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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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Heterologous Gene Expression and Secretion of the Anticoagulant Hirudin in a Methylotrophic Yeast Hansenula polymorpha
Sohn, Jung-Hoon ; Michael-Yu-Beburov ; Choi, Eui-Sung ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 65~72
A heterologous gene expression and secretion system using a methylotrophic yeast, Hansenula polymorpha was developed for the production of anticoagulant hirudin. Hirudin gene was expressed under the control of a strong and inducible methanol oxidase (MOX or AOX) promoter. The mating factor a pre-pro leader sequence of Saccharomyces cerevisiae was employed for hirudin to be secreted into the extracellular medium. Hirudin expression cassette was introduced into three strains of H. polymorpha, A16, HPBl and DLl which have different genetic backgrounds. This expression cassette was stably integrated into the host chromosomal DNA. Biologically active and mature hirudin was efficiently expressed and secreted into the extracellular medium. About 19 mg/L of hirudin was found in the culture supernatant in the case of a two-copy integrant of the strain HPBl under suboptimal culture conditions.
Nucleotide Sequence of a Bacteriolytic Enzyme Gene from Alkalophilic Bacillus sp.
Jung, Myeong-Ho ; Ohk, Seung-Ho ; Yum, Do-Young ; Kong, In-Soo ; Bai, Dong-Hoon ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 73~77
The nucleotide sequence of Bacillus sp. bacteriolytic enzyme gene, lytP and its flanking regions were determined. A unique open reading frame for a protein of Mw. 27, 000, and a putative terminator sequence, were found behind a concensus ribosome binding site located 8 nt upstream from ATG start codon. The primary amino acid sequence deduced from nucleotide sequence revealed a putative protein of 255 amino acid residues with an Mw. of 27, 420. No significant homology could be found between the amino acid sequence of Bacillus sp. bacteriolytic enzyme and that of other cell wall hydrolases.
Selection and Characterization of Catabolite Repression Resistant Mutant of Bacillus firmus var. alkalophilus Producing Cyclodextrin Glucanotransferase
Do, Eun-Ju ; Shin, Hyun-Dong ; Kim, Chan ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 78~85
In order to elucidate the mechanism which regulates the production of cyclodextrin glucanotransferase (CGTase) and to achieve overproduction of CGTase by releasing catabolite (glucose) repression, several catabolite repression resistant mutants were selected from newly screened Bacillus firmus var. alkalophilus H609, after NTG (N-methyl-N -nitro-N-nitrosoguanidine) treatment, using 2-deoxyglucose as a nonmetabolizable analog of catabolite glucose and as a selection marker. Five catabolite repression resistant mutants were selected from about 30, 000 2-deoxyglucose resistant colonies. Relative catabolite repression indices of the selected mutants were in the range of 8~80% assuming 100% for parent strain. The amount of CGTase produced by the mutant strain CR41, which was 250 units/ml, was three times larger than that produced by its parent strain. The mutation seems to have occurred in the regulatory region of CGTase gene and not in the structural region or the glucose transporting system in cell membrane. The enzymatic properties of CGTase excreted from parent and mutant strains were also compared.
Cloning and DNA Sequence of Carboxymethylcellulase (CMCase) Gene from Cellulomonas sp. YE-5
Her, Song ; Kim, Dong-Seob ; Choi, Sun-Jin ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 86~90
CMCase positive clones were screened from Cellulomonas sp. YE-5 and named pCE1, pCE2 and pCE3. Among the positive clones pCE1 was used for this study, because it has the smallest insert and the highest CMCase activity among the 3 clones, and its nucleotide sequence was determined. The CMCase gene in pCE1 was composed of 1071 bp of nucleotides coding 357 amino acids. Computer analysis showed that the pCE1 has 65% sequence homology with the endoglucanase from Cellulomonas fimi.
Nucleotide Sequencing Analysis of a Gene Coding for 3-Isopropylmalate Dehydrogenase of Kluyveromyces fragilis
Hong, Soon-Duck ; Kim, Jong-Guk ; Lee, Dong-Sun ; Woo, Ju-Hyung ; Lee, Sang-Yong ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 91~94
A 3-isopropylmalate dehydrogenase (3-IPMD) gene was cloned from Kluyveromyces fragilis. pJK104 could complement Escherichia coli leuB and Saccharomyces cerevisiae leu2 auxotrophs. The coding region was subcloned and the nucleotide sequence was determined. A 1.8 Kb EcoRI/SphI fragment of pJK104 subcloned in pUC18 could still complement the leuB mutation. An open reading frame of 1164 bp that corresponds to a polypeptide of 387 amino acids was found in the cloned fragment. The homology between the 3-isopropylmalate dehydrogenase of S. cerevisiae and that of K. fragilis was 68.13％ in nucleotides.
