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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 37, Issue 4 - Dec 2009
Volume 37, Issue 3 - Sep 2009
Volume 37, Issue 2 - Jun 2009
Volume 37, Issue 1 - Mar 2009
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Genetic Monitoring of Plant Growth Promoting Rhizobacterium (PGPR), Bacillus subtilis AH18 using Multiplex PCR in Field Soil
Woo, Sang-Min ; Lim, Jong-Hui ; Jeong, Hee-Young ; Kim, Sang-Dal ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 1~9
The genetic monitoring method was developed for the rapid detection of the PGPR and biocontrol agent, B. subtilis AH18 in red-pepper field soil by multiplex PCR using sid, aec and cel gene primers. The monitoring of B. subtilis AH18 in the soil was carried by amplified a 2,3-dihydro-2,3-dihydroxy benzoate dehydrogenase [EC: 1. 3. 1. 28]gene (sid - 794 bp : EF408238) which is a key enzyme of siderophore synthesis, an auxin efflux carrier gene (aec - 1,052 bp : EF408239) and a cellulase gene (cel - 1,582 bp : EF070194). The natural un sterilized soil was inoculated with B. subtilis AH18 to determine the sensitivity (
cfu/g) of multiplex PCR for the rapid dectection and then the strain was monitored successfully in rhizosphere or non-rhizosphere soil of red-pepper cultural soil. At 3 weeks after the treatment, density of the strain was monitored more abundantly in rhizosphere soil.
Isolation of Myxobacteria Carrying Soraphen Biosynthetic Gene Clusters
Lee, Cha-Yul ; Hyun, Hye-Sook ; Cho, Kung-Yun ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 10~16
Polymerase chain reactin (PCR) could be a simple way to screen new microbial strains producing useful secondary metabolites if their biosynthetic genes are known and candidate strains to be screened are available. In this study, we have screened two myxobacterial strains, KYC3047 and KYC3076, carrying genes appeared to be biosynthetic genes of soraphen A, a potent antifungal substance, out of 50 cellulose degrading myxobacteria using PCR. The two strains were identified as Sorangium cellulosum based on morphological, physiological, and molecular biological characteristics. Both of the strains produced substances having strong antifungal activities as expected against Candida albicans, a causative agent of candidiasis, and Colletotrichum acutatum, a causative agent of anthracnose on pepper.
Gene Cloning and Expression of Thermostable DNA Polymerase from Thermus thermophilus HJ6
Seo, Min-Ho ; Kim, Bu-Kyoung ; Kwak, Pyung-Hwa ; Kim, Han-Woo ; Kim, Yeon-Hee ; Nam, Soo-Wan ; Jeon, Sung-Jong ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 17~23
The gene encoding Thermus thermophilus HJ6 DNA polymerase (Tod) was cloned and sequenced. The open reading frame (ORF) of the Tod gene was composed of 2,505 nucleotides and encoded a protein (843 amino acids) with a predicted molecular weight of 93,795 Da. The deduced amino acid sequence of Tod showed 98% and 86% identities to the Thermus thermophilus HB8 DNA pol and Thermus aquaticus DNA pol, respectively, The Tod gene was expressed under the control of the bacteriophage
promoters PR and PL on the expression vector pJLA503 in Escherichia coli strain BL21 (DE3) codon plus. The expressed enzyme was purified by heat treatment,
Q column, and
Sephacryl S-200 HR 26/60 column chromatographies. The optimal temperature and pH for DNA polymerase activity were found to be
and 9.0, respectively. The optimal concentrations of
were 2.5 mM and 1 mM, respectively. The enzyme activity was activated by divalent cations, and was inhibited by monovalent cations. The result of the PCR experiment with Tod DNA polymerase indicates that this enzyme might be useful in DNA amplification and PCR-based applications.
