Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)

M Moresi, Università della Tuscia, Viterbo, Italy

E Parente, Università della Basilicata, Potenza, Italy; and Istituto di Scienze dell’Alimentazione, Avellino, Italy
Ó 2014 Elsevier Ltd. All rights reserved.

Several organic acids are used in a variety of food and nonfood
applications. Table 1 lists the main acids that are produced
commercially by chemical (C) or biotechnological (fermenta-
tion, F, or enzymatic, E) methods or extracted from wine-
making residues (L).
Citric, acetic, lactic, propionic, tartaric, fumaric, and malic
acids are among the most versatile ingredients in the food and
beverage industry because of their valuable properties, such as
solubility, hygroscopicity, acidity, buffering capacity, and
chelation (see Preservatives: Traditional Preservatives – Organic
Acids).
Citric acid accounts for around 80% of the food acidulant
usage, whereas the use of phosphoric or acetic acids is limited,
being almost exclusively utilized in cola soft drinks or in
vinegar (see Vinegar), sauces, and condiments, respectively.
Citric Acid
Citric acid (2-hydroxy-1,2,3-propanetricarboxylic acid: C6H8O7)
is widely distributed in natural raw materials (such as lime,
lemon, and raspberry) and is commercially available in the
monohydrated form (molecular mass of 210.13 Da, relative
density of 1.542 at 20
C, and heat of combustion of
1962 kJ mol1 at 25
C). It is a strong tricarboxylic acid
(TCA; its dissociation constants being K1 ¼ 7.45  104,
K2 ¼ 1.73  105, and K3 ¼ 4.02  107 at 25
C), highly
soluble in water with pleasant acid taste.
Citric acid was first isolated in 1784 by Scheele, who
precipitated it as calcium citrate by adding calcium
hydroxide (lime) to lemon juice. Before 1920, it was almost
exclusively produced in Sicily by pressing lemons: The firm
Arenella (Palermo, Italy) essentially established a monopoly
until the advent of the citric acid fermentation technique in
Belgium (Societè des Produits Organiques de Tirlemont) in
1919 and in the United States (Chas. Pfizer & Co., New
York) in 1923.
About 10 years later, about 80% of the world’s citric acid
was produced by the surface fermentation process. The
submerged fermentation process began to be applied only after
World War II.
From 1950 to 1980, citric acid was mainly used in phar-
maceutical or health products. In fact, in the early 1980s, its
two largest manufacturers were Pfizer and Miles/Bayer, both
suppliers of prescription drugs. Thereafter, as citric acid began
to be used in the food and beverage sector in industrial and
developing countries, its market size experienced significant
growth and several new manufacturers were established in
Europe and North America, as well as in China where several
small-scale fermentation units have produced citric acid from
sweet potatoes or cassava since the 1970s.
In the early 1990s, a few manufacturers gave rise to the so-
called citric acid cartel. The overcharges imposed on US buyers
was estimated in the range of $116–309 million and, on
January 29, 1997, Haarmann & Reimer Corp., a subsidiary of
Bayer AG (D), pled guilty and paid a $50 million criminal fine.
In March 1998, even Archer Daniels Midland Co. (ADM)
agreed to pay $36 million to four citric acid customers that had
opted out of the July 1997 civil class-action antitrust settle-
ment. At that time, the global citric acid capacity was about
840 000 Mg (mega grams) per year with a growth rate of 5%
per year. Afterward, the world citric industry became less
concentrated and numerous new manufactures, especially in
China, as well as Brazil, India, Indonesia, and Thailand, have
entered the market, thus making the formation of cartels less
probable.
Moreover by the early 2000s, almost all citric acid
manufacturing was globally integrated into the corn wet-milling

Table 1 Main organic acids: molecular formulas, world output, production methods, and organisms

Acidulant Chemical formula World output (metric tons) Production methods a,b Organism

Acetic acid (vinegar) C2H4O2 190 000 F 100% Acetobacter aceti

Lactic acid C3H6O3 150 000 F 100% Lactobacillus spp.
Rhizopus spp.
Propionic acid C3H6O2 130 000 C 100% Propionibacterium acidipropionici
Fumaric acid C4H4O4 12 000 C 100% Rhizopus arrhizus
Malic acid C4H6O5 10 000 C 70% –
E 30%
Tartaric acid C4H6O6 28 000 L 100% –
Itaconic acid C5H6O4 15 000 C 100% Aspergillus terreus
Citric acid C6H8O7 1 800 000 F 100% Aspergillus niger
Gluconic acid C6H12O7 87 000 F 100% Aspergillus niger

Note: Because of the lack of published data, the production figures are approximate.
a
Percentage of total production for food uses.
b
F, fermentation; C, chemical synthesis; E, enzymatic synthesis; L, leaching.

