Biotechnology Report
Economic Evaluation of Alternative
Ethanol Fermentation Processes
B. L. Maiorella, H. W. Blanch, and C. R. Wilke
Lawrence Berkeley Laboratory and Department of Chemical Engineering,
University of California, Berkeley, California 94 720
Accepted for Publication December 22, 1983
Eleven alternative fermentation schemes for ethanol
production are compared. Conventional batch, continu-
ous, cell recycle, and immobilized cell processes, as
well as membrane, extraction, and vacuum processes
which remove ethanol from the broth selectively as it is
produced, are considered. The processes are compared
on identical bases using a consistent model for the
yeast metabolism. Both molasses and cellulose hydro-
lyzate are considered as feeds. Optimized ethanol plants,
including feed preparation, fermentation, and product
recovery sections are designed and total costs are
projected.
INTRODUCTION
Many new processes for ethanolic fermentation have
been proposed and studied.'.2 These investigations, how-
ever, generally have been limited to the laboratory scale.
Based on these initial studies, it is now possible to compare
the various processes and to select those most promising
for further larger scale testing. A general fermentative
ethanol production plant facility model is used for the
comparison of alternative fermentation schemes on identi-
cal economic bases; it includes not only direct fermenta-
tion costs, but associated costs for distillation, feed prep-
aration, and stillage handling.
The fermentation process design is central to the overall
plant economic comparison. The fermentation equipment
for a conventional 2 X lo9 L/yr facility described by
Katzen3 makes up 11% of the total fixed capital cost for
the plant and, thus, fermentation has a large direct effect
on overall plant costs. Furthermore, the fermentation
process, by dictating feed sugar concentration and prod-
uct cell and ethanol concentrations, specifies the major
flows and concentrations throughout the remainder of the
plant.
Distillation has been assumed highly energy intensive
and much fermentation development work has been di-
rected toward producing more concentrated alcohol beers
to reduce the distillation cost. A detailed evaluation of a
two-stage vacuum distillation process4 has shown, how-
ever, that while the distillation cost increases rapidly at
Biotechnology and Bioengineering, Vol. XXVI, Pp. 1003-1025 (1984)
0 1984 John Wiley &Sons, Inc.
levels below 6 wt 9'0 ethanol, essentially no further savings
are realized in increasing the feed concentration beyond
9 wt 7 0 .
The costs of auxiliary process steps (feed sterilization,
centrifugation, stillage evaporation, etc.) are often ne-
glected, but these process steps are important, being both
capital and energy intensive. Stillage handling equipment
may contribute 36% of the capital cost of a conventional
batch fermentation plant.3 The energy consumption for
stillage evaporation to produce a dried cattle feed supple-
ment is comparable to the distillation energy requirement.
The cost of these auxiliary process steps is directly related
to the amount of fluid handled, and large savings can be
achieved if the main fermentation step can use concen-
trated feed solutions and produce concentrated products.
If the sugar feed source is dilute (as from most proposed
cellulose hydrolysis processes), then a feed sugar concen-
trator must be added and some of the advantages of fer-
mentation at high feed and product concentration are lost.
The use of a single fermentation plant design model does
aliow the comparison of the various proposed processes on
identical bases. However, since there are alternative fer-
mentation methods, there are also alternative methods for
the other plant process steps. Special cases may exist where
carefully chosen process steps will combine synergistically
to give reduced costs and these special cases may be missed
in this general design comparison. Thus, this comparison
should be considered only as a starting point in the evalua-
tion of fermentation process alternatives.
THE FERMENTATION KINETIC MODEL
The various fermentation processes will be compared on
identical bases using a single fermentation kinetic model
based on data of Bazua for the high rate brewing yeast
Saccharomyces cerevisiae var. a n a r n e n s i ~ .The
~ , ~ fermen-
tation reaction cpn be expressed as:
glucose(S) + nutrients cell\ JS cntilly\t
et hanol(P)
+ cells(X) + by-products (I)
CCC 0006-35921841091003-23$04.00
For the yeast Saccharomyces c e r e v i s i ~ e , the
~ * ~specific
ethanol production rate can be given as:
v - -.
I s 1 r, - -1” P
p”lax
(2)
where v is the specific ethanol production rate (g ethanol
producedlg cells h); S is the substrate (glucose) concen-
tration (g/L); P is the product (ethanol) concentration
(g/L); Y, is the maximum specific production rate (1.85
L/h); C , is the Monod constant = 0.315 (g/L); n is the
toxic power constant (0.36); and P,,, is the maximum
product concentration = 87.5 (g/L). The first term in
brackets is the standard Monod form’ representing the
decrease in fermentation activity under low substrate
(starvation) conditions. The second bracketed term re-
flects the inhibitory effect of ethanol at high concentra-
tiom8 This effect is summarized in Figure 1, which shows
the reduction in specific ethanol productivity with in-
creased ethanol as given by the equation and measured by
Bazua. Fermentation activity is completely halted at 87.5 g
ethanolll.
r I I I I 1
Ethanol Concentrotion (p/f)
Figure 1. Inhibitory effect of ethanol on specific ethanol productivity
by Succhuromyces cerevisiue. Data are from ref. 6.
Figure 2. Overall fermentative ethanol plant flow model for fermentation without sclective ethanol rcmoval.
1004 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984
The cell growth rate I/. (g new cells formed/g cells h) can
be related directly to the ethanol production rate as:
p=Ev (3)
where E is the efficiency of substrate utilization for cell
mass production. For nonstressed cells in the optimized
medium of Bazua, the conversion efficiency is 0.249.
The rate of glucose utilization can be related to the eth-
anol production rate by a yield factor:
Y = 0.434 = AP/M (4)
This value is less than the theoretical yield for the conver-
sion of glucose to ethanol:
1.0gglucose - OSlgethanol+ O.49gCO2 (5)
since a portion of the glucose is utilized in cell mass and
secondary product formation.
THE FERMENTATION PLANT DESIGN MODEL
Two general arrangements of process flows and equip-
ment can be used to describe all of the designs considered.
Figure 2 covers recycle and (with the recycle stream flow
set equal to zero) simple continuous processes. By stagger-
ing harvest times for batch fermentors to provide a con-
tinuous product flow, Figure 2 can also describe a batch
production facility. Figure 3 shows the general case of fer-
mentation with selective ethanol removal. The processes
are operated as follows.
Molasses (the sugar source, at 50 wt 7’0 glucose) and
added nutrients are taken from storage and mixed with
water to the desired feed concentration and sterilized by
direct steam injection in a continuous sterilizer. Other
E rHANOL
STORAGE
 Figure 3. Overall fermentative ethanol plant flow model for fermentation with selective ethanol removal.
feeds can be considered. If a dilute hydrolyzate sugar feed9
is used, then the molasses storage facilities are eliminated
but a multieffect feed evaporator is added to concentrate
the glucose to the desired feed concentration.
The fermentor size and product flows are specified ac-
cording to the fermentation system mass balance and ki-
netic equations for the particular reactor design chosen. It
is important to note the unusual way in which stream flows
are computed. Flows are in kg/h of water in the stream.
All other components are then referenced to the water flow
and reported in g/L of water (not of total solution). This
leads to a considerable simplification, since the nonideal
mixing of sugar, ethanol, and water can be neglected and
the volumetric contribution of the yeast need not be con-
sidered. Mass balances are rigorous on this basis. Some
discrepancy with the kinetic equations is introduced,
though, since these relations are correlated in terms of
grams of ethanol and glucose per liter of clarified (yeast-
free) solution. At the low concentrations found in the fer-
mentors, the two concentration measures are similar and
no problem results.
Filtered air is sparged into the fermentors to maintain
the optimum oxygen concentration. An absorber is used
to recover ethanol from the fermentor vent gas.
For selective ethanol removal processes, the fermenting
beer is cycled through a selective ethanol recovery device
(membrane separator, or flash v e s ~ e l ' ~ ? ~then
to recover a concentrated ethanol product for distillation
and an ethanol depleted beer for recycle to the fermentor.
A portion of the dilute beer is centrifuged. A bleed-the
clarified centrifuge supernatant (containing inhibitors
which are not removed by the selective ethanol removal
device)-is fed to a stripper for initial concentration.
MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT
Stripper vapors and the concentrated ethanol product
steam from the separator are both then fed to a single
vacuum distillation column for final product recovery.
For processes without selective ethanol removal, the
whole beer product is centrifuged. If cell recycle is used,
then a portion of the concentrated yeast cream is returned
to the fermentors to boost the fermentor cell density (and,
hence, the reactor rate). The remaining yeast cream is fed
to a rotary steam tube dryer to produce a dried yeast cattle
feed supplement,16 while the supernatant is fed to the
distillation system (stripper and vacuum column) for eth-
anol product recovery.
The bottom liquid from the stripper contains residual
cells (the centrifuge supernatant is not perfectly clarified),
nonmetabolized feed components, and fermentation by-
products including proteins." This bottom product is
concentrated to 30 wt TOsolids in a multieffect evaporator
and mixed back with the yeast cream feed to the rotary
dryer as a further nutrient supplement. This recycling
eliminates all waste streams, and no special waste disposal
facilities are required.I8
The rotary steam tube dryer is operated at high temper-
ature using 600 psig steam to reduce the drying area. The
ethanol-water vapors evaporated from the yeast (at 50
psig) are condensed in the stillage evaporator to provide a
portion of the evaporation heat. This condensed liquid is
~ ) also fed to the stripper for recovery of residual
ethanol. The last effect of the stillage evaporator generates
1 atm steam. If excess heat is available from the rotary
dryer, a reboiler is used to generate additional low-pressure
steam. It is assumed that low-pressure steam could be used
in auxiliary processes and a small credit is taken for the
low-pressure steam generated.
1005
 The distillation system consists of an atmospheric pres- Table I. Materials. utility, and labor costs (on a mid-1981 basis)
sure stripper followed by a vacuum distillation column to
Ruw muterials
produce 95 wt '70azeotropic alcohol. Vapor reuse meth- $8S/ton
Molasses (50 wt '70 sugar, 95%
ods are used to achieve a high energy efficiency. Figure 4 utilizable)
presents equipment costs and utilities requirements for Nutrients 1 . 1 lollethanol product
distillation as a function of feed ethanol c~ncentration.~ Process water S0.4~/1000gal
The fermentation plant is designed assuming 330 days/ Utilities
600 psig steam $4.41/1000 Ib
yr and 24 h/day operation, producing lo9 L/yr of 95 wt 70 SO psig steam $3.15/1000 Ib
alcohol. Equipment costs are derived from the figures of 1 atm steam exhaust (for credit) $0.76/1000 Ih
Peters and Tirnmerha~s,'~ or from manufacturers quota- Electricity 8.82clkw h
tions,20and updated to a mid-1981 basis (MSI = 717). Cooling water (65°F) 22.6kl1000 gal
Labor, utilities, and raw materials costs (for the mid-1981 Lubor
Operator man hours
period) are summarized in Table I. A conservative credit
(excluding supervision) $10/man h
of 11.4 US cents/kg of 50 wt TOdried yeast
is taken for cattle feed supplement and 75.6C/1000 lb for
low-pressure steam.I9 Finally, an assessment is added to Fino1 Fermentor Ethanol Concentration ( q f l ) *
provide a 15% simple after-tax return on total capital in- 15 65 75 85

