Hyun Myung

Characterization of lead resistant endophytic Bacillus sp. MN3-4 and its potential for promoting lead accumulation in metal hyperaccumulator Alnus firma

The aim of this study was to isolate and characterize endophytic bacteria from the roots of the metal hyperaccumulator plant Alnus firma. A total of 14 bacterial endophytes were isolated from root samples and assayed for tolerance to heavy metals. Isolate MN3-4 exhibited maximum bioremoval of Pb and was subsequently identified as Bacillus sp. based on 16S rRNA sequences. The pH and initial metal concentration highly influenced the Pb bioremoval rate. The growth of isolate MN3-4 was moderately altered in the presence of metals. Scanning electron microscopy, energy dispersive spectroscopy, biological-transmission electron microscopy, and Fourier transform infrared spectroscopy studies revealed that isolate MN3-4 had extracellularly sequestered the Pb molecules with little intracellular accumulation. Isolate MN3-4 did not harbor pbrA and pbrT genes. Moreover, isolate MN3-4 had the capacity to produce siderophores and indoleacetic acid. A root elongation assay demonstrated an increase (46.25%) in the root elongation of inoculated Brassica napus seedlings compared to that of the control plants. Obtained results pointed out that isolate MN3-4 could potentially reduce heavy metal phytotoxicity and increase Pb accumulation in A. firma plants.

Bioleaching characteristics, influencing factors of Cu solubilization and survival of Herbaspirillum sp. GW103 in Cu contaminated mine soil.

This study was aimed at assess the potential of diazotrophic bacteria, Herbaspirillum sp. GW103, for bioleaching of Cu in mine soil. The strain exhibited resistance to As (550mgL(-1)), Cu (350mgL(-1)), Zn (300mgL(-1)) and Pb (200mgL(-1)). The copper resistance was further confirmed by locating copA and copB genes. The survival of the isolate GW103 during bioleaching was analyzed using green fluorescent protein tagged GW103. Response surface methodology based Box-Behnken design was used to optimize the physical and chemical conditions for Cu bioleaching. Five significant variables (temperature, incubation time, CaCO3, coconut oil cake (COC), agitation rate) were selected for the optimization. Second-order polynomials were established to identify the relationship between Cu bioleaching and variables. The optimal conditions for maximum Cu bioleaching (66%) were 30°C, 60h of incubation with 1.75% of CaCO3 and 3% COC at 140rpm. The results of Cu sequential extraction studies indicated that the isolate GW103 leached Cu from ion-exchangeable, reducible, strong organic and residual fractions. Obtained results point out that the isolate GW103 could be used for bioleaching of Cu from mine soils.

Suppression of Phytophthora blight on pepper (Capsicum annuum L.) by bacilli isolated from brackish environment

Abstract Bacteria of the genus Bacillus are well known to possess antagonistic activity against numerous plant pathogens. In the present study, 11 strains of Bacillus spp. were isolated from a brackish environment and assayed for biocontrol activity under in vitro and in vivo conditions. Among the 11 isolates tested, nine isolates effectively inhibited the growth of various plant pathogens, namely Phytophthora capsici, Phytophthora citrophthora, Phytophthora citricola, Phytophthora sojae, Colletotrichum coccodes, Colletotrichum gloeosporioides, Colletotrichum acutatum, Rhizoctonia solani, Fusarium solani, Fusarium graminearum, Pyricularia spp., and Monilina spp. The effective isolates were further screened for suppression of Phytophthora blight of pepper plants under greenhouse conditions. The isolate SB10 exhibited the maximum (72.2%) ability to reduce the disease incidence and increased (32.2%) the vigour index of Capsicum annuum L. plants. Antifungal compounds produced by isolate SB10 were highly thermostable (100°C for 30 min). Matrix-Assisted Laser Desorption Ionization-Time of Flight mass spectrometry of the antifungal compounds revealed three lipopeptide complexes, namely the surfactins, the iturins, and the fengycins, which are well-known antifungal compounds produced by Bacillus spp.

Lead biotransformation potential of allochthonous Bacillus sp. SKK11 with sesame oil cake extract in mine soil

The potential of allochthonous Bacillus sp. SKK11 and sesame oil cake extract for the immobilization of Pb in mine soil was investigated in this study. The isolate SKK11, isolated from a brackish environment and identified as Bacillus sp. based on partial 16S rDNA sequencing, exhibited maximum resistance to Pb (750 mg L−1). Growth kinetic studies revealed that the presence of oil cake extract (2%) increased the biomass of the isolate SKK11. Transmission electron microscopy and X-ray diffraction studies showed that isolate SKK11 transformed Pb either intracellularly or extracellularly. Selective sequential extraction studies showed that the bioremediation decreased 24.9% of the exchangeable fraction in the mine soil in 3 days. However, 75.1% of the exchangeable fraction was not immobilized in the soil. An X-ray diffractogram of the bioremediated soil showed a major decrease (79.0%) in the intensity of the plagioclase mineral peak. Urease, dehydrogenase, amylase, invertase, cellulase, and alkaline phosphatase enzyme activities were increased in the bioremediated mine soil. These results suggest that the isolate Bacillus sp. SKK11 in combination with sesame oil cake extract could be employed for the immobilization of bioavailable Pb in contaminated soil.

Phytoremediation of heavy metals in contaminated water and soil using Miscanthus sp. Goedae-Uksae 1.

