Bioresource Technology 5 !
( 1995 ) 103-109
ELSEVIER 0960-8524(94)00136-7
FERMENTATIVE PRODUCTION OF 2,3-BUTANEDIOL:
A REVIEW
S. K. Garg* & A. Jain
Department of Microbiology, Dr Ram Manohar Lohia University, Faizabad-224 001, U.P., India
(Received 30 September 1994; accepted 12 November 1994)
Abstract
The production of 2,3-butanediol by bacterial species
continues to be of great interest because of its varied
application. Two bacterial species, Bacillus polymyxa
and Klebsiella pneumoniae have demonstrated poten-
tial for butanediol fermentation on a commercial scale.
Klebsiella pneumoniae, owing to its broad substrate and
cultural adaptability, is the most thoroughly investigated
organism. It is generally recommended that natural
resources of 'waste' cellulosic and hemicellulosic sub-
strates be fermented, for the improvement of process
economics. The process of diol fermentation is repor-
tedly influenced by various nutritional and environ-
mental parameters. The high boiling-point characteristic
of 2,3-butanediol poses serious problems in recovery of
diol from fermented slurry. Repeated solvent extraction
and countercurrent-stream stripping have been recom-
mended as the methods of choice for diol recovery.
Key words: Butanediol, fermentation, Klebsiella pneu-
moniae, Bacillus polymyxa, wood hydrolysate, xylose.
INTRODUCTION
In the year 1906, Harden and Walpole investigated the
microbial production of 2,3-butanediol employing
Aerobacter aerogenes ( Klebsiella pneumoniae) and this
was followed by Donker (1926) using Bacillus poly-
myxa. Fulmer et al. (1933) proposed the industrial pro-
duction of the diol for the first time. Attention was
again focused on diol fermentation during wartime due
to the anticipated shortage of 1,3-butadiene.
2,3-Butanediol is an interesting product owing to its
diverse potential uses as a polymeric substance. It is a
colourless and odourless liquid having a very high boil-
ing point (180-184°C) and low freezing point. The
levo isomer ( - 60°C) of butanediol can be used as an
antifreeze agent. However, the commercial application
of diol is not limited to the manufacture of butadiene or
antifreeze. 2,3-Butanediol is a potentially valuable fuel
additive with a heating value of 27 198 J g-1 which
compares favourably with other liquid fuels (methanol
*Author to whom correspondence should be addressed.
103
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22081 J g-l, ethanol 29055 J g-l; Flickinger, 1980).
The presence of ethanol does not affect the usefulness
of 2,3-butanediol as a fuel since an equimolar mixture
of ethanol and diol can provide a combined heating
value of 27660 J g-i (Yu & Saddler, 1982b). Apart
from fuel, butanediol can also be converted to diacetyl
and butan-2-one (methyl ethyl ketone). The diacetyl
formed by catalytic dehydrogenation of the diol is a
highly-valued food additive used as a flavouring agent.
Further, butanediol has a high octane number and can
therefore be used as an octane booster for gasoline or
as high-grade aviation fuel. Condensation of diol to
methyl ethyl ketone (MEK) coupled with subsequent
hydrogenation yields octane isomers that can be used
to produce high quality aviation fuel. MEK can also be
used as a solvent for resins and lequers (Villet, 1981 ).
Esterification of butanediol forms precursors of poly-
urethane foams for use in drugs, cosmetic products,
lotions, ointments and antiperspirants. It has antiseptic
properties and a 0-1% solution will kill most patho-
genic bacteria. Diacetyl is reported to inhibit the
growth of Mycobacterium tuberculosis and many non-
pathogenic organisms more effectively than does
benzoic acid (Underkofler & Hickey, 1954). Butane-
diol has also been shown to have potential application
in the manufacture of printing inks, perfumes, fumi-
gants, moistening and softening agents, explosives and
plasticisers, and has been used as a carrier for pharma-
ceuticals.