vestment as profit.

FERMENTATION PROCESS ECONOMIC

COMPARISON

Batch Fermentation
Conventional fermentation is by batch methods23which
provide the best benchmark for comparison of newer pro-
cess alternatives. The comparison will be made for a typi-
cal lo9 L/yr plant capacity and using molasses feed.
Figure 5 shows the effect of feed sugar concentration on /

batch fermentation residence time and volumetric produc-

tivity. Corresponding alcohol sale prices (for a 15% after-
10 II 12 13 14 15 16 17
tax return on investment) are presented in Figure 6 . For Inlet Sugar Concentration ( ~ 1 9 6 )
all cases, fermentation is to 2.8 g/L residual sugar concen-
tration (sufficient to maintain 90% of the non-feed-limited Figure 5. Batch fermentation volumetric productivity vs. ethanol
fermentation rate). As the feed sugar concentration is in- product concentration [reported as g ethanol/L water (see text)].
creased, the final product cell density increases. This bal-
ances the reduction in specific (per unit cells) ethanol pro- F e r m e n t o r Ethanol Product Concentration (q/tI
55 65 75 85
ductivity at increased ethanol concentration. The residence I I 1 I I -
61 I

Conventional Botch

+-
53 -

51 -
I I 1 I I I I
10 I1 12 13 14 I5 16 17
I n l e t Sugar Concentrotion (wt %)

Figure 6. Ethanol product cost for conventional and agitated high pro-
Figure 4. Equipment costs and utilities requirements for distillation. ductivity batch fermentation systems.

1006 BIOTECHNOLOGY A N D BIOENGINEERING, VOL. 26, SEPTEMBER 1984

time increases only slowly with increasing product concen- I I I I I 1
2[
tration until very near the maximum attainable ethanol
product concentration (87.5 g/L), where inhibition halts
fermentation. The maximum volumetric productivity is I(
for a 16.7 wt 70glucose feed. The minimum ethanol cost
-
also occurs at this feed concentration. A batch fermenta- E
0
tion curve with 16.7 wt % initial glucose concentration
and based on the present kinetic model is shown in Figure 7.
e%
I
cJ.+
c r
o c
An active 10% volume inoculum is used giving a high = o
initial cell density of 2.1 g/L and initial ethanol concentra-
crs
E E
tion of 8.6 g/L. A residence time of 6.54 h is required to 0s
6 6
U V
complete the f e r m e n t a t i ~ n This
. ~ ~ short reaction period -
g % I
justifies the cost of large seed tanks (one-tenth the main eS
fermentor volume) for each fermentor. qs
a
Tables I1 and I11 summarize the equipment specifica-
tions and costs for the optimized batch fermentation case.
Eight 1.27 X 105 L batch fermentors are required in stag-
gered operation with six fermenting while one is emptied
and the last one is filled. The total plant capital cost is
$28.7 million. After taking a credit of 4.78C/L for yeast by-
01
product, 53.0C/L must be charged for the 95 wt YOethanol 2 4 6 8 10 12 14
product to provide a 15% after-tax simple return on in- Time ( h r )

vestment. The manufacturing cost exclusive of raw ma-

Figure 7. Batch fermentation curve for Succhtrromyces crruvisiur var
anamensis, 16.7 wt % glucose feed concentration.

Table II. Batch fermentation purchased equipment summary.

Storuge (2werks)
Molasses 7.86 X l o h L, carbon steel 5 268.000
Ethanol 4.24 X 10' L, carbon steel lh7.000
Yeast 3.23 X lo5 L, carbon stcel, 5 units 247.000
Screw conveyor for yeast delivery 0.40 hp, 100 ft long 8.000
5 690.000
Fermentutiorz
Batch fermentor 1.27 X lo5 L, stainless steel, 8 units,
residence time = 6.54 h $ 591.000
Fermcntor agitator 84 hp, stainless steel, 8 units 433,000
Fermentor cooler stainless steel, 8 units 2x.900
Seed tank 1.27 X l o 4 L, stainlcss steel, 8 units 1h6.000
Seed tank agitator 8.4 hp, stainless steel, 8 units 97,100
Air filter glass fiber, 1200 kg/h air 400
Air compressor 42 hp, 30 psig, stainless steel, 1200 kg/h air 4h.600
Feed-water sterilizer insulated stainless steel pipes plus heat exchangers, I . 13 X los
kg/h water 80, I00
Feed mixing tank 9,450 L. stainless steel 17.600
Feed mixer agitator 12.5 hp. stainless steel I 5,700
Centrifuge 217 hp, 1.17 X los L/h feed ~- 3 19.000
5 I.801.000
Ethuriol recovety
Absorber I in. Raschig rings,
gas rate = 1.05 X lo4 kg/h $ 67.500
Absorber-water sterilizer 3500 kg/h water 13.100
Stillage evaporator 3.73 x 104ft2, effects 1.590.000
Rotary dryer 7140 ft ', 27.4 hp, 1.16 X lo4 kg/h water removed, 3 unitr I.374.000
Distillation 1.15 X los kg/h feed
feed EtOH wt % = 8.0, stripper followed by dirtillation colunin 34 1,000
5 3.380.000

Total purchased equipment 5 5,877,000

Fixed capital investment (4.13 X purchased equipment) $24,270.000
Total capital investment (4.89 X purchased equipment) $28.740.000

MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT 1007

 Table m. Batch fermentation production cost summary.
~~

cost
Item ( S / L product)

Raw niuteriuls
Nutrient solution 1.110
Water 7.4 kg/L 0.099
Molasses 3.7 kg/L 34.700
~

Raw materials subtotal 35.909

Utilities
Power 6.71 X l o h kW h/yr 0.592
Cooling water 6S°F,43.4 kg/L 0.260
Stcam 50 psig, 2.38 kg/L 1.654
Steam 600 psig, 0.92 kg/L 0.896
Operating labor $10/nian h 1.348
Operating supervision 15% operating labor 0.202
Maintenance 6% fixed capital I. 460
Operating supplies 15% maintenance 0.220
Laboratory charges 15% operating labor 0.202
__
Direct production cost 42.743
Depreciation linear, 18 yr, zero salvage 1.348
Property taxes 3% fixed capital 0.730
Insurance 0.7% fixed capital 0.170
Fixed cost 2.248
Plant overhead cost 60% (operating labor + operating supervision + niaintcnancc) __
I ,806
Manufacturing cost ( = direct production + fixed costs + plant ovcrhcnd) 46.797
Administrative expenses 15% operating labor 0.202
Distribution and marketing 5% total production cost 2.474
Total general expenses 2.676
After-tax profit (15% return on investment) 4.311
Income tax 4.311
Yeast credit ($O.l14/kg 50 wt 70)yeast product 4.782(- )
Steam credit (75.6C/1000Ib) 0.268(~)
__
Total product cost 53.045

terials and without the yeast or steam by-product credits is

10.9C/L.
The required batch fermentation time in this example is
lower than is typical for conventional batch fermentation
Fermentor Ethonol Concentration ( 9 h )
facilities. This is partly due to the large inoculum used, 45 55 65 75 85
but is also caused by the lack of agitation and temperature I I I I I I 1 I

regulation in most conventional plants. The fermentation I

rate is reduced by up to 50% without agitation and cool- E
ing.25 Figure 6 also presents ethanol product costs for a
batch plant without agitators. Twice as many fermentors
are required. The final ethanol cost is uniformly higher,
but by only ca. 1.2C/L product.

Simple Continuous Fermentation

Figure 8 shows the effect of glucose concentration on c
e
productivity for the simple continuous reactor at constant Continuous
P Stirred Tonk
2.8 g/L residual glucose concentration in the product. The Fermentor
specific ethanol productivity drops continuously with in- -04 g
creasing sugar feed (and corresponding product ethanol)
concentration. The volumetric productivity, however, ini- 91 107 123 138 153 167
I n l e t Sugar Concentration (wt %)
tially increases as the density of cells (promoting the reac-
tion) increases. This tradeoff gives a maximum volumetric Figure 8. Effect of ethanol product concentration on specific and total
productivity at an intermediate feed concentration of 13.0 volumetric productivity of a continuous stirred tank fcrmentor.

