Supplementary Information
Figure S1. Partial phylogenetic tree of homologs of AhpL demonstrate this AHP group is present in a broad range of bacteria.
Homologs to AhpL (red arrow) are encoded by a diverse group of bacteria including several known Mn(II)-oxidizers (blue arrows). A list of AhpL
homologs was generated via blastp (Altschul et al. 1990) and for simplicity the initial list of
amino acid sequences (62) was trimmed of shorter sequences and of sequences from closely related organisms (based both on 16S rRNA gene sequence and AHP protein sequence). The outgroup (green arrow) represents the closest eukaryotic protein match to the bacterial AHP
cluster (using PSI-BLAST (Altschul et al. 1997) of AhpL and alignment of homologs to the bacterial cluster). Amino acid sequence alignment was conducted using ClustalX (2.1) (Larkin et al. 2007) and the alignment imported into Geneious version 6.1.6 (Biomatters).
1

Table S1. Putative proteins encoded in Locus 1 (Scaffold Accession number: NC_ABCR01000001 )
Start a
End Protein Name Locus Putative Protein Description b
171067 169640 RAZWK3B_01330 EDM72820 hypothetical protein
171227 171994 RAZWK3B_01335 EDM72821 tRNA pseudouridine synthase A
175965 172132 RAZWK3B_01340 EDM72822 heme peroxidase (AhpC)
181188 176068 RAZWK3B_01345 EDM72823 Animal haem peroxidase (AhpB)
182849 181155 RAZWK3B_01350 EDM72824 Animal haem peroxidase (AhpA)
184649 183192 RAZWK3B_01355 EDM72825 Cation/multidrug efflux pump, RND superfamily protein
186252 184567 RAZWK3B_01360 EDM72826 Cation/multidrug efflux pump, RND superfamily protein
187475 186249 RAZWK3B_01365 EDM72827 secretion protein HlyD
188783 187503 RAZWK3B_01370 EDM72828 5-aminolevulinic acid synthase
189770 189150 RAZWK3B_01375 EDM72829 two component transcriptional regulator, LuxR family protein
190490 191317 RAZWK3B_01380 EDM72830 hypothetical protein
193672 191354 RAZWK3B_01385 EDM72831 Hemolysin-type calcium-binding region
194228 193815 RAZWK3B_01390 EDM72832 hypothetical protein
195561 194233 RAZWK3B_01395 EDM72833 type I secretion membrane fusion protein, HlyD family
197426 195576 RAZWK3B_01400 EDM72834 RTX toxins and related Ca2+-binding proteins-like protein
199123 197423 RAZWK3B_01405 EDM72835 type I secretion system ATPase
199578 200666 RAZWK3B_01410 EDM72836 GGDEF domain
200997 201311 RAZWK3B_01415 EDM72837 periplasmic sensor hybrid histidine kinase
201349 201741 RAZWK3B_01420 EDM72838 sensor histidine kinase/response regulator hybrid a – CDS identified on the complementary strand are in italics and start at a higher position than they end b - AHPs are listed in bold and possible proteins associated with TISS are in italics
2

Table S2. Putative proteins encoded in Locus 2 (Scaffold Accession number: NC_ABCR01000012 )
Start a
End Protein Name Locus Putative Protein Description b
71924 71622 RAZWK3B_00390 EDM69961 D-fructose-6-phosphate amidotransferase
72084 72851 RAZWK3B_00395 EDM69962 tRNA pseudouridine synthase A
83708 72989 Animal heme peroxidase (AhpL)
87110 84051 RAZWK3B_00410 EDM69965 probable transporter transmembrane protein
87676 87107 RAZWK3B_00415 EDM69966 Secretion protein HlyD
88326 87685 RAZWK3B_00420 EDM69967 tRNA pseudouridine synthase A
89634 88354 RAZWK3B_00425 EDM69968 5-aminolevulinic acid synthase
90372 90001 RAZWK3B_00430 EDM69969 response regulator receiver
91339 92166 RAZWK3B_00435 EDM69970 hypothetical protein
92259 92690 RAZWK3B_00440 EDM69971 hypothetical conserved protein
92687 93040 RAZWK3B_00445 EDM69972 probable insertion sequence transposase protein
93095 94654 RAZWK3B_00450 EDM69973 probable insertion sequence transposase protein
97039 94721 RAZWK3B_00455 EDM69974 Hemolysin-type calcium-binding region
97595 97182 RAZWK3B_00460 EDM69975 hypothetical protein
98928 97600 RAZWK3B_00465 EDM69976 type I secretion membrane fusion protein, HlyD family
100937 98943 RAZWK3B_00470 EDM69977 RTX toxins and related Ca2+-binding proteins-like protein
102634 100934 RAZWK3B_00475 EDM69978 type I secretion system ATPase
102974 104833 RAZWK3B_00480 EDM69979 Ribosomal-protein-alanine acetyltransferase
104871 105263 RAZWK3B_00485 EDM69980 sensor histidine kinase/response regulator a – CDS identified on the complementary strand are in italics and start at a higher position than they end b - AHPs are listed in bold and possible proteins associated with TISS are in italics. An inserted section different from locus 1 is shaded gray
3

