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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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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 30, Issue 4 - Dec 2002
Volume 30, Issue 3 - Sep 2002
Volume 30, Issue 2 - Jun 2002
Volume 30, Issue 1 - Mar 2002
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Characterization of Bacillus stearothermophilue Cyclodextrin Glucanotransferase that Expressed by Saccharomyces cerevisiae
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 293~297
The cyclodextrin glucanotransferase (CGTase) gene from Bacillus stearothermophilus NO2 was expressed in Saccharomyces cerevisiae 2805 under the adhl promoter. The CGTase was purified from S. cerevisiae 2805/pVT-CGTS. The purified enzyme exhibited a optima of activity around pH 7.0 and
. Thermal stability of the enzyme was increased fairly as compared with the CGTase of B. stearothermophilus NO2. The conversion yield of cyclodextrin (CD) and the production ratio of
-CD from starch were showed similarly aspect to the CGTase of B. stearothermophilus NO2.
Enhanced Activity of Cytidine Deaminase by Gene Family Shuffling.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 298~304
A family shuffling associating PCR-based and in vitro recombination and expression in Escherichia coli cdd mutant was carried out. Two cdd genes encoding cytidine deaminases (CDase) from thermophilic Bacillus caldolyticus and B. stearothermophilus were shuffled. Around 150 viable mutant colonies screened on AB minimal medium without uracil by E. coli cdd complementation were selected for cytidine deaminase assay and 4 candidates (SH1067, SH1077, SH1086, and SH1118) were chosen for the detailed study. The nucleotide sequence analyses of 4 selected mutants revealed that they have several point mutations and recombinations. Surprisingly, the SH 1067 showed 770 fold more specific CDase activity at
than that of T101 from parental B. stearothermophilus.
High-Level Expression of Aspergillus ficuum Acetyl Xylan Esterase Gene in Pichia pastoris,
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 305~311
Acetyl xylan esterase gene (AXE) from Aspergillus ficuum was cloned and its Pichia expression plasmid, pPICZ
C-AXE (4.6 kb), was constructed, in which the AXE gene was under the control of the AOXI promoter and connected downstream of mating factor u-1 signal sequence. The plasmid linearized by Sacl was integrated into the 5'AOXI region of the chromosomal DNA of P. pastoris. In the flask batch culture of P. pastoris transformant on methanol medium, the cell concentration and total AXEase activity reached at 6.0 g-dry cell weight/1 and 77 unit/ml after 36 h cultivation, respectively. In the fed-batch culture employing the optimized methanol and histidine feeding strategy, the cell concentration and total AXEase activity were significantly increased to about 97 g-dry cell weight/1 and 930 unit/ml. Most of AXEase activity (>90%) was found in the extracellular medium and the majority of extracellular protein (>80%) was AXEase enzyme (33.5 kDa). This result means that about 9.8 g/1 of AXEase protein was produced in the extracellular medium.
Molecular Cloning and Nucleotide Sequence Analysis of pyrB Gene Encoding Aspartate Transcarbamylase from Psychrophilic Sporosarcina psychrophilia
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 312~319
The Sporosarcina psychrophilia pyrB gene, which encodes aspartate transcarbamylase (ATcase), was cloned on Sau3AI restriction endonuclease fragment inserted into pUC19 plasmid vector, S. psychrophilia pyrB gene was expressed in Escherichia coli pyrB mutant for the complementation test. The sequence of 2,606 nucleotides including putative pyrB gene was determined. The region contained one full open reading frame (ORf) and two partial ORFs. The deduced amino acid sequence of the second ORF showed 59% identity with that of Bacillus caldolyticus ATCase. The first and third partial ORFs were closely related to the uracil permease (pyrP) and dihydroorotase (pyrC), respectively. Besides, potential terminator, antiterminator, and anti-antiterminator structures were found in the intergenic region between pyrP and pyrB. These results suggested that S. psychrophilia pyrimidine nucleotide biosynthesis genes are clustered as well as other Bacillus sp. Over-expressed product of pyrB encoding ATCase was purified and analyzed by the SDS-PAGE. The purified PyrB protein turned out to be molecular mass of 27 kDa and showed ATCase activity.