1008 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

 Fermentor Ethanol Concentration (g/!) Fermentor Ethanol Product Concentration (g/i)
45 55 65 75 85 45 55 65 75 85
I I 1 I I I l I ~ I I I I I I I I I
Pm=87.5
59t- Simple Continuous Stirred Tank 4
12 Fermentor
lo Total
-0 Equipment Cost
u
c

cost

9 10 II 12 13 14 15 16 17
I n l e t Sugar Concentration (do&)
9I 107 12.3 138 153 16.7
Inlet Sugar Concentration (wt %)
Figure 10. Ethanol product cost for continuous stirred tank reactor
Figure 9. Purchased equipment costs versus ethanol product concen- fermentation versus ethanol product concentration.
tration for the continuous stirred tank fermentor.

reactor volume, thus, does not give the minimum ethanol

wt 70glucose (corresponding to 64 g/L ethanol product). product cost as has been assumed in many previous studies.
Figure 9 shows the change in equipment cost with feed The optimum CSTR operating conditions are summa-
concentration. The minimum fermentor cost, of course, rized as feed concentration of 15.6 wt 70,Pfermof 79 g/L,
occurs for the minimum reactor volume at 13.0 wt 70glu- X,, of 20 g/L, v equal to 0.72 h-l, and volumetric pro-
cose. The minimum total equipment cost and minimum ductivity of 14 g/L h.
total ethanol product cost occur, however, at a much Flows, equipment specifications, and costs for these
higher feed concentration of 15.6 wt 70glucose (Fig. 10). conditions are presented in Figure 11 and Tables IV and
The increased concentration reduces the water to be han- V. Five 1.77 X 105 L fermentors are used. The total capi-
dled in sterilizers, evaporators, and dryers. The minimum tal investment is $27.1 million and the alcohol product

