Byung Taek Oh

Potential use of Pseudomonas koreensis AGB-1 in association with Miscanthus sinensis to remediate heavy metal(loid)-contaminated mining site soil

Endophytic bacteria have the potential to promote plant growth and heavy metal(loid) (HM) removal from contaminated soil. Pseudomonas koreensis AGB-1, isolated from roots of Miscanthus sinensis growing in mine-tailing soil, exhibited high tolerance to HMs and plant growth promoting traits. Transmission electron microscope (TEM) analysis revealed that AGB-1 sequestered HMs extracellularly and their accumulation was visible as dark metal complexes on bacterial surfaces and outside of the cells. DNA sequencing of HM resistance marker genes indicated high homology to the appropriate regions of the arsB, ACR3(1), aoxB, and bmtA determinants. Inoculating mining site soil with AGB-1 increased M. sinensis biomass by 54%, chlorophyll by 27%, and protein content by 28%. High superoxide dismutase and catalase activities, and the lower malondialdehyde content of plants growing in AGB-1-inoculated soil indicate reduced oxidative stress. Metal(loid) concentrations in roots and shoots of plants grown in inoculated soil were higher than those of the controls in pot trials with mine tailing soil. Results suggest that AGB-1 can be used in association with M. sinensis to promote phytostabilization and remediation of HM-contaminated sites.

Remediating TNT-contaminated soil by soil washing and Fenton oxidation

Abstract Past disposal practices at munitions production facilities have generated numerous hectares of TNT-contaminated soil. We previously showed that Fenton oxidation could destroy TNT in soil and water. Our objective was to determine the potential of combining Fenton oxidation with soil washing to remediate TNT-contaminated soil and to test whether plants could be established on washed soil. Aqueous soil washing effectively reduced CH3CN-extractable TNT concentrations but large volumes of water were required to meet remediation goals. This volume was reduced when the wash water temperature was increased to 45°C. Complete destruction of TNT in wash solutions was achieved by Fenton oxidation with greater than 40% mineralized. Combining soil washing with phytoremediation will require plant establishment on the washed soil. We observed no significant reduction in tall fescue (Festuca arundinacea Schreb.) germination or early seedling development in contaminated soil following two wash cycles (soil:H2O, 1:5). This corresponded to reducing CH3CN-extractable soil concentrations from 499 mg TNT kg−1 to approximately 72 mg TNT kg−1. Our results indicate Fenton oxidation can be combined with soil washing for effective abiotic remediation of TNT-contaminated soils. Washed soils may be planted to tall fescue, demonstrating the potential of combining soil washing, Fenton oxidation and phytoremediation into an integrated treatment train for TNT-contaminated sites.

Monascus pigment production by solid-state fermentation with corn cob substrate

Natural pigments are an important alternative to potentially harmful synthetic dyes. We investigated the feasibility of corn cob powder as a substrate for production of pigments by Monascus purpureus KACC 42430 in solid-state fermentation. A pigment yield of 25.42 OD Units/gram of dry fermented substrate was achieved with corn cob powder and optimized process parameters, including 60% (w/w) initial moisture content, incubation at 30°C, inoculation with 4mL of spores/gram of dry substrate, and an incubation period of 7 days. Pigment yield using corn cobs greatly exceeded those of most other agricultural waste substrates. The pigments were stable at acidic pH, high temperatures, and in salt solutions; all important considerations for industrial applications. Our results indicate the viability of corn cob substrate in combination with M. purpureus for industrial applications.

