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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
> Journal Vol & Issue
Microbiology and Biotechnology Letters
Journal Basic Information
Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 26, Issue 6 - Dec 1998
Volume 26, Issue 5 - Oct 1998
Volume 26, Issue 4 - Aug 1998
Volume 26, Issue 3 - Jun 1998
Volume 26, Issue 2 - Apr 1998
Volume 26, Issue 1 - Feb 1998
Selecting the target year
Selective Antimicrobial Effects of Spice Extracts Against Lactobacillus plantarum and Leuconostoc mesenteroides Isolated from Kimchi.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 373~378
This study was performed to investigate antimicrobial effects and their actions of the ethanol extracts of 12 spices against Lactobacillus plantarum and Leuconostoc mesenteroides, which are related to Kimchi fermentation. The cardamon, thyme, and cumin ethanol extracts among 12 spices showed antimicrobial activities against Lac. plantarum, not against Leu. mesenteroides through paper disc method. The growth of Lac. plantarum was inhibited in MRS broth containing each extracts of cardamon (>4%), thyme (>4%) and cumin (>1%). SDS-PAGE and transmission electron micrographs showed that the cell walls and membranes were disrupted and the cytoplasmic components were leaked in strains treated with ethanol extracts.
Purification and Characterization of Natural Antifungal Protein from Astragal Seeds (Astragalus membranaceus L.).
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 379~386
Deterioration of food is in general caused by the presence of microorganisms and chemical compounds of food itself. There exists antimicrobial compound in the food, however, addition of food antiseptics, additives, or physico-chemical processing is a common practice. The safety of artificial chemical antiseptics became a serious public concern, therefore, new natural antiseptic compounds are in need to be developed. We have isolated a new natural antifungal protein (KBS-B2) from Astragal seed through ammonium sulfate precipitation and column chromatography using FPLC Mono-S and Superose 12HR. The purified protein inhibited growth of Candida albicans, and spore germination of food spoiling fungi such as Aspergillus ochraceus, Penicillium expensum, P. digitatum and Botrytis cineria. Antifungal effect of the KBS-B2 protein could be directly assayed by bioautography overlaying the fungal spores on the electrophoresed acrylamide gel. The comparison of N-terminal amino acid sequences of the KBS-B2 with known antifungal protein revealed that had 50% homology to thaumatin and zeamatin like proteins.
Site-speci fic Inactivation o meso-Diaminopimelate-dehydrogenase Gene (ddh) in a Lysine-producing Brevibacterium lactofementum.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 387~392
Brevibacterium lactofermentum, a gram-positive bacteria, has both the diaminopimelate (DAP) pathway and meso-DAP-dehydrogenase (DDH) pathway for L-lysine biosynthesis. To investigate importance of DDH pathway and the related ddh gene in lysine production, we introduced site-specific mutagenesis technique. A 300 bp DNA fragment central to the meso-DAP-dehydrogenase gene (ddh) of B. lactofermentum was used to inactive chromosomal ddh gene via homologous recombination. Southern hybridization analysis confirmed that the chromosomal ddh gene was disrupted by the vector sequence. The B. lactofementum ddh mutant obtained have an inactive DDH pathway. The results reveal that inactivation of the ddh gene in B. lactofermentum leads to dramatic reduction of lysine production as well as decrease of the growth rate, indicating that the DDH pathway is essential for high-level lysine production as well as biosynthesis of meso-DAP.
Tn5 lac Mediated Mutagenesis of Enterobacter sp. B54 Antagonistic to Phytophthora capsici.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 393~399
Enterobacter sp.B54 which shows antagonistic activity to Phytophthora capsici on potato dextrose agar was selected among 112 strains isolated from Korean soil. After Tn5 lac-induced mutants were obtained through Pl :: Tn5 lac mutagenesis, 2 mutants for loss of antibiosis and 1 mutant for increased antibiosis were screened by using in vitro fungal inhibition assay. When the 3 mutants affected in antibiosis were analyzed by southern hybridization with pRZ102 (ColEl :: Tn5) as a probe, its results suggest that Tn5 lac was randomly inserted into different chromosomal sites in these mutants.
