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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Journal of Microbiology and Biotechnology
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Journal DOI :
The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 21, Issue 12 - Dec 2011
Volume 21, Issue 11 - Nov 2011
Volume 21, Issue 10 - Oct 2011
Volume 21, Issue 9 - Sep 2011
Volume 21, Issue 8 - Aug 2011
Volume 21, Issue 7 - Jul 2011
Volume 21, Issue 6 - Jun 2011
Volume 21, Issue 5 - May 2011
Volume 21, Issue 4 - Apr 2011
Volume 21, Issue 3 - Mar 2011
Volume 21, Issue 2 - Feb 2011
Volume 21, Issue 1 - Jan 2011
Selecting the target year
Chemotaxonomy of Trichoderma spp. Using Mass Spectrometry-Based Metabolite Profiling
Kang, Dae-Jung ; Kim, Ji-Young ; Choi, Jung-Nam ; Liu, Kwang-Hyeon ; Lee, Choong-Hwan ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 5~13
DOI : 10.4014/jmb.1008.08018
In this study, seven Trichoderma species (33 strains) were classified using secondary metabolite profile-based chemotaxonomy. Secondary metabolites were analyzed by liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS-MS) and multivariate statistical methods. T. longibrachiatum and T. virens were independently clustered based on both internal transcribed spacer (ITS) sequence and secondary metabolite analyses. T. harzianum formed three subclusters in the ITS-based phylogenetic tree and two subclusters in the metabolitebased dendrogram. In contrast, T. koningii and T. atroviride strains were mixed in one cluster in the phylogenetic tree, whereas T. koningii was grouped in a different subcluster from T. atroviride and T. hamatum in the chemotaxonomic tree. Partial least-squares discriminant analysis (PLS-DA) was applied to determine which metabolites were responsible for the clustering patterns observed for the different Trichoderma strains. The metabolites were hetelidic acid, sorbicillinol, trichodermanone C, giocladic acid, bisorbicillinol, and three unidentified compounds in the comparison of T. virens and T. longibrachiatum; harzianic acid, demethylharzianic acid, homoharzianic acid, and three unidentified compounds in T. harzianum I and II; and koninginin B, E, and D, and six unidentified compounds in T. koningii and T. atroviride. The results of this study demonstrate that secondary metabolite profiling-based chemotaxonomy has distinct advantages relative to ITS-based classification, since it identified new Trichoderma clusters that were not found using the latter approach.
Targeted Gene Disruption and Functional Complementation of Cytochrome P450 Hydroyxlase Involved in Cyclosporin A Hydroxylation in Sebekia benihana
Lee, Mi-Jin ; Han, Kyu-Boem ; Kim, Eung-Soo ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 14~19
DOI : 10.4014/jmb.1009.09026
A cyclic undecapeptide-family natural product, cyclosporin A (CyA), which is one of the most valuable immunosuppressive drugs, is produced nonribosomally by a multifunctional cyclosporin synthetase enzyme complex in a filamentous fungal strain named Tolypocladium niveum. Previously, structural modifications of cyclosporins such as a regionspecific hydroxylation at the
N-methyl leucine in a rare actinomycetes called Sebekia benihana were reported to lead to dramatic changes in their bioactive spectra. However, the reason behind this change could not be determined since a system to genetically manipulate S. benihana has not yet been developed. To address this limitation, in this study, we utilized the most commonly practiced gene manipulation techniques including conjugation-based foreign gene transfer-and-expression as well as targeted gene disruption to genetically manipulate S. benihana. Using these optimized genetic manipulation systems, a putative cytochrome P450 hydroxylase (CYP) gene named CYP506, which is involved in CyA hydroxylation in S. benihana, was specifically disrupted and genetically complemented. The S. benihana
CYP506 exhibited a significantly reduced CyA hydroxylation yield as well as considerable yield restoration by functional complementation of the S. benihana CYP506 gene, suggesting that the genetically manipulated S. benihana CYP mutant strains may serve as a more efficient bioconversion host for various valuable metabolites including CyA.
