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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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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 6, Issue 6 - Dec 1996
Volume 6, Issue 5 - Oct 1996
Volume 6, Issue 4 - Aug 1996
Volume 6, Issue 3 - Jun 1996
Volume 6, Issue 2 - Apr 1996
Volume 6, Issue 1 - Feb 1996
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Microbial Petroleum Desulfurization
KIM, BYUNG-HONG ; PYUNG-KYUN SHIN ; JONG-UK NA ; DOO-HYUN PARK ; SUNG-HO BANG ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 299~308
Cloning and Functional Expression in Escherichia coli of the Polyhydroxyalkanoate Synthase (phaC) Gene from Alcaligenes sp. SH-69
Lee, Il ; Nam, Sun-Woo ; Rhee, Young-Ha ; Kim, Jeong-Yoon ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 309~314
Alcaligenes sp. SH-69 can synthesize poly(3-hydroxybutyrate-co-3-hydroxyvalerate) from a single carbon source such as glucose. To clone the phaC gene from Alcaligenes sp. SH-69, a polymerase chain reaction was performed using the oligomers synthesized based on the conserved regions of the phaC genes from other bacteria. A PCR product (550 bp) was partially sequenced and the deduced amino acid sequence was found to be homologous to that of the phaC gene from Alcaligenes eutrophus. Using the PCR fragment Southern blotting of Alcaligenes sp. SH-69 genomic DNA digested with several restriction enzymes was carried out. To prepare a partial genomic library, about 5-Kb genomic DNA fragments digested with EcoRI, which showed a positive signal in the Southern blotting, were eluted from an agarose gel, ligated with pUC19 cleaved with EcoRI, and transformed into Escherichia coli. The partial library was screened using the PCR fragment as a probe and a plasmid, named pPHA11, showing a strong hybridization signal was selected. Restriction mapping of the insert DNA in pPHA11 was performed. Cotransformation into E. coli of the plasmid pPHA11 and the plasmid pPHA21 which has phaA and phaB from A. eutrophus resulted in turbid E. coli colonies which are indicative of PHA accumulation. This result tells us that the Alcaligenes sp. SH-69 phaC gene in the pPHA11 is functionally active in E. coli and can synthesize PHA in the presence of the A. eutrophus phaA and phaB genes.
Site-specific Disruption of Glyoxylate Bypass and Its Effect in Lysine-producing Corynebacterium lactofermentum Strain
Kim, Youn-Hee ; Lee, Heung-Shick ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 315~320
The role of glyoxylate bypass in a lysine-producing Corynebacterium lactofermentum strain was analyzed. Unlike the wild type, the strain expressed enzymes of glyoxylate bypass during growth in the fermentation broth containing glucose as the carbon source. To evaluate the importance of glyoxylate bypass in the strain, we disrupted chromosomal aceA by using a cloned fragment of the gene. Site-specific disruption of aceA which codes for the isocitrate lyase, the first enzyme of the bypass, was confirmed by Southern blot analysis. The aceA mutant strain completely lost isocitrate lyase activity and ability to grow in a minimal medium containing acetate as the sole carbon source. The mutant strain was similar to its parental strain in growth characteristics and produced comparable amounts of lysine in shake flasks containing glucose as the carbon source. The amount of oxaloacetate accumulated in the fermentation medium was similar for both strains, suggesting that expression of glyoxylate bypass does not necessarily lead to the increase in intracellular oxaloacetate. These data clearly demonstrate that glyoxylate bypass does not function as one of the routes of carbon supply for lysine production in the strain. It appears that the leakiness of the glyoxylate bypass in the strain might be the result of a secondary mutation which arose during previous strain development by random mutagenesis.
Localization of Genes Involved in Exopolysaccharide Biosynthesis in Zoogloea ramigera 115SLR
LEE, SAM-PIN ; OH-SIK KWON ; ANTHONY JOHN SINSKEY ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 321~325
Mutants having altered levels and/or types of EPS in exopolysaccharide biosynthesis were isolated after NTG mutagenesis of Zoogloea ramigera 115SLR. Mutant candidates were classfied with five groups based on the observed characteristics for EPS biosynthesis pattern. The recombinant plasmids pLEX3BS and pLEX3BM were constructed from pEX3B which was previously isolated from genomic DNA of Z. ramigera 115SLR. Plasmid pLEX3BM with a 7.8 kb insert DNA contains an additional 1.8kb DNA fragment which is not present in pLEX3BS containing 13 kb insert DNA. Plasmid pLEX3BM was able to complement the mutation responsible for the changes in morphology of Z. ramigera 115SLR. However, the complementation of EPS negative mutant strains was not successful with pLEX3BM. Plasmid pLEX3BS on the other hand complemented the mutation responsible for the loss of EPS biosynthesis, resulting in the restoration of Z. ramigera 115SLR phenotype. But this plasmid was not able to complement the morphological mutant strain, Z. ramigera 115SLR.
