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REFERENCE LINKING PLATFORM OF KOREA S&T JOURNALS
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Microbiology and Biotechnology Letters
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The Korean Society for Applied Microbiology and Biotechnology
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Volume & Issues
Volume 41, Issue 4 - Dec 2013
Volume 41, Issue 3 - Sep 2013
Volume 41, Issue 2 - Jun 2013
Volume 41, Issue 1 - Mar 2013
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Applications and Prospects of Ionic Liquids in Microbiology and Biochemical Engineering
Ha, Sung Ho ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 1~7
DOI : 10.4014/kjmb.1211.11003
Ionic liquids (ILs) have been widely recognized as environmentally benign solvents. Their unique properties, including negligible vapor pressure, non-flammability, a wide liquid range and their tunable physicochemical properties by proper selection of cations and anions, make them attractive green solvents in a variety of fields such as organic synthesis/catalysis, extraction/ separation, and electrochemistry, amongst others. In this paper, the recent technological developments and their prospects in the application of ILs in microbiology and biochemical engineering, including enzymatic reactions, protein folding/refolding and biomass dissolution, are discussed.
Isolation and Characterization of a Novel Agar Degrading Bacterium, Alteromonas macleodii subsp. GNUM08120, from Red Macroalgae
Chi, Won-Jae ; Lim, Ju-Hyeon ; Park, Da Yeon ; Kim, Mu-Chan ; Kim, Chang-Joon ; Chang, Yong-Keun ; Hong, Soon-Kwang ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 8~16
DOI : 10.4014/kjmb.1208.08001
An agar-hydrolyzing marine bacterium, strain GNUM08120, was isolated from Sargassum fulvellum collected from Yeongil bay of East Sea of Korea. The isolate was Gram-negative, aerobic, motile with single polar flagellum, and grew at 1-10% NaCl, pH 5.0-8.0, and
. G+C content and the predominant respiratory quinone were 46.13 mol% and Q-8, respectively. The major cellular fatty acids were Summed feature 3 (24.5%),
(12.5%). Based on 16S rRNA gene sequence similarity and DNA-DNA hybridization analyses, strain GNUM08120 was identified as a novel subspecies of Alteromonas macleodii, designated Alteromonas macleodii subsp. GNUM08120. Production of agarase by strain GNUM08120 was likely repressed by the effect of carbon catabolite repression caused by glucose. The crude agarase prepared from 12-h culture broth of strain GNUM08120 exhibited an optimum pH and temperature for agarase activity at 7.0 and
, respectively. The crude enzyme produced (neo)agarobiose, (neo)agarotetraose, and (neo)agarohexaose as the hydrolyzed product of agarose.
Use of Human Serum Albumin Fusion Tags for Recombinant Protein Secretory Expression in the Methylotrophic Yeast Hansenula polymorpha
Song, Ji-Hye ; Hwang, Dong Hyeon ; Oh, Doo-Byoung ; Rhee, Sang Ki ; Kwon, Ohsuk ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 17~25
DOI : 10.4014/kjmb.1207.07021
The thermotolerant methylotrophic yeast Hansenula polymorpha is an attractive model organism for various fundamental studies, such as the genetic control of enzymes involved in methanol metabolism, peroxisome biogenesis, nitrate assimilation, and resistance to heavy metals and oxidative stresses. In addition, H. polymorpha has been highlighted as a promising recombinant protein expression host, especially due to the availability of strong and tightly regulatable promoters. In this study, we investigated the possibility of employing human serum albumin (HSA) as the fusion tag for the secretory expression of heterologous proteins in H. polymorpha. A set of four expression cassettes, which contained the methanol oxidase (MOX) promoter, translational HSA fusion tag, and the terminator of MOX, were constructed. The expression cassettes were also designed to contain sequences for accessory elements including His8-tag,
linkers, tobacco etch virus protease recognition sites (Tev), multi-cloning sites, and strep-tags. To determine the effects of the size of the HSA fusion tag on the secretory expression of the target protein, each cassette contained the HSA gene fragment truncated at a specific position based on its domain structure. By using the Green fluorescence protein gene as the reporter, the properties of each expression cassette were compared in various conditions. Our results suggest that the translational HSA fusion tag is an efficient tool for the secretory expression of recombinant proteins in H. polymorpha.