Purification and Characterization of Manganese-Dependent Alkaline Serine Protease from Bacillus pumilus TMS55
Ibrahim, Kalibulla Syed ; Muniyandi, Jeyaraj ; Pandian, Shunmugiah Karutha ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 20~27
DOI : 10.4014/jmb.1009.09001
The purification and characterization of a
-dependent alkaline serine protease produced by Bacillus pumilus TMS55 were investigated. The enzyme was purified in three steps: concentrating the crude enzyme using ammonium sulfate precipitation, followed by gel filtration and cation-exchange chromatography. The purified protease had a molecular mass of approximately 35 kDa, was highly active over a broad pH range of 7.0 to 12.0, and remained stable over a pH range of 7.5 to 11.5. The optimum temperature for the enzyme activity was found to be
. PMSF and AEBSF (1 mM) significantly inhibited the protease activity, indicating that the protease is a serine protease.
ions enhanced the activity and stability of the enzyme. In addition, the purified protease remained stable with oxidants (
, 2%) and organic solvents (25%), such as benzene, hexane, and toluene. Therefore, these characteristics of the protease and its dehairing ability indicate its potential for a wide range of commercial applications.
Amphotericin B Aggregation Inhibition with Novel Nanoparticles Prepared with Poly(
-caprolactone)/Poly(N,N-dimethylamino-2-ethyl methacrylate) Diblock Copolymer
Shim, Yong-Ho ; Kim, You-Chan ; Lee, Hong-Joo ; Bougard, Francois ; Dubois, Philippe ; Choi, Ki-Choon ; Chung, Chung-Wook ; Kang, Dae-Hwan ; Jeong, Young-Il ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 28~36
DOI : 10.4014/jmb.1007.07041
Diblock copolymers composed of poly(
-caprolactone) (PCL) and poly(N,N-dimethylamino-2-ethyl methacrylate) (PDMAEMA), or methoxy polyethylene glycol(PEG), were synthesized via a combination of ring-opening polymerization and atom-transfer radical polymerization in order to prepare polymeric nanoparticles as an antifungal drug carrier. Amphotericin B (AmB), a natural antibiotic, was incorporated into the polymeric nanoparticles. The physical properties of AmB-incorporated polymeric nanoparticles with PCL-b-PDMAEMA and PCL-b-PEG were studied in relation to morphology and particle size. In the aggregation state study, AmB-incorporated PCL-b- PDMAEMA nanoparticles exhibited a monomeric state pattern of free AmB, whereas AmB-incorporated PCL-b- PEG nanoparticles displayed an aggregated pattern. In in vitro hemolysis tests with human red blood cells, AmBincorporated PCL-b-PDMAEMA nanoparticles were seen to be 10 times less cytotoxic than free AmB (5
/ml). In addition, an improved antifungal activity of AmBincorporated polymeric nanoparticles was observed through antifungal activity tests using Candida albicans, whereas polymeric nanoparticles themselves were seen not to affect activity. Finally, in vitro AmB release studies were conducted, proving the potential of AmB-incorporated PCL-b-PDMAEMA nanoparticles as a new formulation candidate for AmB.
Bioconversion of Acrylonitrile to Acrylic Acid by Rhodococcus ruber Strain AKSH-84
Kamal, Ahmed ; Kumar, M. Shiva ; Kumar, C. Ganesh ; Shaik, Thokhir Basha ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 37~42
DOI : 10.4014/jmb.1006.06044
A new versatile acrylonitrile-bioconverting strain isolated from a petroleum-contaminated sludge sample and identified as Rhodococcus ruber AKSH-84 was used for optimization of medium and biotransformation conditions for nitrilase activity to produce acrylic acid. A simple and rapid HPLC protocol was optimized for quantification of acrylic acid, acrylamide, and acrylonitrile. The optimal medium conditions for nitrilase activity were pH of 7.0, temperature of
, agitation of 150 rpm, and inoculum level of 2%. Glycerol as a carbon source and sodium nitrate as the nitrogen source provided good nutritional sources for achieving good biotransformation. Nitrilase activity was constitutive in nature and was in the exponential growth phase after 24 h of incubation under optimal conditions without addition of any inducer. The substrate preference was acrylonitrile and acetonitrile. The present work demonstrates the biotransformation of acrylonitrile to acrylic acid with the new strain, R. ruber AKSH-84, which can be used in green biosynthesis of acrylic acid for biotechnological processes. The nitrilase produced by the isolate was purified and characterized.