Characterization of Antimicrobial Substance Produced by Lactobacillus paraplantarum KNUC25 Isolated from Kimchi
Kim, Ma-Rie ; Lee, Su-Jin ; Seul, Keyung-Jo ; Park, Yu-Mi ; Ghim, Sa-Youl ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 24~32
The KNUC25 strain isolated from over-fermented whole Chinese cabbage kimchi was examined for its physiological characteristics using API 50 CHL system assay and identified as Lactobacillus paraplantarum by analysis of whole-cell protein SDS-PAGE pattern assay and similarity of 16S rDNA sequence. L. paraplantarum KNUC25 had a broad antimicrobial activity spectrum from Gram positive to Gram negative bacteria. Scanning electron micrograph analysis showed that KNUC25 might attack to cell surface of indicator cells and destruction can lead to inhibition of the cell growth. The antimicrobial substance of the KNUC25 strain was stable to various degrading enzymes and at high temperature and not a plasmid-born matter. Resistance to proteolytic enzymes showed that an antimicrobial activity of KNUC25 might not be caused by proteinous substance. Maximum production of antimicrobial substance was the exponential growth phase at
Antiviral Activity of Hot-Water Extract and Its Ethanol Precipitate of Phellinus pini Fruiting Body
Lee, Sae-Mi ; Kim, Sung-Min ; Lee, Yoon-Hee ; Kim, Woo-Jung ; Na, Ye-Seul ; Kim, Hyun-Guell ; Nam, Jae-Hwan ; Shin, Hyun-Dong ; Kwon, Doo-Han ; Park, Yong-Il ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 33~41
The crushed fruiting body of Alaskan Porodaedalea pini (Brot.) Murrill (syn. Phellinus pini) was extracted in boiling water for 4 h with the yield of 20.5% in dry mass. This hot-water extract showed significant antiviral activity by inhibiting the plaque formation in HeLa cells by coxackievirus B3 (CVB3) and also showed highest inhibitory effect against neuraminidase activity among water extracts of various mushrooms. From the water extract, the ethanol precipitate (EP) and supernatant fraction (ES) were obtained through 75% ethanol precipitation with the yield of 43.3% and 28.3% in dry mass, respectively. Whereas ES did not show any detectable level of antiviral activity, EP showed significant dose-dependent inhibition of plaque formation by CVB3 in HeLa cells with an
(50% effective concentration) of 0.45 mg/mL. The cytotoxicity on HeLa cells by EP was relatively low with the
(50% cytotoxic concentration) of 2.25 mg/mL. EP also effectively inhibited neuraminidase activity in a dose-dependent manner showing up to 75% inhibition at 1.7 mg/mL. These results suggest that the hot-water extract and its EP of P. pini fruiting body can be a candidate for the development of a potent broad-range antiviral agent against influenza virus(Flu) as well as CVB3. The major active component of EP was shown to be a heteropolysaccharide-protein complex containing glucose as the main sugar residue with mole percentage of 79.8% and other sugars like galactose (19.2%), xylose (17%), mannose (5.8%), and fucose (4.6%) and a small portion (12.7%, in mass) of protein.
Biological and Antifungal Activity of Herbal Plant Extracts against Candida Species
Kim, Jae-Young ; Yi, Yong-Sub ; Lim, Yoong-Ho ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 42~48
Anticandidial activity of seven herbal extracts, Taraxacum Platycarpum, Houttuyniae Herba, Lonicerae Flos, Anemarrhena Rhizome, Forsythia Fruit, Paeoniae Ratix, and Coptidis Rhizoma, were determined against five different Candida sp. by agar diffusion assay. The concentration of total phenolic compounds of seven herbal extracts ranged from 0.6 to
. The total antioxidant activities showed that Taraxacum Platycarpum and Houttuyniae Herba were 60% in 80% ethanol extract and Lonicera Flower and Paeoniae Ratix were 70, 75%, respectively, in 100% ethanol extract. Coptidis Rhizoma extract showed antifungal activity against non-Candida albicans, C. tropicalis and C. glabrata. The MIC values of a compound separated in TLC from Coptidis Rhizoma extract were 24, and
against C. tropicalis and C. glabrata. The above compound showed the same retention time with berberin in HPLC analysis.