200

M Mn(II) 200

M Mn(II)
+NADH
A B
200

M Mn(II)
+Cu(II)
C
1 2 3 1 2 3 1 2 3
Figure S2. In-gel Mn(II) oxidation assay in HEPES buffer of Roseobacter sp. AzwK-3b soluble proteins.
As in Figure 2, R. sp. AzwK-3b soluble protein extracts from replicate cultures (lanes 2, 3) were separated via SDS-PAGE and assayed for Mn(II) oxidation activity
(Francis et al. 2001, Francis and Tebo 2002). However, unlike in incubations in Tris buffer, an
LBB stained Mn(III,IV) band was observed in incubations conducted in 5 mM HEPES buffer and 200

M Mn(II) without NADH added (A), however, LBB bands were noticeably more intense with the addition of 100

M NADH (B). The addition of CuCl
2
(100

M), which scavenges superoxide, prevented the formation of the Mn(III,IV) band formation (C).
Thus, Mn(II) oxidation was not the result of direct enzymatic activity but was instead initiated by superoxide and the observed proteins were likely involved in the stabilization of Mn(III) and removal of generated H
2
O
2
(Equation 2). These results are in line with previous observations of
superoxide production by HEPES buffer (Keynes et al. 2003),
in vitro oxidation of Mn(II) by R .
AzwK-3b proteins (Figure 1) and the known constraints on superoxide mediated Mn(II)
oxidation (Learman et al. 2013). The observation that HEPES may be a superoxide source to
initiate Mn(II) oxidation suggests that superoxide could also be contributing at least in part to heme peroxidase-mediated Mn(II) oxidation by other organisms.
4

Figure S3. Production and inhibition of superoxide in Roseobacter sp. AzwK-3b culture filtrate proteins.
(Reprinted from Diaz et al., 2013) Proteins from R . AzwK-3b were visible in whole cell extracts and in culture filtrate (Coomassie gel, lanes b and d respectively). Reaction of the protein gel with the superoxide probe NBT under control conditions revealed superoxide production at a single location in the separated whole cell extract (Control gel, lane b). Addition of NADH stimulated superoxide production by whole cell proteins and a single protein band in the cell free filtrate (NADH gel, lanes b and d, respectively). Addition of DPI quenched NADHstimulated superoxide production in whole cell protein extracts and completely inhibited it in the filtrate protein band (NADH+DPI gel, lanes b and d). Protein ladders (all gels, lane a) were prelabeled with stains. Lane description for all gels (labeled in gel 1): protein ladder (lane a), soluble whole cell proteins (lane b), cell-free filtrate proteins (lane d), boiled (denatured) whole cell soluble proteins (lane f) and loading buffer only (lanes c and e).
5

