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Propionic Acid Production via Fermentation Route using Renewable Sources

Article · September 2008

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 Propionic Acid Production via Fermentation Route using
Renewable Sources
Sushil Kumar1 and B.V. Babu*

*Corresponding Author
Dean-Educational Hardware Division & Professor of Chemical Engineering
Birla Institute of Technology and Science (BITS), PILANI – 333031 (Rajasthan), India.
Email: bvbabu@bits-pilani.ac.in; Phone: +91-1596-245073 Ext. 259; Fax: +91-1596-244183
Homepage: http://discovery.bits-pilani.ac.in/discipline/chemical/BVb/
1
Lecturer, Chemical Engineering Group, BITS Pilani- 333031, India Email: skumar@bits-pilani.ac.in

ABSTRACT
Propionic acid is widely used in chemical, pharmaceutical and food industries. The growing
demand of this acid draws attention towards a more economical and energy efficient
technology. The conventional method of recovery of the propionic acid is expensive and
unfriendly to the environment. Producing Propionic acid by fermentation is the most energy
efficient (uses renewable energy source) route. This paper presents a state-of-the-art review
on fermentation technology for the production of propionic acid and various recovery
methods from the fermentation broths. Reactive extraction with specified extractant giving a
higher distribution coefficient has been emerged as a promising technique for the recovery of
acid.
Keywords: Carboxylic Acids; Propionic Acid; Fermentation technology; Recovery; Reactive
Extraction.

INTRODUCTION
There has been a resurgence of interest in large-scale production of fermentation chemicals
due to a sharp increase in petroleum cost in the recent past. So the potential role of a new
energy efficient fermentation technology is becoming increasingly important. The current
economic impact of fermentation chemicals, however, is still limited, because of difficulties
in product recovery. Thus, substantial improvements in the existing recovery technology for
fermentation products are also required for the organic chemical industries. Organic acids -
widely used in the food, pharmaceutical and chemical industries - are important chemicals in
human life. Some of these acids (propionic acid, lactic acid, itaconic acid, tartaric acid, and
citric acid) are obtained by aerobic fermentation of glucose via the glycolytic pathway and
glyoxylate bypass. Corn meal hydrolyzed with amylases was used as the carbon source for
producing acetic acid, propionic acid, and butyric acid via anaerobic fermentation (Atkinson
and Mavituna, 1983). The aerobic fermentation process leading to the formation of organic
acids is one of the best established microbial conversion pathways.

Propionic acid (CH3CH2COOH) is a naturally occurring carboxylic acid. In the pure state, it is
a colorless, corrosive liquid with a sharp and somewhat unpleasant odor. Propionic acid was
first described in 1844 by Gottlieb, who found it among the degradation products of sugar. It
has physical properties intermediate between those of the smaller carboxylic acids, formic and
acetic acid, and the larger fatty acids. It is miscible with water. Like acetic and formic acids,
in gaseous form, it grossly violates the ideal gas law because it does not consist of individual
propionic acid molecules. It also undergoes hydrogen bonded pairing in solution. Chemically,
propionic acid shows the general properties of carboxylic acids, and like most other
carboxylic acids, it can form amide, ester, anhydride, and chloride derivatives. It can react
with chlorine or other halogens to replace one of the hydrogen atoms attached to its carbon
atoms with a halogen atom (Wikipedia, 2008).

Propionic acid is used in the manufacture of herbicides, chemical intermediates, artificial fruit
flavors, pharmaceuticals, cellulose acetate propionate, and preservatives for food, animal feed,
and grain. Its present U.S. market is estimated at 235 million pounds per year (Chemical
Marketing Reporter, 1997) and is growing at 4% annually. Taking into account of the
importance and demand of Propionic acid in the world scenario, this paper aims at reviewing
the production of Propionic acid.

