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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
Microbiology and Biotechnology Letters
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 14, Issue 6 - Dec 1986
Volume 14, Issue 5 - Oct 1986
Volume 14, Issue 4 - Aug 1986
Volume 14, Issue 3 - Jun 1986
Volume 14, Issue 2 - Apr 1986
Volume 14, Issue 1 - 00 1986
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Development of Yeast-Vector System for Eukaryotic Gene Cloning - Optimum Condition for Intact Yeast Cell Transformation and Plasmid Stability in the Transformants -
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 125~131
In order to obtain the optimum conditions for intact yeast cell transformation in the various yeast host-vector systems, 3 yeast plasmid vectors, YRp7, YEpl3 and YIp5 were introduced into 5 yeast hosts, Saccaromyces cervisiae Dl3-1A, DKD-5D, DBY-746, MC-16 and S2022D with various transformation conditions, and plasmid stabilities in all the transformants were also observed. The highest transformation frequencies in all the host-vector system were obtained in the 16 hour Cultured cell (5.4
) treated with 0.1-0.2 M lithium chloride in 0.1 M tris-HCl (pH 7.6), 35% polyethylene glycol 4000, and heat-shocked at 42
for 5 minutes after 60 minutes of induction. The intact cell transformation got more transformation frequency in DKD-5D (YRp7) and DBY-746 (YEpl3) than protoplast transformation, but reverse tendency was observed in DKD-5D (YEp13) and Dl3-lA (YRp7). The transformants, D13-1A (YRp7) and DKD-5D (YRp7) were very unstable in selective medium, with 80 to 85% of the transformants losing the plasmid after 70 generations, but the transformants, DKD-5D (YEpl3) and DBY-746 (YEpl3) were quite stable, with 35% of the transformants losing the plasmid.
Cloning of 17S-Ribosomal RNA Gene from the Hygromycin Resistant Tetrahymena thermophila
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 133~137
17S-ribosomal RNA gene from the hygromycin resistant protozoan Tetrahymena thermophila hmr 3 was cloned on E. coli vector pBR 322 as part of study to work the 17S-rRNA structure and the mechanism of hygromycin resistance. The 17S-rDNA was inserted into the Hind 111 site of pBR 322. The clones having recombinant plasmid were selected by the method of colony hybridization with a 17S-rDNA probe of wild type B1868. The orientation of 17S-rDNA insert was located near the tetracycline resistant gene of pBR 322 in a clone 5-19 with the recombinant plasmid.
Purification and enzyme characteristics of laccase from Ganoderma lucidum
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 139~143
The production media and enzymatic characteristics of laccase from Ganoderma lucidum was investigated. Potato dextrose yeast extract media was proved to be the best for laccase production. The enzyme has optimum pH of 6.45km value of 6.71 mM and appeared to be stable at wide pH range. The enzyme was inactivated partially by methanol and ethanol and totally by sodium azide but not at all by acetone. Also the enzyme purification was performed and the data is given.
Isolation and charaterization of a microbial antihemorrhagic substance on snake venom
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 145~153
For the inactivation of venoms, the chemical methods are generally applied. In the chemical method many works have been carried out with the chemical reagents and immunological antiserums. However, all inhibitory effect of these chemicals acting on snake venomes may well be due not to the specific, but to the nonspecific inhibitory action. Therefore, it is necessary to separate venom into its compositional active proteins and develop specific inhibitor which acts on the each protein. Until now, there have not been any reports about the substance which acts on snake venom as a specific inhibitor. Recently in 1979, we had actually isolated a specific venom inhibitor(ISV) which has a strong inhibitory activity against the proteinase of snake venom of Colubridae. In our experiments described here, a strain of Aspergillus sp., isolated from soil, was able to produce a biological active substance. The partial crystallized substance had a strong inhibitory activity against hemorrhagic action of snake venom of Colubridae. For the inhibitory action of the sample on the lethality of venom, the substance prevented completely the lethal action of the hemorrhagic factor when they were treated with enough amount of the substance. The edema factor of whole venom of Agristrodon bromohoffi brevicaudus was completely inhibited, but those of HR-I and HR-II of Trimeresurus flavoviridis venom were inhibited about 50%, when they were treated with the substance of half amount of venom. On the other hand, from the result of subcutaneous hemorrhage in a rabbit, it was concluded that two kinds of antihemorrhagic substance might be produced by the strain used in this work
Cloning of Bacillus amyloliquefaciens amylase gene using YEp13 as a vector I. Expression of cloned amylase gene in Escherichia coli
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 155~160
-Amylase gene of B. amyloliquefaciens was cloned to E. coli-yeast shuttle vector YEp-13 and expressed in E. coli. Chromosomal DNA of B. amyloliquefaciens was partially digested with Sau3Al and YEp13 plasmid was cleaved with BamH1. The hybrid plasmid, pHA28, was constructed by shotgun method and transformed to E. coli C600 and HB101. The amount of
-amylase produced by transformants of E. coli was about 20% to 30% of that produced by B. amyloli-quefaciens. About 65% of
-amylase produced by transformant was secreted into periplasm and the others were located in cytoplasm.