Microbiological Purification of L-Arabitol from Xylitol Mother Liquor
Jiang, Mingguo ; Wang, Ben ; Yang, Lifang ; Lin, Shuangjun ; Cheng, Hairong ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 43~49
DOI : 10.4014/jmb.1006.06012
As a rare sugar alcohol, L-arabitol can be used in food and can prevent extra fat deposits in the intestinal tract. Commercially, L-arabitol is prepared from pure L-arabinose by hydrogenation, which needs a high temperature and high pressure, leading to a high production cost for Larabitol. Therefore, this study describes a novel L-arabitol production method based on biological purification from the xylitol mother liquor, a cheap and readily available raw material that contains a high concentration of Larabitol. First, a novel Bacillus megaterium strain was screened that can utilize xylitol, sorbitol, and mannitol, yet not L-arabitol. The isolated strain was inoculated into a medium containing the xylitol mother liquor under formulated culture conditions, where a high L-arabitol yield (95%) and high purity (80%) were obtained when the medium was supplemented with 50 g/l of xylitol mother liquor. Upon further purification of the fermentation broth by ion exchange and decolorization, L-arabitol was crystallized with a purity of 98.5%.
Phenotypic and Genotypic Characterization of Salmonella spp. Isolated from Pigs and their Farm Environment in Korea
Lim, Suk-Kyung ; Byun, Jung-Ryul ; Nam, Hyang-Mi ; Lee, Hee-Soo ; Jung, Suk-Chan ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 50~54
DOI : 10.4014/jmb.1007.07027
This study's objective was to determine the prevalence of Salmonella spp. in pigs and their farm environments in Korea, and to investigate the relationship between the strains based on their phenotypic and genotypic characteristics. A total of 36 Salmonella spp. were isolated in this study: 18 isolates from 492 pigs (3.7%) and 18 isolates from 418 (4.3%) farmhouse environmental samples from 16 different pig farms. Of the Salmonella strains isolated from the numerous environmental samples, the highest prevalence was observed in slurry or manure, followed by partitions, farmer's hands, floors, water/nipples, ventilation sources, and feed, respectively. All the Salmonella isolates originating from different farms were genetically distinct. In three farms, however, identical phage types and pulse-field gel electrophoresis patterns were observed among Salmonella isolates from pig feces and environmental samples. This study suggests that environments contaminated with Salmonella could pose an infection risk to pigs on pig farms.
Molecular Genetic Identification of Yeast Strains Isolated from Egyptian Soils for Solubilization of Inorganic Phosphates and Growth Promotion of Corn Plants
Hesham, Abd El-Latif ; Mohamed, Hashem M. ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 55~61
DOI : 10.4014/jmb.1006.06045
Forty yeast strains isolated from soils taken from different locations in Egypt were tested for their P-solubilizing activities on the basis of analyzing the clear zone around colonies growing on a tricalcium phosphate medium after incubation for 5 days at
, denoted as the solubilization index (SI). Nine isolates that exhibited P-solubilization potential with an SI ranging from 1.19 to 2.76 were genetically characterized as five yeasts belonging to the genus Saccharomyces cerevisiae and four non-Saccharomyces, based on a PCR analysis of the ITS1-26S region amplied by SC1/SC2 species-specific primers. The highest P-solubilization efficiency was demonstrated by isolate PSY- 4, which was identified as Saccharomyces cerevisiae by a sequence analysis of the variable D1/D2 domain of the 26S rDNA. The effects of single and mixed inoculations with yeast PSY-4 and Bacillus polymyxa on the P-uptake and growth of corn were tested in a greenhouse experiment using different levels of a phosphorus chemical fertilizer (50, 100, and 200 kg/ha super phosphate 15.5%
). The results showed that inoculating the corn with yeast PSY-4 or B. polymyxa caused significant increases in the shoot and root dry weights and P-uptake in the shoots and roots. The P-fertilization level also had a significant influence on the shoot and root dry weights and P-uptake in the shoots and roots when increasing the P-level from 50 up to 200 kg/ha. Dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 200 kg/ha gave higher values for the shoot and root dry weights and P-uptake in the shoots and roots, yet these increases were nonsignificant when compared with dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha. The best increases were obtained from dual inoculation with yeast strain PSY-4 and B. polymyxa at a P-fertilization level of 100 kg/ha, which induced the following percentage increases in the shoot and root dry weights, and P-uptake in the shoots and roots; 16.22%, 46.92%, 10.09%, and 31.07%, respectively, when compared with the uninoculated control (fertilized with 100 kg/ha).