Isolation of a Specific Antigen Protein on Cell Membrane of Cochlodinium polykrikoides, Red Bloom
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 320~324
To establish a rapidly immunochemical identification on a dinoflagellate, Cochlodinium polykrikoides, a specific antigenic protein as a maker on the cell membrane was isolated. The cell membranes of C. polykrikoides and Gymnodinium sangineum were harvested by centrifugation after osmotic shock. The membrane proteins of both cells were solubilized in 50 mM Na-carbonate contained 1 mM DTT, and separated the proteins on SDS-PACE. Immune-blot on the solubilized membrane proteins of the both cells was performed with antiserum against the solubilized membrane proteins of C. polykrikoides. A 120 kDa membrane protein of C. polykrikoides had remarkablely different antigenicity from that of G. sangineum.
Optimal Production and Characterization of Fibrinolytic Enzyme from Fomitella fraxinea Mycelia.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 325~331
investigated to maximize the production of fibrinolytic enzyme from Fomitella fraxinea mycelia. Among the tested media, Coriolus versicolor medium (CVM) showed the highest production for the enzyme. 2% galactose, 0.6% yeast extract and 0.1%
, and 0.05%
as carbon, nitrogen, phosphorus, and inorganic salt sources resulted in the maximum level of the enzyme activity, respectively. The enzyme production from F. fraxinea was reached to highest level after the cultivation for 10 days at
and pH 9. The enzyme activity of culture supernatant was most active at
and pH 10. The activity of the enzyme was inhibited by phenylmethylsulfonylfluoride and aprotinin, suggesting that it is a serine protease.
Characterization of Extracellular \alpha-galactosidase Produced by Streptomyces sp. YB-4.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 332~338
A strain YB-4 producing the extracellular
-galactosidase was isolated from soil, and has been identified as Streptomyces sp. on the basis of its cultural, morphological and physiological properties. The partially purified
-galactosidase was most active on paranitrophenyl-
-D-galactopyranoside at pH 6.0 and 6
. The enzyme retained 90% of its maximum activity between pH 4.0 and pH 10.0 after pre-incubation for 1 h. The enzyme was able to hydrolyze oligomeric substrates such as melibiose, raffinose and stachyose to liberate galactose residue, indicating that the
-galactosidase of Steptomyces sp. YB-4 hydrolyzed
Antifungal Activities of Equisetin, Zearalenone, and 8'-Hydroxyaearalenone Isolated from Fusarium Species against Plant Pathogenic Fungi.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 339~345
Antifungal substances were isolated from solid cultures of Fusarium equiseti FO-68 obtained from arrowhead and Fusarium sp. FO-510 obtained from egg plant, and then their antifungal activities were investigated against plant pathogenic fungi in vitro and in vivo. An antifungal substance was purifed from rice solid cultures of F. equiseti FO-68 and identified as equisetin. In addition, two antibiotic substances were isolated from solid cultures of Fusarium sp. FO-510 and their chemical structures were determined to be zearalenone and 8'-hydroxyzearalenone. in vitro, equisetin and zearalenone inhibited mycelial growth of most of the plant pathogenic fungi tested, whereas 8'-hydroxyzearalenone hardly inhibited fungal growth. In vitro, equisetin effectively controlled the development of tomato gray mold and tomato late blight. Zearalenone exhibited in vivo antifungal activity against rice blast, rice sheath blight, tomato gray mold, and tomato late blight. However, 8'-hydroxyzearale-none did not control the development of plant diseases except tomato gray mold. This is the first report on the antifungal activities of equisetin, zearalenone, and 8'-hydroxyzearalenone.
Isolation and Enzyme Production of a Neutral Protease-Producing Strain, Bacillus sp. DS-1.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 346~351
A bacterium producing the neutral pretense was isolated from soil, and was identified as Bacillus sp. DS-1 by 16S rRNA sequence comparison and biochemical determinations. The production of protease from Bacillus sp. DS-1 was increased 20% and 30% by the additions of 1% glucose and 1% yeast extract, respectively. The optimum pH and temperature for the protease activity were pH 7.0 and 55
. Bacillus sp. DS-1 produced a metalloprotease as a major protease in culture medium, since the pretense activity in culture supernatant was inhibited by the presence of 1 mM EDTA significantly.
Purification and Characterization of a Maltopentaose-producing Amylase from Bacillus megaterium KSM B-404.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 352~358
An amylase that hydrolyzes starch into maltopentaose as a main product was found in the culture supernatant of a strain of Bacillus megaterium KSM B-404 isolated from local soil. The enzyme was purified 129-fold by ammonium sulfate precipitation, DEAE-Toyopearl and Superdex 75 HR 10/30 column using a FPLC system. The molecular weight of the amylase was determined as about 68 kDa by using SDS-PAGE. Optimum pH and temperature of amylase were found to be
and pH 6.0~7.0, respectively. The enzyme was stable up to
by addition of
and its pH stability was in the range of 6.0~10.0. The activity of enzyme was inhibited by
and maintained by
. EDTA and pCMB also showed inhibitory effect to the enzyme. TLC and HPLC analysis of the products of the enzyme reaction showed the presence of maltopentaose(52%), maltotriose (25%), maltose (11%), glucose, and maltotetraose in the starch hydrolysates.
