LWT - Food Science and Technology 50 (2013) 367e370

Contents lists available at SciVerse ScienceDirect

LWT - Food Science and Technology

journal homepage: www.elsevier.com/locate/lwt

An overview of citric acid production

A.R. Angumeenal*, D. Venkappayya
SASTRA University, Thanjavur 613 401, Tamilnadu, India

a r t i c l e i n f o a b s t r a c t

Article history: This article reviews the present state of research on the conversion of low cost substrates to citric acid by
Received 14 January 2012 fermentation. Fermentation is a powerful incentive for semi-industrialized countries. There is a great
Accepted 22 May 2012 demand for citric acid due to its wide industrial applications and less toxicity. Citric acid can be produced
using less expensive substrates that are renewable too. Plant biomass is one of the desirable raw
Keywords: materials for fermentation due to its availability in abundance. Using natural sources as substrates we
Aspergillus niger
can minimize environmental problems. Always species of Aspergillus and Candida remain the choice of
Candida
candidates for the biosynthesis of citric acid. A concise gist of the various natural sources that can be used
Fermentation
Citric acid
for the production of citric acid along with the necessary fermentation conditions is presented.
Substrates Ó 2012 Elsevier Ltd. All rights reserved.

1. Introduction fruits and pineapple. Pure citric acid is colourless, readily soluble in
water with a molecular weight of 210.14 g/mol. It is biodegradable,
Biotechnology is an interdisciplinary field which is largely oriented ecofriently, economical, safe and a versatile chemical for seques-
towards the industrial applications of microorganisms for the tering, buffering, wetting, cleaning and dispersing. The acid is
conversion of waste to useful products. The number of substances mainly used in the preparation of medicinal citrates, confectionary,
excreted by microorganisms is endless. They are classified as simple soft drinks and effervescent salts. Small quantities are employed in
compounds (eg. lower alcohols, acids etc.) and complex compounds silvering and engraving and in dying and calico printing.
(eg. natural products and cellulosics) or as preliminary products and
compounds evolving from secondary metabolism. Several fermenta-
tions procedures are used for the large scale production of organic
3. Fermentation
chemicals and high energy fuels from renewable sources. Further, the
exploitation of microorganisms for the benefit of mankind was found
Microbiological conversion of organics to cellular material and
to be promising and productive. Mixed culture technology as applied
other useful products is fermentation. Despite being an old tech-
to processes, which relies upon microorganisms for proper func-
nology advances in science have kept this process in the forefront.
tioning, is in its infancy. Much is yet to be explored in terms of
The fermentation process is advantageous as it is based on
sequential transformations of substrates acted upon by a mixed
renewable sources, it facilitates use of waste for productive
culture population of microorganisms. The environmental impetus to
purpose, and useful by-products are formed. It involves very mild
move away from traditional chemical techniques towards biologically
environment friendly conditions and also consumes less energy. It
based production has revealed Aspergilli and Candida as very attractive
also faces some drawbacks some of which are:
cell factories. So an updated review of citric acid production by
Aspergillus and Candida including aspects of economy of available raw
(i) Use of large quantities of water
materials as substrates and their utility is considered very relevant.
(ii) Due to high BOD the waste requires treatment before disposal.
(iii) Infection by foreign microbes can reduce the yield and the
2. Chemistry of citric acid technology is sophisticated.

Citric acid, a tricarboxylic acid (C6H8O7$H2O) is a common
metabolite of plants and animals and is present in juice of citrus
* Corresponding author. Tel.: þ91 4362 264101; fax: þþ91 (0) 4362 2641201.
E-mail address: angumeenal@biotech.sastra.edu (A.R. Angumeenal).
Fermentation is a complex process because several reactions are
integrated into one single bio-reaction for which operating condi-
tions have to be precisely tailored and controlled carefully. Varia-
tions in results often occur even under apparently similar operating
conditions. All the commercial citric acid producers prefer to rely
upon their own experience rather than the so called proven facts.