Updated ahpL DNA sequence
ATGGTTTTCGTGAACACAAACGACCTGAAGCACATCCTCGACCAGATCCGCATCGCAGAACA
GCATGCGGCTGGCACGCCGCTGACTGATCTGGTTGCAAACCCCTTGCTGCCTTATGGCCTGC
GTCTTGTTGACGGCACGTTGAACAATTTGACTCCTGGCCGTGAGGAGTGGGGTTCGGCGGAT
CAGGTCATGCCGCGCCTGCTTGAGTCGACCTTCCTGACGCAGCCTGATGCGGCACATCTGGC
CGATCCGAGCCCGCGTGGTGCGCCGGGTGCCGAGCCCACGTCCTATTTGCAGACTTCCGGCT
CGGTTTATGACGCCGAGCCTCGTGTGATCTCGAACCTGATCGCCGATCAGACCCTGTCCAAC
CCCGCCGTCATCGCGGCGGCGCTCAGCCATGCCGGCATGACTGGACAGGCGATGCTGACCA
CGGCCAATGAAATCGTTCAGGCCTATCAGCGCGTCATCGATGCGCAGGCTGCTGCCGGCAAC
GTCGATCAGGCGCTCGAGCTGCAGCGCCAGGAACTGCAGACAGCGCTGGACACTGCCACTG
CTGAACTGACCGCCGCACAAGGCGATGTCGCGGCCAAGACAACTGCCAAGAATGAGGCCGA
TCAGGCCGTGACGCAGGCTCAGGACACGCTCGACAATGCAGCCGCCACGATGGTCACGCTG
CAAGGTCAGGGCCAGGTTGCCGAAATGCAGGTTGCGTTGAATGCCGCTGTCGCGGCGCATA
CGCAGGCCGAAGCTGATCTCAAGGCGGCGCAGGATGAGCTGTTGCAAGCCGAAAGCGCGGC
CGCATCCATGCTGACCATGCACAATGCGAAACAGACCACCGTCACCAACCTGCAGGGGCAA
AAGGCAACGGCTGATCAGGAGCTGGCCACCGCCGAGGGTTTGCTGTCCGACGCCCGGGCCG
CATTGGCGCTGGAGACCGACGCAGCTGGTGCGGTCACCGAGGCCCAGACGGCGGTTGAAAA
CGCCACCACCGCGGTTAGCAATGCGCAGACGCGGGTCAACGAGCTTGACGCACAGCTCACG
CAGGCTGAGGCGGATCTCGCTGATCTGAACACGCAACTTCAGGCCGCAGAGCAGACCCGCG
ATGCGGCTCAGACACGTGTTACGGCCGCTCAGGAGGCCGCCGCGGCAACCCAGGCGGACCG
CGAGGACGCTCAGGCTCTCTTTGACACAGAGGTGAGCGAGCTTGCCACCGCACAGGCCGAG
ATGCTCAGCGCCCTCACGAATTATATCACCGGTGGATCGCTGGCCGATTTCTCGGTTGCGTCC
GCGGCCTATTCTGCTGCCCGCGAAGAAAAACTGGCCGCCGAAACGGCGCTGAATGATGCAA
TCGACGCGGATGACGAGGCCGATCAGGCGGTCATCGACGAAACCGCCGCTCTCACCACGGC
GCAATCTCGGGTCAGCACCCTGACCACGCAGCGCGATAACGATATGACCACGCGTGACGGC
TTGAGCACCCAGTTGGACGAGGCAAACGGCCTCTTGACCAGCGCGCAAAACACTCTGACAG
CCGCCGAGCAGACACTCCAGACGGCCCAGGACGACAATCAGGCCTATGTCGATGCGCAGAA
CGCCGTTGATCTGGCCCAGGCCGATGTCGATGCGGCTCAAGGGATCGTCGATGGCCTCGTGG
CACAGTTGGGGGTTGCTGAAGGCGAACTTGCAACGCTGAACACCAATCTCGGCAACGCCGA
CGCCGAAGTCACCCGTCTCGAAGGCGTGGTTCAGACCGAAACCACCGATCTGGAAACGGCT
GCAGGTGGTGTGACCACGGCTCAGCAGAATCTTGCGGCGGCGCAGATGGCCGATGCCGCCA
TCCTCGCGCAGCTCGACGTGATTGCAGATGCCACCGTTACGCTCACTGAGGCGACAGCGACG
GCCGCTCAGGCAGCCCTGGATCTGACCGCGGCCGAAGGCGTGGAAGCCACAAAGCAGGTCG
CGGTCGACACCGCCACAAGTGATCTCAATGCCCTGACGGCACCTGGTGCGGCTGAAGCCGCC
CTGGCCGCCGCTCAGGCCACCGCCGCCGAGGCGCAGACCACGCTCGACACGCTGCTGACCA
CCCATAACATCACGATGGACGGCAACAACGTTCTGCTGCCCGACGTGACCCCCGATGAAGGT
CTGTCCGCACCCTACAACTCGTGGATGACGCTGTTTGGTCAGTTCTTCGATCACGGCCTCGAC
CTGGTCGGCAAAGGGGGGAGCGGCACCGTCTACATTCCGCTGCAGCCCGATGATCCGCTTTA
CGATGCGAACTCGCCCACCAACTTCATGGTGCTGACCCGCGCCACCAACCAGCCTGGCCCGG
ATGGTATCCTGGGCACCGCGGACGACATCCGCGAGCATTTCAACAAGACCACGCCTTGGGTC
GACCAGAACCAGACCTACACCTCGCACCCCTCGCATCAGGTGTTCCTGCGCGAATACGATAT
CGACGCCAACGGAAACCCTGTCTCCAACGGCTACCTGCTGCACGGCCAGACCGGCGGCATG
GCCACCTGGGGCGATGTCAAGGCGCAGGCTGCGACCAAGCTGGGCATTCAGCTCAATGACA
GCGACGTGCTCGATGGCCCGCTGCTGGCCACCGACCCCTACGGCAACTTCATCCCCGGCGCC
6