Cloning of a Hemolytic Mosquitocidal Delta-endotoxin Gene (cyt) of Bacillus thuringiensis 73E10-2 (serotype 10) into Bacillus subtilis and Characterization of the cyt Gene Product
Kim, Kwang-Hyeon ; Ohba, Michio ; Kim, Byung-Woo ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 326~330
To illustrate whether a hemolysin in
-endotoxins of Bacillus thuringiensis strain 73E10-2 and subsp. israelensis had immunological identity, a cyt gene of the strain 73E10-2 which encodes a hemolysin was cloned to B. subtilis (transformant 2753). The transformant 2753 containing cyt gene produced the hemolysin which lysed sheep erythrocytes after treatment of proteinase K. The hemolysin was proved also to be toxic against mosquito larvae (Aedes aegypti). The molecular weight of the hemolysin produced from the transformant 2753 was determined to be about 25 kDa by SDS-PAGE and immunoblot. The hemolysin in
-endotoxin of subsp. israelensis and subsp. kyushensis did not react on immunoblot using polyclonal anti-
-endotoxin of the strain 73E10-2, but 70-140 kDa mosquitocidal toxins in
-endotoxin of subsp. kyushuensis reacted.
Molecular Cloning and Expression of the
-Xylosidase Gene (xylB) of Bacillus stearothermophilus in Escherichia coli
Suh, Jung-Han ; Eom, Soo-Jung ; Cho, Ssang-Goo ; Choi, Yong-Jin ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 331~335
-Xylosidase gene (xylB) from Bacillus stearothermophilus was isolated from the genomic library, cloned into pBR322, and subsequently transferred into Escherichia coli HB101. Six out of 10, 000 transformants were selected from the selective LB medium supplemented with p-nitrophenyl-
-L-arabinofuranoside (pNPAf) and ampicillin (
/ml) based on their ability to form a yellow ring around the colony. One of the clones was found to harbor the recombinant plasmid with 5.0 kb foreign DNA, which was identical to the
-L-arabinofuranosidase gene (arfI) previously cloned in this lab, while the other five had 3.5 kb of the foreign DNA. Southern blotting experiments confirmed that the 3.5 kb insert DNA was from B. stearothermophilus chromosomal DNA. A zymogram with 4-methylumbelliferyl-
-L-arabinofuranoside as the enzyme substrate revealed that the cloned gene product was one of the mutiple
-L-arabinofuranosidases produced by B. stearothermophilus. Unlike the arfI gene product, the product of the gene on the insert DNA (xylB) showed an activity not only on pNPAf but also on oNPX suggesting that the cloned gene product could be a bifunctional enzyme having both
Enzymatic Conversion of Glutaryl 7-Aminocephalosporanic Acid to 7-Aminocephalosporanic Acid with an Immobilized Glutaryl 7-Aminocephalosporanic Acid Acylase
SHIN, HAN-JAE ; SEUNG-GOO LEE ; WANG-SIK LEE ; KI-HONG YOON ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 336~339
Glutaryl 7-aminocephalosporanic acid acylase of Pseudomonas sp. SY-77-1 was immobilized with oxiran acrylic beads for the production of 7-aminocephalosporanic acid (7-ACA) from glutaryl 7-aminocephalosporanic acid (GL 7-ACA). The immobilized enzyme maintained its activity at a constant level for 7 days, but lost 30
of its activity after 20 days. Optimal reaction conditions for the synthesis of 7-ACA were found to be
and pH 8.0 using the immobilized enzyme. For the economic production of 7-ACA, substrate and enzyme concentrations were optimized to 60 mM and 0.5 g wet weight per 10
of reaction volume, respectively. Under optimized conditions, 50 mM 7-ACA was produced from 60mM GL 7-ACA within 8 h, resulting in a conversion yield of 83
Isolation and Identification of Staphylococcus sp. from Korean Fermented Fish Products
Um, Mi-Na ; Lee, Cherl-Ho ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 340~346
In order to find out if staphylococci occur in significant numbers in Korean fermented fish products, a total of 40 different fermented fish products were collected from different markets in Korea and analyzed for their physico-chemical and microbiological states. The pH, salt concentration and water activity of the products were measured and the total viable cell count and the number of Staphylococcus grown on mannitol salt agar were determined. The identification of the strains of Staphylococcus were made by API Staph Strip and MIS identification kits, and the physiological properties of the identified strains were further characterized by different conventional methods. The pH, salt content and water activity of fermented fish samples varied widely from 4.8 to 7.1, 7.4-28.7
and 0.77-0.84, repectively, depending on the type of product. The total viable cell count varied from
cfu/ml, and most of the samples had
cfu/ml No correlation was found between the viable cell count and the pH, NaCl concentration and water activity of the samples. Among the 35 colonies identified as Staphylococcus strains by the identification kits, S. xylosus was the most frequently occurring strain marking 17, and S. warneri was 8, S. epidermidis 4 and S. cohnii 2. S. hominis, S. saprophyticus, S. haemolyticus and S. aureus were also identified once each. In some samples (K-3, P-6, K-8, G-5 and G-10), 2-3 different species of Staphylococcus were found. Considering the region of sampling, among the 10 samples from Kunsan 5 were identified as S. warneri, while in the other regions S. xylosus was predominant. Although the physiological characteristics of the identified strains were generally consistent with those in Bergey's Manual, some discrepances were also observed. All the strains were highly salt tolerant, growing in the media containing over 18
NaCl. All the strains except S. aureus (G-11) showed negative in hemolysis activity, plasma coagulation and DNase tests. All the strains including S. aureus (G-11) showed negative in enterotoxin test.