Isolation, Identification and Mutant Development of Butanol Tolerance Bacterium
Jung, Hyesook ; Lee, Jinho ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 26~32
DOI : 10.4014/kjmb.1210.10009
Butanol-resistant bacteria were isolated from butanol solvent. The cell growth of isolated strains declined with increasing concentrations of butanol, and isolated strain BRS02 displayed more resistance to 12.5 g/L of butanol than other isolated strains. In addition, strain BRS251, which was resistant to even higher concentrations of butanol, was developed by the mutation of BRS02 using UV. BRS251 could grow in LB medium containing up to 17.5 g/L of butanol, 32.5 g/L of propanol, or 6 g/L of pentanol, whereas the control strain Escherichia coli was found to be tolerant to 7.5 g/L of butanol, 20 g/L of propanol, or 2 g/L of pentanol. The isolated BRS02, a Gram(+) bacterium seen to have a cocci form under the microscope, grew in 6.5% NaCl. According to biochemical tests, BRS02 can metabolize and produce acid with D-galactose, D-maltose, D-mannitol, D-mannose, methyl-
-Dglucopyranoside, D-ribose, sucrose, or D-trehalose, as carbon sources. Also, this strain showed resistance to bacitracin, vibriostatic agent O/129, and optochin, alongside positive activities for arginine dihydrolase,
-glucosidase, and urease. The BRS02 strain was identified as Staphylococcus sp. by analyses of the 16S rRNA gene, phylogenetic tree, and biochemical tests.
Purification and Characterization of a Fibrinolytic Enzyme Produced by Bacillus amyloliquefaciens HC188
Shin, So Hee ; Hong, Sung Wook ; Chung, Kun Sub ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 33~43
DOI : 10.4014/kjmb.1208.08010
A bacterium producing a fibrinolytic enzyme was isolated from Cheonggukjang. The bacterium was identified as a strain of Bacillus amyloliquefaciens by 16S rDNA analysis and designated as B. amyloliquefaciens HC188. The optimum culture medium appeared to be one containing 0.5% (w/v) maltose and 0.5% (w/v) soytone. Bacterial growth in the optimal medium at
reached the stationary phase after 27 h of incubation and the fibrinolytic enzyme showed optimum activity at 24 h. The enzyme was purified by 20-80% ammonium sulfate precipitation, CM Sepharose fast flow ion exchange chromatography, and Sephacryl S-200HR column chromatography. Its specific activity was 38359.3 units/mg protein and the yield was 5.5% of the total activity of the crude extracts. The molecular weight was 24.7 kDa and the amino acids of the N-terminal sequence were AQSVPYGVSQIKAPA. The fibrinolytic enzyme activity had an optimum temperature of
and an optimum pH of 8.0, and the enzyme was stable in the ranges
and pH 6.0-8.0. Enzyme activity was increased by
but inhibited by
, EDTA, and PMSF. It is suggested that the purified enzyme is a metallo-serine protease.
Production of Antimicrobial Substances by Strains of Myxobacteria Corallococcus and Myxococcus
Shin, Hyejin ; Youn, Jinkwon ; An, Dongju ; Cho, Kyungyun ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 44~51
DOI : 10.4014/kjmb.1210.10011
We prepared culture extracts of 174 Corallococcus and 207 Myxococcus strains isolated in Korea, and compared their antimicrobial activity against Candida albicans, Pseudomonas aeruginosa, and Staphylococcus aureus. The percentage of strains showing antifungal activity was lower in Corallococcus (7.5% [13 of the 174 strains]) than in Myxococcus (51.7% [107 of the 207 strains]). However, the percentage of strains exhibiting antibacterial activity was higher in Corallococcus (12.1% [21 strains]) than in Myxococcus (1% [2 strains]). The culture extracts of 6 Corallococcus strains inhibited both P. aeruginosa and S. aureus and displayed similar high-performance liquid chromatography chromatograms, although the shapes of their fruiting bodies were dissimilar. The rate of production of antibacterial substances was the highest when the strains were cultured in CYS medium for more than 6 days.