CHEMICAL SYNTHESIS OF PROPIONIC ACID

In industry, propionic acid is mainly produced by the hydrocarboxylation of ethylene using
nickel carbonyl as the catalyst given by Eq. 1:

RCH=CH2 + H2O + CO → CH3CH2CO2H (1)

It is also produced by the aerobic oxidation of propionaldehyde. In the presence of cobalt or

manganese ions, this reaction proceeds rapidly at temperatures as mild as 40-50°C given by
Eq. 2:
CH3CH2CHO + ½ O2 → CH3CH2COOH (2)

Large amounts of propionic acid was once produced as a byproduct of acetic acid
manufacture. Current world's largest producer is BASF, with approximately 80 ktpa
production capacity. Propionic acid can also be produced biologically from the metabolic
breakdown of fatty acids containing odd numbers of carbon atoms, and from the breakdown
of some amino acids.

FERMENTATION TECHNOLOGY FOR PROPIONIC ACID

Commercial production of propionic acid is chiefly carried out by chemical synthesis from
petroleum feedstocks, but fermentation is an attractive alternative to produce this acid from
renewable sources. Several carbon sources have been used for this fermentation such as
glucose, xylose, maltose, sucrose and whey lactose (Zhong et al. 1997). The most widely used
carboxylic acids obtained by fermentation process are shown in Table-1.

Table – 1 various carboxylic acids obtained by fermentation process

S.
Name of acid Source Usage
No
whey, potatoes, In biopolymer as a food additive, sweets,
1. Lactic acid
cornstarch, or molasses and soft drinks etc.
2. Citric acid fruits and vegetables As a natural preservative, antioxidant etc
plants, particularly
3. Tartaric acid As an antioxidant in food industries
grapes and tamarinds
As a preservative and to make pesticides,
4. Propanoic acid milk(Gk.protus prion)
pharmaceuticals etc.
In the manufacture of plastics, agricultural
5. Butanoic acid butter (L. butyrum)
chemicals, pharmaceuticals etc.
In polymers, paints, coatings, medicines,
6. Itaconic acid sugar, carbohydrates
cosmetics etc.
plants, fruits and other
7. Gluconic acid In detergent and cleaner industries.
foodstuffs, honey
plant proteins, sprouting In the production of Bio-chemicals,
8. Aspartic acid
seeds, dairy immunoglobulin etc
Propionic acid bacteria have long been used in the dairy industry. These bacteria play an
important role in the development of the characteristic flavor in Swiss-type cheese.
Propionibacteria are Grampositive, nonspore-forming, rod-shaped, facultative anaerobes. Fed-
batch fermentations of glucose by P. acidipropionici ATCC 4875 in free-cell suspension
culture and immobilized in a fibrous-bed bioreactor (FBB) are studied by Suwannakham and
Yang (2005). The enhanced propionic acid production from glucose by P. acidipropionici
was attributed to both a high viable cell density maintained in the reactor and favorable
mutations resulted from adaptation by cell immobilization in the FBB. Blanc and Goma
presented a study of propionic acid production from whey by using Propionibacterium
acidipropionici in batch and continuous fermentation with cell recycle. The highest propionic
acid productivity was 2.14 kg/(m3 h). Biomass concentration and propionic acid productivity
were found to be nine times and six times higher respectively by this route when compared to
the results obtained in batch studies. Despite the great variety of substrates studied for
propionic acid production, glycerol has rarely been considered. However, a better efficiency
of propionic acid production from glycerol could be expected because of its higher reduction
level compared to conventional substrates. A comparative study was carried out in anaerobic
batch cultures of either glycerol or glucose using two propionibacteria strains,
Propionibacterium acidipropionici and Propionibacterium freudenreichii ssp. Shermanii by
Himmi et al. (2000). In all these cases, fermentation end-products were the same and
consisted of propionic acid as the major product, acetic acid as the main by-product and two
minor metabolites, n-propanol and succinic acid. The greater production of propionic acid by
Propionibacteria was obtained with glycerol as carbon and energy sources. P. freudenreichii
ssp. shermanii is scarcely used for glycerol fermentation.

Like most organic acid fermentations, the propionic acid fermentation is inhibited by acidic
pH and propionic acid which is the major fermentation product. The conventional
fermentation technology for propionate production is thus limited by low fermentation rate
and low product concentration. Furthermore, the fermentation is heterogeneous; i.e.,
propionate is produced along with other byproducts. This not only results in a low product
yield but also renders product purification difficult and expensive. Consequently, the
conventional fermentation route for propionic acid production is inefficient and it can not
compete with the petrochemical routes. In order to make the fermentation route economically
viable, it is necessary to develop novel fermentation processes that use highly efficient
bioreactors and separation techniques. The integrated fermentation-separation systems have
been successfully used to reduce end-product inhibition and, thus, to improve the overall
process efficiency.