-Amylase production was maximal when transformants were cultivated for 15hr to 20hr. As the result of agarose gel electrophoresis, pHA28 plasmid was found to be various in its size. This result suggested that pHA28 plasmid was segregated.
Cloning of Bacillus amyloliquefaciens amylase gene using YRp7 as a vector I. Expression of cloned amylase gene in Escherichia coli
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 161~168
A 1.95Kb Sau3Al fragment coding for
-amylase from Bacillus amyloliquefaciens was isolated by the shotgun method using Escherichia coli as a host. The genome of Bacillus amyloliquefaciens was partially digested with the restriction endonuclease Sau3Al and joined to plasmid YRp7 cleaved with the restriction endonuclease BamHI. The
-amylase gene present in a 1.95Kb insert was stably maintained and expressed in Escherichia coli. The amount of
-amylase activity produced by Escherichia coli containing the hybrid plasmid pEA24 was about 65% of the activity produced by the donor Bacillus amyloliquefaciens strain. The properties of
-amylase produced by Escherichia coli were very similar to those produced by Bacillus amyloliquefaciens as based on optimum temperature, pH, and effect of CaCl
concentration. About 70% of the
-amylase produced by Escherichia coli was localized in the periplasmic space, whereas the remaining enzyme was localized in the inner part of the cell.
Cytosine Deaminase of Fungus
;;Takuo Sakai;Kenzo Tonomura;
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 169~174
Cytosine deaminase was partialy purified about 10 fold from the ceil-free extract of Aspergillus fumigatus. The partialy purified enzyme was relatively stable in a pH 5.5 to 8.0, but thermo-unstable. The enzyme activity was found in a pH optimum of 7.0 and temperature optimum of 30 to 35
. The activation energy calculated to be 13,240 cal/mol. The apparent Michaelis constants Km for cytosine was found to be 1.53 mM and the molecular weight was determined to be approximately 32,000. The enzyme was strongly inhibited by 0.1 mM of Hg
, furthermore inhibited by 1mM of ATP, UTP, o-phenanthroline and p-chloromercuribenzoate.
Studies on the Protoplast Formation of Cellulomonas flavigena and its Observations under Scanning Electron Microscope
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 175~179
In order to develope a protoplast fusion of the genus Cellulomonas having high assimilibility of cellulose, the optimum conditions for the protoplast formation of Cellulomonas flavigena NCIB 12901 was investigated and observed by means of Scanning Electron Microscope. The results suggested that the susceptibility of the cell wall by lysozyme treatment on protoplast formation was considerably depend on the cultural periods of the cells. Cells of C. flavigena at mid exponential phase could more efficiently convert to protoplast cells than those at late exponential phase did. The rate of the protoplast formation was 95%, even though the rate was over 99.9% on counting by indirect method after osmotic shock treatment, when cells of the organism at mid exponential phase were treated with lysozyme (400
) for 6 hours and observed by SEM. In the evaluation of protoplast formation of the genus Cellulomonas, direct method of the observation under Scanning Electron Microscope was much more reliable than the counting method of protplasts after osmotic shock treatment. Because defferences between the number of spheroplast and protoplast were not able to be figured out on counting the number of protoplast after osmotic shock treatment.