Optimization of Medium Composition for Lipopeptide Production from Bacillus subtilis N7 using Response Surface Methodology
Luo, Yi ; Zhang, Guoyi ; Zhu, Zhen ; Wang, Xiaohui ; Ran, Wei ; Shen, Qirong ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 52~59
DOI : 10.4014/kjmb.1207.07020
The nutritional requirements for the maximum production of lipopeptides by Bacillus subtilis N7 (B. subtilis N7) were investigated and optimized using response surface methodology (RSM) under shake flask fermentation. A one-factor-at-a-time experimental setup was used to screen carbon and nitrogen sources. A Plackett-Burman design (PBD) was employed to screen the most critical variables for lipopeptides production amongst ten nutritional elements. The central composite experimental design (CCD) was finally adopted to elucidate the composition of the fermentation medium. Statistical analyses (analysis of variance, ANOVA) of the results showed that KCl,
were important components and that their interactions were strong. Lipopeptide production was predicted to reach 709.87 mg/L after a 60 h incubation using an optimum fermentation medium composed of glucose 7.5 g/L, peanut oil 1.25 g/L,
0.75 g/L, monosodium glutamate 6.75 g/L, yeast extract and
(5:3 w/w) 10 g/L, KCl 0.16 g/L,
0.76 mg/L, and an initial pH of 7.0. Lipopeptide production (
mg/L) in the optimized medium confirmed the validity of the predicted model.
Properties and Functions of Melanin Pigment from Klebsiella sp. GSK
Sajjan, Shrishailnath S. ; Anjaneya, O ; Kulkarni, Guruprasad B. ; Nayak, Anand S. ; Mashetty, Suresh B. ; Karegoudar, T.B. ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 60~69
DOI : 10.4014/kjmb.1210.10002
Purified melanin pigment from Klebsiella sp. GSK was characterized by thermogravimetric, differential thermal, X-ray diffraction and elemental analysis. This melanin pigment is structurally amorphous in nature. It is thermally stable up to
and emits a strong exothermic peak at
. Its carbon, hydrogen and nitrogen composition is 47.9%, 6.9% and 12.0%, respectively. It was used to scavenge metal ions and free radicals. After immobilizing the pigment and using it to adsorb copper and lead ions, the metal ion adsorption capacity was evaluated by atomic absorption spectroscopy (AAS) and the identity of melanin functional groups involved in the binding of metal ions was determined by Fourier transform infrared (FT-IR) spectroscopy. Batch adsorption studies showed that 169 mg/g of copper and 280 mg/g of lead were adsorbed onto melanin-alginate beads. The metal ion adsorption capacity of the melanin-alginate beads was relatively significant compared to alginate beads. The metal ion desorption capacity of HCl was greater (81.5% and 99% for copper and lead, respectively) than that of EDTA (80% and 71% for copper and lead, respectively). The ability of the melanin pigment to scavenge free radicals was evaluated by inhibition of the oxidation of 2,2-diphenyl-1-picrylhydrazyl (DPPH) and 2,2'-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) and was shown to be about 74% and 98%, respectively, compared with standard antioxidants.
Effectiveness of Antagonistic Bacterial Metabolites to Control Rhizoctonia solani on Lettuces and Fusarium oxysporum on Tomatoes
Vu, Van Hanh ; Thi, Quyen Dinh ; Rita, Grosch ; Dung, Nguyen Ngoc ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 70~78
DOI : 10.4014/kjmb.1207.07004
Rhizoctonia solani and Fusarium oxysporum cause yield losses in numerous economically important crops. To develop a bio-control agent, cell free extracellular compounds (ECs) of 5 bacterial strains Burkholdria sp. L1, Pseudomonas sp. L4, Pseudomonas chlororaphis VN391, Bacillus subtilis VN21 and Enterobacter sp. VN99 from Vietnamese fields, which reduced levels of R. solani root rot in lettuces and F. oxysporum root rot in tomatoes, were investigated. In a growth chamber, ECs of all antagonists markedly enhanced the biomass of lettuces (10 to 14.1%) and tomatoes (11.38 to 13.88%). In greenhouses, the disease's severity on both crops treated with ECs of the antagonists was reduced significantly and biomass losses in the plants decreased markedly. The reduction level of R. solani root rot in lettuces was 75, 66.7, 50, and 16.7% by ECs of strains L1, L4, VN21 and VN391, respectively. The biomass of lettuces increased markedly by 29.13%, 21.67%, and 23.4% by ECs of strains L1, L4 and VN21, respectively. Similarly, the reduction levels of F. oxysporum root rot in tomatoes was 76.3, 75, 41.7 and 25% by ECs of strain L1, L4, VN21 and VN391, respectively, and the biomass was significantly enhanced by 14.42, 12.7 and 13%, respectively. The ECs of strain L1 exhibited the most effective bio-control agents to suppress R. solani and F. oxysporum.