AACGGCCTGCCGCAGCTTGTCGTGCCGAACCCTGCCTATGTCGAAGGCGGCACCGAGCCTTT
GAATATCCTGATCGAAGGCGATCTGGCCAACCCCGTCGATGCCAGCCAGGCGCTTCGCAACG
GCCATGCCTTCCTCGAGGATATTTCCCACACCGCGTTCCCCAAGGGCATGATCGACCACGAT
CGCAACCCGATGACGCCTGAAATCGAAGTGTTGCCCGATGCCGATACCGACACCGGCAACG
CGATCATGCCGAACATCTTCGGCATGAACGAAACCTACGACAACGAACTGCTGGACCGCCA
CTTCATCGCCGGTGATGGCCGTGGCAACGAAAACTTCGGTCTGACGGCGGTGCACCATGTGT
TCCACACCGAACACAACCGCCAGGCAGCCGAGATGAAGCAGACCATCCTGGACAGTGGCGA
GTTGGCCTTCATCAACGAATGGCTGGCCACCCCGATCACCGAGGCTGACCTGGCCACCGCGA
CCGTCGACACCGTCGAATGGGATGGCGGGCGTCTGTTCCAGGCCGCCAAGTTCACCACCGAG
ATGCAGTATCAGCACCTCGCCTTCGAAGAGTTCGGCCGCACCGTGCAGCCCCAGATCGCGGC
CTTCGGCGTCAATGGCAGTGCCGAGATCGACGGCGCAGTCATGGCCGAATTCGCCCATGTGG
TGTACCGCTTCGGCCACTCGATGCTGACCGAGAACGTTCACACGATGGACCCCAACGGCGTG
AACACCTCGAACGGTCTGATCGAAGCCTTCCTGAACCCGGTCGCCTTCGACCTCGACCAGAC
CCTGACCTCGGATCAGGCGGCAGGTGCCGTGGCCCGTGGCATGTCCCGCGAAACCGGCGCC
AATATCGACGAGTTCATCACCAGCGCCCTGCGCGACAACCTTGTGGGCCTGCCTCTGGACCT
TGCCGCGCTGAACATCGCGCGGGGTCGGGAAACCGGTGTGCCCTCACTCAATGCCGCCCGCG
AGCAGTTCTACTCCGCCACCGGCAGCGAGTTCCTCAAGCCCTATGAAGGCTGGTCGGAATTT
GCCGCGAACCTCAAGAACCCGGCTTCCATCATCAACTTCATCGCGTCCTACGGCACCCACGA
GACCATCGTGAACGCCACGACCGTGGTGGAAAAGCGCGCTGCGGCCACCGATCTGGTTCTG
GGCGGTGATAACGCCCCGGCGGACCGTCTGGACTTCGTCAACGGCACCGGCGCTTGGGCCG
ACGCGGAAACCGGCATCAACGATGTGGAATTCTGGATCGGCGGTCTGGCCGAGGACATTAT
GCCCTTCGGCGGGATGCTCGGTTCCAGCTTCGGATTCGTGTTCCAGCAGCAGATGGAGGCGC
TTCAGAACGGCGACCGTTTCTACTATCTGGCACGGACCGCCGGCATGAACATGATTGCCGAG
TTGGAAAACAACTCCTTTGCGTCGATGATCGTGCGCAACACCGACATCAAGGACGGCGGCG
CGCATATCCCGGCCAACATCTTCTCGTCGATGGATGTCATCCTCGAGGTGGATCAGGCCGTG
CAGTCCATGCCCGACCCGGTGTCCACCGATGTCGATCCGTTCCTTGCCGCCATGGGCACCAC
CATGGTCGAGCGGGCTACGGCCACGGTGGATGCGCCACTGATCGACGGGGTGCGCGAATAC
GACAACTTCCTCAAGTTCAACGGCGGCGAACACGCGGTGCTCGGCGGCACGGATCAGCGCG
ATATCCTCAACGGCGGCCTCGGTGACGATGCGCTCTGGGGCGGCGCGGGCGATGACCTGCTG
ATCGGTGACGCGGGCGTGAACACCCTGCGCGGCGGGGCCGGCAACGACATCCTCAAGGATG
GCGACGATGTCAGCTTCTTGCATGGCGAAGATGGCGATGACGTGATCTCGGCCGGCGGCGGT
ATCGGCGAACTGATGTTCGGTGGCAAGGGCAACGACGCCATCCTCATGGGGTCAGACGATG
CGGCCACGGCGTTCGGCATGGACGGCAACGACTTCATCTATGGCGGTGCCGGTGGCGAGTTC
ATGTTCGGTGGCGAAGGCGATGACTGGATCGAAGGCGGCGACGGCTTCGACATGGTCATGG
GTGACAACGGCGATCCCTTTGGTGGGTCGCGGGTTGTGGGCCATGACGTTCTGGATGGCGGC
GCCAACGACAACGACCTGCTCGGGGAAGGTGGCGACGATATCCTGATCCAGCGCGAGGGCG
TGCATGTCAACGAAGGCGATCTCGGCTTCGACTGGGTGGCGCATAAAGACGCCATGACCGG
CGCCGATATCGACCTCAGCCGTCGTCTGGAAACCAACGAGACGCAGGCCGCACTCCGCGAT
CGCTACGACCTGGTCGAGGCGGCATCGGGCACGCAGTTCGACGATTTCCTCTACGGCGATGA
CCGCGAAGGCGCGGCGCTGCCGGACGGGATTGATCTGCTGGCCAATGACGCGACGCTCTTC
GCCAACGAACTGACCCAGGCTGGTGTTGACCGGATTGCAGGTCTGCGCGAGTTGCTGGCCGA
CATGATGGGGGTCGATCCCGCTGATACCTTCACCGGCGGCAACATCCTGCTCGGTGGTGGCG
GCAGCGACGTGATCCAGGGCCGCGGCGGCGATGACGTTATCGACGGCGACAAGGCGCTCAA
CGTGCGCATCTCGGTCCGCGATCCCCAGGATCCGACCGTGGAACTGCGCTCCATCAACAGCC
7

