Characterization of arsenate transformation and identification of arsenate reductase in a green alga Chlamydomonas reinhardtii

Arsenic （As） is a pervasive and ubiquitous environmental toxin that has created catastrophic human health problems world-wide. Chlamydomonas reinhardtii is a unicellular green alga, which exists ubiquitously in freshwater aquatic systems. Arsenic metabolism processes of this alga through arsenate reduction and sequent store and efflux were investigated. When supplied with 10 μmol/L arsenate, arsenic speciation analysis showed that arsenite concentration increased from 5.7 to 15.7 mg/kg dry weight during a 7-day period, accounting for 18%–24% of the total As in alga. When treated with different levels of arsenate （10, 20, 30, 40, 50 μmol/L） for 7 days, the arsenite concentration increased with increasing external arsenate concentrations, the proportion of arsenite was up to 23%–28% of the total As in alga. In efflux experiments, both arsenate and arsenite could be found in the efflux solutions. Additionally, the efflux of arsenate was more than that of arsenite. Furthermore, two arsenate reductase genes of C. reinhardtii （CrACR2s） were cloned and expressed in Escherichia coli strain WC3110 （？arsC） for the first time. The abilities of both CrACR2s genes to complement the arsenate- sensitive strain were examined. CrACR2.1 restored arsenate resistance at 0.8 mmol/L. However, CrACR2.2 showed much less ability to complement. The gene products were demonstrated to reduce arsenate to arsenite in vivo. In agreement with the complementation results, CrACR2.1 showed higher reduction ability than CrACR2.2, when treated with 0.4 mmol/L arsenate for 16 hr incubation.

Arsenic release from microbial reduction of scorodite in the presence of electron shuttle in flooded soil

Scorodite (FeAsO_(4)·H_(2)O) is a common arsenic-bearing (As-bearing) iron mineral in nearsurface environments that could immobilize or store As in a bound state.In flooded soils,microbe induced Fe(Ⅲ) or As(Ⅴ) reduction can increase the mobility and bioavailability of As.Additionally,humic substances can act as electron shuttles to promote this process.The dynamics of As release and diversity of putative As(Ⅴ)-reducing bacteria during scorodite reduction have yet to be investigated in detail in flooded soils.Here,the microbial reductive dissolution of scorodite was conducted in an flooded soil in the presence of anthraquinone-2,6-disulfonate (AQDS).Anaeromyxobacter,Dechloromonas,Geothrix,Geobacter,Ideonella,and Zoogloea were found to be the dominant indigenous bacteria during Fe(Ⅲ) and As(Ⅴ) reduction.AQDS increased the relative abundance of dominant species,but did not change the diversity and microbial community of the systems with scorodite.Among these bacteria,Geobacter exhibited the greatest increase and was the dominant Fe(Ⅲ)-and As(Ⅴ)-reducing bacteria during the incubation with AQDS and scorodite.AQDS promoted both Fe(Ⅲ) and As(Ⅴ) reduction,and over 80%of released As(Ⅴ) was microbially transformed to As(Ⅲ).The increases in the abundance of arrA gene and putative arrA sequences of Geobacter were higher with AQDS than without AQDS.As a result,the addition of AQDS promoted microbial Fe(Ⅲ) and As(Ⅴ) release and reduction from As-bearing iron minerals into the environment.These results contribute to exploration of the transformation of As from As-bearing iron minerals under anaerobic conditions,thus providing insights into the bioremediation of As-contaminated soil.