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Supplementary Material Integrated 'Omic' Analyses Provide Supplementary Material Integrated ‘omic’ analyses provide evidence that a Ca. Accumulibacter phosphatis strain performs denitrification under micro-aerobic conditions Pamela Y. Camejo1, Ben O. Oyserman1, Katherine D. McMahon1,2, Daniel R. Noguera1,* 1Department of Civil and Environmental Engineering, University of Wisconsin - Madison, Madison, WI, USA 2Department of Bacteriology, University of Wisconsin - Madison, Madison, WI, USA 1 Supplementary Figures 66-26 83.5 83.2 82.7 82.7 82.4 82.4 82.7 83.0 83.1 83.2 83.1 83.1 83.1 83.2 83.2 83.3 83.0 83.7 84.0 83.2 UBA704 9.4 82.4 82.7 82.6 82.5 82.5 82.0 82.6 82.5 82.4 82.7 82.5 82.7 82.3 82.2 82.2 82.2 82.2 82.2 82.5 UW−1 (IIA) 10.9 2.9 84.8 84.9 84.7 84.7 85.1 84.1 85.0 85.0 84.1 83.8 84.0 83.7 83.8 83.8 83.9 83.7 83.9 83.7 SK−01 (IIC) 6.2 1.5 21.7 99.7 93.6 93.6 90.4 83.8 83.8 84.0 83.6 83.5 83.7 83.5 83.8 84.9 84.4 83.5 84.0 83.4 UBA5574 (IIC) 7.2 1.9 25.2 91.0 93.7 93.6 90.5 83.9 83.8 84.0 83.7 83.6 83.7 83.4 83.4 83.4 83.6 83.4 83.6 83.5 HKU-2 (IIC) 6.4 1.6 22.1 70.2 72.3 99.3 89.8 83.9 83.8 83.9 83.6 83.5 83.6 83.2 83.2 83.3 83.2 83.2 83.3 83.3 SK−02 (IIC) 6.6 1.6 22.0 69.8 69.4 77.7 89.8 84.0 83.8 83.9 83.5 83.4 83.7 87.0 87.1 88.0 84.6 84.0 84.6 84.1 BA−91 (IIC) 5.6 1.3 19.8 53.2 54.5 54.4 58.5 84.4 83.7 83.9 83.8 83.6 83.7 83.7 83.7 83.7 84.0 84.9 84.3 83.9 BA−94 (IIF) 13.0 3.0 22.6 16.1 16.5 14.3 15.7 12.5 88.7 88.5 88.4 88.4 88.4 84.1 84.2 84.1 84.0 83.9 83.9 83.8 UBA2327 (IIF) 11.6 2.8 22.9 16.0 16.0 14.3 15.6 12.1 41.4 99.6 88.1 87.7 87.9 83.5 83.7 83.6 83.8 83.5 83.5 83.6 SK−11 (IIF) 9.2 2.0 17.5 11.7 11.5 10.3 11.7 9.5 35.1 78.1 87.8 87.6 87.8 83.6 83.6 83.7 84.0 83.4 83.5 83.7 UBA6585 (IIF) 11.5 2.8 20.8 13.1 13.2 11.3 12.6 9.9 40.9 61.9 54.0 99.9 99.9 83.4 83.4 83.4 83.3 83.1 83.1 83.5 SK−12 (IIF) 11.6 2.7 20.7 12.7 12.8 11.0 12.0 9.6 41.4 62.0 55.1 93.5 99.8 83.3 83.3 83.3 83.1 82.9 82.9 83.4 UBA2315 (IIF) 12.3 3.0 22.4 13.7 13.7 12.0 13.5 10.4 43.4 65.2 57.2 95.9 94.2 83.4 83.4 83.5 83.2 83.1 83.2 83.4 UBA6658 (IA) 12.4 3.4 22.3 16.2 16.2 13.5 18.4 14.1 14.1 16.6 13.7 15.7 14.5 15.4 99.8 99.4 87.5 87.2 87.7 87.5 BA−93 (IA) 11.6 3.1 21.0 15.4 15.0 12.7 17.6 13.1 13.3 15.7 12.7 14.6 13.6 14.2 99.8 99.2 87.7 87.2 87.8 87.5 UW-2 (IA) 10.8 2.7 19.1 15.9 14.0 11.8 17.6 12.6 12.7 14.4 12.3 13.5 12.6 13.3 79.2 83.6 87.9 87.4 87.8 87.5 UW−LDO−IC 7.0 1.5 13.1 10.7 9.7 8.9 12.8 8.7 8.0 9.7 7.7 8.8 8.2 8.7 45.3 47.5 44.0 88.2 88.3 88.7 UBA2783 (IB) 9.1 2.2 16.3 11.3 11.5 10.0 13.3 10.6 9.5 10.9 8.9 11.1 10.3 