Chayacterization of Bacillus polyfermenticus SCD as a Probiotic.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 359~366
Bacillus polyfermenticus SCD which is commonly called as Bisroot strain is being used for functional foods through the treatment of long-term intestinal disorders, since the live strains in the form of active endospores can successfully reach the target intestine in humans. The cells of B. polyfermenticus SCD were treated for 4h in artificial gastric juice (pH 2.0,3.0) and bile acid. Final viability of the strain in artificial gastric Juice (pH 2.0, 3.0) is reached to 62.8% and 81.2% respectively B. polyfermenticus SCD is resistant to antibiotics such as streptomycin, rifampicin, nystatin and ampicilin. B. polyfermenticus SCD is well known supplies the nutrients by synthesizing vitamin
, C and K. B. polyfermenticus SCD produces various digestive enzymes and the enzymes enable to completely digest diets in our body. Above all,
-amylase and pretense activities are very higher than B. subtilis KCTC 1020, about two fold and twenty five fold respectively. B. polyfermenticus SCD is very stable during long-term storage period in phosphate buffers of wide-range pH, solutions of various concentrations of sodium chloride, 5% glucose solution and water.
Optimal Conditions for the Production of Intracellular Cytosine Deaminase from Chromobacterium violaceum YK 391.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 367~372
Cytosine deaminase (cytosine aminohydrolase, EC 220.127.116.11) stoichiometrically catalyzes the hydrolytic deamination of cytosine and 5-fluorocytosine to uracil and 5-fluorouracil, respectively. Optimal medium compositions for production of cytosine deaminase from Chromobacterium violaceum YK 391 were 0.75% soluble starch, 1.5% peptone, 0.1% meat extract, 0.1% yeast extract, 0.01% NaCl, 0.01%
. The optimal pH of medium and incubation temperature were 7.0 and
, respectively. C. violaceum reached stationary phase after 30 hr, and produced a maximum cytosine deaminase (120 units/ml) after 72 h in batch culture.
Production of D-Lactic Acid from DL-Lactonitrile by Pseudomonas sp.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 373~379
By using DL-acetonitrile as enzyme inducer, 90 bacteria were isolated from a field soil. Among the isolated strains, the strain WJ-003 showed the highest activity for production of D-lactic acid from DL-lactonitrile, and was partially identified as Pseudomonas sp. The production condition of D-lactic acid from DL-lactonitrile using resting cells as an enzyme source was optimized as follows: the reaction mixture contained 10 mM of DL-lactonitrile, 20 g of wet cells in 11 of 20 mM potassium phosphate buffer (pH 7.0) and the reaction was carried out at
. After 18 h of reaction, 0.843 g/l of D-lactic acid was produced which corresponded to a conversion ratio of 93.7% and an optical purity of 99.8%. Additionally, when 10 mM of DL-lactonitrile was added once more to the reaction mixture at 14 h, 1.64 g/1 of D-lactic acid was produced after 28 h. In this experiment, the conversion ratio was 91.1% and optical purity 99.8%.
Effects of Xylooligosaccharides on the Growth of Intestinal Microflora.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 380~387
To investigate the effects of xylooligosaccharides on the in vitro growth of intestinal bacteria, various species were cultivated individually on the m-PYF medium containing a carbon source (0.5% w/v) such as xylooligosaccharides, isomaltooligosaccharides, fructooligosaccharides and sucrose, respectively. The health-promoting microorganisms such as Bifidobacterium bifidum, Bifidobacterium infantis, Bifidobacterium longum, Lactobacillus casei and Lactobacillus acidophilus grew more effectively by xylooligosaccharides than by other carbon source, though xylooligosaccharides inhibited the growth of Clostridium perfringens, Bacteroides fragilis, Escherichia coli, Staphylococcus aureus and Salmonella typhumurium. At the mixed culture xylooligosaccharides exerted a preferential stimulatory effects on numbers of the health-promoting microorganisms, while xylooligosaccharides inhibited populations of potential pathogens at relatively low level. Xylooligosaccharides also maintained the acidity of culture with Streptococcus mutans, caries-inducing bacteria, over pH 5.0. These results suggest that xylooligosaccharides selectively promote the growth of the health-promoting microorganisms in human intestine and prevent caries by inhibiting acid production from Streptococcu mutans.