TGTTCGAAATCTCGAACGAACTGCTTGATGGCTCGATCAGCCCCGGCCAGCTGCAGATCGTT
CGGGAAATCCTCGATGCCGGTCAGGAAGGCGATGTGGACACGGCTGTCTACTTCGATGTGCG
GGCCAACTACAACATCCGCACCAACGCCGATGGCAGCGTCACGGTCGATCATATCGCACCG
CCCATCGAAGGCGCGTTCATTGACCCGATCAACGGTCAGCCCAACGTGGAAGGTGGTGACG
GCACCGACCGCCTGACCAATGTCGAAATCCTGCGCTTCGCGGATATGGAGATTGATCTGCGC
ACTGATCCGGCCAGCAACGGGTTGATCCAGGGCACCGATGGCGATGATGTGCTCAATGGCA
GCAACGGTCGCGACGCGATCTTCGCAGGCGCTGGAAATGACGTGATCAACGGCCTTGCCGG
AAATGACGTGATCTTTGGCGAAGGTGGCTCCGACACCATCACATGGAACGCGCCCAACGGC
GGCTACGACGTGGTCGTGGGTCATGGCATCGTCAATGACGGCGGCACCGACATGCTGGTGAT
CAACGGCGATGGCACTGAGGACGGCTTCACGATGTACACCGTGGCAGCTTGGACCGGTCAG
GCCCCTGTCGATCCGTCTTCCGAGATCATCATCACCCGGACAGTGGCCGGTGTGGAGACCGC
GATCATGGAAGTGCGCGGCATCGAAGAGGTCGCAATCAACGGGGTGCAGGGGATCGAAACG
ATCAACGCGGTTGGTGACTTCGCAGGCACCTCGCTGGCGCCCAACACGATCTACGTGAACGG
CACGGCAGGCAGAGAAGTGATCGACTTCTCTGGCTTCACCTCGACCCAGCGTCTTGTGGTGG
ATGCAGGTGACGGTCGCGACGTCATCACCGGTGGCGCAGGCAATGATATCCTGAACGCTGG
TGCGGGAGACGACGAAATCACCTGGAGTGTCGGCGGCGGCAGCGATGTGGTCGATGGCGGG
GCAGGTGAAGACACCTACACCATCAACGGCGATGCGTCTGATGAAACCTTCCGCGTCTACGC
GGCGGACTCCTGGACAGGGGCACCCCTGGCCACCGGAACCGACATCGTCATCACCCGCGAC
GCGGGTGCCGGCGAACAGGTGATCGGTCAGCTTCAGAACATCGAGGAAATCCAGATCAACA
CAGCCGGTGGTGTCGACAACGTCGAGGTGATCGGTGACTTCAACCCGACAGCTCTCAACTTC
AACACCATCGCGGTGACGGGTGATGATGTCAGCGTCGATGTCACGCAGCGTCTGTCCACGCA
CCGCGTGTTCGTTGATGTGGGTGGCAACACACAGGATGTGCCCAAGTCGCTGAATGTGGCGG
GTGTTGTAACCGAAGGCGATGTCTTCCTGATCCCGGTCGATCCGACAACCCTGACCCGGACG
GTCGATGGTCTCACGGTGACCCAGAAGTCGGACAACTTCGTCATGACCTACACGGTCGATGA
CGTGGACACGATCCCGGTTCTGGTCCAGTCCGGCACGCTGATGGCCGACGCTCTGCGGGGTG
TGGCCACTGTCAACGAACAGACCGGCTTGCCGGAAGAAGGGTTCGGGTTCGGTGTGCTGAC
GCGCGGTGCCACCCTTGTGACGGCAGATATGGTGACGGCAGATGATATCGCCGCCCTCAAGT
ACATGGTCGGTGCATCCAACGTCGCGCCGATCTTGCCGCCCGAAGCGGGTGAGCCTGCCGAC
AACGAGCCAGCAGATCTCTTGCAAGAGGTGCGCGATCTTGAAGGTCTGACCAACAATCTCCA
GAACGCGGAGATTTCGGGAGGCGCAACCTTGCCCTTCATGCGTGTCACTGAAGCCCGCTATG
CTGGCATCGGCGAAGACGGTGCAGGCATCATCAACCCCGTCTTTGATAGGCTTGATACCCGG
GATATCTCGAACACCTTGGGCGCTCAGGATCTTGACGCGGCCAAGGCGGCCTCGGCCAACAT
GTTCATGATGTCCTTTGGTCAGTACTTCGACCACGGCCTGACCTTCATTCCGAAGGGCAGCG
GTCCGATGGGTCAAACGCTTGGTCAAATCGAAATCGGTGGGCCTAGCATGGATCGTGGGCC
GATGGCCGACAACCCGGCCGACCTGACCCGCGGTAGCGTGATCGGTTATGATGAGGATGGC
ACGCCGCTGCAGGAAAACATCACCTCGCCGGTGGTTGACCAGAACCAGGTCTATGGCTCGTC
CAGCCTGATCGGTCAGTTGCTGCGTGAAAGCGATGGCGAAATGGGTGTCGGCGCACGCATCT
TGATGGGTGGCGATGATCCTTCGGCAGCACCGGGCTTCAAGATCTTGTCCACGCTGCGTGAA
ACGCTCGATCACCACATCGATGCGGGCACCGTGTTCAAGGGTGCGGGTCTTGGTGAAAACG
GCATGACGCTGCTCGACTACTACCCGGGCTTGCGTGACGCCGATCGCAACTACGATGCCGCA
CTGGTGAAAGAGCTTTCTGGCGACTTCATGGATGAGGGCTGGCCGCTTCTGATCGACACCAA
CCCGTTCATGAACCTGCTCGACCACTTTGTGGGCGGCGATGGGCGCGCGAACGAGAACGTG
GGCCTCACCTCGATGCACACGGTTTGGGCACGTAACCACAACTACCACGTGGACCAGATCAA
CAAACAGGCAGGCCCGGATTTCACCCCCGAACAGCTGTTCCAAGCTGCGCGGATCCTGAAC
8

