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FEMSEC 00331
4. R E S U L T S 0 I
0 4 8 12 16 20 24 28
4.1. Glycerol degradation
Fig. 1 shows the kinetics of glycerol dissimila- Time (days)
Fig. 1. Anaerobic degradation of glycerol by D. alcoholovorans
tion by D. alcoholovorans in the presence of sulfate in pure culture, in the presence of sulfate (a), or in coculture
(Fig. la) and in the presence of M. hungatei (Fig. with M. hungatei (b). I, glycerol; r,, acetate; 0, sulfide; O,
lb). Glycerol dissimilation by D. alcoholovorans in methane.
the presence of sulfate or in association with M.
hungatei led to acetate plus sulfide or methane
production, respectively, and presumably to that
of carbon dioxide. The m a x i m u m specific growth
rate of a pure culture of D. alcoholovorans in the specific growth rate of 0.047 h - 1 and a m a x i m u m
presence of sulfate was 0.22 h - l ; the m a x i m u m cell yield of 9.4 g dry weight/tool glycerol de-
cell yield was approximately 11 g dry w e i g h t / m o l graded were recorded (Table 1). F r o m the results
glycerol degraded (Table 1). In association with in Table 1, the stoichiometry in the presence of
M. hungatei in the absence of sulfate, a m a x i m u m sulfate can be approximated by Eqn. 1 and by
236
Eqn. 2 in the coculture with M. hungatei: In the absence of sulfate, the hydrogen-consum-
CHzOH C H O H CH2OH + 0.75 CO~- ing methanogenic bacterium M. hungatei served
as an alternative acceptor for reducing equivalents
---, CH3COO + H C O f + 0.75 HS
released by D. alcoholovorans from 1,2-propane-
+ 1.25 H + + HzO (1) diol degradation (Fig. 2b). But only propionate
CH2OH C H O H CH2OH was produced in addition to methane and pre-
CH3COO-+ 0.25 HCO 3 + 0.75 CH 4 sumably CO 2. The maximum specific growth rate
+ 1.25 H + + 0.25 H 2 0 (2) of this coculture was 0.005 h - ~. The maximum cell
yield was 2.8 g dry weight/mol 1,2-propanediol
4. 2. 1, 2-Propanediol degradation degraded (Table 1). The following theoretical
In the presence of sulfate, D. alcoholovorans stoichiometry can be proposed:
degraded 1,2-propanediol to acetate, propionate
CH2OH C H O H CH 3 + 0.25 H C O f
and presumably COz with production of sulfide
(Fig. 2a). The acetate/propionate ratio was vari- CH 3 CH 2 COO + 0.25 C H 4 q- 0.75 H +
able from one experiment to the other (see Fig. 2a
+ 0.75 H 2 0 (4)
and Table 1), probably because of the pH or
redox potential of the media used. Apart from
acetate and propionate, no other organic acid was 4. 3. 1,3-Propanediol degradation
detected. This strain did not produce any solvent As expected, 1,3-propanediol degradation led
such as propanol. The maximum specific growth to higher sulfide or methane production than
rate was 0.09 h l. The maximum cell yield was glycerol degradation. At the beginning of incuba-
approximately 7.2 g dry weight/mol 1,2-propane- tion, in the presence of sulfate, 1,3-propanediol
diol degraded (Table 1). As shown in Table 1, D. degradation led only to acetate, sulfide, and pre-
alcoholovorans degraded 1,2-propanediol as fol- sumably CO 2 production (Fig. 3a). After about 4
lows: mmol 1,3-propanediol per litre had been de-
CHzOH C H O H CH 3 + 1.025 SO~- graded, 3-hydroxypropionate was formed as a fur-
ther end-product with an acetate/3-hydroxypro-
0.8 CH3COO + 0.1 C H 3 CH a C O O -
pionate ratio of approximately 3.4. The maximum
+ 1.025 HS +0.975 H + + H 2 0
of 3-hydroxypropionate produced was approxi-
+ 1.1 HCO 3 (3) mately 1.2 mmol per litre (Fig. 3a). The maximum
Table 1
Yield determinations and stoichiometry a for growth of D. ah'oholovorans on glycerol, 1,2-propanediol and 1D-propanediol in the
presence of sulfate (lines 1, 3 and 5) or in coculture with M. hungatei in the absence of sulfate (lines 2, 4 and 6)
Substrate 1 2 3 4 5 6 7 8 9
Substrate OD Cell Cell Cell Acetate Propionate H2S (S) e-
degraded 580 material yield formed (mM) (mM) or recovery L-
(mM) formed (g/mol) expressed CH 4 (M) (%)
(rng/mol) as acetate b (mM)
(mM)
Glycerol 10 0.26 110.6 11.1 2.3 7.9 0 6.5 (S) 96
Glycerol 8.8 0.21 82.7 9.4 1.7 7 0 5.30 (M) 91
1.2-propanediol 10 0.18 72 7.2 1.5 7.7 1.1 6.9 (S) 90
1.2-propanediol 10 0.055 27.5 2.8 0.6 0 10.5 1.5 (M) 102
1.3-propanediol 10 0.28 127.3 12.7 2.6 7.2 0 7.1 (S) 85
1.3-propanediol 10 0.24 102.6 10.3 2.1 7.6 0 7.6 (M) 86
'I
\ j/4 --.
20 _ ~ 5
8
15 0 ~ , I I | i
20
o _ |
0 2 4 6 8 10 12 14 16 18 20 ~ 10
Time (days) i
20 _ ~ 5
b o
0 2 4 6 e 10 12 ~4 16
Time (days)
Fig. 3. Anaerobic degradation of 1,3-propanediol by D. al-
coholovorans in pure culture, in the presence of sulfate (a), or
in coculture with M. hungatei. (b) I , 1,3-propanediol; * ,
presence of sulfate:
° 0 C H 2 O H C H 2 C H 2 O H + SO~
0 2 4 6 8 I0 12 14 16 18 20 CHsCOO-+ HCOj + HS-+ H++ H20
Time (days) (5)
Fig. 2. Anaerobic degradation of 1,2-propanediol by D. al-
coholovorans in pure culture, in the presence of sulfate (a), or Without sulfate and in coculture with M.
in coculture with M. hungatei (b). I , 1,2-propanediol; A, hungatei 1,3-propanediol was first degraded to
propionate; zx, acetate; ~ , sulfide; O, methane. acetate, 3-hydroxypropionate, methane and pre-
238
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