Rhapontigenin Production by Bioconversion and Inhibition of Melanin Synthesis
Jeon, Min ; Lee, Kang-Moon ; Lim, Young-Hee ; Kim, Jeong-Keun ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 49~54
Rhapontin is the glycosylated stilbene compound, and comprising major component of rhubarb root extract. Rhapontin has been used as a raw material of skin-whitening cosmetics in Korea. Rhapontigenin, the aglycone of rhapontin, has been suggested to be more active than its glycosylated form. Therefore, the rhubarb root extract was treated with commercial enzyme, Pectinex to remove glycosylated moiety of rhapontin and rhapontigenin was prepared. The resulting material was analysed and identified as rhapontigenin by proton NMR and MALDI-Mass. Rhapontigenin exhibited tyrosinase inhibitory activity with an
. The tyrosinase inhibitory activity of rhapontigenin was six times higher than that of rhapontin. In melanin biosynthesis inhibition assay using Streptomyces bikiniensis, rhapontigenin showed wider inhibition zone than that of rhapontin. From these results, we expect that rhapontigenin has stronger skin whitening effect than rhapontin and has advantages in cosmetic industry.
Conversion of Ginsenoside
by Ginseng Soil Bacterium Cellulosimicrobium sp. Gsoil 235 According to Various Culture Broths
Na, Ju-Ryun ; Kim, Yu-Jin ; Kim, Se-Hwa ; Kim, Ho-Bin ; Shim, Ju-Sun ; Kim, Se-Young ; Yang, Deok-Chun ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 55~61
Ginseng saponins (a secondary metabolite, termed ginsenosides) are the principal bioactive ingredients of ginseng, and modification of the sugar chains may markedly change the its biological activity. One of soil bacteria having
-glucosidase (to transform ginsenoside
) activity was isolated from soil of a ginseng field in Daejeon. 16S rRNA gene sequence analysis revealed that the isolate belonged to the genus Cellulosimicrobium, with highest sequence similarity (99.7%) to Cellulosimicrobium funkei ATCC BAA-
. The strain, Gsoil 235, could transform ginsenoside
and 3 of un-known ginsenosides by the analyses of TLC, HPLC. By investigating its deglycosylation progress, the optimal broth for,
-glucosidase was nutrient broth (In 48 hours, almost ginsenoside
could be transformed into minor ginsenosides). On the contrary, the optimal broth for growth was determined as trypic soy broth (TSB).
Optimal Conditions for the Laccase Production from Fomitopsis pinicola Mycelia
Park, Naomi ; Park, Sang-Shin ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 62~68
The culture conditions to maximize the production of laccase (EC 184.108.40.206) from Fomitopsis pinicola mycelia were investigated. Among the tested media for the enzyme production, mushroom complete medium (MCM ; 2% dextrose, 0.2% peptone, 0.2% yeast extract, 0.05%
, and 0.05%
) showed the highest activity of the enzyme. To optimize the culture condition for the laccase activity, influence of various carbon and nitrogen sources was investigated in MCM. Among various carbon and nitrogen sources, 2% glucose and 0.4% peptone showed the highest production of the enzyme, respectively. For the phosphorus and inorganic source, 0.05%
were best for the enzyme activity. The enzyme production was reached to highest level after the cultivation for 8 days at
. Native polyacrylamide gel electrophoresis (PAGE) followed by the laccase activity staining using 2, 2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as the substrate was performed to identify the laccase under culture conditions studied. Zymogram analysis of the culture supernatant showed a laccase band with molecular mass of 52 kDa. The optimum pH and temperature for the enzyme activity were
and pH 3.0.