Simultaneous removal of phenol, Cu and Cd from water with corn cob silica-alginate beads

Phenol and heavy metals in petroleum waste are environmental and human health concerns, but physicochemical removal is often cost-prohibitive and can produce toxic secondary products and treatment residues. An environmentally benign alternative combines corn cob silica with alginate and immobilized bacteria into beads for treating contaminated water. The concentration of phenol was decreased >92% by Pseudomonas putida YNS1 on aliginate-silica beads (2%, w/v) after equilibrating for 96h with water containing 214mg phenol/L. GC-MS analysis indicated formation of benzoquinone and other polar products. Beads containing corn cob silica decreased Cu concentrations by 84-88% and Cd by 83-87% within 24h. In a mixture of 114mg phenol, 43mg Cu and 51mg Cd/L, phenol removal (93% within 96h) only occurred with beads containing the silica and bacterial strain. Beads containing corn cob silica removed >97% of the Cu and >99% of the Cd, critical for reducing toxicity to the bacteria. Beads with the immobilized strain removed phenol when zeolite was used instead of corn cob silica, but beads with silica were more effective for Cu and Cd removal. Results show the potential of corn cob silica combined with alginate and immobilized bacteria for removing phenol and heavy metals from contaminated water.

Trichoderma sp. PDR1-7 promotes Pinus sylvestris reforestation of lead-contaminated mine tailing sites.

Vegetation is critical to stabilize and remediate mine tailing sites, but plant growth is often poor due to toxicity from heavy metal(loid)s (HMs). A non-symbiotic endophytic fungus, Trichoderma sp. PDR1-7, isolated from Pb-contaminated mine tailing soil, exhibited both high tolerance to HMs and desirable plant growth-promoting characteristics. PDR1-7 promoted HM solubilization in mine tailing soil and removed significant amounts of Pb and other HMs from liquid media containing single and multiple metals. Pb removal efficiency increased with initial pH from 4 to 6 and with Pb concentration from 100 to 125 mg L(-1). Inoculating soil with PDR1-7 significantly increased nutrient availability and seedling growth, chlorophyll and protein contents, as well as antioxidative enzyme (superoxide dismutase) activity. A decrease in malondialdehyde indicated less oxidative stress. HM concentrations were much higher in Pinus sylvestris roots when PDR1-7 was present. These observations suggest the utility of Trichoderma sp. PDR1-7 for pine reforestation and phytoremediation of Pb-contaminated mine soil.

Aerobic TNT reduction via 2-hydroxylamino-4,6-dinitrotoluene by Pseudomonas aeruginosa strain MX isolated from munitions-contaminated soil

Bioremediation of munitions-contaminated soil requires effective transformation and detoxification of high concentrations of 2,4,6-trinitrotoluene (TNT). Pseudomonas aeruginosa strain MX, isolated from munitions-contaminated soil, aerobically transformed TNT (100 mg/L) in culture medium within 15 h, causing transient accumulation of hydroxylaminodinitrotoluenes (HADNTs). The predominance of 2-hydroxylamino-4,6-dinitrotoluene (2HADNT), as well as 2-amino-4,6-dinitrotoluene (2ADNT) and 4,4′,6,6′ -tetranitro-2,2′ -azoxytoluene (2,2′AZT), indicated preferential reduction of the TNT ortho nitro group. While only 12% of the TNT was transformed to 2ADNT, up to 65% was transformed to tetranitroazoxytoluenes (AZTs), which accumulated as a precipitate. The precipitate was formed by microscopic particles adhering to bacterial cells, which subsequently formed clusters containing lysed cells. Toxicity toward bacteria was primarily attributed to 2ADNT, because pure AZTs preincubated with sterile medium had little effect on the strain. While the culture medium containing TNT exhibited toxicity toward corn (Zea mays L.) and witchgrass (Panicum capillare L.), little phytotoxicity was observed after incubating with P. aeruginosa strain MX for 4 d. Strong binding of HADNTs to soil and low AZT bioavailability may further promote the detoxification of TNT in soil.

Biodegradation of BTEX mixture by Pseudomonas putida YNS1 isolated from oil‐contaminated soil

The presence of mixed contaminants, such as BTEX (benzene, toluene, ethylbenzene and xylene isomers) can affect the biodegradation, fate and environmental impacts of each compound. To understand the influence of interactions among BTEX compounds on their biodegradation, four bacteria were isolated from oil‐contaminated soil and assayed for BTEX biodegradation in vitro. The isolate exhibiting maximum biodegradation was identified as Pseudomonas putida based on the 16S rDNA sequence. The biodegradation of the BTEX compounds was greatly influenced by pH, temperature, and salinity. Substrate mixture studies (binary, tertiary and quaternary) revealed that the presence of toluene increased the biodegradation rate of benzene, ethylbenzene, and xylene.