The production of butanediol from lignocellulosic
wastes has recently been considered as an alternative
approach in the conversion of biomass substrates to
liquid fuels and chemical feedstocks (Flickinger, 1980;
Jansen et al., 1984). Klebsiella pneumoniae is the
organism of choice owing to its ability of utilising all the
major sugars (hexoses, pentoses, certain disaccharides)
and uronic acid derived from the hydrolysates of hemi-
cellulosic and cellulosic materials (Yu et aL, 1982;
Saddler et al., 1983). The sugars present in wood hemi-
celluloses could be efficiently utilised after they had
been released by either acidic (Yu et al., 1982) or enzy-
matic (Saddler et al., 1983) hydrolysis. The present
review provides updated information on the produc-
tion of 2,3-butanediol through microbial fermentation
processes.
104 S. K. Garg, A. Jain
MICROORGANISMS
Several yeasts have the ability to form diol, though the
yields are extremely low, and therefore bacteria are at
present the only organisms of industrial importance in
the butanediol fermentation. A number of species of
the genera Klebsiella, Bacillus, Serratia and Pseudo-
monas have the ability to produce butanediol, but only
two organisms, Klebsiella pneumoniae and Bacillus
polymyxa, have been shown to be potentially useful
(Long & Patrick, 1963). The yield of diol by B. poly-
myxa with all substrates is uniformly lower than that by
K. pneumoniae. Klebsiella pneumoniae, widely dis-
tributed in nature, is stable under a wide range of en-
vironmental conditions and produces double the
amount of diol with much higher yields than does B.
polymyxa (Ledingham & Neish, 1954). The organism
also carries out a rather more complete fermentation,
thereby facilitating product recovery. Klebsiella
pneumoniae grows rapidly in a simple medium
(Anderson & Wood, 1962) and it is able to metabolise
a wide variety of sugars. If genetic improvement of this
organism becomes desirable, its similarity to Esche-
richia coil could prove to be a fortuitous advantage.
Perhaps the most important attribute of K. pneumoniae
related to biomass conversion is its ability to convert all
of the major sugars present in hemicellulose and cel-
lulose hydrolysates into 2,3-butanediol (Yu & Saddler,
1982b). Stock cultures of the valuable strain can be
stored with relative ease. For the aforementioned
reasons, by far the greatest research effort in butane-
diol production has been focused on the use of this
organism.
FACTORS AFFECTING BUTANEDIOL
PRODUCTION
Cultural factors
2,3-Butanediol fermentation is influenced by various
cultural factors: pH, temperature, aeration, agitation
and inoculum.
Effect of p H
The pH is a fundamental parameter in the regulation of
bacterial metabolism and its influence is especially pro-
nounced in the processes involving multiple end-
product formation. The pH of the fermentation
medium affects both the biomass composition and the
nature of the bacterial metabolism, but the molecular
basis of these effects is not very well understood. Pos-
sibly the proton motive force generated across the
plasma membrane in chemiosmosis is affected by the
medium pH value. Hence, in the optimum pH range,
the relative metabolic efficiency of the organism is high
(Pirt, 1975).
As a general rule, alkaline conditions (above pH
6.3-6.5) favour the formation of organic acids, with a
corresponding decrease in the yield of butanediol. In
contrast, under acidic conditions, organic acid syn-
thesis is reduced more than 10-fold, whereas diol pro-
duction increases by 3.7 times. The net result is a
reversal of relative yields, with diol accounting for 67%
of the total solvents produced and acids limited to only
9%. The pH optimum for diol production appears to
be dependent on the substrate from which it is pro-
duced. For most substrates, including wood hydro-
lysate, the optimum pH range has been reported to be
6.0-6.2 (Long & Patrick, 1961; Grover et al., 1990).
Raspoet et al. (1991) have reported maximum butane-
diol production from glucose by Bacillus licheniformis
at pH value 6"0. In contrast, the diol production was
greatest in the pH range of 5"0-5"2 when xylose was
used as a carbon source (Tsao, 1978). There appears to
be a direct correlation between diol and biomass yields.
Jansen et al. (1984) observed the maximum growth
rate and diol yield under aerobic conditions at pH 5"2
when 10% xylose was the substrate. However, no
growth was observed at a pH below 4.2.
