Syntrophomonas wolfei features:

 “Crotonate cultures were prepared in a similar manner substituting equimolar

amounts of crotonate for butyrate. Culture purity was checked by regular
microscopic examination and inoculation of thioglycolate broth (Difco Laboratories,
Inc., Detroit, MI, USA), which does not support the growth of either species.”
“Generation times were calculated from the linear portion of the growth curve by
measuring the time the culture took to double its optical density. Direct microscopic
counts were done using a Petroff-Hausser counting chamber”. Growth was
monitored by spectrophotometry at 600 nm. S. wolfei was grown in co-culture with
M. hungatei and the “best” ratio was when grown with butyrate (1:3) compared with
grown with only crotonate (1:47). After grown in co-culture, S. wolfei was then grown
pure in agar plates, where it appeared as tiny opaque colonies of 1 mm od diameter
(0.2% crotonate). After agar, pure cultres were grown with 0.2% crotonate and 0.3
Na2SO4 and grown was achieved within 360 h. Growth was observed as well without
Na2SO4. Growth was inhibited by the presence of 20% of H2 in the headspace. When
incubated with butyrate, growth wasn’t achieved unless co-cultured with M.
hungatei or D. vulgaris (in the presence of sulfate*). Max absorbance was 0.47 when
co-cultured. Max growth rate of pure S. wolfei was 0.029 1/h with 20-70 mM
crotonate was used. Max PHB percentage obtained when grown pure was 20%.
Gram-negative, slightly helical rods, 0.5 to 1.0 by 2.0 to 7.0 µm with slightly tapered
rounded ends. Most cells occur singly or in pairs with helical chains of three or more
often observed. Multiplication by binary fission. Cells possess two to eight flagella
with a diameter of about 20 nm that are laterally inserted in a linear fashion on the
concave side of the cell about 130 nm or more apart. Under most conditions, cells
usually exhibit only a sluggish twitching motility.(McInerney, Bryant, Hespell, &
Costerton, 1981)(Beaty & McInerney, 1987)
 Grows only with fatty acids of 4 to 8 carbons and produces propionate and acetate.
Medium had originally rumen fluid (difficult stuff to get) but S.wolfei was able to be
grown without it in a “new medium”. Medium was boiled under an 80% N2- 20% CO2
gas phase. Growth of 0.039 1/h and absorbance of 0.5 with rumen fluid. With the
“new medium” the growth rate was 0.023 and absorbance od 0.08; however, adding
lipoic acid, B12, B7 and B1 resulted in a similar growth as with rumen fluid. Max
absorbance was reached at 311 h of incubation. No growth was observed before 192
h. p-aminobenzoic acid stimulated growth as well. The deletion of trace minerals
(except FeSO4 and CoCl2*6H2O) in the defined medium with vitamins didn’t affect
growth. (Beaty & Mcinerney, 1990)
 S. wolfei has two subspecies: wolfei and saponavida. S.wolfei wolfei degrades fatty
acids with 4 to 8 carbons and S. wolfei saponavida degrades fatty acids with 4 to 18
carbons. Best growth rates occur at near neutral pH and with temperatures between
35-37 °C. The use of crotonate as substrate bypasses an unfavorable step of the
oxidation of butyril CoA to crotonyl CoA. For each crotonate oxidized to acetate,
 another crotonate is reduced to butyrate. (Dworkin, Falkow, Rosenberg, Schleifer, &
Stackebrandt, 2006)
 The substrate used to grow S. wolfei had 20 mM sodium crotonate, 0.4 mg/L of
resarzurine, 200 µg/L of lipoic acid and 400 µg/L of thiamine. “Growth was monitored
by determining the optical density (OD) against sterile medium. Prior to
measurement, a few grains of sodium dithionite were added to the cuvettes to keep
resazurine in its reduced state.” The end of exponential growth was achieved at 10
days when co-cultured. (Müller, Schleheck, & Schink, 2009)

 Two thirds of the ATP made go

to the reverse electron
transport and the other third
goes to growth. The reverse
electron transport produces
H2 and formate. Electron
transport by its pili wasn’t
observed since S. wolfei lacks
the genes for that. It lacks the
genes for aerobic and
anaerobic respiration. Stains
for gram negative. Comes
from the philum Firmicutes.
Has motility (*) and probably
has the capability of
sporulating. Produces PHA
while it is in exponential
growth without being stressed
by nutrient limitation, it does it
by converting two molecules
of acetyl-CoA to acetoacetyl-
CoA and then to D(-)-3-
hydroxybutyryl-CoA, or by
using a b-oxidation
intermediate to D(-)-3-
hydroxybutyryl-CoA. The
substrate used to grow S.
wolfei had 20 mM sodium
crotonate. (Sieber et al., 2010)
References

Beaty, P. S., & Mcinerney, M. J. (1990). Nutritional Features of Syntrophomonas-Wolfei.

Applied and Environmental Microbiology, 56(10), 3223–3224.
Beaty, P. S., & McInerney, M. J. (1987). Growth of Syntrophomonas wolfei in pure culture
on crotonate. Archives of Microbiology, 147(4), 389–393.
https://doi.org/10.1007/BF00406138
Dworkin, M., Falkow, S., Rosenberg, E., Schleifer, K.-H., & Stackebrandt, E. (Eds.). (2006).
The Prokaryotes. New York, NY: Springer US. https://doi.org/10.1007/0-387-30744-3
McInerney, M. J., Bryant, M. P., Hespell, R. B., & Costerton, J. W. (1981). Syntrophomonas
wolfei gen. nov. sp. nov., an Anaerobic, Syntrophic, Fatty Acid-Oxidizing Bacterium.
Applied and Environmental Microbiology.
Müller, N., Schleheck, D., & Schink, B. (2009). Involvement of NADH:acceptor
oxidoreductase and butyryl coenzyme A dehydrogenase in reversed electron transport
during syntrophic butyrate oxidation by Syntrophomonas wolfei. Journal of
Bacteriology, 191(19), 6167–6177. https://doi.org/10.1128/JB.01605-08
Sieber, J. R., Sims, D. R., Han, C., Kim, E., Lykidis, A., Lapidus, A. L., … McInerney, M. J.
(2010). The genome of Syntrophomonas wolfei: New insights into syntrophic
metabolism and biohydrogen production. Environmental Microbiology, 12(8), 2289–
2301. https://doi.org/10.1111/j.1462-2920.2010.02237.x
……