Hypocholesterolemic Effect of Lyophilized, Heat-Killed Lactobacillus rhamnosus and Lactobacillus plantarum
Kim, Dae-Weon ; Yang, Dae-Hyeok ; Kim, Sun-Young ; Kim, Kwang-Soo ; Chung, Myun-Gjun ; Kang, Sang-Mo ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 69~74
Lactic acid bacteria (LAB) were well known to enhance the intestinal health of human. For the development of pharmaceutical LAB. it was screened that the LAB with activity lowering the cholesterol in vitro and evaluated the hypocholestrolemic effect of live and heat-killed (HK) LAB on rats. The selected Lactobacillus plantarum CBT 1209 and Lactobacillus rhamnosus CBT 1702 had the deconjugation of bile salts and assimilation of cholesterol micelles activities from laboratory media, The mixture of 1702 and 1209 strains was administrated to the rats with high cholesterol diet. The experiment performed by 4 groups which were control, HCD, LLAB, HKLAB groups. The hypocholesterolemic effect of LAB (strains 1702, 1209) at blood level, the phenomena of AI decreasing through LDL-cholesterol dwindling, was assessed. This effect of 1702 and 1209 was enhanced when it comes to be the HKLAB more the live-LAB, This data means that the Lactobacillus rhamnosus CBT 1702 and Lactobacillus plantarum CBT 1209 were very useful functional ingredient for hypercholesterolemia.
Biosafety of the New Soft Contact Lens Materials in the Fibroblast L-929 Cell Line
You, Young-Hyun ; Nam, Joo-Hyeung ; Kim, Bieong-Kil ; Kim, Soon-Bok ; Moon, Ik-Jae ; Kim, Jong-Pil ; Seu, Young-Bae ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 75~79
In this study, we polymerized new materials for soft contact lens using HEMA (2-hydroxyethyl methacrylate) which is the based-monomer of soft contact lens, EGDMA (ethylene glycol dimethacrylate) as cross linkage agent, and the new additives of monoester or di-ester derived from itaconic acid commercially produced by the fermentation of Asp. itaconicus. New polymer materials for soft contact lens were synthesized with the mixture of HEMA and mono- or diester at different ratios and presented to a good water content and oxygen transmissibility (Dk/L) values. In case of polymerization with HEMA and mono-ester (15%), the water content and oxygen transmissibility of contact lens were found to be good values at 57.6% and 28.5 Dk respectively. The mixture of HEMA and mono-ester is more excellent than HEMA/di-ester in the water content and oxygen transmissibility. The toxicity of new contact lens materials were confirmed in the fibroblast L-929 cell line using a agar overlay test and a growth inhibition test with the extract solution of contact lens.
Antimicrobial Activities of the Anti-acne Compounds from Natural Sources
Kim, Na-Rae ; Lim, Young-Hee ; Park, Sul-Woong ; Nam, Eun-Sil ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 80~84
The in vitro antibacterial activities of anti-acne agents prepared from the extracts of natural sources were investigated against several bacteria including antibiotic-susceptible and -resistant Propionibacterium acnes. SD-1 and SD-2 were prepared with different formulations and they showed strong antibacterial activities. The anti-acne agents completely inhibited the growth of the tested strains at the concentration of 0.5%. There was no difference in antibacterial activity between antibiotic-susceptible and -resistant P. acnes. The inhibitory activities of two agents showed time-dependent manner. In S. aureus, time-kill curve demonstrated 2.8- and 3.4-
-unit killing after 8 h with SD-1 and SD-2, respectively. In P. acnes, time-killing curve demonstrated 5.1- and 6.1-
-unit killing after 24 h with SD-1 and SD-2, respectively. SD-2 showed stronger antimicrobial activity than SD-1. From these results, we expect that SD-1 and SD-2 have strong antibacterial activities and have advantages for treating acne.
Optimal Growth Condition of Pleurotus ostreatus Cultured in the Foodwastes Extracts
Lim, Joung-Soo ; Lee, So-Jin ; Lee, Eun-Young ;
Microbiology and Biotechnology Letters, volume 37, issue 1, 2009, Pages 85~89
For the purpose of the methodological development to convert the food wastes into resources, we have attempted to culture the mushroom hypha, Pleurotus ostreatus. The food-wastes were mixed with distilled water, and the mixture was autoclaved to produce fluid, which was centrifugated and used as the growth media. Concentrations of the food wastes extracts were prepared with 10, 20, 30, 40, and 50%(W/V), and the initial pH were set variously with 4, 5, 6, and 7. These were cultured for 9 days at the temperature of
and the rotation rate of 120 rpm. The result is that the fluid form of the mushroom hypha have been grown best at the concentration of 30% and the optimal pH was 5 and 6.