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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 7, Issue 6 - Dec 1997
Volume 7, Issue 5 - Oct 1997
Volume 7, Issue 4 - Aug 1997
Volume 7, Issue 3 - Jun 1997
Volume 7, Issue 2 - Apr 1997
Volume 7, Issue 1 - Feb 1997
Selecting the target year
-Aminotransferase, the Initial Enzyme of Cephalosporin Biosynthesis in Actinomycetes
Rius, Nuria ; Demain, Arnold L. ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 95~100
Strain Improvement for Enhanced Production of Streptokinase and Streptodornase in Streptococcus sp.
HYUN, HYUNG-HWAN ; YOON-BUM LEE ; KYUNG-HWA SONG ; JI-YOUNG JEON ; HYUNE-HWAN LEE ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 101~106
Strain improvement for the enhanced production of streptokinase and streptodornase in Streptococcus sp. ATCC 12449 was performed. Strain UB111, a hyperproductive mutant which was isolated by use of nitrosoguanidine and selection of colonies with large clear zones on DNase test agar plates supplemented with
ammonium chloride, produced about 3 fold more streptokinase and streptodornase than the wild type when tested in shake flask fermentations. The enhanced production of both streptokinase and streptodornase was achieved by cultivating the mutant in a pH-controlled fermentor containing fermentation medium enriched with yeast extract (
). Under these conditions, the mutant produced 7300 units/ml of streptokinase and 800 units/ml of streptodornase.
Purification and Characterization of Antifungal Chitinase from Pseudomonas sp. YHS-A2
Lee, Han-Seung ; Lee, Hyun-Jung ; Choi, Sung-Won ; Her, Song ; Oh, Doo-Hwan ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 107~113
A strain producing a high amount of chitinase was isolated from soil, identified as Pseudomonas sp., and tentatively named Pseudomonas sp. YHS-A2. An extracellular chitinase of Pseudomonas sp. YHS-A2 was purified according to the procedure of ammonium sulfate saturation, affinity adsorption, Sephadex G-100 gel filtration and Phenyl-sepharose CL-4B hydrophobic interaction column chromatography. The molecular weight of the purified enzyme was estimated to be 55 kDa on SDS-PAGE was confirmed by active staining. Optimal pH and temperature of the enzyme are pH 7.0 and
, respectively, and the enzyme is stable between pH 5.0 and 8.0 and below
. The main products of colloidal chitin by the chitinase were N-acetyl-D-glucosamine and N,N'-diacetylchitobiose both of which were detected by HPLC analysis. The enzyme is supposed to be a random-type endochitinase which can degrade any position of
-l,4-linkages of chitin and chitooligosaccharides. The chitinase inhibited the growth of some phytopathogenic fungi, Fusarium oxysporum, Botrytis cineria, and Mucor rouxii and these antifungal effects were thought to be due to the characteristics of endochitinase.
Characterization of an Endoxylanase Produced by an Isolated Strain of Bacillus sp.
Lee, Jay-J. ; Hahm, Kyoung-Soo ; Lee, Ki-Young ; Lee, Sung-Taik ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 114~120
Microorganisms producing xylanase were screened for the enzymatic production of xylo-oligo saccharides from xylan. One of the bacteria isolated from compost produced an endoxylanase extracellularly. The bacterium was identified as Bacillus sp. according to its taxonomic characteristics examined. Xylanase production reached upto 5 U/ml after 22 h of culture in LB medium at
. The xylanase was purified by ammonium sulfate precipitation and gel filtration. The molecular weight of the xylanase was estimated to be 20,400 by SDS-PAGE. Optimal temperature and pH for the xylanase activity was
and 6.5, respectively. The enzyme was stable at temperatures upto
and pH values from 4 to 10. The xylanase was completely inhibited by the addition of 2 mM mercury ion. Apparent
values for oat spelt xylan were 9.2 mg/ml and 1954 U/mg protein, respectively. For birchwood xylan, the values were 6.3 mg/ml and 1009 U/mg protein. The predominant products of the xylan hydrolysis were xylobiose, xylotriose and xylotetraose, indicating that the enzyme is an endoxylanase. Upto
of the initially added enzyme (2 U/ml) was bound to 50 mg/ml of the insoluble fraction of oat spelt xylan after incubation at
for 30 min.
