ethylene glycol-bis-[
-aminoethyl ether]-N, N,
-tetraacetic acid (EGTA), and ethylenediamine tetraacetic acid (EDTA), but not affected considerably by treatment with other chemical reagents.
Fermentation of MR-387A and B, Novel Aminopeptidase M Inhibitors by Streptomyces sp. SL-387: Phosphate Repression of Inhibitor Formation
YUNG-HEE KHO ; CHUNG, MYUNG-CHUL ; HYO-KON CHUN ; HO-JAE LEE ; CHOONG-HWAN LEE, ; SU-IL KIM ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 213~217
The effect of inorganic phosphate on the fermentative production of aminopeptidase M inhibitors MR-387A and B by Streptomyces sp. SL-387 has been studied. With inorganic phosphate concentrations higher than 0.78 mM, an inverse correlation was found between the maximum inhibitor production and the initial phosphate concentration added. Growth sensitivity of this actinomycete to arsenate, a phosphate analogue, and the use of magnesium carbonate, a phosphate-trapping agent, suggested that the inhibitor formation was under phosphate repression. Exogenous ATP further increased the degree of phosphate interference in both phosphate-repressed and non repressed culture conditions. The use of a phosphate analogue and a protein synthesis inhibitor also suggested that the phosphate itself repressed inhibitor formation.
Induction and Stabilization of Lignin Peroxidase from Phanerochaete chrysosporium
Sang, Byeong-In ; Kim, Yong-Hwan ; Yoo, Young-Je ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 218~223
Veratryl alcohol which has been reported as an inducer for lignin peroxidase showed different effects on the enzyme biosynthesis in Phanerochaete chrysosporium depending on the addition time. Enzyme expression was optimally induced by adding veratryl alcohol when the carbon source began to be depleted. Hydrogen peroxide, to some extent, stimulated production of lignin peroxidase, but beyond a certain concentration, inactivated lignin peroxidase. Tween 80 induced the formation of small pellets, which were resistant to the deactivation by shear stress. Lignin peroxidase production was increased twice compared with that of the control by adopting all the optimal factors in the culture system.
Effect of Galactose Feeding Strategy on Heterologous Human Lipocortin-I Production in the Fed-Batch Culture of Saccharomyces cerevisiae Controlled by the GAL10 Promoter
Chung, Bong-Hyun ; Kim, Byung-Moon ; Rhee, Sang-Ki ; Park, Young-Hoon ; Nam, Soo-Wan ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 224~228
Fed-batch fermentations were conducted to produce human lipocortin-I (LC1), a potential anti-inflammatory agent, from recombinant Sacchromyces cerevisiae carrying a galactose-inducible expression system. The cell growth, expression level of LC1, and the plasmid stability were investigated under various LC1 induction modes performed by three different galactose feeding strategies. Galactoe was fed to induce the expression of LCl from the beginning (initial induction) of culture or when the cell concentration reached 120 OD (mid-phase induction) or 300 OD (late induction). Among the three galactose-induction modes tested, the initial induction mode yielded the best result with respect to a final expression level of LC1. Fedbatch fermentation with initial induction mode produced LC1 at a conentration of 220 mg/l, which corresponded to 1.38- and 1.53-fold increases over those produced by mid-phase and late induction modes.
Studies of Repeated Fed-Batch Fermentation of Cephalosporin C in an Immobilized Cell Bioreactor
Park, Hong-Je ; Khang, Yong-Ho ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 229~233
Acremonium chrysogenum was immobilized in ionotropic gel beads to develop semi-continuous production of cephalosporin C (CPC). Barium alginate beads were more stable than calcium alginate or strontium alginate beads in chemically defined media. The gel stability of Ba-alginate was further increased by cross-linking with polyethyleneimine (PEI). The presence of carboxymethyl cellulose inside Ba-alginate beads did not reduce mass transfer resistance. Ba-alginate microbeads that had little diffusion limitation increased CPC production rate 1.6 fold higher than that of normal beads. CPC fermentation with immobilized cells in Ba-alginate microbeads was performed continuously for 40 days by way of repeated fed-batch operations. Mathematical modeling was developed to describe the repeated fed-batch fermentation system. Results of the computer simulation agreed well with the experimental data, which made it possible to predict an optimal feeding rate that could maximize total CPC productions.
Production of Gericudranins by Hairy Root Culture of Cudrania tricuspidata
Seo, Weon-Taek ; Lee, In-Kyoung ; Yoo, Ick-Dong ; Park , Young-Hoon ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 234~237
Production of new flavanol derivatives with cytotoxic activity, gericudranin A and B, was studied by using hairy root cultures of Cudrania tricuspidata. Schenk and Hildebrandt (SH) medium was chosen for root growth and gericudranin production. After 35 days culture in a half-strength liquid SH medium containing
/l, hairy root growth reached
/I and gericudranin A and B were produced at concentrations of 27mg/l and 21 mg/l, respectively. It was also observed that the contents of gericudranin A and B in hairy root were eight and six times higher than those of cudraniae radix, respectively.
A Simple Method for Recovery of Microbial
by Alkaline Solution Treatment
Lee, In-Young ; Chang, Ho-Nam ; Park, Young-Hoon ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 238~240
A novel and simple purification method for microbial
(PHS) was developed. Sodium hydroxide was found to be efficient for digesting cell materials. Initial biomass concentration, NaOH concentation, digestion time, and incubation temperature were optimized. When 40 g/l of biomass was incubated in 0.1 N NaOH at
for 1 h, PHB purity of 88.4% with a weight average molecular weight (
) of 770,000 and a polydispersity index (PI) of 2.4 was recovered with a yield of 90.8% from the biomass which initially contained PHB of a
of 780,000 and a PI of 2.3.
Increased Cell Surface Hydrophobicity of A Lipopolysaccharide-defective Mutant of Bradyrhizobium japonicum
JAE-SEONG S0 ; PAE, KYEONC-HOON ;
Journal of Microbiology and Biotechnology, volume 5, issue 4, 1995, Pages 241~243
A lipopolysaccharide (LPS) defective mutant of Bradyrhizobium japonicum was characterized in terms of its cell surface hydrophobicity (CSH). By monitoring the kinetics of adhesion to hexadecane the LPS mutant was found to be far more hydrophobic than the wild type strain; the removal coefficients were 4.65
for the mutant, as compared with only 2.40
for the wild type. The possible role of cell surface hydrophobicity of B. japonicum in nodulation process is discussed.