ATCGGTGAATATCAGCAGGTTGTGTTCAACGACTTTGCCGATGCCTTGATCGGTGGCTTGCA
GGGCTCTGGCCGTCATGGGCACAACGAGTATAACCCGAACGCTGATGCCCGCATCAGCCAC
GAGTTTGCCGCCGCCGCCTACCGCTTCGGCCATTCCCAGATCGGCGAAACGATGCTGGTCGA
AAAGGATGGACAGATGGTTGAGCTGCCCCTGATCGACCTGTTCCTGAACCCGACCAGTGATC
CGAATGCCTTCCAGTATGTCGATCGCGAAACAGGTGAATTGGTGACCGGCGAGGCCGCCTTG
GCTGCCTTGGCTCAAATGGACTACGTGCCGCAGGATGGGTACGCTCAATACGGCGTGAACA
GCATCCTCGGCGGTTTGGTTGCACAGCCCTCGGAAGAGGTTGATCTGCAAGTGGTGGATGCG
GTCCGCGACAATCTTGTCCGGGTCAGTGCCGACCTCTTTGCCTTCAACGCTGCACGGGGCCG
CGATTTGGGTCTTGGAACCCTCAACCAGGTCAAGGCCGATCTGGCGGCGTCCACCGACCCCT
ACATCGTGGAAGCGATCGAACAGTCGAACATGAACATGGACCCCTATGCTTCGTGGGATGA
CTTCCAGTGGCGCAACAACCTGTCGGAAACAGAGATCACCAAGTTCAAGACAGCCTACCCG
GATCTGGTTCTGGAAGGTGACGCGATCGCAGCATTCCAGCAGGTCAACCCCGATATCACGCT
GATCCCGGGTGAAAACGGTGCGATGATCGTTCAGGGCATCGACCGGGTCGACATGTGGGTC
GGCGGTTTGGCAGAACCCCATATCGAGGGTGGCGTCGTGGGTCATACCTTCTGGGTGCTGAT
CCATGAACAGCTGGACCGTCTGCAGGAAGGCGACCGGTTCTACTACATCGACCGGATCGGC
GATCTGCCCGTCTACAACAACTTCATCTCCAACCTCACCTTCGGGGATATCGTGACCCGCAA
CACCGGGATGACCGATCTGCCGCTGGATGTGTTCAGCTATGTTCCAGAAGTGCCTGAGGGCG
CAGCTGCAGATGCTGGAAATCAGCAGATCCAGACCGATCAGAACGCAAATCGGACAGATGA
TCCTGCGATCGACACAGCGGATACTGAAGGTGGTGGAACCACGACAGATACCGACGTCACA
GGCGGCGCGGATGATGAGACCGAGGCCGCAGCTACAGAAGGCAACGCTGAGCCAGATCCCC
TGGAGACTGAGGCGAACGCCCTGTCCGAAGGAGCTGTTGAAGGAGGAGTTGACGAAACTGA
ACAACCCGATGCCGATGTGACAGGTTCAACGCTAGAGCAGTCGGCCGAAGAGGAAGAAACC
GAGGAGGATGACGCCACTCAAAACGCTACGGGTGCAGCATCCGGCGGAGAAGACCTGGAG
GCCACGTCTTCTGAAACCAATGAGGAAATGCCTGAAGACAATACTTCCATGTCTCCTGACGA
TGCTGGAGGTGCCGATACAGGCGGCGCGGATGATGAGACCGAAGCCGCAGCTACAGAAGGC
GATAATGAGCCAGATCCCCTGGAGACTGAGGTGAACGCCCTGTCCGAAGGAGCTGGCGAAG
GTGAAGGCGGCGAGGCTGAACAACCTGATGCCGATGTGACAGGTTCAACGCTAGAGCAGTC
GGCCGAAGAGGAAGAAACCGAGGAGGATGACGCCACTCAAAACGCTACGGGTGCAGCATC
CGGTGGAGCAGCCCTGGAGGCCGAAGAGGAAGAAACCGAGGAGGATGACGCCACTCAAAA
CGCTATGGGTGCAGCATCCGGCGGAGCAGACCTGGAGGCCGAAGAGGAAGAAACCGAGGA
GGAAATGCCTGAAGACAACACTTCCATGTCTCCTGACACAGACACCTCGGCCGAGGGGATA
GTCGAAAGCGACACCGATATGGGCGCCCCCCTGGTCGGCGCGGCAGGCGACGACATCCTCA
GCGGGATGGCTGGCGACGACATGCTGGTGGGCAATGCCGGTGACGACATGCTCTTCGGTGG
CGAGGGCCGTGACGACATGCTCGGTGGCGATGGCGCCGACATGCTCTCGGGCGGTGCCGGC
AGCGACCATATCCTCGGCGGCGACGGCGATGACATGATCATCGGCGGAACCGGCTCGGATA
TCCTGATGGGCAATGCAGGCGACGACACGTTCCTTGGAACGAATGGCGACTGCAACGACCT
CATCTACGGTGGCGAAGGGACGGACACGATCGATATGGCGTCGGTCACCTCCGACCTCACG
GTGCGCCTCGGCAATACCGGGACGGATCGTGGCTCGGTCACCACGGAAGAAGGCGGTCGCG
ACACGATCTGGAGCATCGAGAACTTCGTCGGTGGTGCGGGTGACGACACCATCTTCGCCAGC
GACGCGGCCAACGTGCTCGATGGCGGGGATGGCAACGACACCTTTGTGTTCGAAACCGCGG
CCAGCGCGCAGGGCGATCACATCAACGGCCTCAACGCCGGCGATGTCCTGCAGTTCGGCAC
AGGGGCCGACATGTTCGAGCTCACTTGGGGTGACACTGACATGCAGGACGGCTTGTCTTTCT
CCGGAGGAGAAGAAGCCGGTACGACCAAGATCAGTGGAACCATCGAGGATGACCAATTCGA
ACTGACCGTCAGTGGCCGCTTCGACTACGATCAGATCACCTGA
9

