Sulfate, Sulfur Reducing Bacteria, and Homoacetic Acid Bacteria We already considered S oxidation where reduced S was used

d as an electron donor for chemolithotrophic growth. Here we consider organisms that reduce sulfate or sulfur during respiration. Sulfate or sulfur are used as terminal electron acceptors during Dissimilatory Sulfate Reduction o Review assimilatory and dissimilatory processes The use of S or SO4 as a terminal electron acceptor is an obligatory anaerobic form of metabolism where the electron donor can come from a wide variety of substrates including organic compounds (as organic acids, fatty acids, and alcohols) or the electron donor may be H2. The end product of this metabolism is H2S. Use of carbohydrates is not common. The organisms that can do this represent an extremely wide group whose majority falls within the delta Proteobacteria. They are cosmopolitan and may be isolated from soils and anoxic mud where sulfate is available (protein degradation). o Review water heaters Organisms may be characterized by three groupings o Group I sulfate reducers Use lactate, pyruvate, many alcohols, and some fatty acids as electron donors when converting SO4 to H2S Produce acetate as an end product of metabolism o Group II sulfate reducers Use fatty acids (especially acetate) and oxidize substrate completely to CO2 while converting SO4 to H2S Some may grow chemoautotrophically using H2 as the electron donor (acetyl CoA pathway) o Dissimilatory sulfur reducers Organisms that reduce elemental sulfur to sulfide, but cannot reduce sulfate to sulfide SO4 S H2S Use acetate and ethanol are common electron donors Many facultative aerobes reduce So (and other S compounds as thiosulfate, sulfite, DMSO) to H2S (Proteus, Salmonella) BUT dissimilatory S reducers are exclusively anaerobic and S is the only electron acceptor. o A look at the organisms (Source of Table 12.21: Madigan et al. 2003)

Physiology The physiology of SO4 reduction takes place 1) in cytoplasm, where carbon metabolism takes place and 2) at the cell membrane where the energetic reactions occur Some notes: o SO4 is a stable ion and must be activated before use. Activation takes place by the enzyme ATP sulfurylase and uses ATP to create adenosine phosphosulfate (APS). This requires the equivalent of 2 ATPs since the product of this reaction is AMP (Source of Figure 17.38: Madigan et al. 2003) Carbon Flow In the example we will follow, carbon flow proceeds from lactate to acetyl CoA through pyruvate. Acetyl CoA produced may be (depending on the organism) o completely oxidized to CO2, o processed by a modified TCA pathway, or o processed by the Acetyl CoA pathway Reducing power generated in carbon flow is used in the energetics

Energetics o A sequence requiring 8 electrons (generated by carbon flow) o Several forms of bisulfite reductase are known: as desulfoviridin and desulforubidin. They are named according to their absorption spectrum o A look at the energetics linked to carbon flow (Figure 17.39 (adjacent) from Madigan et al. 2003)

Energetics
4H2

2 Lactate
2 NADH + H+

Carbon Flow

lactate dehydrogenase Hydrognease

2 NADH + H+

2 Pyruvate
8e Cyto c

CoA

8 H+

pyruvate-ferrodoxin oxidoreductase CO2

2 Acetyl CoA

Pi

phosphotransacetylase CoA

2 Acetyl P

SO4
ATP ATP sulfurylase

2 ADP 2 ATP acetate kinase

Excreted

Hmc

APS
H+ 2e 5 H+ 2 HSO32e
APS reductase AMP

(Desulfovibrio sesulfurincans)

2 Acetate

8e FeS

HSO3- (bisulfite)
bisulfite reductase 3 H2O (Trithionite)

to...Acetyl CoA Pathway (Group II) Desulfobacterium

Desulfotomaculum

S3O6
H+ 2e

HSO3(Thiosulfate)

to Modified TCA Desulfobacter Pathway (G roup I)

S2O3
2 H+ 2e

HSO3-

HS-

The Acetyl CoA pathway (Source of Figure 13.4: White 2000) o This is an important pathway for many organisms and assumes as much importance as the TCA cycle in others. o The pathway is used to oxidize acetate and for autotrophic growth using CO2. o We will consider this pathway in one form or another when looking at the sulfate reducers, the homoacetogens, and the methanogens. In the Sulfate reduces we have two aspects of the cycle to consider o Oxidation of acetate to CO2 and Autotrophic fixation of CO2 o Oxidation of Acetate -------------------- = Production of acetate by non-acetate oxidizing sulfate reducers -------------------- = Production of CO2 by acetate oxidizing sulfate reducers

Autotrophic function (Source of Figure 13.4: White 2000) A series of reactions result in the reduction of CO2 to CH3 bound to tetrahydrofolic acid (THF) The CH3 is transferred to a corrinoid enzyme (CO-E) [Corrinoid enzymes are proteins containing B12 derivatives as prosthetic groups. B12 is a cobalt containing coenzyme]. CH3 is transferred to carbon monoxide dehydrogenase (CODH) which also catalyses the conversion of CO2 to CO A second CO2, which becomes the carbonyl carbon (CO) bound to the enzyme is reduced by CODH. This will eventually become the carboxyl carbon (COOH) of acetate. Electrons to do this come from H2 CODH condenses CH3 and CO to form CH3COa bound acetyl group Bound acetyl group reacts with CoA~SH to form acetyl CoA which will be acted upon by pyruvate synthase (a ferredoxin-linked enzyme) which will add a CO2 to create pyruvate. The pyruvate can be incorporated into cell material.

Sulfur metabolism in those cells utilizing acetate by the TCA cycle (Desulfobacter) (Source of Figure 13.9: White 2000) Initial step is activation of acetate to acetyl CoA at the expense of succinyl CoA. This is an energy saving stepnormally to make acetyl CoA from acetate requires ATP (acetate + ATP + CoA Acetyl CoA) Citrate lyase cleaves acetyl CoA and conserves the energy to ATP via substrate-level phosphorylation

CoA transferase

Acetate

The Acetogens (Source of Figure 13.4: White 2000) These are obligate anaerobes utilizing CO2 as the terminal electron acceptor and they produce acetate as the only product of anaerobic respiration. The overall reaction looks like CH3COO- + 4H2O 4H2 + H+ + 2HCO3 Electrons for reduction of CO2 come from H2 (chemolithotrophic growth) or other C1 compounds, sugars, organic acids, alcohols, amino acids, and certain nitrogen bases (chemoorganotrophic growth). CO2 is converted to acetate via the acetyl CoA pathway A diverse group of organism are capable of this type of metabolism

References:
Madigan, M.T., J.M. Martinko, and J. Parker. 2003. Brock: Biology of Microorganisms. Prentice Hall.

White, D. 2000. The physiology and biochemistry of prokaryotes. Oxford University Press, New York.