11.0 54.7 55.8 50.6 58.2 99.5 91.0 BA−92 (IB) 9.3 2.3 16.1 11.7 11.4 9.7 13.6 9.9 9.2 10.5 9.0 10.5 9.8 10.2 57.4 59.6 53.0 60.2 99.4 91.0 HKU-1 (IB) 11.2 2.8 19.6 14.0 14.4 11.5 15.7 11.6 12.1 13.9 11.1 13.1 12.5 12.7 64.1 65.3 58.7 67.2 81.6 78.0 66-26 UBA704 UW-2 (IA) HKU-1 (IB) BA−93 (IA) UW−1 (IIA) BA−92 (IB) BA−94 (IIF) SK−11 (IIF) SK−12 (IIF) HKU-2 (IIC) SK−01 (IIC) SK−02 (IIC) BA−91 (IIC) UW−LDO−IC UBA6658 (IA) UBA2783 (IB) UBA2327 (IIF) UBA6585 (IIF) UBA2315 (IIF) UBA5574 (IIC) Average 80 84 88 92 96 100 Minimum 0 10 20 30 40 50 60 70 80 90 100 Nucleotide Fraction Identity (ANI) Aligned Figure S1. Comparison of the genome-wide average nucleotide identity and alignment percentage, of Accumulibacter-like genomes. The heatmap shows the average nucleotide identity (red low section of matrix) and the minimum percentage of the two genomes that aligned (blue upper section). 2 Figure S2. Schematic illustration of denitrifying genes loci in different genomes of Accumulibacter. Changes in the position of homologous genes are indicated by lines, the arrows show direction of transcription. Genes are drawn to scale. Proteins with unknown function are colored grey and transposases black. End of a scaffold is indicated with a red line. aa trans, aminoacid carrier family; nar, nitrate reductase; ppiC, peptidyl-prolyl cis-trans isomerase C; uspA, universal stress protein A, ppp, protein phosphatase; dinJ, DNA-damage-inducible protein J, nir, nitrite reductase; nsrR, Nitric oxic-sensitive repressor; nor, nitric oxide reductase; ccoG, type cbb3 cytochrome oxidase biogenesis protein; menA, 1,4-dihydroxy-2-naphthoate octaprenyltransferase; cysG, siroheme synthase; yqfl, ATP/GTP-binding protein; sco1, cytochrome c oxidase biogenesis protein; nos, nitrous oxide reductase; cyt, cytochrome; resA, cytochrome c-type biogenesis protein; fpr, Ferredoxin— NADP reductase; yicL, carboxylate/amino acid/amine transporter; malE, carboxylate/amino acid/amine transporter; kefA, potassium efflux system protein; lysE, L-lysine exporter family protein; luxO, two-component signal response regulator. *Genome lacks one of the scaffolds. 3 Comamonadaceae Cupriavidus basilensis (CP010536) Pseudomonadaceae Acidovorax sp. KKS102 (CP003872) Acidovorax sp. P4 (CP021366) Burkholderiaceae 100 100 Acidovorax sp. NA3 (CP021361) Rhodocyclaceae (other than Accumulibacter) 75 85 Acidovorax sp. NA2 (CP021359) Acidovorax sp. RAC01 (CP016447) 100 97 Acidovorax sp. T1 (CP021648) Acidovorax sp. JS42 (CP000539) 100 100 Acidovorax ebreus (CP001392) Alicycliphilus denitrificans (CP002657) Pseudomonas denitrificans (CP004143) 