B

i
o
m
a
s
s

o
r

O
i
l

(
g
/
L
)
0
10
20
30
Biomass
SCO
ECO
B
i
o
m
a
s
s

o
r

O
i
l

(
g
/
L
)
0
10
20
30
40
50
Biomass
SCO
ECO
Lucas McNea & Robert W. Nicol
University of Guelph Ridgetown Campus, Ridgetown ON Canada N0P 2C0
Centre for Agricultural Renewable Energy and Sustainability, www.ridgetownc.com/cares
ABSTRACT
Thin stillage and crude glycerol are produced during
manufacture of ethanol and biodiesel respectively. Although
there is some demand for these biofuel co-products, their full
value has not been realized due to a saturated distillers grains
market and due to the low purity of crude glycerol. Our goal is
to use these substrates in a novel bioconversion application
based on the newly described oleaginous fungus Galactomyces
geotrichum. Thin stillage was found to have a pH of 4.8 and
potential fungal inhibitors, organic acids and residual ethanol,
were found to be low at 0.3%. Crude glycerol was found to
have a pH of 9.4 and be composed of 29.7% residual oils, 28%
glycerol and 26% methanol. Quantity of elements was
generally low except for potassium, originally from the
potassium hydroxide catalyst, which was found at a
concentration of 28,000 g g
-1
. Galactomyces geotrichum was
cultured in a range of biofuel co-product concentrations and
found to produce 12.5 to 41.0 g L
-1
biomass, and 2.1 to 50.1 g/L
of oil. Galactomyces geotrichum is a promising microorganism
for valorizing biofuel co-products and work on scaling-up this
process will continue.
Corn
Thin
Stillage*
Stillage
EtOH
CDS
WDG
Crude
Glycerol*
Bio-
diesel
Vegetable
Oil
Figure 1: Simplified ethanol production process.
Thin stillage is the mostly liquid fraction of the
whole stillage and was used in this study.
Figure 4: Biomass and oil production by Galactomyces geotrichum cultivated
for 5 (A) or 3 (B) days in mixtures of biofuel co-products. There is 29 g/L of
residual oil in the co-products, so total oil above this level, as seen in CG +
100% TS, represents a net gain. TS=thin stillage, CG=50 g/L crude glycerol,
SCO=single cell oil, ECO=extracellular oil (recovered from spent medium).
Figure 3: Bioconversion of mixtures of thin stillage from ethanol
production and crude glycerol from biodiesel production. Galactomyces
geotrichum converts these biofuel co-products to single cell oil, a new
biodiesel feedstock or potentially useful in higher value applications.
Figure 2: Simplified biodiesel production process.
Crude glycerol is a product of the transesterification of
the triacylglycerol molecule and was used in this study.
distillation
centrifugation
+
evaporation
+
DDGS
MeOH
KOH
Thin
Stillage
Crude
Glycerol
Bioconversion
Single
Cell
Oil
100%
TS
70%
TS
CG +
10%
TS
CG +
70%
TS
CG +
100%
TS
100%
TS
CG +
70%
TS
A B
Fatty acid Amount (%)
16:0 12.51
16:1 0.89
18:0 3.87
18:1 35.46
18:2 37.39
18:3 3.35
20:0 0.37
20:1 0.44
Table 1: Fatty acid composition
of the single cell oil by GC-FID.
SUMMARY
Galactomyces geotrichum was able to grow
in untreated thin stillage and crude glycerol

this fungus readily produced biomass, but
only produced a net gain of oil in undiluted
thin stillage plus 50 g/L crude glycerol

the single cell oil contains very little
omega-3 fatty acids, but is suitable for
biodiesel production