Elucidating Multiple Electron-Transfer Pathways for Metavanadate Bioreduction by Actinomycetic Streptomyces microflavus.

While microbial reduction has gained widespread recognition for efficiently remediating environments polluted by toxic metavanadate [V(V)], the pool of identified V(V)-reducing strains remains rather limited, with the vast majority belonging to bacteria and fungi. This study is among the first to confirm the V(V) reduction capability of Streptomyces microflavus, a representative member of ubiquitous actinomycetes in environment. A V(V) removal efficiency of 91.0 ± 4.35% was achieved during 12 days of operation, with a maximum specific growth rate of 0.073 d-1. V(V) was bioreduced to insoluble V(IV) precipitates. V(V) reduction took place both intracellularly and extracellularly. Electron transfer was enhanced during V(V) bioreduction with increased electron transporters. The electron-transfer pathways were revealed through transcriptomic, proteomic, and metabolomic analyses. Electrons might flow either through the respiratory chain to reduce intracellular V(V) or to cytochrome c on the outer membrane for extracellular V(V) reduction. Soluble riboflavin and quinone also possibly mediated extracellular V(V) reduction. Glutathione might deliver electrons for intracellular V(V) reduction. Bioaugmentation of the aquifer sediment with S. microflavus accelerated V(V) reduction. The strain could successfully colonize the sediment and foster positive correlations with indigenous microorganisms. This study offers new microbial resources for V(V) bioremediation and improve the understanding of the involved molecular mechanisms.

Woodchip-sulfur based mixotrophic biotechnology for hexavalent chromium detoxification in the groundwater.

This study investigated groundwater hexavalent chromium (Cr(VI)) decontamination by a biological permeable reactive barrier (bio-PRB), where a woodchip-elemental sulfur [S(0)] based mixotrophic process was established. 89.0 ± 0.27% of Cr(VI) was removed from the synthetic groundwater after 72 h at a concentration of 50 mg/L during the preliminary batch experiment. The impact of geochemical and hydrodynamic conditions Cr(VI) removal was investigated in the bio-PRB over 225 days. Although elevated Cr(VI) (i.e., 75 mg/L), addition of nitrate and short hydraulic retention time reduced the Cr(VI) removal, 87.2 ± 2.09% of Cr(VI) removal was accomplished. Characterization of the solids indicated that the soluble Cr(VI) was converted and immobilized as the insoluble trivalent chromium [Cr(III)]. 16S rRNA gene based sequencing results suggested that micromolecules produced by woodchip cellulose hydrolyzing- and sulfur oxidizing bacteria were further used by functional Cr(VI) removal bacteria (e.g., Geobacteraceae and Pseudomonas). The extracellular protein and humic-like substances in extracellular polymeric substances (EPS) can bind toxic Cr(VI) through carboxyl and hydroxyl groups, and reduce Cr(VI) in an enzymatic manner. The preliminary finding of this study provided a potential way to utilize agricultural waste for in-situ Cr(VI) contaminated-groundwater remediation.

Novel Pathway for Vanadium(V) Bio-Detoxification by Gram-Positive Lactococcus raffinolactis.

Whereas prospects of bioremediation for a vanadium(V) [V(V)]-contaminated environment are widely recognized, reported functional species are extremely limited, with the vast majority of Gram-negative bacteria in Proteobacteria. Herein, the effectiveness of V(V) reduction is proved for the first time by Lactococcus raffinolactis, a Gram-positive bacterium in Firmicutes. The V(V) removal efficiency was 86.5 ± 2.17% during 10-d operation, with an average removal rate of 4.32 ± 0.28 mg/L·d in a citrate-fed system correspondingly. V(V) was bio-reduced to insoluble vanadium(IV) and distributed both inside and outside the cells. Nitrite reductase encoded by gene nirS mainly catalyzed intracellular V(V) reduction, revealing a previously unrecognized pathway. Oxidative stress induced by reactive oxygen species from dissimilatory V(V) reduction was alleviated through strengthened superoxide dismutase and catalase activities. Extracellular polymeric substances with chemically reactive hydroxyl (-OH) and carboxyl (-COO-) groups also contributed to V(V) binding and reduction as well as ROS scavenging. This study can improve the understanding of Gram-positive bacteria for V(V) bio-detoxification and offer microbial resources for bioremediation of a V(V)-polluted environment.

Multiple pathways of vanadate reduction and denitrification mediated by denitrifying bacterium Acidovorax sp. strain BoFeN1.

Contamination of aquifers by a combination of vanadate [V(V)] and nitrate (NO3-) is widespread nowadays. Although bioremediation of V(V)- and nitrate-contaminated environments is possible, only a limited number of functional species have been identified to date. The present study demonstrates the effectiveness of V(V) reduction and denitrification by a denitrifying bacterium Acidovorax sp. strain BoFeN1. The V(V) removal efficiency was 76.5 ± 5.41 % during 120 h incubation, with complete removal of NO3- within 48 h. Inhibitor experiments confirmed the involvement of electron transport substances and denitrifying enzymes in the bioreduction of V(V) and NO3-. Cyt c and riboflavin were important for extracellular V(V) reduction, with quinone and EPS more significant for NO3- removal. Intracellular reductive compounds including glutathione and NADH directly reduce V(V) and NO3-. Reverse transcription quantitative PCR confirmed the important roles of nirK and napA genes in regulating V(V) reduction and denitrification. Bioaugmentation by strain BoFeN1 increased V(V) and NO3- removal efficiency by 55.3 % ± 2.78 % and 42.1 % ± 1.04 % for samples from a contaminated aquifer. This study proposes new microbial resources for the bioremediation of V(V) and NO3-contaminated aquifers, and contributes to our understanding of coupled vanadium, nitrogen, and carbon biogeochemical processes.

Dynamics of vertical vanadium migration in soil and interactions with indigenous microorganisms adjacent to tailing reservoir.

Severe vanadium pollution in deep soil through surface infiltration during mining activities has been particularly concerned, but little is known about vanadium migration dynamics in vertical soil profile. Indigenous microorganisms widely exist in soil, however, their functions and suffered impacts during vertical vanadium migration have rarely been investigated. In this study, 100 cm height columns were constructed with undisturbed soil around vanadium tailing reservoir were constructed to describe vertical vanadium transport process and corresponding interactions between vanadium and indigenous microorganisms. 91 d continuous leaching with pentavalent vanadium [V(V)] showed that V(V) gradually downward migrated. Soil microorganisms slowed down vertical V(V) migration rate by transferring V(V) to insoluble tetravalent vanadium. Enriched Gemmatimonadaceae and Actinobacteria were identified to contribute to microbial V(V) transformation. Co-existing nitrate weakened the soil's ability to intercept V(V) via electron competition. Microbial communities were reshaped by vanadium during leaching, while enzyme activities increased slightly due to vanadium stimulation. This work advances the understanding of vertical vanadium migration characteristics in soil, which is essential to risk management and effective remediation of vanadium-polluted sites.