caa a22 4500 605501440 CHVBK 20210128100600.0 cr unu---uuuuu 210128e20150901xx s 000 0 eng 10.1007/s00253-015-6606-1 doi (NATIONALLICENCE)springer-10.1007/s00253-015-6606-1 Elucidating carbon uptake from vinyl chloride using stable isotope probing and Illumina sequencing [Elektronische Daten] [Fernanda Paes, Xikun Liu, Timothy Mattes, Alison Cupples] Vinyl chloride (VC), a known human carcinogen, is a common and persistent groundwater pollutant at many chlorinated solvent contaminated sites. The remediation of such sites is challenging because of the lack of knowledge on the microorganisms responsible for in situ VC degradation. To address this, the microorganisms involved in carbon assimilation from VC were investigated in a culture enriched from contaminated site groundwater using stable isotope probing (SIP) and high-throughput sequencing. The mixed culture was added to aerobic media, and these were amended with labeled (13C-VC) or unlabeled VC (12C-VC). The cultures were sacrificed on days 15, 32, and 45 for DNA extraction. DNA extracts and SIP ultracentrifugation fractions were subject to sequencing as well as quantitative PCR (qPCR) for a functional gene linked to VC-assimilation (etnE). The gene etnE encodes for epoxyalkane coenzyme M transferase, a critical enzyme in the pathway for VC degradation. The relative abundance of phylotypes was compared across ultracentrifugation fractions obtained from the 13C-VC- and 12C-VC-amended cultures. Four phylotypes were more abundant in the heavy fractions (those of greater buoyant density) from the 13C-VC-amended cultures compared to those from the 12C-VC-amended cultures, including Nocardioides, Brevundimonas, Tissierella, and Rhodoferax. Therefore, both a previously identified VC-assimilating genus (Nocardioides) and novel microorganisms were responsible for carbon uptake. Enrichment of etnE with time was observed in the heavy fractions, and etnE sequences illustrated that VC-assimilators harbor similar Nocardioides-like etnE. This research provides novel data on the microorganisms able to assimilate carbon from VC. Springer-Verlag Berlin Heidelberg, 2015 SIP nationallicence Vinyl chloride nationallicence etnE nationallicence Nocardioides nationallicence Brevundimonas nationallicence Tissierella nationallicence Rhodoferax nationallicence Paes Fernanda Department of Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut Liu Xikun Department of Civil and Environmental Engineering, University of Iowa, 52242, Iowa City, IA, USA aut Mattes Timothy Department of Civil and Environmental Engineering, University of Iowa, 52242, Iowa City, IA, USA aut Cupples Alison Department of Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7735-7743 0175-7598 99:18<7735 2015 99 253 https://doi.org/10.1007/s00253-015-6606-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 research-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-015-6606-1 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- Paes Fernanda Department of Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 700 1- Liu Xikun Department of Civil and Environmental Engineering, University of Iowa, 52242, Iowa City, IA, USA aut NATIONALLICENCE 700 1- Mattes Timothy Department of Civil and Environmental Engineering, University of Iowa, 52242, Iowa City, IA, USA aut NATIONALLICENCE 700 1- Cupples Alison Department of Civil and Environmental Engineering, Michigan State University, 48824, East Lansing, MI, USA aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/18(2015-09-01), 7735-7743 0175-7598 99:18<7735 2015 99 253