0023-6438/$ e see front matter Ó 2012 Elsevier Ltd. All rights reserved.
doi:10.1016/j.lwt.2012.05.016
368 A.R. Angumeenal, D. Venkappayya / LWT - Food Science and Technology 50 (2013) 367e370
3.1. Types of fermentation
The development of the processes for citric acid fermentation
can be divided into three phases. In the first phase citric acid
production was confined to species of Penicilium and Aspergillus
using stationary or surface culture conditions. The beginning of the
second phase consisted of the development of submerged
fermentation processes for citric acid production using Aspergillus.
The third stage, which is of recent origin, involves the development
of solid state culture, continuous culture and multistage fermen-
tation techniques for citric acid production.
3.2. Biochemistry of citric acid
The products of carbohydrate metabolism form the building
blocks of many aerobic and anaerobic microorganisms. The main
intermediate of carbohydrate metabolism is citric acid. In certain
microorganisms such as Aspergillus niger, under specific environ-
mental conditions, citric acid is produced as an over flow product
due to the faulty operation of the tricarboxylic acid cycle (TCA). TCA
is an intermediate cycle involving the terminal steps in the
conversion of carbohydrates, proteins and fats to carbon dioxide
and water with concomitant release of energy for the growth,
movement, luminescence etc. Studies on the enzyme content of
A. niger in relation to citric acid accumulation have indicated the
vital role played by the TCA cycle in fermentation. Biological
formation of citric acid is purely enzymatic. The merit of citric acid
fermentation depends on the regulation of the synthesis of each
one of the enzymes (involved in the TCA cycle) and its activity is
under various control mechanisms such as the cofactors associated
with the enzymes. Metal ions form a part of cofactors and in turn
controlling trace element concentration can regulate the enzyme
activity. Under suitable environmental conditions different species
of Candida can also produce citric acid.
3.3. Microbial strains
Starting from fungi to yeast, a few bacteria are also suitable for
citric acid production. The fermentation capabilities of the selected
strains are sometimes improved using mutagens (Pandey, Soccol,
Rodriguez-Leon, & Nigem, 2001: pp. 113e126; Soccol,
Vandenberghe, Rodrigues, & Pandey, 2006; Vendenberghe et al.,
1999). Mutation is a process by which the characteristics of a strain
are improved physiologically and morphologically which results in
improving the process kinetics. Under such conditions membrane
permeability of the microbe is also altered. Studies on branch
formation in filamentous fungi have shown that mutations with
various chemicals cause alterations in the cell wall composition and
changes in physical conditions which cause a marked effect on the
frequency of branching.
A. niger is a common black mold which can act as a citric acid
producing cell factory (Haq, Khurshid, Ashraf, Qadeer & Rajoka,
2001). Other species of Aspergillus namely, Aspergillus aculeatus,
Aspergillus awamori, Aspergillus carbonaries, Aspergillus wenti,
Aspergillus foetidus (Papagianni, 2007) were also found to produce
citric acid in appreciable amounts. Among the yeast Saccaromicopsis
lipolytica (Rane & Sims, 1993; Wojtatowics, Marchin & Ericksen,
1993), Candida tropicalis (Kapelli, Muller, & Fiechter, 1978), Candida
oleophils, Candida guilliermondi (Angumeenal, Kamalakannan,
Prabhu & Venkappayya, 2003a, 2003b), Candida parapsilosis (Omar
& Rehm, 1980), C. citroformans (Uchio, Maeyashiki, Kikuchi, &
Hirose, 1975) were good citric acid producing candidates. Bacterial
species were also used earlier for citric acid fermentation. However,
among all A.niger remains the best choice for citric acid production
due to its ease of handling, its ability to ferment a variety of raw
materials and yield high yield (Vandenberghe, Soccol, Pandey, &
Lebeault, 1999). The production of citric acid also depends on an
appropriate strain, along with aeration, carbon source, nitrogen and
phosphate, pH and trace elements. The morphology of the producer
microorganism also plays a role in controlling and augmenting citric
acid formation (Belen Max, Salgado, Rodriguez & Dominguez, 2010).