Updated AHPL Protein Sequence
VFVNTNDLKHILDQIRIAEQHAAGTPLTDLVANPLLPYGLRLVDGTLNNLTPGREEWGSADQVM
PRLLESTFLTQPDAAHLADPSPRGAPGAEPTSYLQTSGSVYDAEPRVISNLIADQTLSNPAVIAAA
LSHAGMTGQAMLTTANEIVQAYQRVIDAQAAAGNVDQALELQRQELQTALDTATAELTAAQG
DVAAKTTAKNEADQAVTQAQDTLDNAAATMVTLQGQGQVAEMQVALNAAVAAHTQAEADL
KAAQDELLQAESAAASMLTMHNAKQTTVTNLQGQKATADQELATAEGLLSDARAALALETDA
AGAVTEAQTAVENATTAVSNAQTRVNELDAQLTQAEADLADLNTQLQAAEQTRDAAQTRVTA
AQEAAAATQADREDAQALFDTEVSELATAQAEMLSALTNYITGGSLADFSVASAAYSAAREEK
LAAETALNDAIDADDEADQAVIDETAALTTAQSRVSTLTTQRDNDMTTRDGLSTQLDEANGLLT
SAQNTLTAAEQTLQTAQDDNQAYVDAQNAVDLAQADVDAAQGIVDGLVAQLGVAEGELATL
NTNLGNADAEVTRLEGVVQTETTDLETAAGGVTTAQQNLAAAQMADAAILAQLDVIADATVTL
TEATATAAQAALDLTAAEGVEATKQVAVDTATSDLNALTAPGAAEAALAAAQATAAEAQTTL
DTLLTTHNITMDGNNVLLPDVTPDEGLSAPYNSWMTLFGQFFDHGLDLVGKGGSGTVYIPLQPD
DPLYDANSPTNFMVLTRATNQPGPDGILGTADDIREHFNKTTPWVDQNQTYTSHPSHQVFLREY
DIDANGNPVSNGYLLHGQTGGMATWGDVKAQAATKLGIQLNDSDVLDGPLLATDPYGNFIPGA
NGLPQLVVPNPAYVEGGTEPLNILIEGDLANPVDASQALRNGHAFLEDISHTAFPKGMIDHDRNP
MTPEIEVLPDADTDTGNAIMPNIFGMNETYDNELLDRHFIAGDGRGNENFGLTAVHHVFHTEHN
RQAAEMKQTILDSGELAFINEWLATPITEADLATATVDTVEWDGGRLFQAAKFTTEMQYQHLAF
EEFGRTVQPQIAAFGVNGSAEIDGAVMAEFAHVVYRFGHSMLTENVHTMDPNGVNTSNGLIEAF
LNPVAFDLDQTLTSDQAAGAVARGMSRETGANIDEFITSALRDNLVGLPLDLAALNIARGRETG
VPSLNAAREQFYSATGSEFLKPYEGWSEFAANLKNPASIINFIASYGTHETIVNATTVVEKRAAAT
DLVLGGDNAPADRLDFVNGTGAWADAETGINDVEFWIGGLAEDIMPFGGMLGSSFGFVFQQQM
EALQNGDRFYYLARTAGMNMIAELENNSFASMIVRNTDIKDGGAHIPANIFSSMDVILEVDQAV
QSMPDPVSTDVDPFLAAMGTTMVERATATVDAPLIDGVREYDNFLKFNGGEHAVLGGTDQRDI
LNGGLGDDALWGGAGDDLLIGDAGVNTLRGGAGNDILKDGDDVSFLHGEDGDDVISAGGGIGE
LMFGGKGNDAILMGSDDAATAFGMDGNDFIYGGAGGEFMFGGEGDDWIEGGDGFDMVMGDN
GDPFGGSRVVGHDVLDGGANDNDLLGEGGDDILIQREGVHVNEGDLGFDWVAHKDAMTGADI
DLSRRLETNETQAALRDRYDLVEAASGTQFDDFLYGDDREGAALPDGIDLLANDATLFANELTQ
AGVDRIAGLRELLADMMGVDPADTFTGGNILLGGGGSDVIQGRGGDDVIDGDKALNVRISVRDP
QDPTVELRSINSLFEISNELLDGSISPGQLQIVREILDAGQEGDVDTAVYFDVRANYNIRTNADGS
VTVDHIAPPIEGAFIDPINGQPNVEGGDGTDRLTNVEILRFADMEIDLRTDPASNGLIQGTDGDDV
LNGSNGRDAIFAGAGNDVINGLAGNDVIFGEGGSDTITWNAPNGGYDVVVGHGIVNDGGTDML
