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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 36, Issue 4 - Dec 2008
Volume 36, Issue 3 - Sep 2008
Volume 36, Issue 2 - Jun 2008
Volume 36, Issue 1 - Mar 2008
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Real-Time PCR for Quantitative Detection of Bovine Parvovirus during Manufacture of Biologics
Lee, Dong-Hyuck ; Lee, Jung-Hee ; Kim, Chan-Kyong ; Kim, Tae-Eun ; Bae, Jung-Eun ; Kim, In-Seop ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 173~181
Bovine blood, cell, tissue, and organ are used as raw materials for manufacturing biologics such as biopharmaceuticals, tissue-engineered products, and cell therapy. Manufacturing processes for the biologics have the risk of viral contamination. Therefore viral validation is essential in ensuring the safety of the products. Bovine parvovirus (BPV) is one of the common bovine pathogens and has widely been known as a possible contaminant of biologics. In order to establish the validation system for the BPV safety of biologics, a real-time PCR method was developed for quantitative detection of BPV contamination in raw materials, manufacturing processes, and final products. Specific primers for amplification of BPV DNA were selected, and BPV DNA was quantified by use of SYBR Green 1. The sensitivity of the assay was calculated to be
. The real-time PCR method was validated to be reproducible and very specific to BPV. The established real-time PCR assay was successfully applied to the validation of Chinese hamster ovary (CHO) cell artificially infected with BPV. BPV DNA could be quantified in CHO cell as well as culture supernatant. Also the real-time PCR assay could detect
of BPV artificially contaminated in bovine collagen. The overall results indicated that this rapid, specific, sensitive, and robust assay can be reliably used for quantitative detection of BPV contamination during manufacture of biologics.
Gene Cloning and Expression of Trehalose Synthase from Thermus thermophilus HJ6
Kim, Hyun-Jung ; Kim, Han-Woo ; Jeon, Sung-Jong ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 182~188
A hyperthermophilic bacteria (strain HJ6) was isolated from a hot springs located in the Arima-cho, Hyogo, Japan. The cells were long-rod type (
in diameter. The pH and temperature for optimal growth were 6.5 and
, respectively. Phylogenetic analysis based on the 16S rDNA sequence and biochemical studies indicated that HJ6 belonged to the genus Thermus thermophilus (Tt). The gene encoding the Trehalose synthase (TS) was cloned and sequenced. The open reading frame (ORF) of the TtTS gene was composed of 2,898 nucleotides and encoded a protein (975 amino acids) with a predicted molecular weight of 110.56 kDa. The deduced amino acid sequence of TtTS showed 99% and 83% identities to the Thermus caldophilus TS and Meiothermus ruber TS, respectively. TtTS gene was expressed in Escherichia coli cells, and the recombinant protein was purified to homogeneity. The optimal temperature and pH for Trehalose synthase activity were found to be
and 7.5, respectively. The half-life of heat inactivation was about 40 min at
. The maximum trehalose conversion rate of maltose into trehalose by the enzyme increased as the substrate concentration increased, and reached 55.7% at the maltose concentration of 500 mM, implying that the enzyme conversion was dependent of the substrate concentration.
Molecular Cloning of a Putative Gene Encoding Phospholipase B (plbA) from Aspergillus nidulans
Hong, Sa-Hyun ; Cho, Eun-Min ; Song, Seung-Eun ; Eom, Chi-Yong ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 189~194
The phospholipase B (PLB) families are enzymes sharing phospholipase (PL), lysophospholipase (LPL) and lysophospholipase-transacylase (LPTA) activities. In this study, we report the putative gene encoding phospholipase B (plbA) containing lipase motifs was cloned for the first time from the filamentous fungus, Aspergillus nidulans. plbA was isolated from A. nidulans genomic DNA library using a PCR-amplified probe, which is designed on the basis of sequence information derived from the conserved lipase regions of various PLBs. The deduced product of plbA is of 626 amino acids. From the assigned sequence, PlbA showed 72% identity with Penicillium notatum PLB but have low similarity with phospholipase A of other organisms.
Evaluation of Antimicrobial, Antithrombin, and Antioxidant Activity of Aerial Part of Saxifraga stolonifera
Sohn, Ho-Yong ; Ryu, Hee-Young ; Jang, Yu-Jin ; Jang, Han-Su ; Park, Yu-Mi ; Kim, Sa-Youl ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 195~200
Saxifraga stolonifera (Saxifragaceae) is a perennial herbaceous plant growing in Korea, China, Japan and Russia. The aerial part has been used as herbal medicine for treatment of pneumonia, frostbite, inflammation and microbial infection. In this study, fresh juice and methanol extract were prepared from the aerial part of S. stolonifera, and their antimicrobial, antithrombin, and antioxidant activity were evaluated, respectively. The fresh juice showed weak antimicrobial activity against Proteus vulgaris, Escherichia coli O157:H7 and Candida albicans with ignorable DPPH scavenging activity. But, the methanol extract showed strong antioxidant activity (
) with minor, broad-range antimicrobial activity. Antithrombin activities were not observed in fresh juice and extract, up to 1.5 mg/mL. Sequential organic solvent fractionation of methanol extract showed that
of ethylacetate and the butanol fraction were 6.9 and
, respectively, that is comparable with vitamin C or butylated hydroxytoluene. Analysis of component in extract and fractionates suggested that the antioxidants in fractions are diverse and the active substances have glycosylated phenolic structure. Our results suggest that the aerial part of S. stolonifera could be used as the natural source of potential antioxidant.