The Growth Promoting Effect of Bifidobacterium bifidum by Combination of Natural Products Bearing Antioxidative Capacity
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 388~394
The growth of B. bifidum was promoted by natural products bearing antioxidative capacity and mixed combined two, three and four kinds of them. B. bifidum was expressed a good growth by Nelumbo nuclfera gaertner, Corni fructus, Beiamcanda chinensis, alone, and two mixed combinations were composed of Nelumbo nuclfera gaertner and Corni fructus, Nelumbo nuclfera gaertner and Beiamcanda chinensis, Nefumbo nuclfera gaertner and Theae folium, three mixed combinations were oraganized with Nelumbo nuclfera gaertner, Corni fructus and Theae folium, Nelumbo nuclfera gaertner, Corni fructus and Beiamcanda chinensis, Nelumbo nuclfera gaertner, Theae folium and Beiamcanda chinensis, and four mixed combinations were formed with Nelumbo nuclfera gaertner, Theae folium, Beiamcanda chinensis aud Corni fructus, Theae folium, Beiamcanda chinensis, Corni fructus and Glycyrrhizae radix, Nelumbo nuclfera gaertner, Corni fructus, Teae folium and Glycyrrhizae radix, and Nelumbo nuclfera gaertner, Corni fructus, Beiamcanda chinensis and Glycyrrhizae radix. These few mixed combinations promoted cell growth by 2.6 times than that of control, and its antioxidative capacity was also 5.6 times higher, and the ratio of elimination of hydroxyl radical was more than 80% in each dilution rate. As these combinations of natural products could activate some parts of body, they might be applied to pharmaceuitcal applications, functional foods, also expected to multifunctional fermentative beverages.
Degradation Characteristics of Methyl Ethyl Ketone and Methyl Isobuthyl Ketone by Pseudomonas putida KT-3.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 395~401
Methyl ethyl ketone (MEK) and methyl isobutyl ketone (MIBK) have been widely used as solvents in various industries. Biodegradation of MEK and MIBK by Pseudomonas putida KT-3, which could utilize MEK or MIBK as a sole carbon source, was characterized, and the cosubstrate interaction in MEK/MIBK mixture was also studied. Within the range of initial MEK concentration (from 0.5 to 5.5 mM), an increased substrate concentration increased the specific degradation rate of MEK by P putida KT-3 (from 3.15 to 10.58 mmol/g DCW
h), but the rate sightly increased at 11.0 mM of initial MEK concentation (11.28 mmol/g DCW
h). The similar degradation rates of MIBK (4.69-4.92 mmol/g DCW
h) were obtained at more than 3.0 mM of initial MIBK concentation. Kinetic analysis on the degradation of MEK/MIBK mixture by P. putida KT-3 showed that MEK or MIBK acted as a competitive inhibitor. Maximum degradation rate (
), saturation constant (
) and inhibition constant (
) were as follows:
=12.94 mmol/g DCW
Lowering Effects in Plasma Cholesterol and Body Weight by Mycelial Extracts of Two Mushrooms: Agaricus blazai and Lentinus edodes.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 402~409
The effects of protein-bound polysaccharides (A-PBP and L-PBP) that were extracted from the mycelia of two edible mushrooms, namely Agaricus blazai and Lentinus edodes, on serum cholesterol and body weight were investigated in mice and female volunteers. Six groups of Male Balb/c mice were fed six kinds of diet supplement- solutions composed of L-PBP, A-PBP, chitosan, and other fiber constituents, for 30 days under the normal diet. Ninety female volunteers were also supplemented for 8 weeks with six kinds of capsules including control and five test groups as the same manners (two times a day, 4 capsules). From 12 days after feeding of L-PBP (Group I) and A-PBP (Group II), the weight of mice began to reduce as compared with control, whereas that of Group III fed chitosan was decreased 15 days after feeding. Group W and Group V which were fed mixture of L-PBP, A-PBP, chitosan, and other dietary fiber, were more significant in lowering weight. After 4 weeks of the supplementation in women, their serum LDL-cholesterol level and body weights in Group I and II were reduced, but Croup 111 taken with chitosan capsule showed weaker effect than Group I and II. After 8 weeks, LDL-cholesterol content in the sera of Group I (132.5 mg/dL) and II(131.5 mg/dL) was decreased to ideal level (125.4 and 122.8 mg/dL) for healthy blood vessel. In the case of Group W supplemented with mixture of L-PBP, A-PBP, and chitosan, the weight-reduction effect (11.8%) and hypocholesterolemic effect (11.0%) was most significant, indicating their synergistic action. These data suggested that the weight-controlling and hypolipidemic effect of L-PBP and A-PBP was involved, at least in part, in absorption of cholesterol as their role of dietary fiber, as well as cholesterol metabolism.