In Vitro Antagonistic Activity Evaluation of Lactic Acid Bacteria (LAB) Combined with Cellulase Enzyme Against Campylobacter jejuni Growth in Co-Culture
Dubois-Dauphin, Robin ; Sabrina, Vandeplas ; Isabelle, Didderen ; Christopher, Marcq ; Andre, Thewis ; Philippe, Thonart ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 62~70
DOI : 10.4014/jmb.1007.07006
The antibacterial effects of nine lactic acid bacteria (LAB) against Campylobacter jejuni were investigated by using agar gel diffusion and co-culture assays. Some differences were recorded between the inhibition effects measured with these two methods. Only two LAB, Lb. pentosus CWBI B78 and E. faecium THT, exhibited a clear anti- Campylobacter activity in co-culture assay with dehydrated poultry excreta mixed with ground straw (DPE/GS) as the only growth substrate source. It was observed that the supplementation of such medium with a cellulase A complex (Beldem S.A.) enhanced the antimicrobial effect of both LAB strains. The co-culture medium acidification and the C. jejuni were positively correlated with the cellulase A concentration. The antibacterial effect was characterized by the lactic acid production from the homofermentative E. faecium THT and the lactic and acetic acids production from the heterofermentative Lb. pentosus CWBI B78. The antagonistic properties of LAB strains and enzyme combination could be used in strategies aiming at the reduction of Campylobacter prevalence in the poultry production chain and consequently the risk of human infection.
Biodegradation of Diazinon by Serratia marcescens DI101 and its Use in Bioremediation of Contaminated Environment
Abo-Amer, Aly E. ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 71~80
DOI : 10.4014/jmb.1007.07024
Four diazinon-degrading bacteria were isolated from agricultural soil by using an enrichment technique. The biochemical analysis and molecular method including RFLP indicated that these isolates were identical, and one strain designated DI101 was selected for further study. Phylogenetic analysis based on 16S rDNA sequencing indicated that the strain DI101 clearly belongs to the Serratia marcescens group. The ability of the strain to utilize diazinon as a source of carbon and phosphorus was investigated under different culture conditions. The DI101 strain was able to completely degrade 50 mg/l diazinon in MSM within 11 days with a degradation rate of 0.226
. The inoculation of sterilized soil treated with 100 mg/kg of diazinon with
CFU/g DI101 resulted in a faster degradation rate than was recorded in non-sterilized soil. The diazinon degradation rate by DI101 was efficient at temperatures from 25 to
and at pHs from 7.0 to 8.0. The degradation rate of diazinon was not affected by the absence of a phosphorus supplement, and addition of other carbon sources (glucose or succinate) resulted in the slowing down of the degradation rate. The maximum degradation rate (
) of diazinon was 0.292
and its saturation constant (
) was 11 mg/l, as determined by a Michaelis-Menten curve. The strain was able to degrade diethylthiophosphate-containing organophosphates such as chlorpyrifos, coumaphos, parathion, and isazofos when provided as a source of carbon and phosphorus, but not ethoprophos, cadusafos, and fenamiphos. These results propose useful information for the potential application of the DI101 strain in bioremediation of pesticide-contaminated environments.
Evaluation of Anti-Phytoplasma Properties of Surfactin and Tetracycline Towards Lime Witches' Broom Disease Using Real-Time PCR
Askari, N. ; Jouzani, Gh. Salehi ; Mousivand, M. ; Nazari, A. Hagh ; Abbasalizadeh, S. ; Soheilivand, S. ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 81~88
DOI : 10.4014/jmb.1007.07047
The anti-phytoplasma activities of surfactin (derived from Iranian native Bacillus subtilis isolates) and tetracycline towards Candidatus "Phytoplasma aurantifolia", the agent of lime Witches' broom disease, were investigated. HPLC was used to quantify the surfactin production in four previously characterized native surfactin-producing strains, and the one producing the highest amount of surfactin (about 1,500 mg/l) was selected and cultivated following optimized production and extraction protocols. Different combinations of purified surfactin and commercial tetracycline were injected into artificially phytoplasmainfected Mexican lime seedlings using a syringe injection system. An absolute quantitative real-time PCR system was developed to monitor the phytoplasma population shifts in the lime phloem during 3 months following the injections. The results revealed that the injections of surfactin or tetracycline had a significant inhibitory effect on Candidatus "P. aurantifolia". However, the combined treatment with both surfactin and tetracycline (1:1) resulted in the highest inhibition due to a synergic effect, which suppressed the phytoplasma population from about
to less than 10 phytoplasma units/g plant tissue.
Rapid Multiplex PCR Assay for the Simultaneous Detection of the Brucella Genus, B. abortus, B. melitensis, and B. suis
Kumar, Sanjay ; Tuteja, Urmil ; Sarika, Kumari ; Singh, Dhirendra Kumar ; Kumar, Ashok ; Kumar, Om ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 89~92
DOI : 10.4014/jmb.1007.07051
The routine identification and differentiation of Brucella species is a time-consuming and labor-intensive process, which frequently places personnel at risk of laboratory-acquired infection. Here, we describe the development of a rapid multiplex PCR assay for the confirmation of presumptive Brucella isolates. The assay was able to identify and differentiate major human pathogens, namely B. abortus, B. melitensis, and B. suis, in a single test of less than an hour and a half.