Temperature
The efficiencies of bioprocesses become strictly tem-
perature-dependent owing to the strong dependence of
enzymatic activity and cellular maintenance upon tem-
perature. In general, the optimum temperature for the
bacterial fermentation of butanediol is reported to be
in the range of 30-35°C (Perlman et al., 1944). Since
production of butanediol is generally accepted to be a
growth-associated phenomenon, the condition for
maximum product formation must approximately be
the same as that of maximum biomass yield. Grover et
al. (1990) have reported a correlation between butane-
diol and ethanol production and growth of K. pneu-
moniae cultured in wood hydrolysate at pH 6'0 and
30°C. The yield of butanediol was 26-27% during
44-50 h; the maximum ethanol yield of 9.8% was at
44 h. The total reducing sugars utilised at 50 h was
88"5%. The total viable count reached a maximum of
101° cfu ml- 1 at 24 h and then remained constant up to
38 h, declining thereafter (Fig. 1 ). Further utilisation of
sugars may have been prevented by the accumulation
of inhibitory products, including ethanol and organic
acids, in the fermenting slurry. The effect of different
temperatures on growth rate could be predicted in
terms of the activation energy required for growth.
Above the optimum temperature, cell degradation
probably becomes dominant over the growth process;
with suboptimal temperature, the regulation of meta-
bolism may fail (Pirt, 1975).
Aeration
Although 2,3-butanediol is a product of anaerobic
metabolism, aeration has been shown to increase its
production by K. pneumoniae (Ledingham & Neish,
1954). Laube et al. (1984a) observed that less xylose
substrate was used and less diol accumulated in sealed
flasks and in unsealed stationary cultures of B. poly-
myxa than was the case in open, agitated, flasks.
Perhaps elimination of the evolved CO2 under aerated
conditions was the factor responsible for superior
xylose utilisation and diol production. Excessive aera-
 Production of 2,3-butanediol: a review
60 12
50 10
8 8
00
30 6 6~0
20 ~-4
~ -
I0 2 -2
0 !0 20 30 40 50
T i m e (h)
Fig. 1. Production of butanediol (o) and ethanol (zx);
residual sugar (A) pattern at different time intervals; growth
curve as colony forming units (c.f.u.) ml- ~(o) by Klebsiella
pneumoniae at pH 6, 30°C during 50 h shake-flask fer-
mentation. From Grover et aL (1990) with permission.
tion, however, was detrimental to diol formation since
the diol yield was higher in cultures shaken at 125 rpm
than in those shaken at 300 rpm. During the fermenta-
tion of five-carbon sugars, or especially when high sub-
strate concentrations are encountered, diol production
has been shown to be most efficient under aerobic con-
ditions. In contrast, Motwani et aL (1993) obtained
similar levels of 2,3-butanediol from water hyacinth
fermented by Klebsiella oxytoca under both static
(anaerobic) and agitated conditions, although the
former took relatively longer to achieve the maximum
level of diol. However, Ramachandran et aL (1990)
observed that increasing lactose concentrations did not
inhibit cell growth of Klebsiella oxytoca but indirectly
influenced 2,3-butanediol yield and productivity by
affecting specific oxygen availability to the cell, as
optimum oxygen-transfer rates are needed for maxi-
mum productions, yields and productivities at different
initial lactose concentrations. Further, 2,3-butanediol
inhibited specific growth-rate but had less effect on
specific rate of product formation. However, at high
lactose concentration, inhibition of growth by 2,3-
butanediol was reduced. Motwani et al. (1993) noted a
reduction in the maximum butanediol concentration
for periods of agitation longer than 20 h; this was attri-
buted to the diversion of more substrate towards bio-
mass formation because of the availability of more
oxygen. Jansen et al. (1984) also found that oxygen
supply favoured cell mass formation at the expense of
butanediol. Motwani et al. (1993) have noted low 2,3-
butanediol production from water hyacinth at a high
HRT, and this was attributed to the possible utilisation