VINGDGTEDGFTMYTVAAWTGQAPVDPSSEIIITRTVAGVETAIMEVRGIEEVAINGVQGIETINA
VGDFAGTSLAPNTIYVNGTAGREVIDFSGFTSTQRLVVDAGDGRDVITGGAGNDILNAGAGDDEI
TWSVGGGSDVVDGGAGEDTYTINGDASDETFRVYAADSWTGAPLATGTDIVITRDAGAGEQVI
GQLQNIEEIQINTAGGVDNVEVIGDFNPTALNFNTIAVTGDDVSVDVTQRLSTHRVFVDVGGNTQ
DVPKSLNVAGVVTEGDVFLIPVDPTTLTRTVDGLTVTQKSDNFVMTYTVDDVDTIPVLVQSGTL
MADALRGVATVNEQTGLPEEGFGFGVLTRGATLVTADMVTADDIAALKYMVGASNVAPILPPE
AGEPADNEPADLLQEVRDLEGLTNNLQNAEISGGATLPFMRVTEARYAGIGEDGAGIINPVFDRL
DTRDISNTLGAQDLDAAKAASANMFMMSFGQYFDHGLTFIPKGSGPMGQTLGQIEIGGPSMDRG
PMADNPADLTRGSVIGYDEDGTPLQENITSPVVDQNQVYGSSSLIGQLLRESDGEMGVGARILM
GGDDPSAAPGFKILSTLRETLDHHIDAGTVFKGAGLGENGMTLLDYYPGLRDADRNYDAALVK
ELSGDFMDEGWPLLIDTNPFMNLLDHFVGGDGRANENVGLTSMHTVWARNHNYHVDQINKQA
GPDFTPEQLFQAARILNIGEYQQVVFNDFADALIGGLQGSGRHGHNEYNPNADARISHEFAAAA
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YRFGHSQIGETMLVEKDGQMVELPLIDLFLNPTSDPNAFQYVDRETGELVTGEAALAALAQMDY
VPQDGYAQYGVNSILGGLVAQPSEEVDLQVVDAVRDNLVRVSADLFAFNAARGRDLGLGTLNQ
VKADLAASTDPYIVEAIEQSNMNMDPYASWDDFQWRNNLSETEITKFKTAYPDLVLEGDAIAAF
QQVNPDITLIPGENGAMIVQGIDRVDMWVGGLAEPHIEGGVVGHTFWVLIHEQLDRLQEGDRFY
YIDRIGDLPVYNNFISNLTFGDIVTRNTGMTDLPLDVFSYVPEVPEGAAADAGNQQIQTDQNANR
TDDPAIDTADTEGGGTTTDTDVTGGADDETEAAATEGNAEPDPLETEANALSEGAVEGGVDETE
QPDADVTGSTLEQSAEEEETEEDDATQNATGAASGGEDLEATSSETNEEMPEDNTSMSPDDAGG
ADTGGADDETEAAATEGDNEPDPLETEVNALSEGAGEGEGGEAEQPDADVTGSTLEQSAEEEET
EEDDATQNATGAASGGAALEAEEEETEEDDATQNAMGAASGGADLEAEEEETEEEMPEDNTSM
SPDTDTSAEGIVESDTDMGAPLVGAAGDDILSGMAGDDMLVGNAGDDMLFGGEGRDDMLGGD
GADMLSGGAGSDHILGGDGDDMIIGGTGSDILMGNAGDDTFLGTNGDCNDLIYGGEGTDTIDM
ASVTSDLTVRLGNTGTDRGSVTTEEGGRDTIWSIENFVGGAGDDTIFASDAANVLDGGDGNDTF
VFETAASAQGDHINGLNAGDVLQFGTGADMFELTWGDTDMQDGLSFSGGEEAGTTKISGTIEDD
QFELTVSGRFDYDQIT
Supplemental Information References
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Learman, D. R., B. M. Voelker, A. S. Madden and C. M. Hansel (2013). "Constraints on superoxide mediated formation of manganese oxides." Front Microbiol 4: 262.
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