IAA production by Bacillus sp. JH 2-2 promotes Indian mustard growth in the presence of hexavalent chromium

Bacillus sp. strain JH 2‐2, isolated from the rhizosphere of plants at a multi‐metal contaminated mine site, has the potential to reduce Cr(VI) to Cr(III) and promote plant growth by reducing Cr toxicity and producing IAA. The minimum inhibitory concentration of Cr(VI) to Bacillus sp. JH 2‐2 was 1000 mg L−1 and the strain reduced 99% of 10 mg Cr(VI) L−1 to Cr(IV) within 24 h. Lower Cr(VI) stress (10 mg L−1) stimulated IAA production, but much less IAA was produced at 30 or 50 mg Cr(VI) L−1. Inoculation with Bacillus sp. JH 2‐2 increased the length of Brassica juncea L. roots by 364% and stems by 735% in the presence of 10 mg Cr(VI) L−1 from those of uninoculated control plants. These findings suggest potential use of Bacillus sp. JH 2‐2 to promote phytoremediation of soil contaminated with Cr(VI).

Fluorinated TiO2 as an ambient light-activated virucidal surface coating material for the control of human norovirus

We evaluated the virucidal efficacy of light-activated fluorinated TiO₂ surface coatings on human norovirus and several surrogates (bacteriophage MS2, feline calcivirus (FCV), and murine norovirus (MNV)). Inactivation of viruses on surfaces exposed to a common fluorescent lamp was monitored and the effects of UVA intensity, temperature, and fluoride content were assessed. Destruction of RNA and capsid oxidation were evaluated for human norovirus inocula on the F-TiO₂ surfaces, while contact with the F-TiO₂ surface and exposure to residual UVA radiation of 10 μW cm(-2) for 60 min resulted in infectivity reductions for the norovirus surrogates of 2-3 log₁₀. Infectivity reductions on pristine TiO₂ surfaces in identical conditions were over 2 orders of magnitude lower. Under realistic room lighting conditions, MS2 infectivity declined below the lower detection limit after 12h. Reductions in RNA were generally low, with the exception of GII.4, while capsid protein oxidation likely played a larger role in infectivity loss. Inactivation of norovirus surrogates occurred significantly faster on F-TiO₂ compared to pristine TiO₂ surfaces. The material demonstrated antiviral action against human norovirus surrogates and was shown to effectively inhibit MS2 when exposed to residual UVA present in fluorescent room lighting conditions in a laboratory setting.

Pseudomonas fluorescens JH 70-4 promotes Pb stabilization and early seedling growth of Sudan grass in contaminated mining site soil

A bacterial strain (JH 70-4) exhibiting plant growth promoting characteristics (indoleacetic acid production and 1-aminocyclopropane-1-carboxylate deaminase activity), as well as heavy metal(loid) (HM) tolerance and Pb precipitation, was isolated from HM-contaminated soil at an abandoned mine site. The bacterium was identified as Pseudomonas fluorescens based on 16S rDNA sequencing. The JH 70-4 strain induced precipitation of Pb as PbS nanoparticles, confirmed by X-ray diffraction. Solution pH, incubation time, and Pb concentration influenced removal and PbS formation. Inoculating contaminated soil with JH 70-4 decreased Pb availability; exchangeable Pb decreased while organic- and sulphide-bound Pb increased. The toxicity characteristic leaching procedure showed a 65% decrease in Pb in leachate 60 d after inoculating soil with JH 70-4. Shoot and root lengths of Sudan grass grown in the inoculated soil were greater than in the uninoculated soil. Findings suggest that microbial Pb fixation is a viable strategy for remediating soil and promoting plant growth for phytostabilization of contaminated sites.