Synthesis and Optimization of Cholesterol-Based Diquaternary Ammonium Gemini Surfactant (Chol-GS) as a New Gene Delivery Vector
Kim, Bieong-Kil ; Doh, Kyung-Oh ; Bae, Yun-Ui ; Seu, Young-Bae ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 93~99
DOI : 10.4014/jmb.1008.08012
Amongst a number of potential nonviral vectors, cationic liposomes have been actively researched, with both gemini surfactants and bola amphiphiles reported as being in possession of good structures in terms of cell viability and in vitro transfection. In this study, a cholesterol-based diquaternary ammonium gemini surfactant (Chol-GS) was synthesized and assessed as a novel nonviral gene vector. Chol-GS was synthesized from cholesterol by way of four reaction steps. The optimal efficiency was found to be at a weight ratio of 1:4 of lipid:DOPE (1,2-dioleoyl-L-
- glycero-3-phosphatidylethanolamine), and at a ratio of between 10:1~15:1 of liposome:DNA. The transfection efficiency was compared with commercial liposomes and with Lipofectamine, 1,2-dimyristyloxypropyl-3-dimethylhydroxyethylammonium bromide (DMRIE-C), and N-[1-(2,3-dioleoyloxy)propyl]-N,N,N-trimethylammonium chloride (DOTAP). The results indicate that the efficiency of Chol-GS is greater than that of all the tested commercial liposomes in COS7 and Huh7 cells, and higher than DOTAP and Lipofectamine in A549 cells. Confirmation of these findings was observed through the use of green fluorescent protein expression. Chol-GS exhibited a moderate level of cytotoxicity, at optimum concentrations for efficient transfection, indicating cell viability. Hence, the newly synthesized Chol-GS liposome has the potential of being an excellent nonviral vector for gene delivery.
Validation of a Real-Time RT-PCR Method to Quantify Newcastle Disease Virus (NDV) Titer and Comparison with Other Quantifiable Methods
Jang, Juno ; Hong, Sung-Hwan ; Kim, Ik-Hwan ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 100~108
DOI : 10.4014/jmb.1006.06006
A method for the rapid detection and quantification of Newcastle disease virus (NDV) produced in an animal cell culture-based production system was developed to enhance the speed of the NDV vaccine manufacturing process. A SYBR Green I-based real-time RT-PCR was designed with a conventional, inexpensive RT-PCR kit targeting the F gene of the NDV LaSota strain. The method developed in this study was validated for specificity, accuracy, precision, linearity, limit of detection (LOD), limit of quantification (LOQ), and robustness. The validation results satisfied the predetermined acceptance criteria. The validated method was used to quantify virus samples produced in an animal cell culture-based production system. The method was able to quantify the NDV samples from mid- or late-production phases, but not effective on samples from the early-production phase. For comparison with other quantifiable methods, immunoblotting, plaque assay, and tissue culture infectious dose 50 (
) assay were also performed with the NDV samples. The results demonstrated that the real-time RT-PCR method is suitable for the rapid quantification of virus particles produced in an animal cell-culture-based production system irrespective of viral infectivity.
Bioethanol Production from the Hydrolysate of Rape Stem in a Surface-Aerated Fermentor
Yeon, Ji-Hyeon ; Lee, Sang-Eun ; Choi, Woon-Yong ; Choi, Won-Seok ; Kim, Il-Chul ; Lee, Hyeon-Yong ; Jung, Kyung-Hwan ;
Journal of Microbiology and Biotechnology, volume 21, issue 1, 2011, Pages 109~114
DOI : 10.4014/jmb.1008.08001
In this study, we investigated the feasibility of producing bioethanol from the hydrolysate of rape stem. Specifically, the most ideal yeast strain was screened, and the microaeration was performed by surface aeration on a liquid medium surface. Among the yeast strains examined, Pichia stipitis CBS 7126 displayed the best performance in bioethanol production during the surface-aerated fermentor culture. Pichia stipitis CBS 7126 produced maximally 9.56 g/l of bioethanol from the initial total reducing sugars (about 28 g/l). The bioethanol yield was 0.397 (by the DNS method). Furthermore, this controlled surface aeration method holds promise for use in the bioethanol production from the xylose-containing lignocellulosic hydrolysate of biomass.