An undisturbed metabolic flow of carbohydrates through
glycolysis is required to get a high citric acid accumulation by
A. niger. Initial intracellular pH in spores should be as high as
possible if a high rate of metabolic flow through hydrolysis is
desired for high yields of citric acid. But the pH of the culture may
change depending upon microbial metabolic activities. Nature of
substrate and production technique has influence on pH. A pH of
2.0 was optimum for molds to produce citric acid (Srivatsava, & De,
1980), whereas a pH in the range of 5e6 is favoured with molasses.
When working with yeasts, initial pH values of above 5 are
required. It is reported that below pH 5, citric acid production
decreases and accumulation of other products like erythritol, ara-
bitol and mannitol is favoured. Transport of citrate across cell
membrane is also affected (Anastasisdis & Rehm, 2005; Karasu,
Yalcin, Bozdemir & Ozhas, 2010).
The optimum temperature for citric acid formation will differ
based on strain and medium composition (Shuler, Kargi, 2002).
Hence it is necessary to optimize and determine the appropriate
favourable temperature before scaling up the process. However, the
optimum temperature reported for yeast lies between 22 and 35  C.
A.niger and other fungi require an optimum temperature of
25e30  C (Corolla & Kennedy, 2001). An increase in temperature
beyond 30  C has been found to increase oxalic acid accumulation
irrespective of the fermentation conditions. Therefore, mainte-
nance of favoured temperature is very much essential for effective
citric acid biosynthesis.
3.4. Medium composition
Fermentation of a substrate to citric acid is directly related to the
quality and quantity of the sugar source. The nature of the substrate
which in turn depends on the carbon source will also have
a marked influence on the metabolic activity of the microbial
strains. Among the synthetic substrates tried so far, sucrose was the
best candidate followed by glucose, fructose and galactose
(Vandenberghe et al., 1999). To make citric acid production cost-
effective nowadays a lot of plant and fruit sources rich in sugar
content are used as substrates. They happen to be renewable
sources making the fermentation process more economical.
Molasses is the effluent got from sugar industry and is the non
crystallisable residue remaining after sucrose isolation. It is reported
to be in sugar and hence a good substrate for citric acid fermentation
(Adham, 2002; Cevrimli, Kariptas, & Ciftci, 2009; El Aasar, 2006).
However, while using such substrates suitable treatment proce-
dures are needed to make them suitable for fermentation.
Some of the natural sources available may contain nitrogen and
phosphate and so their supplementation may not be required. But
their presence is required at optimum levels. Otherwise citrate
formation will be affected. Nitrogen can be supplied through KNO3
or NH4NO3 and phosphate by adding K2HPO4 or KH2PO4 in limited
quantities.
3.5. Metal ions
Fungi and yeast require trace quantities of metal ions (hence
called as trace elements) for their growth. Presence of magnesium
and zinc are essential for the microbial growth (Angumeenal,
Kamlakanna, Prabhu & Venkappayya, 2002). The metal ions are
expected to cause a mutational change in the species and also
 A.R. Angumeenal, D. Venkappayya / LWT - Food Science and Technology 50 (2013) 367e370
a change in the metabolic role in which the microorganism is
involved. The mechanism by which the microelements play a part
in fungal nutrition is obscure though a few of them form a part of
several enzymatic systems or a constituent of certain pigments.
Along with nitrates, phosphates and sulphates A. niger can utilize
certain minerals as nutrients during the biosynthesis of citric acid.
Thus in a given microbial system several mechanisms of uptake
may operate simultaneously and or/sequentially. Metal ions like
cadmium, chromium, lead and molybdenum were found to be good
stimulants for citric acid fermentation using A.niger in a glucose
containing medium. The intermediates of the microbial metabo-
lism in such a condition were found to interact with the cell wall
components like the lipid bilayer, protease and chitin e glucan. The
secondary metabolites in the fermentation medium are to be
analysed to explain the cell activity in more detail. Growth kinetics
of heavy metal adapted A.niger during citric acid accumulation was
evaluated as a part of the fermentation study undertaken. Among
the metal ions added to the medium during fermentation the order
of growth yield is found to be Mo > Cr > Pb > Cd and the citric acid
production was high with cadmium as stimulant (Angumeenal &
Venkappayya, 2004).
3.6. Substrates
Molasses for long considered as a waste product of the sugar
industry is now termed as a by e product due to its low price
compared to other sugar sources and the presence of minerals,
organic and inorganic compounds. Molasses is used in the
production of alcohol, organic acid and single cell proteins.