L-Lysine Production by Amplification of the ddh Gene in a Lysine-producing Brevibacterium lactofementum.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 400~405
The ddh gene encoding meso-DAP-dehydrogenase (DDH) involved in the dehydrogenase pathway is essential for high-level lysine production in Brevibacterium lactofermentum. To investigate the effect of the ddh gene amplification on lysine production by B. lactofementum, we constructed two E. coli -B. lactofermentum shuttle vector, pEB1 and pEB2. The recombinant plasmids, pRK1 and pRK2, carrying the ddh gene were introduced into B. lactofermentum by electroporation. The specific activity of DDH by amplification of the ddh gene was increased 7-fold, and also L-Lysine production of B. lactofermentum strains harboring recombinant plasmids were 18∼20% higher than that of the control.
Expression and Secretion of Trichodema Endoglucanase in Saccharomyces cerevisiae.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 406~412
The endoglucanase gene, egl6, of Trichoderma sp. was connected with the yeast ADH1 promoter, and the resultant plasmid, pVT-C4, was introduced into three S. cerevisiae host strains (YNN27, 2805, and SEY2102). Among each 80 transformants, the cell growth and expression level of endoglucanase were compared in test-tube cultivation, and three respective transformants for each host cells showing the highest expression level and cell growth were selected. When three recombinant yeast cells were batchwise cultivated for 48 hr in flask, the total activities of endoglucanase expressed were about 1140 unit/l with 2805/pVT-C4, 1020 unit/l with SEY2102/pVT-C4, and 590 unit/l with YNN27/pVT-C4. Irrespective of host strain, about 80% of the expressed endoglucanase was detected in the extracellular medium. In addition, it was also found that the recombinant enzyme was secreted into the culture medium as two major forms of lightly and heavily glycosylated proteins.
Purification and Characterization of Aryl Acylamidase from Pseudomonas sp.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 413~419
Aryl acylamidase [EC 126.96.36.199] present in an acetaminophen-assimilating Pseudomonas sp. has been purified to a homogeneity using series of ammonium sulfate fractionation, DEAE-Sephacel anion exchange, Phenyl-Sepharose CL-4B hydrophobic, and Sephadex G-100 gel-permeation chromatography. The molecular weight, which was estimated by gel-permeation filtration and sodium dodecyl sulfate polyacylamide gel electrophoresis, was about 57 kDa and 56 kDa, respectively, indicating that this enzyme is a monomeric protein. The optimum pH was 10.5 and the optimum temperature was 40
. After incubation of the enzyme at 50
for 30 min, residual activity of the enzyme was 34% compared to its original activity. The Km values for acetaminophen and 4'-nitroactanilide were 0.10 mM and 0.11 mM, respectively.
Characterization of a Glutamyl Aminopeptidase from Bacillus licheniformis NS115.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 420~426
An extracellular glutamyl aminopeptidase (EC 188.8.131.52) producing bacterium was isolated from soil and identified as Bacillus licheniformis based on its morphological and physiological characteristics. The aminopeptidase was purified to homogeneity by ammonium sulfate precipitation, Phenyl Sepharose, Resource Q, and Superose 12 column chromatographies. The specific activity of the purified aminopeptidase was 9.2 unit/mg for glutamyl p-nitroanilide with 17.6 purification folds. The purified aminopeptidase had an estimated molecular mass of 64 kDa consists of two different subunits (42 kDa and 22 kDa), and its isoeletric point was 5.2 measured by isoelectric focusing. The optimum pH and temperature of the aminopeptidase were 8.0 and 55
, respectively. The aminopeptidase was inhibited by EDTA and 1,10-phenanthroline, suggesting it be a metalloenzyme. Comparing with other aminopeptidase, the enzyme showed relatively high activity against peptide having glutamic acid as N-terminal.