Role of Unstable Phenanthrene-Degrading Pseudomonas species in Natural Attenuation of Phenanthrene-Contaminated Site
Prakash, Om ; Lal, Rup ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 79~87
DOI : 10.4014/kjmb.1207.07011
An unstable yet efficient phenanthrene-degrading bacterium strain Ph-3 was isolated from a petroleum-contaminated site at the Mathura Oil Refinery, India. The strain was identified as Pseudomonas sp. using a polyphasic approach. An analysis of the intermediates and assays of the degradative enzymes from a crude extract of phenanthrene-grown cells showed a novel and previously unreported pattern of 1, 2-dihydroxy naphthalene and salicylic acid production. While strain Ph-3 lost its phenanthrene- degrading potential during successive transfers on a rich medium, it maintained this trait in oligotrophic soil conditions under the stress of the pollutant and degraded phenanthrene efficiently in soil microcosms. Although the maintenance and in vitro study of unstable phenotypes are difficult and such strains are often missed during isolation, purification, and screening, these bacteria constitute a substantial fraction of the microbial community at contaminated sites and play an important role in pollutant degradation during biostimulation or monitored natural attenuation.
Antibiofilm Activity of Scutellaria baicalensis through the Inhibition of Synthesis of the Cell Wall (1, 3)-
Kim, Younhee ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 88~95
DOI : 10.4014/kjmb.1209.09003
Candida biofilms are self-organized microbial communities growing on the surfaces of host tissues and medical devices. These biofilms have been displaying increasing resistance against conventional antifungal agents. The roots of Scutellaria baicalensis have been widely used for medicinal purpose throughout East Asia. The aim of the present study was to evaluate the effect of S. baicalensis aqueous extract upon the preformed biofilms of 10 clinical C. albicans isolates, and assess the mechanism of the antibiofilm activity. Its effect on preformed biofilm was judged using an XTT reduction assay and the metabolic activity of all tested strains were reduced (
%) at MIC values. The S. baicalenis extract inhibited (1, 3)-
-D-glucan synthase activity. The effect of S. baicalensis on the morphology of C. albicans was related to the changes in growth caused by inhibiting glucan synthesis; most cells were round and swollen, and cell walls were densely stained or ruptured. The anticandidal activity was fungicidal, and the extract also arrested C. albicans cells at
. The data suggest that S. baicalensis has multiple fatal effects on target fungi, which ultimately result in cell wall disruption and killing by inhibiting (1, 3)-
-D-glucan synthesis. Therefore, S. baicalensis holds great promise for use in treating and eliminating biofilm-associated Candida infections.
Antibiotic Resistance and Bacterial Biofilm Formation by Staphylococcus aureus Strains Isolated from Various Foods
Lee, Joo-Young ; Wang, Hae-Jin ; Shin, Dong-Bin ; Cho, Yong-Sun ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 96~104
DOI : 10.4014/kjmb.1208.08012
Staphylococcus aureus is a major human pathogen that produces a wide array of toxins, leading to a number of adverse symptoms. We examined 275 strains of Staphylococcus aureus isolated from various foods between 2006 and 2008 for antimicrobial susceptibility. At least 259 (94.2%) of the tested strains showed antibiotic resistant properties, and 106 (40.7%) of them showed multiple antibiotic resistance. Eleven of the tested strains were resistant to oxacillin and mec A-positive. Moreover, oxacillin-resistant strains were significantly more likely to be multi-drug resistant (p < 0.01). Of the 275 isolates tested, 24.4% were noted as being positive for slime production and 30.5% were positive for biofilm assay. Antibiotic resistance was not associated with a significantly higher prevalence of biofilm formation. Twenty strains were classified using the DiversiLab system. Most of the strains could be classified into 2 clusters and 4 unique types. All 10 mec A-positive strains (cluster I) were grouped together into the same sub-cluster. Cluster II (6 strains) was not found to be resistant to oxacillin in this study. Although the prevalence of methicillin-resistant S. aureus in food is currently low, the risk of its transmission through the food chain cannot be disregarded.