CONVENTIONAL SEPARATION TECHNIQUES

Fermentation technology for the production of organic acids in particular has been known for
more than a century and acids have been produced in the form of aqueous solutions. These
bioconversions and recovery from fermentation broth are severely inhibited by the products.
The growing importance of biological production, expressed with new routes and increasing
production rates, asks for adapted downstream processing for product separation. Several
separation methods such as liquid extraction, ultra filtration, reverse osmosis, electro-dialysis,
direct distillation, liquid surfactant membrane extraction, anion exchange, precipitation,
adsorption, etc. have been employed to remove carboxylic acids (Kumar and Babu, 2008).

Above-mentioned methods have some advantages and disadvantages. Conventional calcium

precipitation method is simple and reliable but it is expensive and unfriendly to the
environment as it consumes lime and sulphuric acid. It also produces a large quantity of
calcium sulphate sludge as solid waste. Electrodialysis and dialysis have good potential and
have advantage of simultaneous separation, but they have the problem of membrane fouling
which requires frequent cleaning of the dialyzer. Electrodialysis gives a higher extent of
propionic acid separation but with increased power and energy consumption. Adsorption or
ion exchange process requires regeneration of ion exchange resin and adjustment of feed pH
to increase the sorption efficiency requiring large amount of chemicals. In reverse osmosis,
there is a tendency to form emulsion and complexity of operation. Distillation is the well-
established and reliable technology but it has its drawback for there is formation of high
boiling esters and dimmers. The main advantage of liquid surfactant membranes over other
separation techniques is the large surface area available for mass transfer, which results in a
fast rate of separation. Several drawbacks of these membranes are hindering implementation,
complexity of operation and swelling in liquid surfactant membrane (Wasewar et al. 2004).
REACTIVE EXTRACTION: EXTRACTIVE FERMENTATION
Reactive extraction with a specified extractant giving a higher distribution coefficient has
been proposed as a promising technique for the recovery of carboxylic and hydroxyl-
carboxylic acids. This method is advantageous for alcohol and organic fermentations. Some
of the advantages are increased reactor productivity, ease in reactor pH control without
requiring base addition, and use of a high-concentration substrate as the process feed to
reduce process wastes and production costs. This method may also allow the process to
produce and recover the fermentation products in one continuous step and reduce the
downstream processing load and the recovery costs.

Conventional solvents such as alcohols, ketones, ethers, and aliphatic hydrocarbons are not
efficient when applied to dilute, carboxylic acid solutions because of the low aqueous activity
of carboxylic acids resulting in low distribution coefficients as shown in Table 2.
Organophosphoric derivatives such as tri-n-octyl phosphine oxide (TOPO), tri-n-butyl
phosphate (TBP) and long-chain, aliphatic amines such as lauryl-trialkylmethylamine
(Amberlite LA-2), tri-n-octylamine, tri-iso-octylamine, tri-n-(octyl-decyl)-amine (Alamine
336) and quaternary alkylammonium salt (Aliquat 336) are effective extractants for separation
of carboxylic acids from dilute aqueous solution (Kumar et al, 2008 and Kertes and King,
1986). Generally, the amine extractants are dissolved in a diluent, an organic solvent that
dilutes the extractant. It controls the viscosity and density of the solvent phase. Reactive
extraction strongly depends on various parameters such as the distribution coefficient, degree
of extraction, loading ratio, complexation equilibrium constant, types of complexes (1:1, 2:1,
etc.), rate constant of carboxylic acid-extractant reaction, properties of the solvent (extractant
and diluent), type of solvent, temperature, pH and acid concentration.
Table 2. Distribution coefficients (KD) of carboxylic acids between the water and organic
solvents at 25 oC
Carboxylic Acid Solvent KD
Propionic acid n-Hexane 0.005
Cyclohexane 0.006
Benzene 0.043
Toluene 0.034
Xylene 0.03
Carbon tetrachloride 0.015
Chloroform 0.11
Nitrobenzene 0.16
Diethyl ether 1.75
Diisopropyl ether 0.80
Methylisobutyl ketone 2.15
Cyclohexanone 3.30
n-Butanol 3.20
n-Pentanol 2.95
Lactic acid Diethyl ether 0.10
Diisopropyl ether 0.04
Methylisobutyl ketone 0.14
n-Octanol 0.32
Succinic acid Diethyl ether 1.50
Methylisobutyl ketone 0.19
n-Butanol 1.20
Fumaric acid Diethyl ether 1.50
Methylisobutyl ketone 1.40
Maleic acid Diethyl ether 0.15
Methylisobutyl ketone 0.21
Itaconic acid Diethyl ether 0.35
Methylisobutyl ketone 0.55