of butanediol by K. oxytoca because of the substrate
(reducing sugars) limitation. Utilisation of 2,3-butane-
diol by Klebsiella sp. has also been observed by Yu and
Saddler (1982a); this was shown by an increase in 2,3-
butanediol concentration at a lower hydraulic retention
105
112 time (HRT) when the residual sugars were higher and
hence not limiting. Afschar et al. (1991) also reported
10 that K. oxytoca fermented 2,3-butanediol at an
extremely high rate once the reducing sugars in the
medium were exhausted, and this could be prevented
by complete inhibition of K. oxytoca by the addition of
u.... 15 g ethanol 1-1 and switching off aeration. Decreasing
the oxygen supply increases the butanediol yield, but it
g. decreases the overall conversion rate due to the lower
,..1
4 cell concentration formed. The effect of aeration is
probably of considerable importance in diol fermenta-
2 tion, especially because it provides agitation also. This
stirring action increases the efficiency of fermentation
by continuously exposing new substrate to the culture
0 and disseminating the metabolic end products through-
out the medium (Long & Patrick, 1963). Oxygen is the
terminal electron acceptor in the production of ATP
via oxidative phosphorylation. The ATP subsequently
provides energy required in many of the reactions of
cell synthesis, maintenance and product formation;
butanediol formation is thus mediated by the presence
of oxygen in the culture environment.
Inoculum
The size of inoculum used in the bioconversion of
either glucose or xylose does not appear to have any
significant effect on the final yield of butanediol. Yu
and Saddler (1982a) observed that the initial fermenta-
tion rate of a high concentration of xylose was in-
fluenced by the inoculum size of K. pneumoniae, but
the eventual diol yield was not altered. Laube et al.
(1984a) reported that increasing the inoculum size
from 2"5 to 5.0% did not improve xylose-substrate
utilisation, or diol production, by B. polymyxa.
Acclimatisation of cultures has been shown in several
cases to be extremely beneficial for the improvement of
both fermentation rates and diol yields. Perlman (1944)
obtained greater yield of diol using cultures of K. pneu-
moniae which had been acclimatised to a wood-
hydrolysate substrate. He also observed that it was
necessary to supplement the substrate with yeast
extract or malt-sprouts extract if high sugar and
unadapted cultures were used. These additives were
not required when the cultures were acclimatised
(Grover et al., 1990). The authors reported that the
supplementation of wood-hydrolysate medium with
malt extract, yeast extract, beef extract or (NH4)2SO4
gave only slight improvements in butanediol produc-
tion without affecting ethanol yield. Clearly the pre-
sence of these nutrient supplements did not improve
the removal, or prevent the accumulation, of inhibitory
materials in the medium, as had been suggested by
Long and Patrick (1963). Adaptation of K. pneu-
moniae to high substrate concentration prior to ino-
culation proved effective in the aerobic dissemination
of 150 g1-1 xylose medium (Yu & Saddler, 1983).
Perhaps the greatest benefits of inoculum acclimatisa-
tion are to be obtained in the bioconversion of natural
substrates. Concentrated wood-hydrolysates and agri-
cultural-residues are traditionally difficult to utilise.
106 S. K. Garg, A. Jain
While the inhibitory agents found in these materials
have not as yet been fully identified (Mes-Hartree &
Saddler, 1983), breakdown products of pentose sugars
(furfural, hydroxymethyl furfural, etc.) and lignin are
believed to be the principal compounds. Frazer and
McCaskey (1991) have evaluated the effect of acetate,
sulphate, furfural and phenolic compounds, the
potential microbial inhibitors commonly present in
acid-hydrolysed hardwood, on conversion of D-xylose
to 2,3-butanediol by K. pneumoniae. The high solute
concentrations that are characteristic of these sub-
strates may, also, be a major factor in the inhibition of
such poorly osmotolerant species as K. pneumoniae.