Solid Fermentation of Medicinal Herb Using Phellinus baumii Mycelium and Anti-thrombin and Anti-oxidation Activity of its Methanol Extract
Shin, Yong-Kyu ; Jang, Han-Su ; Kim, Jong-Sik ; Ryu, Hee-Young ; Kim, Jong-Kuk ; Kwun, In-Sook ; Sohn, Ho-Yong ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 201~208
To produce bioactivity-strengthen medicinal herbs, the 36 medicinal herbs which have antioxidation or blood circulation activity, were solid fermented using Phellinus baumii mycelium. Most of medicinal herbs, except Chrysanthemum indicum (flower), Zizyphus jujuba Miller (fructus), Aconitum koreanum R. Raymond (root), Magnolia denu-data (flower), and Polygonatum sibiricum Redt (root bark), showed good fermentation at
for 20 days under 90% of relative humidity. The poor fermentations of the herbs could be explained by lack of nutrient, structural rigidity, and the content of antifungal substance. After fermentation, the average water content of herbs were increased to
, but the average pH and average methanol extraction ratio were slightly decreased to
, respectively. The analysis of thrombin inhibition and DPPH scavenging activity of the methanol extracts of herbs showed that thrombin inhibition activities of the fermented Drynaria fortunei Kunze, Melia azedarach var. japonica, Prunus persica and Orostachys japonicus, and DPPH scavenging activities of the fermented Polygala tenuifolia, Scrophularia buergeriana, Angelica dahurica, Drynariafortunei Kunze, Cyperus rotundus, and Boschniakia rossica were increased as compared with those activities of non-fermented its cognate herbs. Our results suggest that the production of bioactivity-strengthen medicinal herbs is possible by solid fermentation of Phellinus baumii mycelium, as fermented Drynaria fortunei Kunze showed increased antioxidant and thrombin inhibitory activities than those of non-fermented herbs.
Characterization of Viable But Nonculturable Condition of Escherichia coli Induced with Copper
Ku, Hyung-Keun ; Park, Sang-Ryoul ; Kim, Sook-Kyung ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 209~214
VBNC (Viable but nonculturable) state is an adaptive response of cells in adverse environments, which lead cell not grow on routine nutrient agar. In this study, we induced VBNC in Escherichia coli using copper and verify the characterization of it. After treatment of copper, we didn't detect any cells via plate cultivation, namely, colony forming unit (CFU) was zero. However, we identified the existence of VBNC by staining live cells with Live/Dead BacLight bacterial viability kit and counting them through flow cytometry. Then we isolated genomic DNA and RNA from VBNC-induced cells and analyzed the stability of them. Degradation of RNA is more severe than that of DNA and RNA is degraded as specific fragments. In addition, we showed the morphology of VBNC cell by Bio-Transmission Electron Microscope (Bio-TEM). VBNC cell showed impaired periplasmic space and inner and outer membrane were separated and the amount of cytosol were significantly decreased.
Characteristics of Rahnella aquatilis Strain AY2000 for an Anti-Yeast Substance Production
Kang, Min-Jung ; Lee, Bok-Kyu ; Kim, Kwang-Hyeon ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 215~220
Rahnella aquatilis AY2000 has an unique characteristic which produces an anti-yeast substance (AYS). The AYS of the strain AY2000 was always secreted on agar plate, however, its activity in liquid culture was labile upon storage of the medium. In this paper, cultural conditions of the strain AY2000 for the AYS production were investigated in liquid culture, and minimal inhibitory concentration (MIC) against Saccharomyces cerevisiae was determined for the AYS activity. MIC of the AYS cultured in PYG broth at
for 24 hr was
, however, that in MYCS (pH 5.5) broth at the same condition was
. The activity of the AYS had increased rather in MYCS broth excluded
-citrate than in the same broth contained
-citrate, and MIC of the AYS produced in MYCS broth without
. When the strain AY2000 was maintained in MYCS broth without
-citrate but added
, the activity of the AYS had increased and its MIC was
. MIC of the AYS was
after the strain AY2000 was cultured in MYCS broth containing
-citrate, however, its MIC was
after 48-60 hr culture in the same broth.