Cascaval and Galaction (2004) reported the reactive extraction of mono-, di- and
polycarboxylic acids using Amberlite LA-2 dissolved in diluents. The design of an amine
extraction process requires equilibrium data for the acid-amine (solvent) system used.
Equilibria for propionic acid extraction using trioctyl amine in various solvents such as
hexanol, butyl acetate, and petroleum ether have been determined by Wasewar and Pangarkar
(2006). Kumar et al. (2006) studied the recovery of propionic acid using reactive extraction.
Aliquat 336 is employed as extractant to recover the acid from aqueous solution using
reactive extraction. Various researchers examined the use of the organophosphine oxides
(Cyanex@921 and Cyanex@923) and organophasphates (tri-n-butyl phosphate) for the
extraction of formic, acetic and propionic acids from aqueous solutions.
Extractive fermentation has been applied in the production of propionic acid as a means of
overcoming end-product inhibition. Several extractive fermentation systems also have been
studied for organic acid production. Extractive fermentation removes the inhibitory, acidic
product from the reactor. It, therefore, provides better pH control on the reactor and results in
higher reaction rates. Also, products are present in relatively pure and concentrated forms.
Thus, savings in the downstream recovery and purification costs can be realized. However, all
prior extractive fermentation studies dealt with homo fermentative products, such as lactic,
acetic, and citric acids.

An extractive fermentation process was developed by Lewis and Yang to produce propionate
from lactose. The bacterium Propionibacterium acidipropionici was immobilized in a spiral
wound, fibrous matrix packed in the reactor. Propionic acid is the major product from lactose
fermentation, with acetic acid and carbon dioxide as byproducts. Propionic acid is a strong
inhibitor to this fermentation. Solvent selection for extractive fermentation for propionic acid
was conducted with three solvent systems by Zhong et al. (1997). Among them, the solvent
containing 2-octanol exhibited the highest partition coefficient in acid extraction, but it was
also toxic to propioni bacteria. An extractive fermentation process using an amine extractant
and a hollow-fiber membrane extractor to selectively remove propionic acid from the
fermentation broth was developed to produce propionate from lactose (Jin and Yang 1998).
Compared to the conventional batch fermentation, the extractive fermentation had a much
higher productivity (1 g/(l.h) or 5-fold increase), higher propionate yield (up to 0.66 g/g or
more than 20% increase), higher final product concentration (75 g/L or higher), and higher
product purity (90%). The improved fermentation performance can be attributed to the
reduced product inhibition and a possible metabolic pathway shift to favor more propionic but
less acetic and succinic acid production.

4. CONCLUSION
The growing demand of propionic acid draws attention to the improvement of a conventional
process for its production. Commercial production is mainly carried out by chemical synthesis
from petroleum feedstocks, but fermentation is an attractive alternative route to produce this
acid from renewable resources. It is important to have an efficient and sustainable process for
recovery of acid from the fermentation broth. Although commercial processes for acid
recovery are based on the classical method of separation and this separation followed by final
purification stages account for up to 50% of the production costs. The productivity of these
fermentation processes can be significantly increased by in-situ recovery of propionic acids
from fermentation broths by reactive extraction. Extractive fermentation process has many
advantages over the conventional propionic acid fermentation, including improved
productivity, propionate yield, final product concentration, and product purity.

REFERENCES:
Atkinson, B. and Mavituna, F. (1983). Biochemical Engineering and Biotechnology
Handbook The Nature Press-MacMillan, London.
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Available at http://en.wikipedia.org/wiki/Category:Carboxylic_acids.
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