The organic and inorganic components present in molasses may
inhibit the fermentation process and hence need to be treated by
suitable methods to make it suitable for citric acid production. Some
of the commonly followed procedures include treatment with
ferrocyanide (Baby & Saad, 1996), sulphuric acid, tricalcium phos-
phate, tricalcium phosphate with HCl, tricalcium phosphate with
HCl followed by sephadex fractionation (Kundu, Panda, Majumdar,
Guha, & Bandyopadhyay, 1984). Molasses became more efficient
by treatment with ammonium oxalate followed by treatment with
diammonium phosphate. Molasses treated by this method was
found to serve as a better substrate in producing citric acid
compared to other methods commonly practiced (Angumeenal &
Venkappayya, 2005). The above molasses medium on supplemen-
tation with selective metal ions as stimulants made citric acid
fermentation more successful.
Agroeindustrial wastes are frequently used as substrates in
fermentation as an inexpensive source. Apple pomace (Hang &
Woodams, 1984), carob pod (Roukas, 1998), carrot waste (Garg &
Hang, 1995), coffee husk (Shankaran & Lonsane, 1994), corn cobs
(Hang & Woodams, 1998), grape pomace (Hang &. Woodams, 1985),
kiwi fruit peel (Hang &. Woodams, 1987), kumara (Lu, Brooks &.
Maddox, 1997), orange waste (Aravantinos- Zafiris, Tzia, Orea
poulou, & Thomopoulos, 1994), date syrup (Roukas & Kotzekidou,
1997, Moataza, 2006), pineapple waste (Tran l.i.Sly 1998), banana
extract (Sassi, Ruggeri, Specchia, & Gianetto, 1991) potato chips
waste (Sanat, & Korish, 2007), and pumpkin were tried successfully
as substrates for citric acid formation. Pumpkin, either as a single or
a mixed substrate with molasses is known to produce good quan-
tities of citric acid (Majumdar, Khalid, Munshi, Alam, & Or-Rashidet,
2010).
A waste from jackfruit was found to be a good and economical
substrate for citric acid fermentation. Artrcocarpus heterophyllus
(Jack fruit) is a large tree grown in tropical countries and is one of the
common fruits in South India. The fruiting perianths (bulbs), seeds
and rind constitute 29, 12 and 59% of the ripe fruit, respectively. The
rind portion includes the carpel fibre which holds the fruity portion
369
intact. Chemical analysis of this carpel fibre indicates the presence of
sugars and minerals and hence was used as a substrate for citric acid
production. It was treated chemically to extract the reducing sugar
present. Batch fermentation using A. niger was followed and the
results indicate that jackfruit carpel fibre can serve as a substrate for
citric acid production (Angumeenal & Venkappayya, 2005a, 2005b,
2005c). When this substrate is completely analysed and the limiting
substances identified, steps can be taken to remove them and make
it a more efficient substrate for citric acid fermentation.
Tuber crops belonging to the family Araceae namely Colocassia
antiquorum, Aponogetannatans and Amorphophallus campanulatus
are cultivated in large quantities for their edible portion. These
tubers were chemically treated and utilized as substrates for
citrate production by fermentation using A. niger by batch
fermentation. Their fermentation capabilities were improved by
adding trace elements (cadmium, molybdenum, chromium and
lead) in optimum quantities (Angumeenal et al., 2003a, 2003b).
While using A. campanulatus as a substrate along with citric acid,
succinic acid was also produced in high amounts. In fact, succinic
acid produced was higher than citric acid. This is due to the
increased activity of aconitase in the later stages of fermentation.
Hence this substrate can be further explored for succinic acid
production using some growth promoters. The potential of A.
campanulatus in producing citric acid was enhanced by the addi-
tion of metal ions.