Expression of Human Heavy-Chain and Light-Chain Ferritins in Saccharomyces cerevisiae for Functional Foods and Feeds
Han, Hye-Song ; Lee, Joong-Lim ; Park, Si-Hong ; Kim, Jae-Hwan ; Kim, Hae-Yeong ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 221~226
To produce human ferritins in yeast, human H-chain and L-chain ferritins were amplified from previously cloned vectors. Each amplified ferritin gene was inserted into the pYES2.1/V5-His-TOPO yeast expression vector under the control of the GAL1promoter. Western blot analysis of the recombinant yeast cells revealed that H-and L-chain subunits of human ferritin were expressed in Saccharomyces cerevisiae. Atomic absorption spectrometry (AAS) analysis demonstrated that the intracellular content of iron in the ferritin transformant was 1.6 to 1.8-fold higher than that of the control strain. Ferritin transformants could potentially supply iron-fortified nutrients for food and feed.
Antifungal Activity of Bacillus sp. AM-651 Against Phytophthora capsici
Lee, Jung-Bok ; Shin, Jeong-Hak ; Jang, Jong-Ok ; Shin, Kee-Sun ; Choi, Chung-Sik ; Kim, Kun-Woo ; Jo, Min-Sub ; Jeon, Chun-Pyo ; Kim, Yun-Hoi ; Kwon, Gi-Seok ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 227~232
Biological antagonists of Phytophthora capsici were isolated from soil in Gyeongbuk, Korea. Among the isolated bacteria, a Bacillus sp. was identified from l6S rDNA sequence analysis and named Bacillus sp. AM-651. Bacillus sp. AM-65l strain which can strongly a antifungal activity against Phytophthora capsici. Culture conditions for the maximum production of the antagonistic substance were optimized. The production of antibiotic were high on modified Davis mineral medium pH 7 at
. The medium for highest production of the agonistic substance optimized. It is composed the best activity on glucose,
at 0.5%, 0.1%, and 0.7%, respectively. By time course of culture solution selected Bacillus sp. AM-65l, the culture solution after 48hrs had strongly growth inhibition rate against P. capsici. And culture solution of Bacillus sp. AM-651 was stable within a pH range
and temperature range
. Bacillus sp. AM-651 cultured broth shown fungal growth inhibitory activity against B. sorokiniana, B. cinerea, R. solani avove and beyond P. capsici and comparatively showed a high activity against C. gloeosporioides, B. dothidea, B. cinerea and F. graminearum by agar diffusion method.
Solubilization of Sewage Sludge by Inoculation of Lactic Acid Bacteria
Yang, Hyun-Sang ; Lee, Jung-Eun ; Lee, Eun Young ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 233~239
A new approach to the solubilization of excess activated sludge by the inoculation of lactic acid bacteria was studied to reduce the amount of sludge produced in the activated sludge treatment process. Aerobic microorganism in sludge was lysed in anaerobic condition and the cytoplasmic substance eluted was utilized as a carbon source by lactic acid bacteria. On the basis of sludge solubilization efficiency, Lactobacillus brevis and Leuconostoc mesenteroides subsp mesenteroides were selected the best candidates among five kinds of Lactobacillus sp. and seven kinds of Leuconostoc sp. The sludge solubilization efficiency by heterofermentative lactic acid bacteria was more efficient than that of homofermentative bacteria. Initial value of soluble COD (sCOD) was 1050 mg/L at the initial inoculation time increased to 3070 mg/L (192% solubilization) at 96 h of the incubation time. The inoculation of lactobacillus brevis to the sludge resulted in 2824% increase in sCOD value after 96 h of incubation than the control experiment. Leuconostoc mesenteroides subsp mesenteroides showed 152% increase of solubilization and 30% increase of S-COD/T-COD on 96 h of incubation time. Considering the increase of S-COD by the inoculation of Leuconostoc sp. on 24 h, 10% inoculation of lactic acid bacteria to the sludge was most effective.
Protective Effects of Fermented Aloe vera on Carbon Tetrachloride-induced Hepatotoxicity in Sprague-dawley Rats
Lim, Byung-Lak ;
Microbiology and Biotechnology Letters, volume 36, issue 3, 2008, Pages 240~245
Aloe vera extract was fermented by Lactobacillus casei. The ability of fermented Aloe vera (FAV) as an antioxidant to protect against
-induced oxidative stress and hepatotoxicity in rats was investigated. The rats were administered orally with various doses of FAV with 50, 100 mg/kg for 14 consecutive days. For this study, we not only tested activity of various plasma enzymes (AST, ALT), which are used as indicators of liver disease, but also checked those change of liver components such as superoxide dismutase and catalase activity. Pretreatment of FAV for two weeks significantly reduced the elevated plasma enzyme activities induced by
. Pretreatment of FAV also restored the hepatic enzyme, malonedialdehyde (MDA) formation. The results indicate that FAV has a protective effect against acute hepatotoxicity induced by the administration of
in rats, and that the hepatoprotective effects of FAV may be due to both the inhibition of lipid peroxidation and the increase of antioxidant activity.