Selective Isolation and Characterization of Schwanniomyces castellii Mutants with Increased Production of a-Amylase and Glucoamylase
Ryu, Yeon-Woo ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 95~98
This study was carried out to isolate and characterize the mutant strains of Schwanniomyces castellii NRRL Y-2477. Mutants were prepared with the treatment of ethyl methane sulfonate. 2-deoxy-D-glucose resistant mutants were isolated and two mutants were selected based on their high production of amylolytic enzymes and their ability to ferment starch. The mutants selected had higher a-amylase and glucoamylase activities than the wild type strain from several other carbon sources. Especially, it was revealed that mutant strain M-9, when cultured in the presence of glucose as a sole carbon source, shows relatively high activities of a-amylase and glucoamylase compared to those of the wild type strain. In result, this mutant strain can be considered as a constitutive producer of amylolytic enzymes. To compare the ethanol production ability of wild type strain and of mutant strains selected, an alcohol fermentation was carried out using 100 g/l soluble starch. Mutant strain M-9 did not improve the direct alcohol fermentation of starch, despite its excellent amylolytic activities performance. On the other hand, mutant strain M-6 produced 37.9 g/l (4.8%, v/v) ethanol by utilizing about 82% of substrate.
Isolation of Soil Bacteria Secreting Raw-Starch-Digesting Enzyme and the Enzyme Production
Sung, Nack-Moon ; Kim, Keun ; Choi, Sung-Ho ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 99~107
Two strains (No. 26 and 143) of bacteria which secrete both pectinase and raw-starch-digesting amylase simultaneously, were isolated from various domestic soil samples. The two bacteria were identified as Pasteurella ureae judging by their morphological and physiological characteristics. The optimal culture conditions for the production of raw-starch-digesting enzyme by the Pasteurella ureae 26 were using
as the nitrogen source at
with the pH of 7.5, and 15 of C/N ratio. Since the enzyme was produced only when raw or soluble starch was used as a carbon source, but not when glucose or other sugars was used, the enzyme was considered to be an inducible enzyme by starch. Thin layer chromatography of the hydrolyzed product of starch by the raw-starch-digesting enzyme of the strain No. 26 showed that glucose, maltose and other oligosaccharides were present in the hydrolyzates, and therefore the enzyme seemed to be
. The enzyme had adsorbability onto raw com starch in the pH range of 3 to 9.
Characterization of a New Acidophilic Acetobacter sp. Strain HA Isolated from Korean Traditional Fermented Vinegar
CHUN, HONG-SUNG ; SUNG-JUN KIM ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 108~114
A new strain of acidophilic, acetogenic bacterium, Acetobacter sp. strain HA was isolated by selective enrichment from the traditionally fermented rice wine vinegar in Korea. It was a gram-negative, non-motile short rod and oxidized acetate and lactate. The optimal temperature and pH for growth were
and 4.0, respectively. The strain HA differed from other Acetobacter species by growing well on methanol, xylitol, inositol, dulcitol, D-xylose, L-arabinose, and D-mannose as sole sources of carbon and energy. The isolated strain HA did not produce
-pyrones from glucose and did not produce ketone bodies from glycerol. The quinone system used in this study was an ubiquinone-9 isoprene unit. The guanine-plus-cytosine content of the DNA was 50.7 mol%, and the major cellular fatty acids were
P(3HB) Accumulation in Alcaligenes eutrophus H16(ATCC 17699) under Nutrient-Rich Condition and Its Induced Production from Saccharides and Their Derivatives
Song, Jae-Jun ; Shin, Yong-Chul ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 115~122
Poly(3-hydroxybutyrate)(P(3HB)) accumulation under nutrient-rich condition with various amounts of
was systematically investigated. The results of the electron-microscopy and the solvent extraction showed that the P(3HB) accumulation is unavoidable even under nutrient-rich condition. This indicates that in a two-step culture of Alcaligenes eutrophus H16, the researches should be careful in interpreting the data of polyhydroxyalkanoates(PHAs) accumulation in terms of the carbon-source fed in the second step because the two-step culture product contains the P(3HB) produced under nutrient-rich condition. The polyester production capability in a two-step batch culture of A. eutrophus H16(ATCC 17699) was also investigated using various saccharides and their derivatives such as glucose, fructose, gluconic acid, glucaric acid, sorbitol, lactose, galactose, and mannose. The polyesters synthesized were characterized by 500 MHz
spectroscopy, intrinsic viscosity
measurement in chloroform and differential scanning calorimetry(DSC). 500 MHz
analysis showed that all polyesters synthesized generally contained 1~2 mol% of 3HV. Another finding is that the glucose utilization can be increased by changing the autoclaving procedure of the substrate to enhance the P(3HB) production yield up to 46 wt% of P(3HB) in dry cells.
