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Delft University of Technology Desulfonatronospira Sulfatiphila Sp Delft University of Technology Desulfonatronospira sulfatiphila sp. Nov., and Desulfitispora elongata sp. nov., two novel haloalkaliphilic sulfidogenic bacteria from soda lakes Sorokin, Dimitry Y.; Chernyh, Nikolai A. DOI 10.1099/ijsem.0.001640 Publication date 2017 Document Version Accepted author manuscript Published in International Journal of Systematic and Evolutionary Microbiology Citation (APA) Sorokin, D. Y., & Chernyh, N. A. (2017). Desulfonatronospira sulfatiphila sp. Nov., and Desulfitispora elongata sp. nov., two novel haloalkaliphilic sulfidogenic bacteria from soda lakes. International Journal of Systematic and Evolutionary Microbiology, 67(2), 396-401. [001640]. https://doi.org/10.1099/ijsem.0.001640 Important note To cite this publication, please use the final published version (if applicable). Please check the document version above. Copyright Other than for strictly personal use, it is not permitted to download, forward or distribute the text or part of it, without the consent of the author(s) and/or copyright holder(s), unless the work is under an open content license such as Creative Commons. Takedown policy Please contact us and provide details if you believe this document breaches copyrights. We will remove access to the work immediately and investigate your claim. This work is downloaded from Delft University of Technology. For technical reasons the number of authors shown on this cover page is limited to a maximum of 10. International Journal of Systematic and Evolutionary Microbiology Desulfonatronospira sulfatiphila sp. nov., and Desulfitispora elongata sp. nov., the two novel haloalkaliphilic sulfidogenic bacteria from soda lakes --Manuscript Draft-- Manuscript Number: IJSEM-D-16-00918R1 Full Title: Desulfonatronospira sulfatiphila sp. nov., and Desulfitispora elongata sp. nov., the two novel haloalkaliphilic sulfidogenic bacteria from soda lakes Short Title: Desulfonatronospira sulfatiphila sp. nov., and Desulfitispora elongata sp. nov. Article Type: Note Section/Category: New taxa - Proteobacteria Keywords: soda lakes, sulfidogens, haloalkaliphilic, Desulfonatronospira, Desulfitispora Corresponding Author: Dimitry Y Sorokin, Ph.D., Dr.Sci. Winogradsky Institute of Microbiology RAS Moscow, NA RUSSIAN FEDERATION First Author: Dimitry Y Sorokin, Ph.D., Dr.Sci. Order of Authors: Dimitry Y Sorokin, Ph.D., Dr.Sci. Nikolai A Chernyh, PhD Manuscript Region of Origin: RUSSIAN FEDERATION Abstract: Two novel haloalkaliphilic bacteria with dissimilatory sulfidogenic metabolism were recovered from syntrophic associations obtained from anaerobic sediments of hypersaline soda lakes in Kulunda Steppe (Altai, Russia). Strain ASO3-2T was a member of a sulfidogenic syntrophic association oxidizing acetate at extremely haloalkaline conditions, and was isolated in pure culture using formate as electron donor and sulfate as electron acceptor. It was identified as a new member of the genus Desulfonatronospira within the Deltaproteobacteria. In contrast to the two known species of this genus, the novel isolate was able to grow with formate as electron donor and sulfate, as well, as with sulfite as electron acceptor. Strain Acr1 was a minor component in a soda lake syntrophic association converting benzoate to methane and acetate. It became dominant in a subculture fed with crotonate. While growing on crotonate, Acr1 formed unusually long cells filled with PHA-like granules. Its metabolism was limited to fermentation of crotonate and pyruvate and the ability to utilize thiosulfate and sulfur/polysulfide as e-acceptor. Strain Acr1 was identified as a new member of the genus Desulfitispora in the class Clostridia. Both isolates were obligately haloalkaliphilic with extreme salt tolerance. On the basis of phenotypic and phylogenetic analyses, the novel sulfidogenic isolates from soda lakes are proposed to form two new species: Desulfonatronospira sulfatiphila sp. nov. (ASO3-2T = DSM 100427= UNIQEM U993T) and Desulfitispora elongata sp. nov. (Acr1T = DSM 29990 = UNIQEM U994T). Powered by Editorial Manager® and ProduXion Manager® from Aries Systems Corporation Manuscript Including References (Word document) Click here to download Manuscript Including References (Word document) ASO3-2_Acr1_Sorokin016_rev.doc 2 Desulfonatronospira sulfatiphila sp. nov., and Desulfitispora elongata 3 sp. nov., the two novel haloalkaliphilic sulfidogenic bacteria from 4 soda lakes 5 6 Dimitry Y. Sorokin1,2, Nikolai A. Chernyh1 7 8 9 10 11 1 Winogradsky Institute of Microbiology, Research Centre of Biotechnology, Russian Academy of 12 Sciences, Leninskii Avenue 33/2, 119071 Moscow, Russia 13 14 2 Department of Biotechnology, Delft University of Technology, Van der Maasweg 9, 2629 HZ Delft, 15 The Netherlands 16 17 18 19 20 21 22 Author for correspondence: 23 D.Y. Sorokin 24 e-mail: [email protected]; [email protected] 25 26 27 28 Running title: Desulfonatronospira sulfatiphila sp. nov., and Desulfitispora elongata 29 sp. nov. 30 31 32 33 The GenBank/EMBL/DDBJ accession number for the 16S-rRNA gene sequences of strains 34 ASO3-2T and Acr1T are KP223255 and KP657487; the numbers of DsrB gene/protein 35 sequences of strains ASO3-2T and Acr1T are KF835251 and KP939039. 