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. 2003 Aug;69(8):4628–4638. doi: 10.1128/AEM.69.8.4628-4638.2003

TABLE 2.

Biochemical characteristics of purified chloroethene RDasesa

Enzyme Organism Molecular mass (kDa)
Localizationb Apparent Km (μM)
Sp act (nkat/mg)
Cofactor(s)
SDS-PAGE Gel filtration PCE TCE PCE TCE
PCE-RDase Dehalococcoides ethenogenes 51 NDc m ND 342 Corrinoid, Fe/Sd
Desulfitobacterium sp. strain PCE-1 48 ND m ND 92 Corrinoidd
PCE/TCE-RDase Dehalospirillum multivorans 57 58 s 200 240 2,640 ∼2,200 1 Corrinoid, 8Fe/8Sf
Dehalobacter restrictus 60 164/71g m 20 24 250 338 1 Corrinoid, 2[4Fe/4S]h
Desulfitobacterium hafniense strain PCE-S 65 198 m 10 4 650 690 1 Corrinoid, 8Fe/8Sf
Desulfitobacterium hafniense strain TCE1 59 ND m ND ND 190 157 Corrinoidd
Desulfitobacterium sp. strain Y51 58 67 s 106 535 2.7 13.5 Corrinoidd
Clostridium bifermentans strain DPH-1 35 70 s 12 1.2 Corrinoidd
TCE-RDase Dehalococcoides ethenogenes 61 ND m ND 200 Corrinoid, Fe/Sd
pH opti- mum Temp optimum (°C) Oxygen sensitivity half-life (min) Inhibitors Other substrates Compounds not dechlorinated N-terminal amino acid sequence Reference(s)
NRe NR NR 1-Iodoethane, sulfite, dithionite, cyanide, azide NR TCE, 1,1-DCE, cis-1,2-DCE, trans-1,2-DCE, VC NR 19
NR NR NR 1-Iodopropane, N2O, sulfite TCE cis-1,2-DCE GQESESAIVXFAVQXV 42
8 42 120 1-Iodopropane, sulfite, cyanide, nitrite, N2O, EDTA, chlorinated methanes trans-1,3-DCP, 2,3-DCP, 1,1,3-TCP 1,1-DCE, cis-1,2-DCE, trans-1,2-DCE, HCA, 1,1,1-TCA, 3-CB, 3,4-DCB, 4-CP GVPGANAAEKEKNAA EIRQQFAMTAGS 25-27
8.1 NR 280 1-Iodopropane CT, HCA, 1,1,2,2-TeCA, 1,1,1,2-TeCA, 1,1,2-TCA, (1,1,1-TCA) 1,1-DCE, cis-1,2-DCE, trans-1,2-DCE ADIVAPITETSEFPYKV DAK 33, 34; this study
7.2 50 50 1-Iodopropane, cyanide, azide, sulfite, EDTA NR CT, 1,1-DCE, cis-1,2-DCE, trans-1,2-DCE ADIVAPITETSEFPYKV DAK 21, 22
NR NR NR NR NR cis-1,2-DCE, chlorophenols ADIVAPITEXTEFPYPV 42
7.2 37 330 1-Iodopropane, sulfite HCA, PCA, 1,1,2,2-TeCA, 1,1,1,2-TeCA 1,1-DCE, cis-1,2-DCE, trans-1,2-DCE, 1,1,1-TCA, 1,1,2-TCA ADIVAPITETSEFPYKV DAK 41
7.5 35 1,200 1-Iodopropane 1,1-DCE, cis-1,2-DCE, trans-1,2-DCE, 1,1,1-TCA, 1,1,2-TCA VC AEVYNKDGNKLDLYGK VDGLHYFSNDT 29
NR NR NR 1-Iodopropane, sulfite, dithionite, cyanide, azide, Cu2+, Zn2+ 1,1-DCE, cis-1,2-DCE, trans-1,2-DCE, (VC), (VB), 1,2-DCA, 1,2-DBA PCE, 1,1,2,2-TeCA KDVDDLLSAGKALEGD HANKVNNHPWW 18, 19
a

Abbreviations for compounds: VC, vinyl chloride; DCP, dichloropropene; TCP, trichloropropene; CT, carbon tetrachloride; HCA, hexachloroethane; PCA, pentachloroethane; TeCA, tetrachloroethane; TCA, trichloroethane; DCA, dichloroethane; VB, vinyl bromide; DBA, dibromoethane; CB, chlorobenzoate; DCB, dichlorobenzoate; CP, chlorophenol.

b

m, membrane fraction; s, soluble fraction.

c

ND, not determined.

d

Determined by photo-reversible inhibition of the reduced enzyme by iodo-alkanes.

e

NR, not reported.

f

The corrinoid was determined by photo-reversible inhibition of the reduced enzyme by iodo-alkanes and was quantified by analysis of cobalt content and by extraction of the corrinoid from purified enzyme, followed by spectroscopic analysis. The 8Fe/8S was quantified by atomic absorption spectroscopy and by standard procedures for acid-labile sulfide.

g

The molecular mass depended on the detergent used; it was 164 kDa with Triton X-100 (micelle mass, 90 kDa), and it was 71 kDa with OGP (micelle mass, 8 kDa).

h

The corrinoid was determined by photo-reversible inhibition of the reduced enzyme by iodo-alkanes and by EPR spectroscopy and was quantified by analysis of cobalt content and by extraction of corrinoid from purified enzyme, followed by spectroscopic analysis. The presence of two [4Fe/4S] clusters was determined by EPR spectroscopy, and the iron and sulfur contents were quantified by atomic absorption spectroscopy and by standard procedures for acid-labile sulfide.