Purification and Characterization of an Inulin Fructotransferase from Flavobacterium sp. LC-413
Cho, Chul-Man ; Lee, Sang-Ok ; Hwang, Ji-Sook ; Jang, Kyung-Lip ; And, Tae-ho ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 121~126
A bacterial strain LC-413, producing an extracellular inulin fructotransferase (depolymerizing) which converts inulin into di-D-fructofuranose dianhydride (DFAIII), was isolated from soil. Inulin fructotransferase from the isolate identified as a strain Flabobacterium sp. was purified from the culture broth by ammonium sulfate precipitation, followed by column chromatograpies on DEAE-Toyopearl 650 M and phenyl-Toyopearl 650 M. The purified enzyme gave a single band on an electrophoretic disc-gel. The molecular weight of the enzyme was estimated to be 44, 000 Da by SDS-polyacrylamide gel electrophoresis, and 45, 000 Da by gel filtration, suggesting the monomeric state of the enzyme. The isoelectric point of the enzyme was about pH 4.5. The optimal pH and temperature for the enzyme reaction were 6.0 and
, respectively. The purified enzyme digested inulin into di-D-fructofuranose-l, 2': 2, 3'-dianhydride, confirming the enzyme was an inulin fructotransferase (inulinase II).
Effects of Amino Acids, Carbohydrates and Phosphorus Sources on Growth and Alkaline Phosphatase Activity of the Marine Cyanobacterium Anabaena sp. Strain CA
Singh, Jeet Bahadur ; Vyas, Deepak ; Kumar, Har Darshan ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 127~131
Alkaline phosphatase (APase) was found to be inducible in Anabaena sp. strain CA Growth was less than control in presence of most amino acids except glycine and serine, but most amino acids enhanced APase activity. Highest APase activity was recorded in tyrosine supplemented culture followed by hydroxyproline, cystein, valine and glutamic acid. Threonine supplemented material showed lowest APase level (1.8 nmol/mg protein/min). Lactose, glucose, sodium pyruvate and succinate stimulated growth but not APase activity. APase activity was high in the presence of sucrose, mellibiose, mannitol, arabinose, maltose and sorbose, even though the growth in these supplements was less than in control. Organic phosphate sources supported good growth of the organism. Best growth occurred in presence of inorganic phosphate, adenosine diphosphate, fructose 1,6-diphosphate or ribulose 1,5-diphosphate, followed by other phosphorus sources tested. APase activity in presence of any of the organic phosphate sources was 3 to 5 fold low as compared to phosphate limited culture. Also, there was no APase activity in cultures grown on inorganic phosphate. These data indicate that most amino acids and a few carbohydrates (sucrose, mellibiose, arabinose and sorbose) are suitable for APase production. Lactose, glucose, pyruvate or succinate may be used as a carbon source during photoheterotrophic growth of the cyanobacterium. Glycine and serine are preferred nitrogen sources for its growth. Phosphate repressible APase activity has been found in Anabaena sp. strain CA.
Design and Expression of High Nutritional Peptide (HEAAE) in E. coli
Kim, Jae-Ho ; Lee, Chang-Kook ; Hong, Bum-Shik ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 132~137
A novel protein (HEAAE, High Essential Amino Acid Encoding Protein), rich in essential amino acids (
of total), was designed and constructed in our laboratory. The designed peptides were analyzed by SYBLE and stable secondary and tertiary structures were predicted. The monomeric form (HEAAE-1) of the protein consists of 20 amino acid residues with four additional amino acids comprising a potential
-turn (HEAAE-4). Size exclusion analysis demonstrated that the monomer is self-aggregates in aqueous solution to form higher ordered multimeric structures, which are very reminiscent of natural plant storage proteins. The DNA encoding this amino acid sequence was synthesized, and from this monomeric gene fragment (heaae-1), the stable tetrameric form of the gene (heaae-4) was generated by subcloning into the E. coli expression vector pKK223-3. A clear 6 kDa polypeptide band corresponding to the molecular weight of the dimeric form (HEAAE-2) was detected. The smeared band which appeared around the molecular weight corresponding to HEAAE-4 of 11 kDa suggested that the tetramer form of this protein might be processed into smaller size products.