Nutritional factors
Substrate concentration
In most studies, it seems that the usual concentrations
of carbohydrates employed were in the range of
5-10%. It is probable that these rather low concentra-
tions were caused, in part, by the fact that as the sugar
concentration in a raw material is increased, accom-
panying toxic substances are also increased. The latter
substances are particularly apparent in wood hydro-
lysates and various types of molasses (Long & Patrick,
1963; Frazer & McCaskey, 1991). Yu and Saddler
(1992a;b) reported the bioconversion of sugars
present in wood hemicelluloses to diol by K. pneu-
moniae grown on high initial sugar concentrations (up
to 10%) in the presence of acetic acid. High test/invert
molasses was found to be a good substrate for the pro-
duction of 2,3-butanediol by K. pneumoniae, and the
yield could be further increased by increasing the cell
mass (Afschar et al., 1991).
Under finite aeration, significant inhibition of both
diol formation and sugar utilisation has been observed
at carbohydrate levels exceeding 50 g1-1 (Yu &
Saddler, 1983). While this trend was particularly
evident when xylose was the carbon source, the effect
was also noted in studies utilising D-glUCOSe, L-ara-
binose, D-galactose, D-mannose, and D-cellobiose.
Improvement in diol yield, in contrast, was observed
with increasing glucose concentration when K. pneu-
moniae was grown in an anaerobic environment
(Sablayrolles & Goma, 1984). Conflicting reports exist
regarding the effect of initial xylose concentration upon
diol yield (Laube et al., 1984a; Jansen et al., 1984).
This might be attributed to the choice of the organisms.
The influence of xylose concentration on diol fer-
mentation thus appears to be species specific. Diol
yields by B. polymyxa were improved by increasing the
substrate concentration and up to 6"4 g1-1 was
obtained with 4 and 6% xylose; no appreciable amount
of xylose was fermented or diol produced with 10%
substrate (Laube et al., 1984a). The hydrogen fluoride
saccharification of wood chips and the post hydrolysis
of the reversion products were carried out when using
K. pneumoniae for the production of butanediol and
ethanol (Hardt & Lamport, 1982; Yu et al., 1984). The
sugar constituents found in hemicelluloses were tested
under best conditions for diol production from xylose
and with an inoculum of B. polymyxa grown on xylose.
The monomers and cellobiose were readily fermented,
with more than 90% of the substrate being used, and
yielded significantly more diol than xylose. However,
diol yields from soluble starch were similar to thoSe
from xylose (Laube et al., 1984a).
Product concentration
Butanediol fermentation has a competitive advantage
over ethanol or acetone-butanol fermentations, in
which the end products are considerably more toxic
(Yu & Saddler, 1982a). The major influence of high
butanediol concentration is on biomass formation and
not on product yield. It appears, therefore, that butane-
diol at high concentrations strongly inhibits bacterial
growth but has very little effect upon the metabolic
pathways leading to its formation (Jansen et al., 1984;
Laube et al., 1984a; Sablayrolles & Goma, 1984). This
explains why Grover et al. (1990) could find maximum
growth of K. pneumoniae at 24 h which remained
almost constant up to 38 h and declined thereafter,
whereas butanediol production continued up to 50 h.
A butanediol concentration of up to 100 g 1-1 had no
effect on the specific rate of its production (Yu &
Saddler, 1985). Zeng and Deckwer (1991) have pro-
posed a model for multiproduct inhibition of Entero-
bacter aerogenes growth in which ethanol was
identified as a strong inhibitor, in addition to acetic
acid and 2,3-butanediol, in butanediol production.
Medium supplements
Culture media must contain all the essential nutrients
for the growth and maintenance of the particular
organism for which they are formulated. In addition to
carbon and nitrogen, a minimal medium may include
vitamins, trace elements, etc.; the selection of which
may, in part, be determined by the nature of the desired
fermentation products.