Root Colonization and Quorum Sensing of the Antagonistic Bacterium Pseudomonas fluorescens 2112 involved in the Red-pepper Rhizosphere
Jung, Byung-Kwon ; Kim, Yo-Hwan ; Kim, Sang-Dal ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 105~111
DOI : 10.4014/kjmb.1210.10006
Biofilm formation of multifunctional plant growth promoting rhizobacterium (PGPR), Pseudomonas fluorescens 2112 is necessary for P. fluorescens 2112 to have a positive impact on the rhizosphere of red-pepper. This study investigated whether signal molecules of the quorum sensing AHLs are produced in order to confirm biofilm formative ability. Through the use of Petri dish bioassays a blue circle formed evidence of AHLs. It was confirmed that P. fluorescens 2112 produced six-carbon-chain-long AHLs by TLC bioassay. The bacterial density of P. fluorescens 2112 on the top and bottom of pepper plant roots was estimated as
CFU/g root, respectively. P. fluorescens 2112 exist more with high-density of
CFU/g soil at a depth of 1 cm but at a low-density of
CFU/g soil at a depth of 5 cm, from the surface of rhizosphere soil. In addition, biofilm formation of P. fluorescens 2112 on the epidermises and the tips of the red-pepper roots were confirmed visually by SEM. Thus, the production of AHLs by P. fluorescens 2112 brings about quorum sensing signaling and the formation of biofilm on the roots which has a positive effect on economically important crops such as red-pepper by additionally producing a variety of antifungal substances and auxin.
Lipase Inhibitory Mode of Dieckol Isolated from Eisenia bicyclis Ethanol Extract
Jung, Seul-A ; Kim, Koth-Bong-Woo-Ri ; Kim, Dong-Hyun ; Cho, Ji-Young ; Kim, Tae-Wan ; Ahn, Dong-Hyun ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 112~118
DOI : 10.4014/kjmb.1210.10003
This study was performed to investigate the possible use of Eisenia bicyclis (EB) ethanol extract to inhibit activity against lipase. In tests, the lipase inhibitory activity of EB ethanol extract was noted as being 43, 27, and 24% at concentrations of 5, 2.5, and 1 mg/ml, respectively. Isolation was carried out by liquid and liquid extraction, silica-gel column chromatography, and HPLC. The results showed that the lipase inhibitory activity of the ethyl acetate (EA) fraction from EB ethanol extract exhibited the strongest lipase inhibitory activity with an
value of 1.31 mg/ml. The EA fraction was separated using silica-gel column chromatography and we obtained 22 sub-fractions. Amongst them, the EA1 fraction showed the highest lipase inhibitory activity with an
value of 0.54 mg/ml. Eight peaks were obtained from the EA1 fraction by HPLC. Fraction 5 also showed a strong lipase inhibitory activity with an
value of 0.37 mg/ml. The fraction 5 was identified as dieckol and the inhibition pattern analyzed from Lineweaver-Burk plots revealed a non-competitive inhibitor. These results suggest that EB has potential as a natural anti-obesity agent.