Purification and Properties of Alkaline Pretense from Xanthomonas sp. YL-37
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 427~434
An alkaline protease was 4-fold purified, yielding 2.3% of recovery by ammonium sulfate precipitation, CM-cellulose column chromatography and Sephadex G-100 column chromatography. The purified enzyme was estimated to be monomeric with molecular weight of about 62,000 from polyacrylamide gel eletrophoresis (PAGE) and sodiumdodecylsulfate polyacrylamide gel electrophoresis (SDS-FAGE). The optimal pH and temperature of the alkaline pretense activity were 11.0 and 50
, respectively, exhibiting high stability at pH value from 6.0 to 11.0 at 50
for 30 minute. The alkaline pretense was activated by MnSO
, and was inhibited by CuSO
, EDTA and EGTA. Also, the enzyme was found to be a metaloenzyme requiring Mn
2+/ as cofactor. The NH
-terminal amino acid of alkaline protease was alanine. The Km and Vmax values of this enzyme for casein was 4.0 mg/
and 5,500 unit/
Optimization of Betacyanin Production by Red Beet (Beta vulgaris L.) Hairy Root Cultures.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 435~441
Optimal conditions for the production of natural color, betacyanin were investigated by varying light intensity, C/N ratio, concentrations of phosphate and kinds of elicitors. Batch cultivation was employed to characterize cell growth and betacyanin production of 32 days. The maximum specific growth rate,
max/, was 0.3 (1/day) for batch cultivation. The maximum specific production rate, q
p/, was enhanced 0.11 (mg/g-cell/day) at 3 klux. A light intensity of 3 klux was shown to the best for both cell growth and betacyanin production. The maximum specific production rate was 0.125 (mg/g-cell/day) at 0.242 (1/day), the maximum specific growth rate. The dependence of specific growth rate on the light lintensity is fit to the photoinhibition model. The correlation between
p/ showed that the product formation parameters,
p/ were 0.3756 (mg/cell) and 0.001 (mg/g-cell/day), respectively. The betacyanin production was partially cell growth related process, which is different from the production of a typical product in plant cell cultures. In C/N ratio experiment, high carbon concentration, 42.1 (w/w) improved cell growth rate while lower concentration, 31.6 (w/w) increased the betacyanin production rate. The
max/ and q
p/ were 0.26 (1/day) and 0.075 (mg/g-cell/day), respectively. Beta vulgaris L. cells under 1.25 mM phosphate concentration produced 10.15 mg/L betacyanin with 13.46 (g-dry wt./L) of maximum cell density. The production of betacyanin was elongated by adding 0.1
M of kinetin. This also increased the cell growth. Optimum culture conditions of light intensity, C/N, phosphate concentration were obtained as 5.5 klux, 27 (w/w), 1.25 mM, respectively by the response surface methodology. The maximum cell density, X
max/, and maximum production, P
max/, in optimized conditions were 16 (g-dry wt./L), 12.5 (mg/L) which were higher than 8 (g-dry wt./L), 4.48 (mg/L) in normal conditions. The
max/ and q
p/ were 0.376 (1/day) and 0.134 (mg/g-cell/day) at the optimal condition. The overall results may be useful in scaling up hairy root cell culture system for commercial production of betacyanin.
Synthesis of Transglucosylated Xylitol Using Cyclodextrin Glucanotransferase and Its Stimulating Effect on the Growth of Bifidobacterium.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 442~449
Several transglucosylated xylitols were synthesized using intermolecular transglucosylation reaction of cyclodextrin glucanotransferase (CGTase) and their bifidogenic effects were investigated. The CGTase from Thermoanaerobacter sp. showed the highest transglycosylation activity on xylitol compared to those obtained from other strains. Extruded starch was identified to be the most suitable glucosyl donor for transglucosylation reaction on xylitol molecule by CGTase. The optimum reaction conditions for transglucosylation were also studied using extruded starch as a glucosyl donor. The transglucosylated xylitols were purified by activated carbon column chromatography with ethanol gradient elution from 0 to 18%, and their chemical structures were analyzed by fast atom bombardment mass spectrometer,
13/C-nuclear magnetic resonance spectrometer, and enzyme digestion method. Two transglucosylated xylitol, F-I and F-II, which had one or two glucose molecules attached to maternal xylitol by
-1,4-linkage, were mainly obtained. F-II showed increased stimulation effect on the growth of Bifidobacterium breve compared to xylitol, indicating the possibility utilized as a new functional alternative sweetners having bifidogenic effects.