Effect of Acasia (Robinia pseudo-acasia) Flower on the Physiological Functionality of Korean Traditional Rice Wine.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 410~414
In order to develop a Korean traditional rice wine which by acasia flower added alcohol fermentation was investigated by addition of 5%, 10'h, 15% nuruk and 10% acasia into the wine mash. The maximum amount of ethanol (16.4%) was obtained when 10% acasia flower and 15% nuruk were added in cooked rice for the fermentation by Saccharomyces cerevisiae at
for 20 days. The overall acceptability and physiological functionalities of the rice wine prepared by addition of different concentration (5-50%) of acasia flower into mash were investigated and compared. The A-15 rice wine which was brewed by addition of 15% acasia flower into mash showed the best acceptability. Its angiotensin-converting enzyme inhibitory activity and tyrosinase inhibitory activity were 80.3% and 94.2%, respectively. The electron-donating ability (23.4%) and nitrite scavenging activity (21.5%) were also higher than those of traditional rice wine.
Fermentation Characteristics of Kimchi Supplemented with Cheese.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 415~419
The replacement effects of cheese far salted and fermented fish on growth of lactic acid bacteria, fermentation velocity and sensory characteristics of Kimchi were investigated. In both control and cheese Kimchi, the total viable cell count of lactic acid bacteria was increased rapidly during the initial 2 days of fermentation. From 3 days after preparation, Kimchi added with cheese showed higher number of lactic acid bacteria than control Kimchi. The pH of Kimchi decreased rapidly after a small increase at the first day of fermentation, reaching 4.18-4.33 at the third day of fermentation, and the pH was slightly lower in Kimchi added with cheese than in control. Proximate analysis of Kimchi added with cheese was slightly higher in moisture and lower in crude protein and fat than control Kimchi. Sensory evaluation of the Kimchi fermented for 3 days showed that the Kimchi added with 3 or 5% of cheese had higher scores of appearance, flavor and overall taste than the control Kimchi.
Isolation and Identification of Anticariotic Compound from Sophora flavescens Ait.
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 420~424
The purpose of this study is to investigate anticariotic activity of the ethyl acetate soluble extract of Sophora flavescens Ait. for the prevention of dental caries and glucosyltransferase activity caused by Streptococcus mutans. The fraction 5-4-3 showed strong growth inhibition activity against Streptococcus mutans (MIC, 3.13
/ml). The glucosyltransferase activity of the active fraction 5-4-3 inhibited the formation of glucan and showed 77% of the antiproliferative effect at 100
/ml (P<0.05). Two flavanones, (2S)-2'-methoxy kurarinone (1) and (+)-kurarinone (2), were isolated from the active fraction 5-4-3 of the ethyl acetate soluble extract of S. flavescens Ait. Their structures were elucidated using spectroscopic methods.
Saccharomyces cerevisiae KNU5377 with Multiple Stress Tolerance and its Potential as a Worldwide On-site Industrial Strain for Alcohol Fermentation
Paik, Sang-Kyoo ; Ingnyol Jin ; Yun, Hae-Sun ; Park, Sae-Hun ; Shin, Seong-Chul ; Kim, Jae-Wan ; Shin, Ki-Sun ; Lee, Jung-Sook ; Park, Yong-Ha ;
Microbiology and Biotechnology Letters, volume 30, issue 4, 2002, Pages 425~429
Saccharomyces cerevisiae KNU5377 was examined to assay the recovering capacity against heat and other stressors. Along with a particular fermentation ability that is able to produce ethanol even at high temperature such as
with a comparable rate to the fermentation at
, this strain also exhibited higher viability than a reference strain owing to its own thermotolerance that conferred the survival after the severe heat shock at
for 30 minutes. Furthermore, this strain showed outstanding tolerances against
, ethanol and some chemical compounds. But, especially due to the thermotolerance, this strain has been suspected of other species of yeast. However, ITS (internally transcribed spacer) 1 and 2 sequencing data confirmed this strain was a typical strain of S. cerevisiae. The outstanding tolerances to various environmental stressors Indicate this S. cerevisiae KNU5377 is enough to use both as an on-site potential strain for world-wide alcohol fermentation industry and as a model strain for researches into the routes to acquire the tolerance to various stressors.