Yeast-extract constitutes an excellent source of
nitrogen as well as other growth factors, although the
excessive cost involved would be prohibitive in its use
on an industrial scale. A nitrogenous compound which
is both cheap and adequate for diol fermentation is
urea, which has been used with such diverse substrates
as hydrolysed wheat mashes and wood hydrolysates. In
general, K. pneumoniae and related species are less
demanding in their nutritional requirements and give
satisfactory diol fermentation in media which contain
sugar and inorganic salts (Long & Patrick, 1963). How-
ever, Nilegaonkar et al. (1992) reported that addition
of peptone/beef extract as medium supplement
enhanced butanediol production (47 g per 100 g
glucose) by Bacillus licheniformis so much that the
yield was higher than those previously reported for K.
oxytoca (37 g per 100 g glucose) and Bacilluspolymyxa
(24 g per 100 g glucose). Laube et al. (1984a) reported
that with increasing wheat extract concentrations, from
0"3 to 1"2% (w/v), xylose utilisation and diol produc-
 Production of 2,3-butanediok a review
tion by B. polymyxa improved. Surprisingly, this effect
was not evident when 1% glucose was the substrate.
However, when 5% glucose was present, diol yields
with 1% yeast extract were significantly higher than
those with 0"5% yeast extract. Adding 1% (w/v) malt
extract to the wood-hydrolysate medium slightly
enhanced 2,3-butanediol production by K. pneu-
moniae without changing ethanol yield (Grover et al.,
1990).
However, to make the diol fermentation more
economical, searching for an inorganic replacement for
at least a portion of yeast extract would have potential
impact. Certain metallic ions have been found to
improve the conversion efficiencies. The principal
stimulatory trace metals were found to be Fe 2+ and
Mn 2+, which gave maximum stimulation of butanediol
fermentation at concentrations of 40 and 1"7 /~M,
respectively (Laube et al., 1984b). Supplementation of
medium containing 0-5% yeast extract with Fe 2+, Mn 2+
or Fe 2+ plus Mn 2+ resulted in yields of 8.7, 12.6 and
13"0 g 1-1, respectively, as compared to a control yield
of 6-6 g 1- ~ by B. polymyxa. The yield in the presence
of 1.5% yeast extract (17.8 g 1-l) was not, however,
achieved by the supplementation of the medium with
these former factors. Murphy and Stranks ( 1951 ) have
shown phosphate to stimulate diol production. The
addition of an optimum concentration of phosphate
(78/v~) in the form of potassium phosphate or sodium
phosphate to a medium containing 0-5% yeast extract
was found to improve the conversion efficiencies,
resulting in an average diol yield of 13"8 g 1-1 from 5%
glucose. Thus, the possible effects on diol yields and
glucose metabolism were not due to a cationic effect
but were truly an effect of phosphate, most likely func-
tioning to stimulate the complete metabolism of the
bacterium. Through combination of these stimulatory
factors (Fe2+, Mn z+, phosphate), the resulting yield of
butanediol ( 15.4 g 1-1 ) was 89% of that obtained in the
presence of 1"5% yeast extract (Laube et al., 1984b).
Thus, considering yields and rates, low (0"5%) yeast
extract medium, supplemented with phosphate, Mn 2+,
Fe 2+, could adequately replace 1-5% yeast extract
medium. Frazer and McCaskey (1991) studied the
effect of components of acid-hydrolysed hardwood on
conversion of D-xylose to 2,3-butanediol by K. pneu-
moniae. Of the various potential inhibitors evaluated
for butanediol production, acetate at 1-6% (w/v) stimu-
lated its production, whereas sulphate up to 0.2% (w/v)
did not affect growth but reduced butanediol yield by
approximately 30%. Similar growth results were
obtained with furfural concentrations up to 0-2% (w/v),
but butanediol yields were slightly higher. However,
phenolic compounds (syringealdehyde and vanillin),
even at low concentrations of 0"05 and 0"10% (w/v),
were found to be inhibitory for butanediol formation
and growth, respectively.
Water activity
Water activity (aw), which is related to osmotic pressure
and varies inversely with solute concentration, is
107
another important variable affecting 2,3-butanediol
production (Jansen & Tsao, 1983). It is dependent
upon the molar concentration and the activity coef-
ficient of each solute present in an aqueous solution
(Pirt, 1975). Increasing the solute concentration
decreases the water activity of a solution. In organisms
such as K. pneumoniae, which are known to possess
relatively weak osmotolerance, water activity may be
an extremely important environmental consideration.