36 2 2 Two novel haloalkaliphilic bacteria with dissimilatory sulfidogenic metabolism 3 were recovered from syntrophic associations obtained from anaerobic sediments 4 of hypersaline soda lakes in Kulunda Steppe (Altai, Russia). Strain ASO3-2T was 5 a member of a sulfidogenic syntrophic association oxidizing acetate at extremely 6 haloalkaline conditions, and was isolated in pure culture using formate as 7 electron donor and sulfate as electron acceptor. It was identified as a new 8 member of the genus Desulfonatronospira within the Deltaproteobacteria. In 9 contrast to the two known species of this genus, the novel isolate was able to grow 10 with formate as electron donor and sulfate, as well, as with sulfite as electron 11 acceptor. Strain Acr1 was a minor component in a soda lake syntrophic 12 association converting benzoate to methane and acetate. It became dominant in a 13 subculture fed with crotonate. While growing on crotonate, Acr1 formed 14 unusually long cells filled with PHA-like granules. Its metabolism was limited to 15 fermentation of crotonate and pyruvate and the ability to utilize thiosulfate and 16 sulfur/polysulfide as e-acceptor. Strain Acr1 was identified as a new member of 17 the genus Desulfitispora in the class Clostridia. Both isolates were obligately 18 haloalkaliphilic with extreme salt tolerance. On the basis of phenotypic and 19 phylogenetic analyses, the novel sulfidogenic isolates from soda lakes are 20 proposed to form two new species: Desulfonatronospira sulfatiphila sp. nov. 21 (ASO3-2T = DSM 100427= UNIQEM U993T) and Desulfitispora elongata sp. nov. 22 (Acr1T = DSM 29990 = UNIQEM U994T). 23 24 3 2 Our recent research into syntrophic oxidation of volatile fatty acids (VFA) at 3 extremely haloalkaline conditions in anaerobic sediments of hypersaline soda lakes 4 resulted in discovery of several highly enriched associations oxidizing VFA either 5 with sulfate as electron acceptor and forming sulfide (Sorokin et al., 2014) or without 6 sulfate at methanogenic conditions (Sorokin et al., 2016). An association, oxidizing 7 acetate in presence of sulfate as e-acceptor at extreme salinity of up to 3.5 M total 8 Na+ was purified to two components consisting of a novel lineage of acetate-oxidizing 9 clostridium 'Ca. Syntrophonatronum acetioxidans' and its sulfate-reducing partner, 10 strain ASO3-2, identified as a member of the genus Desulfonatronospira. This genus 11 of extremely haloalkaliphilic SRB has previously been found in hypersaline soda 12 lakes and is characterized by its ability to grow chemolithoautotrophically by 13 dismutation of sulfite and thiosulfate, while growth with sulfate was only possible in 14 presence of organic e-donors, such as lactate (Sorokin et al., 2008). Another 15 syntrophic association obtained from soda lakes, along with two dominant organisms, 16 participating in benzoate conversion to methane and acetate (Sorokin et al., 2016), 17 also contained a minor bacterial component with unusually long cells. This organism 18 was apparently feeding on some intermediates of benzoate conversion and was finally 19 isolated using crotonate as substrate. 20 21 This paper is describing the properties of the novel isolates from the soda lake 22 syntrophic associations and suggest to place them into two new species within the two 23 genera of sulfidogenic bacteria, Desulfonatronospira and Desulfitispora, previously 24 found in soda lakes. 25 4 2 The two syntrophic associations which served as the source of novel isolates were 3 obtained from anoxic sediments in hypersaline soda lakes in the Kulunda Steppe 4 (south-western Siberia, Altai, Russia; sampled in July 2010 and 2011) (Sorokin et al., 5 2015; 2016). The brines had salinities from 120 to 300 g l-1, a pH from 10.1 to 10.4 6 and a total soluble carbonate alkalinity from 0.8 to 3.4 M. 7 8 The mineral sodium carbonate-based medium with pH 10 and 0.6 M-4 M total Na+ 9 used for the enrichment and growth experiments, the anaerobic cultivation technique, 10 and the measurements of pH/salinity growth profiles was similar to those described 11 previously (Sorokin et al., 2011). The incubation temperature was 30oC. Electron 12 donors were used at concentration of 10-50 mM and electron acceptors at 13 concentrations of 5 (nitrate, nitrite, selenite, selenate, arsenate, arsenite) or 20 (sulfate,
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