Plant Capacity 13OsIObE/yr 95% EtOH Product

Water S p s q Steorn(0I) W: Water

E Ethanol
,%
-? STERILIZER , ~

Y Yeast
CO2 (9.6) S Sugar
Flows In lo3 K g h ETHANOL
STORAGE

~~~~ ~

DRYER

600-psig

1
Steom (I1 7 )
YEAS? I Water (12 9)
STORAGE
( 5) 1 -
1 1 I I
II I
STRIPPER
w. 1.0
Y. 0.1
S : 0.4 2
~
j a t e r (88.8) g3
s: 0.4 .
50-psig Steam (17.0)

Figure 11. Flow diagram for the optimized CSTR fermentor plant design.

MA IORELLA, BLANCH , AN D W I LKE: BIOTECH N0LOGY REPORT 1009

 Table IV. Simple CSTR purchased equipment summary

CO\t
Item (US dollar\)

Stonigr (2 weeks)
Molasses 7.86 X 10" L, carbon $tee1 $ 268,000
Ethanol 4.24 X lo6 L, carbon steel I 67,000
Yeast 3.23 X los L, carbon steel, 5 units 247.000
Screw conveyor for yeast delivery 0.40 hp, 100 ft long 8,000
!3 h0O ,000
Furnieii tot ion
Fermentor 1.77 X lo5 L, stainless steel, 5 units b 443,500
Fermentor agitator 117 hp, stainless steel, 5 units 3.36.000
Fermentor cooler 438 ft2, stainless steel, 5 units 2 1 .000
Air filter glass fiber, 792 kg/h air .1 I0
Air compressor 27.6 hp, 30 psig, 792 kg/h air 4 I..300
Fccd-water steriliier insulated stainless steel pipes plus heat exchanger\.
1.23 X lo5 kg/h water 00. I00
Feed mixing tank 1.03 X lo4 L, stainlcss steel I 8. 400
Feed mixer agitator 13.6 hp. stainless steel I h ,000
Cenlrifuge 237 hp, I .26 X lo5 L/h fccd 342.000
$ I ..309,000
Ethunol recovery
Absorber 1 in. Raschig rings,
gas rate = 1.02 X lo4 kg/h .6 hS.000
Absorber-water sterilizer 3340 kg/h water 12.800
Stillage evaporator 4.08 X lo4 f t 2 , 5 effects 1 ,670,000
Rotary dryer 7180 f t 2 , 27.6 hp, 1.17 X 10' kg/h water rcmovcd, 3 units I ,458,000
Distillation 1.25 X los kg/h feed,
feed EtOH wt 70 = 7.4, stripper followed by distillation column 345.000
!3 3.551 ,000

Total purchased equipment $:5.550.000

Fixed capital investment (4.13 X purchased equipment) $22.920.000
Total capital investment (4.89 X purchased equipment) !327.140,000

cost is 51.3C/L. The manufacturing cost exclusive of raw
materials and without by-product credits is 9.8C/L.
CSTR With Recycle
Recycle of cells to increase the CSTR cell density in-
creases productivity and reduces costs.26Figure 12 shows
the effect of centrifuge cell recycle on ethanol cost at vary-
ing sugar concentrations. The recycle ratio is defined as
the ratio of recycle stream flow (L water/h) to total feed
flow to the fermentors (recycle plus new feed). The recycled
yeast density is 180 g yeast/L water. Limiting the fermentor
cell density to 100 g yeast/L water (to maintain Newtonian
flow and not interfere with mixing27)the maximum recycle
ratio (RR) is 0.6.
The optimum operating conditions are: feed concentra-
tion of 16.7 wt 70,an RR of 0.5, Pfer,of 85.6 g/L, X,,, of
100 g/L, v equal to 0.42 h-I, and volumetric productivity
of 42.5 g/L h. Flows, equipment specifications and costs
for this optimum case are summarized in Figure 13 and
Tables VI and VII. Two 1.47 X 1@ L fermentors are
used. The total capital investment is reduced to $23.7 mil-
lion and the ethanol product cost is 49.lC/L (8.64c/L
manufacturing cost exclusive of raw materials).
The minimum ethanol cost occurs for a very high etha-
nol fermentation product concentration of 85.6 g/L with
specific productivity reduced 75% by end-product inhibi-
tion. This high level of inhibition can be tolerated because
of the high fermentation cell density. The fermentor size is
reduced compared to the simple CSTR case, making the
fermentor costs less significant. The reduction in auxiliary
equipment cost at reduced water flow then dominates and
shifts the optimum toward higher feed and product con-
centrations.
A special case of a CSTR with cell recycle is the partial
recycle reactor proposed by Hough, Gough, and Davis.2s
Product is drawn from the fermentor through a baffled
settler/separator built into the reactor. A portion of the
cells is retained, and the cell density in the fermentor is in-
creased (for the less active yeast strain used by Hough) by
a factor of 2.54. No added centrifugation equipment is
needed. It is unlikely that this high separator efficiency
could be achieved with a very active yeast with rapid C 0 2
generation and the associated turbulence but, if we apply
this separation factor and the present kinetic model, a
considerable cost saving results compared to the simple
CSTR. Table VIII compares the simple and partial recy-
cle stirred tank reactors. The volumetric productivity is
increased from 14.1 to 35.9 g/L h. The ethanol product

1010 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

Table V. Simple CSTR production cost summary.

Item cost

H ( i w riiirrrriiils
Nutrient solution 1.110
Water 8.16 kg/L 0 . 1 OY
Molasses 3.7 kg/L 34.700
~

Raw material subtotal 35.919

ut ilit i p s
Power 6.98 X 10'kW h/yr 0.610
Cooling watcr 65'F, 44.1 kg/L 0.204
Steam 50 psig, 2.58 kg/L 1.790
Steam 600 psig, 0.93 kg/L 0.90 1
Operating labor $10/nian h 0.844
Opcrating supervision 15% operating labor 0.127
Maintenance 6% fixed capital 1.376
Opcrating supplics 15% maintenance 0.206
Laboratory charges 15"/0 operating labor ~
0.127
Direct production cost 42. I70
Depreciation linear, 18 yr, zero salvage 1.274
Property taxes 3oiU fixed capital 0. 688
lnsurancc 0.7% fixed capital ~
0.101
Fixed cost 2.123
Plant overhcad cost 60% (operating labor + operating supervision + main!cn;incc) 1.408
+ fixed costs + plant overhead)
~

Manufacturing cost (= direct production 45.701

Administrative expenses 15% operating labor 0 . 127
Distribution and marketing 5'/0 total production cost 2.412
Total general expenses 2.539
After-tax profit (15% return on investment) 4.07.3
Income tax 4.073
Yeast credit ($0.114/kg 50 wt %I) yeast product 4.807( - )
Steam credit 75.6C/1000 Ib ~
0.293( ) ~

Total product cost 51.286

cost is reduced from 51.3 to 49.8e/L. This is comparable

to the savings for the optimum centrifuge recycle case.

65 75 85
I I I I I I
- The use of CSTR fermentors arranged in series has been

a:
57 -
C S T R with Cell Recycle suggested to overcome end-product inhibition effect^.^^.^^
- For two reactors, the first is operated at an intermediate
ethanol product concentration and high corresponding
5s 55-
-
specific productivity. The second reactor, producing the
c
v) -
RR=O - final high concentration ethanol product, is left with less
0
u sugar to convert and the overall productivity may be in-
53- -
0
3
creased.
a
: Table IX shows the effect of intermediate (first reactor)
- ethanol concentration for two series CSTRs at various feed
c

I-
sugar concentrations. For low feed sugar and intermediate
ethanol concentrations (i.e., 12.3 w t 70glucose and 35 g/L
- ethanol), the total reactor volume can actually be increased
49
-I I I I
0.5
by using series reactors as compared to a single CSTR.

MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT 1011

 Plant Capaclty lCOalO"#/yr 95% EtOH Product

Figure 13. Flow diagram for the optimized CSTR fermentor with centrifuge cell recycle.

Table VI. CSTR with centrifuge cell recycle purchased equipment summary.

Cost
Item (US dollars)

Storage (2 weeks)
Molasses 7.85 X 10' L, carbon steel $ 268,000
Ethanol 4.24 X IO'L, carbon steel 167.000
Yeast 3.21 X los L, carbon steel, 5 units 247.000
Screw conveyor for yeast delivery 0.40 hp, 100 ft long 8.000
$ 690.000
Fermentation
Fermen tor 1.47 X 10' L, stainless steel, stainless steel, 2 units $ 160,200
Fermentor agitator 97 hp, stainless steel, 2 units 119,200
Fermentor cooler I 100 ft2, stainless steel, 2 units I 1,260
Air filter glass fiber, 1200 kg/h air 400
Air compressor 42 hp, 30 psig, 1200 kg/h air 46,600
Feed-water sterilizer insulated stainless steel pipes plus heat exchangers, I . 13 X 10'
kg/h water 86,100
Feed mixing tank 9450 L, stainless steel 17,600
Feed mixer agitator 12.5 hp, stainless steel 15,700
Centrifuge 233 hp, 1.17 X lo5 L/h feed, 2 units 674.000
$ 1,131,000
Ethanol recovery
Absorber 1 in. Raschig rings,
gas rate = 1.05 X l o 4 kg/h $ 67,500
Absorber-water sterilizer 3500 kg/h water 13,100
Rotary dryer 7620 f t 2 , 29 hp, 8280 kg/h water removed, 2 units 1.008,000
Distillation 1.18 X lo5 kg/h feed,
feed EtOH wt 70 = 7.8
stripper, stainless steel, followed by distillation column,
carbon steel 342,000
Stillage evaporator 3.73 x lo4 ft2, 5 effects 1,590,000
$ 3,020,600

Total purchased equipment S 4,842.000

Fixed capital investment (4.13 X purchased equipment) $20.000,000
Total capital investment (4.89 X purchased equipment) $23,680,000

1012 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

 Table VII. CSTR with recycle-production cost summary.

Nutrient solution 1.110

Water 7.4 kg/L 0.099
Molasses 3.7 kg/L 34.700
~

Raw materials subtotal 35.909

Utilities
Power 5.70 X lob kW h/yr 0.503
Cooling water 65°F. 43.6 kg/L 0.26 I
Steam 50 psig, 2.65 kg/L 1.842
Steam 600 psig. 0.66 kg/L 0. h38
Operating labor $IO/nian h 0.725
Operating supervision 15% operating labor 0.109
Maintenance 6% fixed capital 1.200
Operating supplies 15% maintenance 0.180
Laboratory charges 15% operating labor ~
0.109
Direct production cost 41.476
Depreciation linear, 18 yr. zero salvage 1.111
Property taxes 3% fixed capital 0.600
Insurance 0.7% fixed capital 0.140
Fixed cost 1.852
Plant overhead cost 60% (operating labor + operating supei-vision f maintenance) I .220
direct production + fixed costs + plant ovcrhcnd)
~

Manufacturing cost (= 44.548

Administrative expenses 15% operating labor 0.109
Distribution and marketing 5 % total production cost ~
2.350
Total general expenses 2.459
After-tax profit (ISTOreturn on investment) 3.55.3
Income tax 3.553
Yeast credit 4.782( - )
Steam credit 0.268( - )
Total product cost 49.063

Table Vm. Partial recycle fermentor.

Total
Volumetric Fermentor Manufacturing product
Pfmm Xferm productivity cost cost" cost
(g/L) (g/L) (g/L h) (SIOS) (OIL) (C/L)

Partial recycle fermentor 79.1 50.0 35.9 3.29 8.8 49.8

Simple CSTR 79.1 19.7 14.1 8.01 9.8 51.3

aThis is exclusive of raw materials and yeast credit.