Cloning of Acetyl CoA Carboxylase (fabE) in Escherichia coli
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 181~186
A defective lambda transducing phase carrying acetyl CoA carboxylase gene (fabE) from Escherichia coli chromosome (72 min on the current linkage map) has been isolated. A restriction map of the chromosomal region from defective transducing phage was established by digestion with combination of the restriction enzymes. No cleavage site for the enzyme EcoRI was found in this region. Restriction fragments were cloned from defective transducing phage into high copy number plasmid vector pACYC184 to generate hybrid plasmids which were capable of complementation of fabE temperature sensitive mutation. We show here that the fabE gene is located on a 3.4 megadalton Bam HI-SalI fragment with a HindIII site, which lies within the 7.4 megadalton BglIIfragment, by complementation analysis.
The inoculation effect of R. japonicum on the nodulation and nitrogen fixation activity in Glycine max with the different kinds of soil.
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 187~192
The inoculation effect of the highly nitrogen fixing strains of R. japonicum were tested in the 3 kinds of soil with different cultural history onto Gycine max cv. Jang-yeob. The nodulation and nitrogen fixation activity in 3 test soils all showed the great increase in inoculated group compared to the non-inoculated group. The plant dry weight of the in-oculated groups were increased about 10% than that of the non-inoculated groups. The numerical index of the increase in total nitrogen fixation activity were 238% in the pre-cultivated, 266% in the immatured and 157% in the matured soil and these results suggested the clear effect of inoculation. Among the strains tested, R. japonicum R214 and Rl38 showed the excellent inoculation effect.
Continuous Alcohol Fermentation by Cell Recycling Using Hollow Fiber Recycle Reactor
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 193~198
Improvement of productivity in ethanol fermentation was attempted using a hollow fiber bioreactor (HFR) where Saccharomyces cerevisiac var. ellipsoideus cells were recycled to achieve a high yeast concentration. Industrial wort was used as the fermentation media without supplying any additional nutrients. The performances in hollow fiber recycle reactor (HFR) were compared with those of batch and continuous cultures. In a continuous culture with 11
P and 15
P wort media final ethanol concentrations were 4.71% and 5.82% (v/v) and yields 86.2% and 78.6% respectively when the dilution rate (D) was 0.1 h
, in contrast, the ethanol concentration and productivity in HFR were 7.64%(v/v) and 6.1g/l/h at D=0.1h
P media. When the dilution rate was increased to 0.2 h
, the concentration and the Productivity were 7.62% (v/v) and 12.2g/l/h. At D=0.3h
the sugar was completely consumed and the productivity was 18.1g/l/h. This correponds to 4 times that in continuous system and 16.3 times that in the batch system performed in comparable conditions.
Continuous Ethanol Fermentation using Immobilized Yeasts
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 199~203
A tubular tormentor was prepared by packing the wood chips and pumping the yeast solution of Saccharomyces formosensis in a tubular column. Investigations to characterize the ethanol fermentation in the immobilized cell tubular fermentor and to compare such a fermentors with other type fermentors were undertaken. Ethanol productivity of 24.4g EtOH/
.hr has been obtained from glucose substrate. This productivity is higher or compared favourably with that reported in immobilized bio-reactors.
Biochemical characterization of Bacillus thuringiensis, 23 serovars
Lee, Hyung-Hoan ; Park, Mi-Yeoun ; Lee, Chang-Woon ;
Microbiology and Biotechnology Letters, volume 14, issue 2, 1986, Pages 205~208
The 23 serovars of Bacillus thuringiensis strain were commonly gram-positive and motile, formed endotoxin crystals, produced acid and alkali in the KIA media, and acid from glucose, hydrolyzed starch, and reduced nitrate but did not produce H
S, oxidase and indole, did not decompose lysine, ornithine, phenylalanine, malonate, lactose, dulcitol, adonitol, inositol, sorbitol, arabinose, raffinose, rhamnose, maltose, and xylose. Eighteen serovars were positive in the MR tests and 15 in the VP tests. Four serovars used citrate. Five serovars produced urease, 5
from glucose, 2 DNase, and 15 lecithinase. Twelve serovars decomposed arginine, 11 did sucrose, 2 manitol, and 9 salicin Serovar tohokuensis did not hemolyze, but the others did.