Selection of Acid-tolerant and Hetero-fermentative Lactic Acid Bacteria Producing Non-proteinaceous Anti-bacterial Substances for Kimchi Fermentation
Kim, Hye-Rim ; Lee, Jong-Hoon ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 119~127
DOI : 10.4014/kjmb.1211.11005
Twenty-three strains of Leuconostoc species and 45 strains of Weissella species inhibiting the growth of Lactobacillus sakei, one of the most populous lactic acid bacteria in over-ripened kimchi, were isolated from kimchi in our previous study. Among these hetero-fermentative 68 strains, Leuconostoc mesenteroides CK0128, Weissella cibaria CK0633, and W. cibaria KK0797 exhibited a relatively high survival rate in MRS medium, which was adjusted to pH 4.3 using an acid mixture consisting of acetic and lactic acids, and produced a large amount of exopolysaccharides. The culture supernatants of 3 strains were fractionated by a molecular weight cutter and lyophilized. The fractions with a molecular weight smaller than 3,000 Da showed antagonistic activity against Staphylococcus aureus and Lb. sakei. The anti-bacterial substances were very stable to heat treatments (
, 15 min) and active at acidic conditions below pH 5.
-Amylase, lipase, and proteolytic enzymes (proteinase K and pepsin) did not affect their activities. These non-proteinaceous anti-bacterial substances inhibited the growth of several food pathogens.
Characterization of the Salmonella typhi Outer Membrane Protein C
Toobak, Hoda ; Rasooli, Iraj ; Gargari, Seyed Latif Mousavi ; Jahangiri, Abolfazl ; Nadoushan, Mohammadreza Jalali ; Owlia, Parviz ; Astaneh, Shakiba Darvish Alipour ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 128~134
DOI : 10.4014/kjmb.1207.07009
Salmonella enterica serovar typhi, a Gram-negative food-borne pathogen, causes typhoid fever in humans. OmpC is an outer membrane porin of S. typhi expressed throughout the infection period. OmpC is potentially an attractive antigen for multivalent vaccines and diagnostic kit designs. In this study we combined in silico, in vitro and in vivo approaches to analyze various aspects of OmpC's antigenic properties. The conserved region, in addition to secondary and tertiary structures, and linear B cell epitopes, were predicted. A number of results obtained from in silico analyses were validated by experimental studies. OmpC was amplified, cloned and then expressed, with the recombinant protein then being purified. BALB/c mice were immunized by purified denatured OmpC. The titer of antibody was raised. Results of challenges with the pathogen revealed that the immunity is non-protective. Most of the theoretical and experimental results were in consensus. Introduced linear B cell epitopes can be employed for the design of diagnostic kits based on antigen-antibody interactions.
Antioxidative, and Inhibitory Activities on Melanogenesis of Vitex negundo L. Leaf Extract
Kim, A Reum ; Park, Su Ah ; Ha, Ji Hoon ; Park, Soo Nam ;
Microbiology and Biotechnology Letters, volume 41, issue 1, 2013, Pages 135~144
DOI : 10.4014/kjmb.1211.11004
The aim of this study was to evaluate various aspects of Vitex negundo L. leaf extract, such as the antioxidative activity, tyrosinase inhibitory effects, and inhibitory activities on
-MSH induced melanogenesis, and active component analysis. The DPPH (1, 1-diphenyl-2-picrylhydrazyl) scavenging activities (
) of the ethyl acetate fraction and aglycone fraction of V. negundo L. leaf extract were
, respectively. A luminol-dependent chemiluminescence assay revealed that the reactive oxygen species (ROS) scavenging activity (
) of the aglycone fraction of V. negundo L. leaf extract on ROS generated in an
system was the most prominent at
. The protective effects of the extracts fractions of V. negundo L. leaf against the rose-bengal sensitized photohemolysis of human erythrocytes were increased in a concentration dependent manner (
). In particular, there were greater protective effects of the aglycone fraction on the cellular membrane than that of the fat-soluble antioxidant (+)-
-tocopherol. The inhibitory effects (
) on mushroom tyrosinase were the highest for the ethyl acetate fraction (
). The inhibitory effect on
-MSH induced melanogenesis in B16 melanoma cells was 41.80% at
of ethyl acetate fraction. Active component analyses by TLC, HPLC and LC/ESI-MS revealed luteolin and isoorientin. These results indicate that V. negundo L. leaf extract can be used as an antioxidant for ROS scavenging. Particularly, the luteolin and isoorientin of the ethyl acetate fraction may be applicable to new whitening cosmetics because of its inhibitory effect on mushroom tyrosinase and
-MSH induced melanogenesis in B16 melanoma cells.