not necessarily give reduced reactor cost (i.e., 12.3 wt "70
glucose comparing 35- and 40-g/L intermediate ethanol).
For high final ethanol product concentrations (16.7
wt "70 glucose feed case and 85.6 g/L ethanol final prod-
uct), the series reactor arrangement can result in a sub-
stantial 41% reduction in reactor volume as compared to
the simple CSTR. The corresponding savings in total prod-
uct cost is, however, very small.
The perforated plate tower fermentor has been sug-
gested as a simple means to provide several fermentors in
series.31 A distillation column is divided into a series of
fermentation zones separated by perforated plates and
fitted with a single agitator shaft for all. Reaction takes
place progressively as beer ferments at the top level and se-
quentially overflows down to lower levels.
Figure 14 shows the variation in product ethanol cost
with sugar feed concentration in a perforated plate tower
fermentor with four equivolume stages, along with results
for the optimized two vessel series CSTR and simple
CSTR systems. The four-stage tower is inferior to the sim-
ple CSTR for all but the most concentrated of feeds. This
occurs because, while initial stages show very high specific

MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT 1013

 Table M. Two-stage series CSTR fermentors.

Inlet Final Intermediate Total

sugar EtOH EtOH Fermentor Fermentor Manufacturing product
concentration concentration concentration volume cost cost" cost
(wt 010) (g/L) (g/L) ( l o " L) ($105) (C/L) (C/L)
~ ~~

12.3 59.5 35 8.66 7.90 11.26 55.0

40 8.22 8.27 11.32 55.1
45 7.82 7.47 11.22 54.9
50 7.50 7.26 11.15 54.8
55 7.26 7.02 11.13 54.7
simple CSTK 7.65 7.36 11.20 54.9
15.3 76.9 45 8.07 7.59 9.53 51.3
55 7.45 7.23 9.50 51.2
60 7.24 6.50 9.41 51.0
65 7.10 7.05 9.47 51.1
70 7.15 7.02 9.46 51.1
simple CSTR 8.40 7.76 9.58 51.4
16.7 85.6 50 10.12 9.93 9.59 51.5
60 9.00 8.70 9.48 51.1
70 8.26 8.28 9.40 51 .o
80 8.76 8.54 9.4.3 51.1
simple CSTK 13.92 12.55 10.04 52.4

'This is exclusive of raw materials and yeast credit.

ethanol productivity, the cell density for the low initial Fino1 Fermentor Ethanol Concentration ( g h )
stage sugar conversions is low and the volumetric produc-
tivity suffers.
Minimum product costs for the perforated plate tower,
simple CSTR, and two vessel series CSTR systems, re-
5at \
spectively, are 52.3, 51.3, and 51 .OC/L, compared to the
optimal simple CSTR recycle case cost of 49.lC/L. The
perforated plate tower and series CSTR costs would also
be reduced if these reactors were used in combination with
centrifuge cell recycle.

Plug Flow Fermentor

In the plug flow f e r m e n t ~ r ,cell
~ ~ mass
. ~ ~ and ethanol
concentration both increase continuously along the reactor
length. Cells must be recycled to the reactor inlet to initiate 5 0 5
I0 II 12 13 14 15 16 17
the reaction. A large recycle (RR = 0.2 of centrifuged I n l e t Suqor Concentration (wt%)
yeast cream, giving 36 g/L initial cell density) is used to
overcome the problem of low volumetric productivity as- Figure 14. Ethanol product costs for series CSTR and perforated plate
sociated with low cell density at the reactor entrance. This column fermentor systems.
also insures a high reaction rate with much C 0 2 evolution
to prevent yeast settling. The agitation caused by C 0 2evo- Table X. Ideal plug flow reactor ethanol product costs.
lution will cause backmixing, but this effect is not consid-
ered in the reactor model. Backmixing increases the reac- Product
tion rate for a simple autocatalytic reaction and decreases Feed Residence ethanol Fermentor Product
concentration time concentration cost cost
it for an end product inhibited reaction. The actual effect
(wt 010) (h) (g/L) ($10~) (OIL)
on this combined type reaction will depend on the relative ~~ ~~ ~

backmixing rates of cells and ethanol. 13.0 0.83 63.9 1.87 52.5
For the idealized plug flow model assumed, Table X 13.8 0.90 68.2 1.88 50.4
gives the ethanol product cost at various feed concentra- 14.5 0.98 72.5 1.89 50.0
tions. The minimum cost of 49.lC/L at 16.7 wt 70glucose 15.3 1.06 76.9 1.91 49.7
16.0 1.16 81.2 1.94 49.4
feed (8.6C/L manufacturing cost exclusive of feed) is com- 16.7 1.30 85.6 1.99 49.1
parable to the minimum costs for the CSTR with recycle.

1014 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

Immobilized Cell Tower Fermentors
To achieve high cell densities without the need for large
centrigues, yeast have been immobilized by binding in
polymer gels or on a solid s ~ p p o r t . A ~ ~simple
- ~ ~ alterna-
tive is retention of cells at high density by settling in a tower
f e r m e n t ~ r . The
~ ~ . APV
~ ~ tower fermentor is an upright
shell topped by a large diameter settling head fitted with
special baffles which separate the gas bubble flow. The
upward liquid velocity in this settling head is less than the
average yeast cell settling velocity and most living yeast are
retained in the fermentor. Dead lysed yeast are washed
out (and can be used as cattle feed supplement). Green-
shields and Smith37report a 70-g/L cell density for an in-
dustrial APV tower fermentor. Even higher cell densities
(80 g/L) can be achieved in a Raschig ring packed tower
where the rings provide quiescent zones for yeast reten-
t i ~ nbut , ~some
~ reactor volume is then lost to the volume
taken up by packing.
While the cell density is maintained at a constant high
level throughout a tower fermentor, the liquid ideally
moves in plug flow mode, and high specific ethanol pro-
ductivity is maintained through most of the reactor. In
practice; some backmixing of the liquid should be expected
due to agitation from COZ evolution. Flocculent yeast with
good settling characteristics are generally used in tower
fermentors and these typically have somewhat lower in-
trinsic reaction rates than the high rate brewers yeast used
in establishing the present kinetic model. Diffusion limi-
tations in dense yeast flocks might also reduce reaction
rates, and this is not considered in the reactor model. To
prevent bypassing, fermentation may have to be occasion-
ally interrupted to sparge the reactor with a rapid gas
stream to redistribute cells.39 This has also been ignored
in the analysis.
Using the idealized reactor model, APV and packed
tower fermentors were designed with cell densities of 70
and 80 g/L, respectively. Figure 15 gives the ethanol prod-
uct cost for the tower fermentor systems compared to the
optimum CSTR designs with and without cell recycle.
Only one curve is presented for the two tower fermentor
types (APV and packed column) as both types give almost
identical final product costs (differing by less than O.lc/L).
Because of the assumed plug flow nature of the reactor,
the minimum product cost occurs at a very high feed con-
centration (16.7 wt 70glucose). The optimum fermenta-
tion conditions for the APV tower fermentor are a feed
concentration of 16.7 wt 70,Pexit of 85.6 g/L, X,,,, of 70
g/L, a residence time of 0.98 h, and volumetric produc-
tivity of 86.9 g/L h.
Stream compositions, equipment specifications, and
costs are presented for the APV tower fermentor in Figure
16 and Tables XI and XII. A single tower 9.2 ft in diam-
eter and 76.4 ft tall is used. The total capital investment is
$23.6 million and the minimum product cost is 48.7c/L
(8.3c/L manufacturing cost exclusive of raw material),
which is comparable to the optimum case for the CSTR
with recycle.
MAIORELLA, BLANCH, AND WI LKE: BIOTECHNOLOGY REPORT
Fermentor Ethonol Concentration (g/P)
%
=
I1 12 13 14 15 16 17
Inlet Sugar Concentration (wt %I
Figure 15. Ethanol product cost for the tower system versus ethanol
product concentration.
Selective Ethanol Removal Processes
Fermentation processes with selective ethanol removal
have been proposed to allow very high productivity by
eliminating end product inhibition.26Only the flashferm
process (with ethanol removal by flashing in an auxiliary
flash vessel) has been tested sufficiently to allow a detailed
evaluation including design and costing of the separation
equipment. Here, costs are calculated for the selective
membrane," extractive fermentation,I2,l3and flash fer-
m e n t a t i ~ n ' ~processes,
.'~ excluding the cost of the separa-
tor. The savings found as compared to the best methods
without selective removal then establishes the value of a
separator of the specified characteristics.
Figure 17 illustrates flows for a selective ethanol re-
moval system. In this system, fermenting beer is cycled
through a separator which recovers concentrated ethanol
and recycles ethanol depleted beer back to the fermentor
for further reaction. End-product inhibition is reduced
and the fermentation rate for a given feed sugar concen-
tration is increased.
Selective membrane separations, ethanol extractors, or
the flashferm flash vessel can all be modeled by correctly
specifying K , the separation device effectiveness. Param-
eter K is the ratio of ethanol concentration (g ethanol/L
water) in the concentrated product stream (see 4 in Fig.
17) to its concentration in the dilute stream (see 5 in Fig.
17) leaving the separator. Approximate K values for vari-
ous separator types are presented in Table XIII.
The bleed-to-feed ratio (BFR) is the ratio of dilute etha-
nol product (flows 8 and 11 in Fig. 17) exhausted from the
system through the centrifuge, to the new feed flow (1 in
Fig. 17). This ratio is helpful in establishing the mass bal-
ance relations but is also important in computing the
1015
 Plant Capacity 1001106P/yr 95 % €?OH Product
W Water
E Ethanol
Y Yeast
CO2 (96)
+ W 104
S Sugor
F b f s Inlo3 Kg/hr ETHANOL
A r (091
E 89 STORAGE

STRIPPER

50psq Steam (16 7 )

Figure 16. Flow diagram for the optimized tower fermentor plant design.

Table XI. APV tower fermentor equipment rummary.

('051

Item ( U S doll:lr\)

Storqy (2 weeks)
Molasses 7.85 X IO'L, carbon steel $ 2h8.000
Ethanol 4.24 X 10' L, carbon steel lh7.000