An Optimization of Flavonoid Production from the Suspension Culture of Scutellaria baicalensis Georgi Cells
SEO, WEON-TAEK ; YOUNG-HOON PARK ; TAE-BOO CHOE ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 347~351
Flavonoid production by suspended cells of Scutellaria baicalensis Georgi was studied and the medium was optimized for cell growth and baicalin production. In SH medium the flavonoid production was not closely associated with the cell growth. A modified SH medium, FPM, was therefore designed for enhanced baicalin production. In FPM, both cell growth and baicalin production were increased by 1.5 times and 1.67 times than in the original SH medium, respectively. The increases could be attributed to the increased metabolic activities involved in the flavonoid biosynthesis as represented by enhanced activities of phenylalanine ammonia-lyase.
Salting-out Effects on the Partition of Proteins in Aqueous Two-phase Systems
KIM, CHAN-WHA ; CHO KYUN RHA ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 352~357
The partition of proteins in the salt-rich phase of polyethylene glycol (pEG)/salt aqueous two-phase systems is limited by the salting-out effects of salt. The logarithm of the concentration of proteins partitioned in the salt-rich phase decreases linearly with increases in the concentration of salt in the salt-rich phase (salting-out). Therefore, the partition of a given protein in the salt-rich phase of aqueous two-phase systems can be estimated from the salting-out constant. The slope of the solubility line (salting-out con-stant) for a given protein is determined by the type of salt in the two-phase systems.
Extraction of Docosahexaenoic Acid (DHA) from Lyophilized Thraustochitrium sp.
CHO, JOONG-HOON ; GWI-SUK HEO ; JI-WON YANG ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 358~360
Solvent extraction, soxhlet method, and supercritical fluid extracion were considered, respectively, as the method of choice for the recovery of DHA from lyophilized Thraustochitrium sp., and the results of corresponding extraction were compared. Supercritical fluid extraction seems to be the most appropriate process with respect to time, process simplicity, and extractant intoxicity.
Terminal Nucleotide Sequences in the Double-stranded RNA Genome Segments of Infectious Pancreatic Necrosis Virus DRT Strain
Chung, Hye-Kyung ; Park, Hong-Chul ; Ichiro Uyeda ; Masamichi Isogai ; Lee, Hyung-Hoan ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 361~363
The terminal regions of the double-stranded RNA (dsRNA) genome segments of infectious pancreatic necrosis virus (IPNV) DRT strain were sequenced. The dsRNAs, which were
-labelled at their 3'-termini by incubation with [
]pCp and T4 RNA ligase, were separated by 5
polyacrylamide gel electrophoresis, and the segments A and B of IPNV-DRT were sequenced by two-dimensional gel electrophoresis. The 5'-terminal sequences of the IPNV-DRT plus strand from two genome segments were found to have the same conserved nucleotide (5'-CGG(C/A)A-), but the 3'-terminal sequences -CCCCAGGCG-3' and -CGGACCCCG-3' were found in the plus strand from segments A and B, respectively. The inverted oligonucleotide sequences of 3'-terminal of between segments A and B were found and they differ from those of other IPNVs.
Nucleotide Sequence of 16S rRNA Gene from Streptomyces melanosporofaciens 7489
LEE, DONG-SUN ; SUNG-OUI SUH ; SEON-KAP HWANG ; TAEG-KYU KWON ; TAE-HO KIM ; WOO-CHANG SHIN ; SOON-DUCK HONG ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 364~365
A region encoding the 16S rRNA was cloned by PCR from Streptomyces melanosporofaciens 7489 and sequenced by the chain-termination dideoxy sequencing method. A phylogenetic tree constructed by sequence alignment of 24 Streptomyces species suggests that there is little evolutionary distance between this strain and Streptomyces rimosus.
Inhibition Mode of DNA Topoisomerase by Dibutyl Phthalate
Lee, Dong-Sun ; Hong, Soon-Duck ;
Journal of Microbiology and Biotechnology, volume 6, issue 5, 1996, Pages 366~367
Dibutyl phthalate induced topoisomerase Ⅰ mediated DNA relaxation comparable to that of camptothecin, and topoisomerase Ⅱ mediated DNA relaxation equipotent to that of 4'-(9-acridinylamino) methanesulfon-m-anisidide (m-AMSA). The relaxation activities of dibutyl phthalate were dose-de-pendent and nearly as potent as those of camptothecin and m-AMSA.