3.7. Co - culture studies
Sometimes when two competitors are present the bioprocess
becomes more successful. Problems may also arise where one
strain can behave antagonistic to the other. Initial testing methods
should always be performed to confirm the coexistence of the
strains in the culture medium. Presence of cadmium, lead, molyb-
denum and chromium in a glucose containing medium can produce
citric acid in good quantities and the growth kinetics of such
a heavy metal adopted microbial strain was analysed (.Angumeenal
& Venkappayya, 2004) Co-culture fermentation was experimented
with glucose as substrate using A. niger and Candida guilliermondii
in the presence of trace elements. For comparison single culture
fermentation was also done with A. niger in the presence of the
same trace elements under identical experimental conditions.
Results obtained reveal that mixed culture fermentation with
A. niger and C. guilliermondii can produce citric acid in higher
quantities than with single culture, namely A. niger (Angumeenal
et al., 2003a, 2003b) The metabolic changes in A. niger remain
unaffected while performing co-culture fermentation as inferred
from the protein and lipid estimations study.
4. Conclusion
Due to the wide application of citric acid, its production by
fermentation continues to be of interest for extensive study
(Berovic & Legisa, 2007; Cevrimli, Yasar, & Kariptas 2010;
Darouneh, Alavi, Vosoughi, Arjmand, Seifkordi & Rajabi, 2009;
Dhillon, Brar, Verma, & Tyagi, 2011a, 2011b; Hanapi, Alam, &
Karim, 2011; Papagianni, 2007). Over a period of years a lot of
substrates were introduced for citric acid fermentation in pilot
scale so as to enable the industry to scale it up and increase the
production to meet the demand for citric acid. At laboratory level
scientists try introducing new substrates and methods and
confirm their potential with respect to various aspects. The
substrates from economical sources can of course reduce the cost
of production but in terms of limiting substances in them and
their removal, it needs extensive attention to make the process
more successful. It is now realized that conversion of industrial
370 A.R. Angumeenal, D. Venkappayya / LWT - Food Science and Technology 50 (2013) 367e370
waste with the microorganisms to value added products is prof-
itable provided the process control strategies are carefully moni-
tored and controlled.
References
Adham, N. Z. (2002). Attempts at improving citric acid fermentation by Aspergillus
niger in beet-molasses medium. Bioresource Technology, 284, 97e100.
Anastasisdis, S., & Rehm, H. J. (2005). Continuous production of citric acid secretion
by a high specific pH dependent active transport system in yeast Candida
oleophila, ATCC 20177. Electronic Journal of Biotechnology, 8, 26e42.
Angumeenal, A. R., & Venkappayya., D. (2004). Growth kinetics of heavy metal
adapted Aspergillus niger during citric acid biosynthesis. Journal of Scientific and
Industrial Research, 63, 610e613.
Angumeenal, A. R., & Venkappayya., D. (2005a). Effect of transition metal ions on
the metabolism of Aspergillus niger in the production of citric acid with
molasses as substrate. Journal of Scientific and Industrial Research, 64, 125e128.
Angumeenal, A. R., & Venkappayya., D. (2005b). Artrocarpus heterophyllus e
a potential substrate for citric acid biosyntheis using Aspergillus niger. LWT e
Food Science and Technology, 38, 89e93.
Angumeenal, A. R., & Venkappayya., D. (2005c). Bioconversion of Amorphophallus
campanulatus to citric acid by Aspergillus niger e effect of metal ions on
fermentation, modeling studies and correlarion of theoretical and experimental
parameters. Indian Journal of Biotechnology, 4, 246e250.
Angumeenal, A. R., Kamlakannan, P., Prabhu, H. J., & Venkappayya, D. (2002). Effect
of transition metal cations and anions on the production of citric acid by
Aspergillus niger. Journal of the Indian Chemical Society, 9, 508e512.
Angumeenal, A. R., Kamalakannan, P., Prabhu, H. J., & Venkappayya, D. (2003a).
Effect of transition metal cations on the production of citric acid using mixed
cultures of Aspergillus niger& Candida guilliermondii. Journal of the Indian
Chemical Society, 80, 903e906.
Angumeenal, A. R., Kamalakannan, P., Prabhu, H. J., & Venkappayya, D. (2003b).
Bioconversion of Colocassia antiquorum and Aponogetannatans to citric acid by
Aspergillus niger e effect of metal ions and kinetics. Journal of Scientific and
Industrial Research, 62, 447e452.