Yeast 3.21 X los L, carbon steel, 5 units 247.000
Screw conveyor for yeast delivery 0.40 hp, 100 ft long 8,000
$ h90.000
Fernirrircrtivrr
Tower fermentor stainless steel, 9.2 ft diameter 76.4 ft height S 255,000
Fermentor cooler 2190 f t 2 , stainless steel 7,000
Air filter glass fiber, 1200 kg/h air 400
Air compressor 42 hp, 30 psig, 1200 kg/h air 4h,600
Feed-water sterilizer insulated stainless steel pipes plus heat exchangers, 1.13 X lo5
kg/h water 8h, 100
Feed mixing tank 9,450 L, stainless steel 17.h00
Mixer agitator 12.5 hp, stainless steel 15,700
Centrifuge 217hp, 1.17 X 1OSL/hfeed 3 1 9,000
$ 747,000
Ethuriol recovry
Absorber 1 in. Raschig rings,
gas rate = 1.05 X lo4 kg/h $ 67.500
Absorber-water sterilizer 3500 kg/h water 13,100
Stillage evaporator 3.73 x lo4 ft2, 5 effects 1,590.000
Rotary dryer 7140ft2, 27.4 hp, 1.16 X los kg/h water removed, 3 unit, I .374,000
Distillation 1.15 X los kg/h feed,
feed EtOH wt '70= 8.0. stripper followed by distillation colunln 34 1.000
$ 3,386,000

Total purchased equipment $ 4,823,000

Fixed capital investment (4.13 X purchased equipment) $ 19.920,OOO
Total capital investment (4.89 X purchased equipment) $2.3 .580. 000

1016 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

 Table XII. Tower fermentor production cost summary.

Run! rituteriuls
Nutrient solution 1.110
Water 7.4 kg/L 0.099
Molasses 3.7 kg/L 34.700
Raw material subtotal 35.909
Utilities
Power 2.62 X loh kW h/yr 0.231
Cooling water 65°F. 43.4 kg/L 0.260
Steam 50 psig. 2.38 kg/L 1.654
Steam 600 psig. 0.92 kg/L 0.896
Operating labor $lO/nian h 0.685
Operating supervision 15% operating labor 0.103
Maintenance 6% fixed capital 1.195
Operating supplies 15% maintenance 0.179
Laboratory charges 15% operating labor 0. I03
Direct production cost 41.215
Depreciation linear, 18 yr, zero salvage 1.107
Property taxes 3% fixed capital 0.598
Insurance 0.7% fixed capital 0.139
Fixed cost 1.844
Plant overhead cost 60% (operating labor + operating supervision + maintenance) 1.190
Manufacturing cost ( = direct production + fixed costs + plant ovcrhcad) 44.249
Administrative expenses 15% operating labor 0. I03
Distribution and marketing SYOtotal production cost ~
2.334
Total general expenses 2.437
After-tax profit (15% return on investment) 3.537
Income tax 3.537
Yeast credit 4.782( ~)
Steam credit ~
O.268( - )
Total product cost 48.710

Concentrated
Product
buildup of toxic secondary components which are not re-
moved in the separator concentrated ethanol product
stream. In the flashferm process, for instance,15 the sepa-
rator operates by boiling away ethanol at reduced pres-
sure. The cell growth efficiency has been found to diminish
in flash fermentation and this has been explained by the
buildup of inhibitors at low bleed-to-feed ratios. Nonvola-
tile feed components and fermentation by-products are
I t not boiled up. The steady-state concentrations of these
Cell Recycle 4
.
0
components is inversely proportional to the bleed-to-feed
Figure 17. Mass balance relations for fermentation with cell recycle ratio and at small values of BFR, the kinetic equations
and selective ethanol removal.
must be modified to account for the toxic effects of these
concentrated secondary inhibitor^.^^
Table XIII. Selective ethanol removal device effectiveness factors. Acetic acid was found to be the most important inhibi-
tor.40The production rate of acetic acid is approximately
Separator type Effectiveness, K" Reference
0.5170of the ethanol production rate, and the concentra-
Selective membrane 1 .5-4 10, 11 tion of acetic acid in a selective ethanol removal reactor
Solvent extraction 2-8 12, 13 which does not remove acetic acid in the concentrated eth-
Flash fermentation 8-12 14, 15 anol product stream is given as:
aK is the ratio of ethanol concentration (g/L water) in the separator
concentrated product stream to its concentration in the dilute product
0.0051 (hourly ethanol production rate)
A(g/L) = (6)
stream. bleed rate (L/h)

MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT 1017

 I I I I with a bleed-to-feed ratio of 0.2. Balancing this, however,
10 -0.418A + 0 . 0 s O A 2 - 0 . 0 0 6 0 A3
=0.988
€0 is the large cost savings in auxiliary equipment (evapora-
tors, dryers, etc.) as less and less of the product ethanol is
removed in the dilute form.
The effect of changing the fermentor cell concentration
is shown in Table XV (with cost exclusive of the separator).
Very low cell densities reduce the fermentor productivity
0 2 1 Acetic Acid I n h i b i t i o n
of Cell Growth and increase costs, but above 80 g/L cells, no further savings
OL I 1 I I are realized. The savings in fermentor and rotary dryer costs
0 2 4 6 8 10
A = g/f Acetic Acid
are just balanced by the increased cost of centrifugation.
The effect of varying fermentor ethanol concentration is
Figure 18. By-product (acetic acid) inhibition of cell production ef- shown in Table XVI. Contrary to the initial assumption,
ficiency. the lowest overall alcohol product cost is for high fermen-
tor ethanol concentrations where end product inhibition is
significant. This result is correct though. As with the case
The effect of acetic acid concentration on cell mass pro- for the CSTR with recycle, the high cell density makes the
duction is shown in Figure 18. This inhibition can be fermentor costs small and of less significance to the total
modeled as: product cost. High specific productivity is sacrificed to al-
low the production of very concentrated product flows
- -
Eo
- 0.989 - 9.418A + 0.0901A2 - 0.006006A3 (7) which require smaller less costly auxiliary equipment, and
a cost savings results. The distillation equipment cost sav-
This relationship has been incorporated into the fermen- ings results from the reduction of the stripper column size
tation kinetic model to show the effect of by-product inhi- for the dilute bleed stream. Higher fermentation ethanol
bition in selective ethanol removal systems. concentrations also make for a simple separation and the
Table XIV shows the effect of varying bleed-to-feed ra- cost of the selective removal device (not included in this
tio. The separation factor, K = 6, corresponds to a sepa- analysis) should also be least at high fermentation ethanol
rator using liquid-liquid contacting to extract ethanol concentrations.
with an extractant which is selective for ethanol over water. Table XVII compares results from selective membrane
Low bleed-to-feed ratios increase the level of toxic by- ( K = 2), solvent extraction ( K = 6), and flash fermenta-
product buildup and reduce the production of cells. The tion ( K = 10) cases. There is a large reduction in cost (ex-
cattle feed by-product credit is reduced from 4.8C/L clusive of the separator) for the K = 6 case as compared to
ethanol produced for a standard CSTR to only 1.9C/L K = 2, and a small further savings at K = 10. This simply
ethanol produced for the extractive separation process indicates that a separator which can produce a more con-

Table X N . Effect of bleed-to-feed ratio (BFK) on sclcctivc rcnioval fermentation product

cost, with K = 6,Xler,,,= 100 g/L, and Plern,= 50 g/L.

Acetic acid Cell growth Yeast lnlet sugar Product

concentration inhibition product concent rat ion coct "
BFK (g/L) (E/E,) ( I 0' kg/yr) (wt 010) (C/L)

0.2 4.78 0.392 1.57 30.2 44.5

0.4 1.91 0.476 1.94 25.8 45.9
0.6 0.96 0.666 2.75 20.7 47.1

"This cost is exclusive of separator cost. Reduction comparcd to the optimum recycle CSTK
case (49.IC/L) indicates the value of a separator of the specified characteristics.

Table XV. Effect of cell concentration (X,,,.")) on selective removal fernientalion product
cost, where K = 6,(extractive fermentation), BFR = 0.3,and PI,,.,,,= 50 g/L.

lnlet sugar Fernientor Centrifuge Rotary dryer Product

X,,,,, concentration cost cost cost cost a
(g/L) (wt 70) ($103 ($105) ($103 (C/L)

80 28.1 1.34 0.83 5.01 45.1

100 28.1 1.18 1.01 4.87 45.1
120 28.1 1.07 1.36 4.60 45.1

"This cost is rxclusive of separator cost. Reduction compared to the optimum recycle CSTK
case (49.la/L)indicates the value of a separator of the specified characteristics.

1018 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

 Table XVI. Effect of fermentor ethanol concentration (Pi,,,,,) on selective removal fermentation product cost. where K = 6
(extractive fermentation), BFR = 0.3, and Xtelm= 100 g/L.

Inlet sugar Fermentor Centrifuge Evaporator Dryer Distillation Product

Pt,,,, concent rat ion cost cost cost cost cost cost "
(g/L) (wt 7 0 ) ($105) ($10~) ($105) ($105) ($107
~ ~~

30 19.0 1.09 1.69 7.50 7.34 4.82 47.9

50 28.1 1.18 1.01 5.70 4.87 3.28 45.1
80 38.4 1.64 0.53 4.35 5.02 2.90 43.9

"This cost is exclusive of separator cost. Reductions compared to the optimum recycle CSTR case (49.lclL) indicate the
value of a separator of the specified characteristics.

Table XW. Effect of separator efficiency ( K ) on selective removal fermentation product cost, where BFR = 0.3 and Pi,,,,, =
50 g/L.

Inlet sugar Fermentor Centrifuge Evaporator Dryer Distillation Product

X,,,, concentration cost cost cost cost cost cost
K (g/U (wt 7 0 ) ($105) ($105) ($105) ($105) ($105) (PlL)

2 80 12.9 1.37 3.12 9.72 7.64 3.49 47.3

6 80 28.1 I .34 0.83 5.71 5.01 3.39 45.1
6 100 28.1 1.18 1.01 5.70 4.87 3.28 45.1
10 loo 38.7 1.18 0.50 4.32 5.05 3.31 44.5

aThis cost is exclusive of separator cost. Reduction compared to the optiniuni recycle CSTR case (49.14lL) indicates the value of a
separator of the specified characteristics.

centrated ethanol product from a dilute stream is more
valuable.
Figure 19 and Tables XVIII and XIX present flows,
equipment specifications, and costs for the optimum sol-
vent extraction (I( = 6) case. The conditions for this case
are a separation factor ( K )of 6, inlet sugar concentration
(wt %) of 38.4, X,,,, of 100 g/L, Pferm of 80 g/L, v equal to
0.69 h-', and volumetric productivity of 68.7 g/L h.