The aim of this study is to characterize the heavy metal phytoremediation potential of Miscanthus sp. Goedae-Uksae 1, a hybrid, perennial, bio-energy crop developed in South Korea. Six different metals (As, Cu, Pb, Ni, Cd, and Zn) were used for the study. The hybrid grass effectively absorbed all the metals from contaminated soil. The maximum removal was observed for As (97.7%), and minimum removal was observed for Zn (42.9%). Similarly, Goedae-Uksae 1 absorbed all the metals from contaminated water except As. Cd, Pb, and Zn were completely (100%) removed from contaminated water samples. Generally, the concentration of metals in roots was several folds higher than in shoots. Initial concentration of metals highly influenced the phytoremediation rate. The results of the bioconcentration factor, translocation factor, and enrichment coefficient tests indicate that Goedae-Uksae 1 could be used for phytoremediation in a marginally contaminated ecosystem.

Eco-friendly approach towards green synthesis of zinc oxide nanocrystals and its potential applications

In the present study, we investigated a novel green route for synthesis of zinc oxide (ZnO) nanocrystals using Prunus × yedoensis Matsumura leaf extract as a reducing agent without using any surfactant or external energy. Standard characterization studies were carried out to confirm the obtained product using UV–Vis spectra, SEM–EDS, FTIR, TEM, and XRD. In addition, the synthesized ZnO nanocrystals were coated onto fabric and leather samples to study their bacteriostatic effect against odor-causing bacteria Brevibacterium linens and Staphylococcus epidermidis. Zinc oxide nanocrystal-coated fabric and leather showed good activity against both bacteria.

Effect of heavy metals on acdS gene expression in Herbaspirillium sp. GW103 isolated from rhizosphere soil.

This study aimed to understand the influence of heavy metals on 1‐aminocyclopropane‐1‐carboxylate deaminase activity (ACCD) and acdS gene expression in Herbaspirillium sp. GW103. The GW103 strain ACCD activity decreased in cells grown in a medium supplemented with Pb and As, whereas cells grown in medium supplemented with Cu showed increase in enzyme activity. The GW103 strain produced 262.2 ± 6.17 μmol of α‐ketobutyrate per milligram of protein per hour during ACC deamination at 25 °C after 24 h incubation. Using a PCR approach, an acdS coding‐gene of 1.06 kbp was amplified in isolate GW103, showing 92% identity with Herbaspirillum seropedicae SmR1 acdS gene. Real time quantitative polymerase chain reaction results indicate that the acdS expression rate was increased (7.1‐fold) in the presence of Cu, whereas it decreased (0.2‐ and 0.1‐fold) in the presence of As and Pb.

Production and characterization of bacterial cellulose by Leifsonia sp. CBNU-EW3 isolated from the earthworm, Eisenia fetida

A total of five bacterial strains were isolated from earthworm, Eisenia fetida and examined for bacterial cellulose (BC) production in Hestrin-Schramm medium (HS). Among the five strains tested, CBNU-EW3 exhibited excellent BC production and was identified as Leifsonia sp. by 16S rDNA sequence analysis. BC production by Leifsonia sp. CBNU-EW3 was optimum at pH 5, 30°C, and with glucose and yeast extract as carbon and nitrogen sources, respectively, according to 15 day-long experiments. (XRD) analysis of the dried pellicle indicated that the BC was partially crystalline type I. Fourier transform infrared spectroscopy (FT-IR) analysis showed that the obtained pellicle contained the same functional groups as typical BC. Field emission scanning electron microscopy (FESEM) images showed that the BC micro-fibril matrix consisted of a flat surface with large pore size and cellulose aggregation.

Inhibitory effects of Euphorbia supina on Propionibacterium acnes -induced skin inflammation in vitro and in vivo

BackgroundEuphorbia supina (ES) plant has been used as treatment for inflammatory conditions. The antibacterial effect and the anti-inflammatory mechanism of ES for Propionibacterium (P.) acnes-induced inflammation in THP-1 cells and acne animal model remain unclear. Therefore, the objective of the present study was to determine the antibacterial and anti-inflammatory activities of ES against P. acnes, the etiologic agent of skin inflammation.MethodThe antibacterial activities of ES were tested with disc diffusion and broth dilution methods. Cytotoxicity of ES at different doses was evaluated by the MTT assay. THP-1 cells were stimulated by heat-killed P. acnes in the presence of ES. The pro-inflammatory cytokines and mRNA levels were measured by ELISA and real-time-PCR. MAPK expression was analyzed by Western blot. The living P. acnes was intradermally injected into the ear of BLBC/c mice. Subsequently, chemical composition of ES was analyzed by liquids chromatography-mass spectrometry (LC-MS).ResultES had stronger antibacterial activity against P. acnes and inhibitory activity on lipase. ES had no significant cytotoxicity on THP-1 cells. ES suppressed the mRNA levels and production of IL-8, TNF-a, IL-1β in vitro. ES inhibited the expression levels of pro-inflammatory cytokines and the MAPK signaling pathway. Ear thickness and inflammatory cells were markedly reduced by ES treatment. Protocatechuic acid, gallic acid, quercetin, and kaempferol were detected by LC-MS analysis in ES.ConclusionsOur results demonstrate antibacterial and anti-inflammatory activities of ES extract against P. acnes. It is suggested that ES extract might be used to treatment anti-inflammatory skin disease.