Aravantinos- Zafiris, G., Tzia, C., Orea poulou, V., & Thomopoulos, C. D. (1994).
Fermentation of orange processing waste for citric acid production. Journal of
Science of Food and Agriculture, 65, 117e120.
Baby, E. A., & Saad, M. (1996). Production of citric acid by a heavy metal adapted
Aspergillus niger NRRL 595. African Journal of Mycology and Biotechnology, 4,
59e69.
Belen, M., Salgado, J. M., Rodriguez, N., & Dominguez, S. C. (2010). Biotechnological
production of citric acid. Brazilian Journal of Microbiology, 41, 802e875.
Berovic, M., & Legisa, M. (2007). Citric acid production. Biotechnology Annual
Reviews, 13, 303e343.
Cevrimli, B. S., Kariptas, E., & Ciftci, H. (2009). Effects of fermentation conditions on
citric acid production from beet molasses by Aspergillus niger. Asian Journal of
Chemistry, 21, 3211e3218.
Cevrimli, B. S., Yasar, A., & Kariptas, E. (2010). Effect of o-cresol, resorcinol and
hydroquinone on citric acid productivity by Aspergillus niger in stirred
fermentor. Asian Journal of Chemistry, 22, 699e704.
Corolla, A., & Kennedy, K. J. (2001). Optimisation of citric acid production from
Candida lipolytica Y095 using n- paraffin. Journal of Biotechnology, 89, 27e40.
Darouneh, E., Alavi, A., Vosoughi, M., Arjmand, M., Seifkordi, A., & Rajabi, R. (2009).
Citric acid production: surface culture versus submerged culture. African Journal
of Microbiology Research, 3, 541e545.
Dhillon, G. S., Brar, S. K., Verma, M., & Tyagi, R. D. (2011a). Recent advances in citric
acid bio-production and Recovery. Food and Bioprocess Technology, 4, 505e529.
Dhillon, G. S., Brar, S. K., Verma, M., & Tyagi, R. D. (2011b). Apple pomace ultrafil-
tration sludge - a novel substrate for fungal bio-production of citric acid:
optimisation studies. Food Chemistry, 128, 864e871.
El Aasar, S. A. (2006). Submerged fermentation of cheese whey and molasses for
citric acid production by Aspergillus niger. International Journal of Agriculture and
Biology, 4, 463e467.
Garg, N., & Hang, Y. D. (1995). Microbial production of organic acids from carrot
processing waste. Journal of Food Science and Technology, 32, 119e121.
Hanapi, N. H. M., Alam, M. Z., & Karim, M. I. A. (2011). Sewage sludge as a renewable
resource for citric acid production: optimization of supplementary nutrients.
Journal of Appled Science, 11, 2163e2170.
Hang, Y. D., & Woodams, E. E. (1984). Apple pomace; a potential substrate for citric
acid production by Aspergillus niger. Biotechnology Letters, 6, 763e764.
Hang, Y. D., & Woodams, E. E. (1985). Grape pomace; a novel substrate for microbial
production of citric acid. Biotechnology Letters, 7, 253e254.
Hang, Y. D., & Woodams, E. E. (1987). Microbial production of citric acid by solid
state fermentation of kiwi fruit peel. Journal of Food Science, 52, 226e227.
Hang, Y. D., & Woodams, E. E. (1998). Production of citric acid from corn cobs by
Aspergillus niger. Bioresoure Technology, 6, 251e253.
Haq, I. U., Khurshid, S., Ashraf, A. H., Qadeer, M. A., & Rajoka, M. L. (2001). Mutation
of Aspergillus niger for hyperproduction of citric acid from black strap molasses.
World Journal of Microbiology and Biotechnology, 17, 35e37.
Kapelli, O., Muller, M., & Fiechter, A. (1978). Chemical and structural alterations at
cell surface of Candida tropicalis induced by hydrocarbon substrate. Journal of
Bacteriology, 133, 952e958.
Karasu Yalcin, S., Bozdemir, M. T., & Ozhas. (2010). Effects of different fermentation
conditions on growth and citric acid production kinetics of two Yarrowia lip-
olytica strains. Chemical and Biochemical Engineering Quarterly, 24, 347e360.