Compared to the optimum CSTR process with recycle,
the stillage evaporator size is reduced from 37,300 to 3380
ft2and the rotary dryer from 15,240 to 7690 ft2. The distil-
lation equipment costs are similar. A single 1.82 X lo5 L

Plant Capacity l0011o6P/yr 95 % EtOH Product

Air (0 6 )
Flaws in lo3 Kg/hr ETHANOL
Concentraled Product STORAGE
w. 2 34 ABSORBER
S: 2 3 . 4 ~ M o l a s s e s
Nutrients
CENTRIFUOE
Steam
(9.61
1 - . I , p-- \ / I I -I IF.04 I I I

VACUUM
DISTILLATION
COLUMN
Air
(08)
I T
w 48
E 03
I atm Steom (I 7)
(191

50 psq Steam ( 2 31
S03

Figure 19. Flow diagram for the optimized selective ethanol removal fermentation proces plant design

MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT 1019

 Table XVIII. Continuous fermentation with selective ethanol removal. purchased equipnicnt wmmary (wlcctivc ethanol
separation device not includcd).

co\t
Item (US dollar\)

Storcrgc, (2 wrrksl
Molarses 7.85 X I O h L, carbon steel $ 268,000
Ethanol 4.24 X 10' L, carbon stccl 107.000
Yeast 2.95 X lo5 L. carbon steel. 2 units Y3.200
Screw conveyor for yeast delivery 0.20 hp. 100 ft long 5.320
s $33.500
Ferrirrvrtrrtiorr
Fermentor 1.82 x 10' L. stainless stcel 89.200
Fermentor agitator 120 hp, stainlcss stccl 67,800
Fermentor cooler 2190 ft', stainless steel 6.")
Air filter glass fiber, 818 kg/h air 3 10
Air compressor 28.6 hp. 30 psig. 818 kg/h air 41.200
Feed-water stcrilircr insulated stainless steel pipes plus hcat exchangers, 3.37 X 10'
kg/h water 44.200
Feed mixing tank 2810 L, stainless steel 8.930
Feed mixer agitator 3.7 lip, stainless stccl 7.080
Centrifuge 21.5 hp. 1.43 X lo4 L/h feed 52,800
3 318.500
Ethurrol rc'covqv
Absorber 1 in. Raschig rings,
gas rate = 1.02 X 104kg/h $ 65, 100
Absorber-water sterilizer 3350 kg/h water 12,700
Rotary dryer ',
7690 f t 29.3 hp, stainless stccl,
4180 kg/h water rcmovcd 502.000
Rotary-dryer overhead reboiler
to recover excess hcat 138 f t 2 . carbon stcel 6,300
Distillation 1.04 X lo4 kg/h. 6 wt "/I, EtOH dilute feed to stripper. stainIc\s
steel, then combined with concentratcd product (20.5 wt 'XI
EtOH) and fed t o distillation colunin. (carbon steel) 200.000
Stillagc evaporator 3380 ft2. 5 effects, stainless steel 435 .ooo
$ 1..311,100

Total purchased equipment 5 2,1h3.000

Fixed capital investment (4.1.3 X purchased equipment) s 8.Y30.000
Total capital investment (4.8 X purchased equipment) s 10,580.000

fermentor is used. The total capital investment (excluding
the separator) is reduced to only $10.6 million, compared
to $23.7 million for the optimum CSTR with cell recycle
case. The final ethanol product cost is reduced from 49.1
to 43.9C/L (again exclusive of the separator).
This savings of 5.2c/L ($5.2 million/yr) represents the
value of an extractive separator of the specified character-
istics. Up to $2.4 million (purchased equipment cost) could
be paid for this separator (assuming no labor or utilities
charges) while still achieving a savings in the final product
cost compared to the optimum CSTR with cell recycle but
no selective ethanol removal.
ECONOMICS WITH CELLULOSE
DERIVED SUGARS
Introduction
Production of glucose b y hydrolysis of cellulosic wastes
is under development as a means to expand the supply of
glucose for ethanol production. The feedstocks considered
have ranged from municipal waste paper4' to crop resi-
d u e and ~ wood grown on energy plantation^.^^ The
~ ~even
cellulose is decomposed to the monomer glucose units by
either a ~ i dor~e n~ ~,y r~n a~t i chydrolysis
~,~~ methods. Both
procedures are very capital intensive and the costs of col-
lecting and pretreating most residues is also high. There-
fore, the cost of cellulosic-derived sugars is presently higher
than the cost of sugar from conventional sources.
A further problem with most cellulose hydrolysis pro-
cesses is that the sugar solutions produced are quite dilute
(typically 30-SO g glucose/L). Molasses solution, by con-
trast, is SO wt 70glucose. Hydrolysis of starchy feedstocks
like corn or manioc can produce 20-25 wt 7'0 glucose so-
lutions. The cost of concentrating cellulose hydrolysis
derived sugar solutions prior to fermentation can counter-
balance the savings in stillage evaporation costs for con-
centrated feeds and shift the optimum fermentation con-
ditions toward lower feed and product concentrations.

1020 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

 Table XIX. Selective ethanol removal production cost summary.

Cart
Item ( C / L pl.0duct)

H I I W riirr/(virrls
Nutrient soh t ion 1.110
Water 1.1 kg/L 0.015
Molasses 3.7 kg/L 34.700
Raw materials subtotal 35.825
Utilities
Power 1.49 X 10'kW h/yr 0.131
Cooling water 65'F, 46.2 kg/L 0.277
Steam 50 psig. 1.54 kg/L 1.072
Steam 600 psig, 0.33 kg/L 0.318
Operating labor $10/man h 0.487
Operating supervision 15% operating labor 0.073
Maintenance 6% fired capital 0.536
Operating supplies 15% maintenance 0.080
Laboratory charges 15% operating labor 0.073
__
Direct production cost 38.872
Depreciation linear, 18 yr, zero salvage 0.496
Property taxes 3% fixed capital 0.268
Insurance 0.7% fixed capital 0.063
Fixed cost 0.827
Plant overhead cost 60% (operating labor + operating supervision + niaintenancc) 0. 658
Manufacturing cost (= direct production + fixcd costs + plant ovcrhead) 40.357
Adniinistrativc expenses 15% operating labor 0.073
Distribution and marketing 5% total production cost 2.128
Total general expenses 2.201
After-tax profit (15% return on investment) 1.581
Income tax 1.587
Yeast credit 1.744(- )
Steam credit 0.049( - )
__
Total product cost 43.939

I 1 1
Perez4*presents detailed designs and cost estimates for 68 -
the production of glucose from corn stover residue by en- With Cool Generated

zymatic hydrolysis. Costs can be put on a consistent basis - 66-

with the current fermentation economic evaluation by v)
0

updating the capital cost estimates to mid-1981 basis 64-

With Woste Generoted
(MSI = 717) and requiring a 15% simple after-tax return <f 62-
0
Steorn
on investment. Using the suggested cost of $25/ton for
collected corn stover, the cost of a 35 g/L glucose solution -
0

is 52.5c/kg glucose. Multieffect evaporation is then used

to concentrate the sugar solution for fermentation.
-6 60-
3

51 58-
Figure 20 presents the cost of sugar feed as a function of L

0,
concentration. Hydrolysis of corn stover leaves a burnable
56-
lignin residue which (depending on the hydrolysis yields)
can be sufficient to provide heat for the complete hydroly-
-
s
10

54-
sis and evaporation process. The cost of steam is then re-
duced from $3.15/1000 Ib for coal-generated steam to 52 -
$0.756/1000 lb for waste-generated steam (the capital asso- I I 1

MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT 1021

35 g/L hydrolysis solution requires the evaporation of 19.6 Ethanol Product Concentrotion ( q / l )
kg water/kg glucose or 446,000 kg water/h for a lo9 L/yr 50 60 70 80 90
I I I I I I I I
alcohol plant. The purchased cost of a five-effect evapora-
tor system of this capacity is $6.75 million. Steam (3.91 kg) 1401
is used to concentrate each kilogram of glucose. These
capital investment operating costs are in agreement with
recent data published by General Mills for sugar concen-
tration costs in the food i n d ~ s t r y . ~The
' incremental cost
of concentrating feeds beyond 15 wt 70glucose is small as
the amount of added water to be evaporated for further
concentration is small.

CSTR and CSTR with Recycle a

Figure 21 presents total costs for ethanol produced
from corn stover derived glucose as a function of the fer-

Ethonol Product Concentrotion (g/f

I301
CSTR with Centrifuge Cell Recycle,
RR=05
1
1
I4*k 1 \
1
10
I
I1 12
1
13
1
14
1
I5
1
16
Inlet Supor Concentrotion (weight %)
I I
17
]

Figure 22. Fermentation economics with glucose from corn stover hy-
146- drolysis (using waste-generated steam).

-
5- 144-
- In
mentor sugar feed concentration. The feed sugar costs are
0 as given by Figure 20 for evaporation using coal generated
c
0 steam. The feed sugar storage capacity is reduced from 2
142-
e
a
weeks to 1 day.
- The minimum ethanol product cost for the CSTR sys-
'
0
c

140- CSTR with Centrifuge Cell Recycl:,

RR = 0 5
tem is $1.42/L. As with the molasses feed, cell recycle re-
duces the final cost, in this case to $1.4O/L. Feed evapora-
tion costs now partly offset the savings in reduced stillage
evaporation costs and the optimum feed concentration for
the CSTR with recycle is reduced from 16.7 to 15.2 wt 70
10 II 12 13 14 15 16 17 glucose.
Inlet Sugor Concentration (weight %) Figure 22 presents results for the case where waste-
generated steam is used throughout (for the fermentation
Figure 21. Fermentation economics with glucose from corn stover hy-
drolysis (using coal-generated steam).
plant as well as for feed sugar concentration).

Table XX. Selective ethanol removal fermentation with dilute corn stover hydrolyzate sugar feed, with K = 6. BFR = 0.3, and Xtcr,,,=
loo g/L.
Waste-generated Coal-generated
steam steam
Stillage Sugar
Inlet sugar Fernientor evaporator concentrator Steam Product Steam Product
Pf,,,, concentration cost cost cost cost cost cost cost"
(g/L) (wt '70) ($10") ($109 ($109 (C/L) ($/I-) (C/L) ($/L)

50 28.1 1.18 5.70 87.0 2.05 1.290 8.50 1.375

60 31.8 1.27 5.21 88.0 2.00 I .288 8.29 1.372
70 35.3 1.39 4.76 88.5 1.98 1.287 8.18 1.370
80 38.4 1.64 4.35 89.0 1.96 1.287 8.I3 1.368

aThis cost is exclusivu of separator cost. Reduction compared to the optimum recycle CSTR case ($1.32/L for waste-generated steam
and $1.40/L for coal-generated rteam) indicates the value of a separator of the specified characterirtics.

1022 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984

Selective Ethanol Removal Fermentation
Table xx presents costs for a selective ethanol removal
(extractive fermentation) process using corn stover derived
glucose. As in the molasses feed case (Table XVI), the
minimum cost is for high concentration feed and products.
