Isolation and characterization of a novel Dehalobacter species strain TCP1 that reductively dechlorinates 2,4,6-

trichlorophenol

Shanquan Wang,† Weijie Zhang,‡ Kun-Lin Yang,‡ and Jianzhong He*,†

† Department of Civil and Environmental Engineering‡ Department of Chemical and Biomolecular Engineering

National University of Singapore, Singapore 117576

Figure S1. DGGE characterization of dechlorinating bacteria present in culture TCP1 after 21 times of serial dilution. The PCR products were obtained by using the universal bacterial primer set, 341FGC and 518R.

Dehalobacter

Dehalobacter16S rRN

A

5 µm

Figure S2. Morphology of Dehalobacter sp. strain TCP1 as observed with fluorescence microscopy. Cells were stained with fluorescent dye Syto-9 (5 µM).

Dehalobacter sp. TCP1

1

Dehalobacter sp. AD14-PCE

2

Figure S3. RpoB DGGE characterization of Dehalobacter bacteria present in culture TCP1 (lane 1) and culture AD14-PCE (lane 2) with Dehalobacter genus-specific rpoB gene targeted primers, DebrpoBFGC and DebrpoBR.

Figure S4. Sequence heterogeneity of the five 16S rRNA genes in strain TCP1 when excluding the insertion sequences (from base 255-1653).

16S-1 (JX999720)16S-2 (JX999721)16S-3 (JX999722)16S-4 (JX999723)16S-5 (JX999724)

16S-1 (JX999720)16S-2 (JX999721)16S-3 (JX999722)16S-4 (JX999723)16S-5 (JX999724)

Figure S5. Phylogenetic tree of identified putative chlorophenol reductive dehalogenase gene - debcprA. Phylogenetic analyses are conducted in MEGA4 (Tamura et al. 2007).

0.2

debcprA (Dehalobacter sp. TCP1, JX999725)

putative rdhA (Dehalobacter sp. DCA, 13831363)

cprA (Desulfitobacterium chlororespirans, AAL84925)

pceA (Desulfitobacterium hafniense Y51, YP519072)

cprA (Desulfitobacterium sp. PCE-1, AF259790)

cprA (Desulfitobacterium hafniense DCB-2, AY013365)

cprA (Desulfitobacterium hafniense Y51, YP517388)