The problem associated with the bioconversion of
complex substrates such as starch or wood-hydro-
lysates may, therefore, result, in part, from the high
solute-concentrations of such feedstocks. At a water
activity of 0-985, growth of K. pneumoniae is reduced
to one-half its optimal level. Water activity may also
explain why the butanediol process is more difficult
with a natural, complex source of carbohydrate. In
addition to possible presence of toxic compounds, the
total solute concentration in these complex carbon-
sources is likely to be higher because of the presence of
non-carbohydrates; these cause a lower water activity,
which may be responsible for lower metabolic rates
(Jansen & Tsao, 1983).
Recovery of 2,3-butanediol
The major difficulties in recovering butanediol are due
to its high boiling point, its great affinity for water and
the presence of dissolved- and solid-constituents of fer-
mentation mashes. The high boiling-point renders
simple vacuum distillation particularly useless. Before
the distillation temperature is attained, the dissolved
constituents concentrate into a thick tarry mass which
effectively binds and retards vaporisation of the diol
(Underkofler & Hickey, 1954).
Repeated solvent extraction has been used as a
method of recovery, with some success but primarily
on a laboratory scale. A number of solvents are suit-
able for this purpose, including ethyl acetate, diethyl
ether and n-butanol (Johnson, 1944). Tsao (1978)
found that 75% of the diol present in a fermentation
broth could be recovered by a single extraction with
diethyl ether; recovery of the co-products, acetoin,
ethanol and biacetyl, was found to be 65, 25 and
75-90%, respectively. The most practical method of
diol recovery appears to involve countercurrent stream
stripping.
FUTURE PROSPECTS
By contrast with the older fermentations, 2,3-butane-
diol is a relative newcomer to the field, and as such has
yet to attain importance as a commercial product by
way of developing suitable methods of production of
this interesting chemical.
The production of butanediol from lignocellulosic
wastes has recently been considered as an alternative
approach in the conversion of biomass substrates to
liquid fuels and chemical feedstocks. However, most of
the processes considered only the cellulose fraction;
the hemicellulose and lignin were generally ignored. If
108 S. K. Garg, A. Jain
such a process of butanediol production is to be econo-
mically attractive, the efficient utilisation of all the
available constituents in the cellulose and hemi-
cellulose, as well as the lignin, of the biomass must be
achieved.
No recent research has been directed towards one of
the greatest problems of the butanediol fermentation,
that of economical recovery of the product from the
fermentation mash. Solvent-extraction methods have
been used with a little success on a laboratory scale.
The most practical method to date appears to involve
countercurrent stream-stripping for diol recovery.
Finding a novel and economical method for butanediol
recovery would certainly help develop the process on
an industrial scale.
Klebsiella pneumoniae, owing to its broad substrate
spectrum and cultural adaptability, is by far the most
sought-after organism for the commercial production
of 2,3-butanediol. The advent of genetic and protein-
engineering techniques has opened up new vistas in
the field, through which efficient strains suitable for
industries could be developed. Furthermore, the genes
from such a strain could be transplanted into some
other suitable organisms which are still better adapted
for the efficient conversion of agro-industrial wastes to
assets like 2,3-butanediol.
Various researchers have demonstrated the utility of
fixed-film-reactor technology for the production of
2,3-butanediol (Chua et aL, 1980; Lee & Maddox,
1986). The need for recirculation is obviated owing to
the high cellular concentration in fixed-film systems.
Thus a low hydraulic-retention time can be attained by
using such systems, leading to high productivity and
hence enhanced cost-effectiveness. A limited success
has so far been achieved for the butanediol production
by immobilised cultures. However, Motwani et al.
(1993) have reported encouraging results with anaero-
bic-fixed-film and upflow-anaerobic-sludge-blanket
reactor systems for the continuous production of 2,3-
butanediol. The process could be suitably improved
further so as to have commercial viability.
REFERENCES
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