Breeding of Aspergillus oryzae for the Alkaline Pretense Overproducing Strain.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 450~455
Aspergillus oryzae M-2-3 strain (argB
-/) was transformed with pTAalp plasmid which was constructed for expression of the alkaline pretense gene, alpA, and 16 transformants were selected on arginine minus medium. When these transformants were tested for productivity of alkaline proteases using agar plate containing skim milk, the halo was observed around each colony of transformants, but not observed around the host strain in this condition. Southern analysis showed that the pTAalp plasmid having alpA gene was integrated into the chromosome of the host strain. The highest level of alkaline protease production was obtained in the culture filtrate of the transformant No. 14, which was estimated to 80-90% of total secreted proteins, and the enzyme activity was 64-450 times higher than those of host strain and industrial strain. Total nitrogen content and the digestion rate in soybean Koji extracts were also increased to 1.5 times in Aspergillus oryzae transformant No. 14.
Enzymological Characteristics and Identification of Useful Fungi Isolated from Traditional Korean Nuruk
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 456~464
For the standardization and quality improvement of traditional Korean Nuruk, 10 strains of fungi, which were isolated from Nuruks and showed good productivity of the saccharogenic and dextrinogenic enzymes, acid and flavor, were selected and their enzymological characteristics and identification were carried out. Aspergillus spp. and Rhizopus sp. showed a high liquefying activity without regard to cultivation time, whereas the majority of strains except for Rhizopus sp. had decreasing saccharifying activity in proportion to the increase in cultivation time. Aspergillus spp. No.17-2, No.17-6 and Rhizopus sp. No.18-1 showed high liquefying and saccharifying activity after 15 and 30 day cultivation. The optimum temperature of most of these saccharogenic and dextrinogenic enzymes was from 40
, and their optimum pH was extensive between pH 3 and pH 11. But Penicillium spp.(2 strains) and Rhizopus sp. showed low activity under the alkalic and acidic conditions. Among these isolated strains, 5 strains which had shown the high productivity of materials were identified as Aspergillus oryzae NR3-6 and Aspergillus oryzae NR17-6, Aspergillus penicilloides NR12-1, Penicillium expansum NR7-7 and Rhizopus oryzee NRl8-1, respectively. Five kinds of mixed culture were carried out and all of them showed a better productivity of saccharogenic and dextrinogenic enzymes than single culture. These results indicate that it is possible to make traditional Korean liquors of good quality by using these fungi.
Measurement and Acceleration of Biodegradation in Soil.
Microbiology and Biotechnology Letters, volume 26, issue 5, 1998, Pages 465~469
The quantitative and rapid method for measuring the biodegradation of polymer materials in soil was developed. In this study, cellophane film was used as a model biodegradable polymer and the biodegradation was assayed by measuring the amount of glucose which was produced by a hydrolysis reaction using HCl after collecting the film from soil. Cellophane film was degraded 41.2% in 4 months during winter while it was degraded 76.5% in 2 months during summer. It means that biodegradation in soil is affected by environmental conditions. The biodegradation was also measured in an incubator (30
, humidity 50-55%) to exclude the environmental variations. Cellophane film was degraded 94% in that condition in 40 days. The biodegradation showed the first order kinetics and the rate constant was 0.067 (1/day). Acceleration of the biodegradation in soil was also studied. We added cultured soil microorganisms or nutrients such as N, P, and S into the soil. While the addition of microorganisms showed the temporary increase of rate constant, the addition of nutrients not only showed the increase of rate constant from 0.096 (1/day) to 0.21 (1/day) but also maintained the effect continuously.