Kundu, S., Panda, T., Majumdar, S. K., Guha, B., & Bandyopadhyay, K. K. (1984).
Pretreatment of Indian cane molasses for increased production of citric acid.
Biotechnology and Bioengineering, 26, 1114e1121.
Lu, Brooks, M. J. D., & Maddox, I. S. (1997). Citric acid production by solid state
fermentation in a packed bed reactor using Aspergillus niger. Enzyme Microbial
Technology, 21, 392e397.
Majumdar, L., Khalid, I., Munshi, M. K., Alam, K., & Or-Rashid, H. (2010). Citric acid
production by Aspergillus niger using molasses and pumpkin as substrates.
European Journal of Biological Sciences, 2, 1e8.
Moataza, M. S. (2006). Citric acid production from pretreated crude date syrup by
Aspergillus niger NRRL595. Journal of Applied Sciences Research, 2, 74e79.
Omar, S. H., & Rehm. (1980). Physiology and metabolism of alkane oxidizing and
citric acid producing strains of Candida parapsilosis. Applied Microbiology and
Biotechnology, 11, 42e49.
Pandey, A., Soccol, C. R., Rodriguez-Leon, J. A., & Nigem, P. (2001). Production of
organic acids by solid state fermentation in solid state fermentation in biotech-
nology, fundamentals and applications. New Delhi, India: AsiaTech Publishers,
Inc.
Papagianni, M. (2007). Advances in citric acid fermentation by Aspergillus niger:
biochemical aspects, membrane transport and modeling. Biotechnology
Advances, 25, 244e263.
Rane, K. D., & Sims, K. A. (1993). Production of citric acid by Candida lipolytica, Y-
1095; effect of glucose concentration on yield and productivity. Enzyme
Microbial Technology, 15, 646e651.
Roukas, T., & Kotzekidou, P. (1997). Pretreatment of date syrup to increase citric acid
production. Enzyme and Microbial Technology, 21, 273e276.
Roukas, T. (1998). Carob pod, A new substrate for citric acid production by Asper-
gillus niger. Applied Biochemistry and Biotechnology, 74, 43e53.
Sanat, E. I.-S., & Korish, M. (2007). Utilisation of potato chips waste for production of
high value products: production of citric acid by solid state fermentation. Alex
Journal of Food Science & Technology, 4, 19e26.
Sassi, G., Ruggeri, B., Specchia, V., & Gianetto, A. (1991). Citric acid production by
Aspergillus niger with banana extract. Bioresource Technology, 37, 259e269.
Shankaran, V. S., & Lonsane, B. K. (1994). Coffee husk and inexpensive substrate for
production of citric acid by Aspergillus niger in a solid state fermentation. World
Journal of Microbiology and Biotechnology, 10, 165e168.
Shuler, M. L., & Kargi, F. (2002). Bioprocess engineering, basic concepts (2nd ed.).
Prentice Hall, Inc.
Soccol, C. R., Vendenberghe, L. P. S., Rodrigues, C., & Pandey, A. (2006). New
perspectives of citric acid production and applications. Food Technology and
Biotechnology, 44, 141e149.
Srivatsava, A. S., & De, S. K. (1980). Effect of some cultural conditions on microbial
citric acid fermentation. Indian Journal of Agricultural Chemistry, 13, 39e44.
Tran, C. T., & l.i.Sly, D. A. (1998). Mitchel, selection of a strain of Aspergillus niger for
the production of citric acid from pine apple waste in solid state fermentation.
World Journal of Microbiology and Biotechnology, 14, 399e404.
Uchio, R., Maeyashiki, I., Kikuchi, K., and Hirose, Y. (1975). Citric acid production by
yeast, Jp.Kokai, 76, 63, 059.
Vandenberghe, L. P. S., Soccol, C. R., Pandey, A., & Lebeault, J. M. (1999). Review -
microbial production of citric acid. Brazilian Archives of Biology and Technology,
42, 263e276.
Wojtatowics, M., Marchin, G. L., & Ericksen, L. E. (1993). Attempts to improve strain
A-10 of Yarrow lipolytica for citric acid production from n-parrafins. Process
Biochemistry, 28, 453e460.