100 Pseudomonas citronellolis (CP015878) 100 Pseudomonas aeruginosa (CP008869) 100 Pseudomonas chlororaphis (LT629761) 100 Pseudomonas balearica (CP007511) 100 Pseudomonas stutzeri (CP003725) 100 Pseudomonas xanthomarina (LT629970) 100 Thauera terpenica (ATJV01000000) 100 100 Thauera chlorobenzoica (CP018839) 83 Thauera phenylacetica (AMXF01000000) 100 Thauera aminoaromatica (AMXD01000000) 100 100 Thauera sp. SWB20 (JTDM01000000) 100 Thauera sp. MZ1T (CP001281) 93 Thauera sp. DNT-1 (BAZL01000000) Azoarcus sp. CIB (CP011072) 95 100 Azoarcus aromaticum (CR555306) Burkholderia pseudomallei (CP009150) Burkholderia vietnamiensis (CP013456) 100 Burkholderia cenocepacia (CP013451) 100 100 100 Burkholderia stabilis (AP018113) 86 Burkholderia contaminans (CP009745) 100 53 Burkholderia lata (CP013392) Burkholderia ambifaria (CP001027) 99 Burkholderia sp. MSMB0856 (CP013429) 100 Burkholderia diffusa (CP013364) 87 Burkholderia territorii (CP013367) Accumulibacter UBA704 100 Accumulibacter UW-LDO-IC Type I 6 6 Accumulibacter BA-91 100 Accumulibacter SK-01 85 93 Accumulibacter HKU-2 Type II 100 Accumulibacter SK-02 Accumulibacter UBA5574 9 8 Burkholderia phymatum - plasmid pBPHY01 (CP001045) 100 Burkholderia sp. HB1 (CP012193) 100 Burkholderia sp. CCGE1003 (CP002218) 0.04 Figure S3. Neighbor-joining phylogenetic tree based on the full-length nucleic acid sequences of homologs to narG found in the genome of Accumulibacter and annotated genomes in NCBI. Bootstrap values are shown in the tree branches based on 1000 bootstrap replicates. The scale bar represents the number of nucleotide substitutions per site. 4 Cupriavidus taiwanensis (CU633750) Dechloromonas agitata (NZ KI912532) 100 Dechloromonas aromatica nirS-2 (CP000089) 81 Azoarcus sp. CIB nirS-2 (CP011072) 100 Thauera aminoaromatica (AMXD01000000) 87 Thauera terpenica (ATJV01000000) 98 Thauera chlorobenzoica (CP018839) 69 Thauera humireducens nirS-2 (CP014646) 100 Accumulibacter 66-26 nirS-1 98 Accumulibacter 66-26 nirS-2 Accumulibacter HKU-1 nirS-1 Accumulibacter UW-LDO-IC nirS-1 61 Accumulibacter BA-92 nirS-1 100 52 57 Accumulibacter UBA2783 nirS-1 Type I Accumulibacter UW-IA nirS-1 98Accumulibacter UBA6658 nirS-1 100 Accumulibacter BA-93 nirS-1 Accumulibacter 66-26 nirS-3 100 Accumulibacter 66-26 nirS-4 100 Accumulibacter UW-1 IIA nirS-1 51 Accumulibacter SK-12 nirS-1 100 Accumulibacter UBA2315 nirS-1 92 78 Accumulibacter UBA6585 nirS-1 Type II Accumulibacter UBA2327 nirS-1 99 Accumulibacter SK-11 nirS-1 100 Pseudomonas Genus Thauera phenylacetica (AMXF01000000) 100 Thauera sp.
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