Polyesters Biosynthesis of Alcaligenes eutrophus H16(ATCC 17699) from Various Mono- and Dicarboxylic Acids and Diols
Song, Jae-Jun ; Shin, Yong-Chul ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 123~128
The polyesters (polyhydroxyalkanoates; PHAs) production capability in a two-step cultivation of Alcaligenes eutrophus H16(ATCC 17699) was investigated by using various organic carbon sources. The carbon sources used included linear $C_2~C_10$ monocarboxylic acids,
dicarboxylic acids, crotonic acid, and several linear vicinal and
-diols. The polyesters synthesized were characterized by 500 MHz
spectroscopy, intrinsic viscosity
measurement in chloroform and differential scanning calorimetry (DSC). The PHAs synthesis data showed that the use of C-odd (
) monocarboxylic acids resulted in poly(3-hydroxybutyrate-co-3-hydroxyvalerate)(P(3HB-co-3HV) (3HV content ranging 40 to 70 mol%) while the use of
substrate gave the copolyester containing only 4 mol% of 3HV. All culture products obtained on
dicarboxylic acids gave exclusively P(3HB). 500 MHz
analysis showed that all polyesters synthesized generally contained 1~2 mol% 3HV even for the unrelated substrates such as the carboxylic acids with even number of carbon. When
-diols with even number of carbon were used as substrates, 4-hydroxybutyrate(4HB) was inserted into the polyester chain composed of P(3HB-co-4HB). Vicinal diols were generally not utilized by the bacterium for polyester production.n.
Organic Acid Composition and Flavor Characteristics of Lactic Acid Fermented Cereal Beverages
Yi, Do-Youn ; Kim, Gi-Myung ; Lee, Ki-Young ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 129~133
The effect of different compositions of organic acids on the flavor profile of 10% sugar solution was investigated by the response surface methodology, and the results were used to evaluate the flavor characteristics of lactic acid fermented cereal beverages. A mixture of extruded rice flour (10%) and soymilk (7.8% dry matter) was fermented with Leuconostoc mesenteroides (Sikhae). Depending on the substrate pretreatments, for example, the malt or amylase digestion and the proteolytic enzyme hydrolysis, the sugar and organic acid composition of the product varied. The organic acid composition of the fermented beverages was in the ranges of 0.44-0.55% lactic acid, 0.05-0.09% acetic acid and 0.07-0.09% citric acid, while that of commercial apple juice was 1.59% malic acid and 0.49% acetic acid. The flavor profiles of fermented beverages added with 10% sucrose were compared to those of apple juice and a model mixture containing 0.48% citric acid, 0.39% lactic acid and 0.12% acetic acid in 10% sugar solution. The QDA diagram of fermented beverages approached to that of apple juice, when the substrate was digested by amylase but not by protease.
Induction of a Mutant, Monascus anka 732Y3 from Monascus anka KFCC 11832 and its Morphological Observations
Kim, Jun-Sung ; Choi, Kee-Hyun ; Choi, Jang-Yoon ; Lee, Yoon-Soo ; Chang, Young-Youl ;
Journal of Microbiology and Biotechnology, volume 3, issue 2, 1993, Pages 134~138
Monascus anka 732Y3 was induced from Monascus anka KFCC 11832 (IFO 4478, ATCC 16360) by ultra-violet light irradiation. The growth of this new fungus is frequently more dependent on sexual propagation than asexual propagation, compared with that of its parental strain, M. anka KFCC 11832. Less conidia than those of M. anka KFCC 11832 were observed by a microscope. The optical density of the red pigments (
) produced by M. anka 732Y3 was 157, which was about 10 times higer than that of M. anka KFCC 11832. Such high production of the red pigments by the mutant could be explained by the following observations.