The savings in stillage handling, centrifugation, and feed
sterilization costs is larger than the added cost of feed
concentration.
For the coal-generated steam case, the minimum final
product cost from extractive fermentation (exclusive of
the extractor) is $1.37/L, compared to $1.40/L for the
CSTR with recycle. This savings is less than for the mo-
lasses feed case, but indicates that a selective ethanol re-
moval fermentation may be advantageous even when pro-
cessing dilute hydrolysis sugar solution feeds.

Table XXI.

Fci-nicntor
Feed sugar Fermentor ccII ethanol Specific Volunictric
concentration concentration conccn~ration productivity. I) ~)roducti\ity
Process (wt ' I n ) (g/L)" (Y/L) (g/h g cell) ( g / l , 11)

Batch 16.7 initial 2.1 8.6 1.78 11.8

final 21.3 85.6 0.42
CSTR 15.6 19.7 79.1 0.72 14.1
Recycle CSTK 16.7 100 85.h 0.42 42.5
Partial recycle 15.6 50.0 79.1 0.72 35.9
2 series CSTR 16.7 first 17.4 70.0 1.03 17.9
second 21.3 85.6 0.42 9.0
Perforated plate column 16.9 21.6 86.9 9.0
(final) (final)
Plug flow 16.7 inlct 36.0 17.1 1.71 52.7
outlet 53.0 85.6 0.42
APV tower 16.7 70.0 0 I .us 86.0
(inlet)
85.6 0.42
(outlet)
Selective membrane' 12.9 80.0 50.0 I .23 08.4
Extractive 38.4 100 80.0 0.69 68.7
Flashb 38.7 100 50.0 1.23 I23

"Concentrations arc in grams per liter water.

hConditions were not optiniiLed for minimum cost.

Table XXU.

Purchased equipment cost

Ethanol Utilities
+ Fi 11 ii I
stillage Cooling I.nhor Yea51 protluct
Storage Fermentation recovery Total Steam wntcr Elcctricity (worker\/ credit co\t"
Process ($103 ($10") ($10") ($10') (kg/L) (kg/L) (lo" k W h/yr) shift) (e/L) (Cl L)

Batch 6.90 18.01 33.86 58.77 3.30 43.4 6.71 I(>. 15 4.8 53.05
CSTK 6.90 13.09 35.51 55.50 3.52 44.1 0.08 10.65 4.8 51.29
Kecycle CSTK 6.90 11.31 30.21 48.42 3.31 43.6 5.70 9.15 4.8 4S.06
Partial recycle 6.90 8.31 35.50 50.71 3.52 44.1 4.27 9.15 4.8 49.76
2 series CSTR 6.90 13. I3 34.67 54.70 3.30 43.4 6.63 11.15 4.8 50.96
Perforated plate column 6.90 19.51 34.56 60.97 3.26 43.4 7.73 10.15 4.8 52.M
Plug flow 6.90 8.17 34.04 49. I I 3.30 43.6 3.03 9. I5 1.8 1').I I
APV tower 6.90 7.47 33.86 48.23 3.30 43.4 2.62 8.65 4.R 48.71
Selective membraneb 5.87 6.27 22.62 34.76 3.39 49. I 2.75' 7. IS 2.9 47.26
Extractive 5.33 3.18 13. I I 21.63 I .87 46.2 I.4YC 0. IS 1.7 4.1.94
Flashb 5.29 2.64 14.50 22.43 2.52 4h.6 I .02' t>, 15 1.7 44.46

"This cost is exclusive of the separation.

"Conditions were not optimized for minimum cost.
'Utilities are exclusive of those required for the separator.

MAIORELLA, BLANCH, AND W I L G : BIOTECHNOLOGY REPORT 1023

CONCLUSIONS
Tables XXI and XXII summarize results for the 11 dif-
ferent fermentation processes evaluated with molasses as
the feed. Raw material costs make up 70% of the final
ethanol sale price, but substantial savings are still possible
when improved methods are used superseding traditional
batch fermentation.
High cell density is important in reducing fermentation
High overall productivity can be maintained even at
high ethanol concentration and the cost of auxiliary equip-
ment can thus be r e d u ~ e d . ~The
’ continuous flow stirred
tank fermentor with cell recycle, plug flow, and tower fer-
mentors show similar cost savings compared to batch fer-
mentation. The plug flow and tower fermentor analyses
neglected backmixing and the tower fermentor analysis
assumed that a flocculent (settling) yeast could achieve
high specific ethanol productivities. Results for these two
cases are, therefore, less certain than for the CSTR case
with recycle.
Selective ethanol removal fermentation processes offer
even greater savings as more concentrated feeds and prod-
ucts can be handled, further reducing the cost of auxiliary
equipment. Very expensive separation devices can be jus-
tified and research should be continued in this area. It
should be noted, however, that much of this savings could
also be achieved by employing stillage recycle in combina-
tion with conventional fermentation processes.
When a dilute sugar solution derived from cellulose hy-
drolysis is used as the feed, sugar concentration costs
partly offset the savings in reduced fermentation cost as-
sociated with handling less water through the remainder
of the plant. The optimum feed sugar concentration to a
CSTR with recycle is slightly reduced. Selective ethanol
removal methods (exclusive of separator cost) offer smaller
cost savings, but may still prove advantageous, depending
on the separator cost.
References
I . B. L. Maiorella, C. R. Wilke, and H. W. Blanch, Adv. Biochem.
Eng., 20,43 (1981).
2. C. R. Wilke and B. L. Maiorella, “High Productivity Anaerobic
Fermentation with Dense Cell Culture,” in Advances in Biotech-
nology. Volume 1 (Pergamon, New York, 1981).
3. R. Katzen, “Grain Motor Fuel Alcohol Technical and Economic
Assessment Study,” U.S. National Technical Information Service,
NO. 061-000-00308-8, 1978.
4. B. L. Maiorella, “Vacuum Ethanol Distillation Technology,” in
The Ethanol Separation Handbook, L. Douglas, Ed. (Solar Energy
Research Institute, Golden, CO, 1984).
5. C. D. Bazua, “The Effect of Alcohol Concentration on the Kinetics
of Ethanol Production by Saccharomyces cerevisiae,” M.S. thesis,
Department of Chemical Engineering, University of California,
Lawrence Berkeley Laboratory, Berkeley, CA, 1976, No. LBL-4423.
6. C. D. Bazua and C. R. Wilke, Biotechnol. Bioeng. Symp., 7 , 105
(1977).
7. J. Monod, Ann. Rev. Microbiol., 3, 371 (1949).
8. 0. Levenspiel, Biotechnol. Bioeng., 22, 1671 (1980).
9. C. Wilke, B. Maiorella, A. Sciamanna, K. Tangnu, D. Wiley, and
1024 BIOTECHNOLOGY AND BIOENGINEERING, VOL. 26, SEPTEMBER 1984
H. Wong, Enzymatic Hydrolysis of Cellulose-Theory and Applicu-
tions (Haigh and Hochland, Manchester, England, 1983).
10. H. Gregor and T. Jefferies, Ann. N. Y. Acad. Sci., 326, 273 (1979).
11. H. Gregor, “Membrane Processes of Separation and Concentration
in Biomass Harvesting, Production and Refining,” Proceedings of
the 3rd Annual Biomass Energy Systems Conference, US Depart-
ment of Energy, Solar Energy Research Institute, Golden, CO,
1980.
12. H. W. Wang, F. M. Robinson, and S . S. Lee, Biotechnol. Bioeng.
Symp., 11, 555 (1981).
13. T. K. Murphy, H. W. Blanch. and C. R. Wilke, Process Biochem.,
17(6), 6 (1982).
14. G. Cysewski and C. R. Wilke, Biotechnol. Bioeng., 19, 1125 (1977).
15. B. L. Maiorella, H. W. Blanch, and C. R. Wilke, “Rapid Ethanol
Production via Fermentation,” report No. LBL-10219, Lawrence
Berkeley Laboratory, Berkeley, CA, 1979.
16. S. P. Prescott and C. G. Dunn, Industrial Microbiologv, 2nd ed.,
(McGraw-Hill, New York, 1949).
17. B. L. Maiorella, H. W. Blanch, and C. R. Wilke, “Ethanol/Water,
Physical and Chemical Properties for Separation Process Design,”
in The Ethanol Separation Hundhook, L. Douglas, Ed. (Solar En-
ergy Research Institute, Golden, CO, 1984).
18. B. L. Maiorella, H. W. Blanch, and C. R. Wilke, Process Uiochem.,
18(4), 5 (1983).
19. M. S. Peters and K. D. Timmerhaus, Plunt Design and Economics
for Chemical Engineers (McCraw-Hill, New York, 1980).
20. Manufacturer’s data, Alfa Lava1 Co., May 14, 1981.
21. R. Hutton, Ed., “Impacts of Applied Genetics,” U.S. Office of
Technology Assessment, U . S . Government Printing Office, Wash-
ington, DC, 1981.
22. D. Milton, “Gasohol Economic Feasibility Study,” Energy Research
and Development Center, University of Nebraska, Lincoln, NB,
July, 1978.
23. B. L. Maiorella, “Ethanol,” in Coniprrherzsive Biotechtzology. Vol-
ume 2, M. Moo-Young, Ed. (Pergamon, Oxford, 1984).
24. F. J. Castillo, M. E. Izaguirre, V. Michelena, and B. Moreno, Bio-
technol. Lett., 4, 567 (1982).
25. H. R. Bilford, R. E. Scalf, W. H. Stark, and P. J. Kolachov, Ind.
Eng. Chem., 34, 1406 (1942).
26. G. R. Cysewski and C. R. Wilke, Biotechnol. Bioeng., 20, 1421
(1978).
27. M. Reuss, D. Josic, M. Popovic, and W. K. Brown, Eur. J. Appl.
Microbiol. Biotechnol., 8(3), 167 (1979).
28. J. S. Hough, P. E. Cough, and A. D. Davis, J. fnst. Brew, 68,478
(1962).
29. T. K. Ghose and R. D. Tyagi, Biotechnol. Bioeng., 21, 1401 (1979).
30. K. Rosen, Process Biochem., 13, 25 (1978).
31. E. Falch and E. Gaden, Biotechnol. Bioeng., 11, 927 (1969).
32. H. R. Bungay, Biosources Dig., 2(1), 27 (1980).
33. M. Moreno and G. Goma, Biotechnol. Lett., 1,483 (1979).
34 P. Linko and Yu-Yen Linko, “Continuous Ethanol Fermentation by
I
Immobilized Biocatalyst,” Proceedings of the 6th Enzyme Engi-
-
neering Conference, Kashikoiima, Japan, September, 1981.
35. T. K. Chose and K. K. Bandyopadhyay, Biotechnol. Bioeng., 22,
1489 (1980).
36. W. Griffith and A. Compere, Dev. Ind. Microbiol., 17, 241 (1976).
37. R. Greenshields and E. Smith, Chem. Eng., 249, 182 (1971).
38. I. G. Prince and J. P. Barford, Biotechnol. Lett., 4, 621 (1982).
39. 0. C. Sitton and J . L. Gaddy, “Design and Performance of an Im-
mobilized Cell Reactor for Ethanol Production,” presented at the
72nd AIChE meeting, San Francisco, CA, November, 1979.
40. B. L. Maiorella, H. W. Blanch, and C. R. Wilke, Biotechnol. Bio-
eng., 25, 103 (1983).
41. R. D. Yang and C. R. Wilke, Appl. Polym. Symp., 28, 175 (1975).
42. J. Perez, C. R. Wilke, and H. W. Blanch, “Economics of Sugar Pro-
duction with Trichoderma Reesei Rutgers C-30,” Lawrence Berkeley
Laboratory, report No. LBL-11489, presented at the Second Chemi-
cal Conference of the North American Continent, Las Vegas, NV,
August, 1980.
43. H. R. Bungay, Adv. Biochem. Ertg., 20, 1 (1981).
44. W. Brenner, B. Rugg, and C. Rogers, Clean Fuels Biomuss Wastes
Symp., 1, 201 (1977).
45. H. E. Grethlein, J. Appl. Chrm. Biotechnol., 28, 296 (1978).
46. M. M. Chang, T. Y. C. Chow, and G. T. Tsao, Adv. Biochem.
Etig., 20, 15 (1981).
47. T. A. Renshaw, S. F. Sapakie, and M. C. Hanson, Chem. Eng.
Prog., 78(5), 33 (1982).
48. H. H. Topiwala and B. Khosrovi, Biotechrtol. Bioeng., 20, 73
(1978).
49. E. J. Del Rosario, K. J. Lee, and P. L. Rogers, Biotechnol. Bioeng.,
21, 1477 (1979).

MAIORELLA, BLANCH, AND WILKE: BIOTECHNOLOGY REPORT 1025