Expression of de novo Designed High Nutritional Peptide (HEAAE) in Tobacco
Kim, Jae-Ho ; Lee, Chang-Kook ; Hong, Bun-Shik ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 138~143
We have designed and constructed a gene encoding novel high essential amino acid encoding protein(HEAAE). The resultant DNA fragment was tested for in vitro and in vivo expression and then cloned into plant expression vector pBI121, under the control of the cauliflower mosaic virus 35S promoter. Agrobacterium tumefaciens, strain LBA4404, was subsequently transformed with this new construct and Nicotiana tabacum var. Xanthi transgenic plants were obtained. DNA analysis by Southern procedure confirmed the presence of the multi-copy number of genes in the transformed plants. Analysis of RNA and protein synthesized in these transgenic plants demonstrated the stable expression of this gene.
An Outer Membrane Protein Preparation as a Vaccine against Pseudomonas aeruginosa Infection
Park, Wan-Je ; Cho, Yang-Je ; Ahn, Dong-Ho ; Jung, Sang-Bo ; Lee, Na-Gyong ; Kim, Hyun-Su ; Hahm, Kyung-Soo ; Kim, Yu-Sam ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 144~150
We developed a simple and efficient method to prepare a Pseudomonas vaccine of outer membrane (OM) proteins free from lipopolysaccharide (LPS). A three step purification process including extraction, ultrafiltration and ultracentrifugation effectively removed LPS from the OM protein fraction. Approximately 2 mg of the OM proteins was obtained from 1 g of wet cell. LPS contaminant in the vaccine preparation was less than 0.003% (w/w) of protein and protease activity was not detectable. To achieve a wide range of protection, OM proteins prepared from four attenuated P. aeruginosa strains were mixed in equal amounts and used as a vaccine, which elicited in rabbits a high titer of antibody reactive to all of the seven Fisher types. The antisera from the immunized rabbit had a strong reactivity to vaccine proteins larger than 25 kDa. In a burned mouse infection model, immunization with the vaccine significantly enhanced bacterial clearance in the Pseudomonas infected skin. The vaccination also provided mice an excellent protection against Pseudomonas infection (11, 16). Data on antigenicity, mutagenicity, acute, subacute toxicity and pharmacological tests confirmed the safety of the vaccine (1, 3, 10, 12, 17). These data demonstrate that this method can be applied to manufacture a bacterial vaccine of OM proteins with safety and prophylactic efficacy at a practical low cost.
Hydrolysis of Olive Oil by Lipase, Immobilized on Hydrophobic Support
Jung, Ju-Young ; Yun, Hyun-Shik ; Kim, Eun-Ki ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 151~156
Two commercially available lipases, Lipase OF (non-specific lipase from Candida rugosa) and Lipolase 100T (1, 3-specific lipase from Aspergillus niger), were immobilized on insoluble hydrophobic support HDPE (high density polyethylene) by the physical adsorption method. Hydrolysis performance was enhanced by mixing a non-specific Lipase OF and a 1, 3-specific Lipolase 100T at a 2 : 1 ratio. The results also showed that the immobilized lipase maintained its activity at broader temperature (
) and pH (4-8) ranges than soluble lipases. In the presence of organic solvent (isooctane), the immobilized lipase retained most of its activity in upto 12 runs of hydrolysis experiment. However, without organic solvent in the reaction mixture, the immobilized lipase maintained most of its activity even after 20 runs of hydrolysis experiment.
A Partial Nucleotide Sequence of Chitin Synthase (CHS) Gene from Rice Blast Fungus, Pyricularia oryzae and Its Cloning
Hwang, Cher-Won ; Park, In-Cheol ; Yeh, Wan-Hae ; Takagi, Masamchi ; Ryu, Jin-Chang ;
Journal of Microbiology and Biotechnology, volume 7, issue 2, 1997, Pages 157~159
A 340-bp chitin synthase gene(CHS) fragment was cloned from the genomic DNA of Pyricularia oryzae using a PCR process with two primer DNAs corresponding to highly conserved sequences within fungal CHS genes. The entire DNA nucleotide sequences of the cloned DNA fragment were determined and analyzed. The amino acid sequences deduced from the nucleotide sequence of the amplified DNA fragment showed 86% homology to that of the Aspergillus fumigatus CHSE gene (9). Using this PCR-amplified DNA, about 2.3 kb of including the PCR fragment of CHSE gene was cloned from genomic library.