804 Encyclopedia of Food Microbiology, Volume 1 http://dx.doi.org/10.1016/B978-0-12-384730-0.00111-7

 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)
industry either by acquisition (Pfizer and Miles/Bayer were
bought by ADM and Tate & Lyle, respectively) or by new process
development (Cargill).
In the years 1987–89, US list prices for citric acid anhydrous
remained unchanged at $1.79 kg1. By late 1989, the list price
reduced to $1.65 kg1, while in the fall 1990 it was as low as
$1.39 kg1. Then, thanks to the “citric acid conspiracy” in the
years 1993–96, the citric acid cartel accomplished its main goal
of raising and keeping list price at $1.87 kg1. Then, it lowered
from $1.76 kg1 in November 1994 to $1.54 kg1 in the early
1997.
Thereafter, the severe competition resulted in selling prices
of anhydrous citric acid decreasing to $0.70–$0.80 kg1, thus
forcing the smaller manufacturers, unable to benefit from the
economy of scale, to exit the business. As a consequence, the
panorama of organic acid manufacturers has profoundly
changed over the last decade. In 2010, China approximately
accounted for more than 50% of global citric acid production
capacity, while Europe and North America covered the 19
and 24%, respectively, and as much as 65–70% of global
consumption.
Several substrates are used as fermentation substrates
depending on the local availability. Maize starch is mainly used
in the United States and China, while sugarcane or sugar beet
molasses prevail in the Brazilian and Indian or European
markets, respectively.
Cellulosic materials are currently unused in citric acid
production, even if there are projects to assess the technical
feasibility of such feedstock materials in the citric acid
industry.
From January 2008 to January 2009, export prices of
Chinese (anhydrous) citric acid oscillated in the range of US
$0.7–$0.8 kg1; thereafter, they steadily increased to reach
a peak of $1.1 kg1 in June 2011, as a direct result of the
increase in the market prices for agricultural raw materials,
particularly corn. The present economic crisis in Europe and the
United States has newly reduced the market prices of (anhy-
drous) citric acid to US $0.70$0.96 kg1 depending on the
amount ordered.
In conclusion, the global citric acid production capacity
reached almost 1.8 million metric tons (Mg) in 2010, while
it was about 1.5  106 Mg in 2005.
Organisms and Metabolic Pathways Involved
Several molds (Penicillium spp., Aspergillus niger, Aspergillus
wentii, Trichoderma viride; see Aspergillus and Penicillium and-
Talaromyces: Introduction), yeasts (Yarrowia lipolytica, Candida
guillermondii), and bacteria (Arthrobacter) produce citric acid
from a variety of substrates (glucose, sucrose, n-alkanes), but
industrial processes have been developed only for the
production of citric acid from sugars (glucose, sucrose) with
A. niger and from sugars and n-alkanes with yeasts. Industrial
strains are not freely available, but citric acid–producing
strains (A. niger, NRRL 2270, NRRL 599, ATCC 11414, ATCC
9142; Y. lipolytica ATCC 20346, ATCC 20390, NRRL Y-7576,
NRRL Y-1095) can be obtained from international culture
collections.
Metabolic pathways involved in citric acid overproduction
by A. niger are shown in Figure 1.
805
A high flux through the glycolysis, decreased activity of TCA
cycle reactions that degrade citrate, and an anaplerotic reaction
to replenish the oxaloacetate (OAA) used for the synthesis of
citrate are all essential (see Metabolic Pathways: Release of
Energy (Aerobic)). Key regulatory steps in the process include
glucose transport and phosphorylation, citric acid export from
the mithocondria and cell, phosphofructokinase, pyruvate
carboxylase (PC), citrate synthase (CS), and a-ketoglutarate
dehydrogenase (KDH).
The metabolic changes necessary for citric acid over-
production in A. niger are induced by high sugar concentration,
low pH, and manganese (Mnþ2) deficiency. Other factors (i.e.,
phosphate and nitrogen concentrations, high dissolved oxygen
(DO) concentration, trace metals), however, are important.
Very low concentrations of Mn2þ (<10 mg m3) are critical.
They result in decreased activity of the pentose phosphate
pathway and increased glycolytic flux, increased intracellular
NHþ 4 pool and turnover of nucleic acids and proteins, changes
in membrane lipid composition and cell wall composition,
and morphological changes.
Improvement of strains for citric acid production tradi-
tionally has been carried out by mutagenesis and screening.
Overexpression of proteins critical to acidogenesis (hexokinase,
glucose carrier) or inactivation of genes encoding enzymes that
produce allosteric inhibitors of hexokinase has been attempted,
but with limited success, in the additional production of citric
acid. It has been postulated that the activity of seven glycolytic
enzymes needs to be increased to obtain increased production
of citric acid. The availability of the complete genome sequence
of A. niger is likely to allow for the design of overproducing
mutants.
Citric acid overproduction in yeast is relatively insensitive to
trace metals concentration and is triggered by nutrient (N, S, P,
or Mg) limitations coupled with a high rate of glucose
utilization, which results in a high adenosine triphosphate/
adenosine monophosphate ratio and, in turn, in inactivation of
nicotinamide adenine dinucleotide (NADþ)-specific isocitrate
dehydrogenase (IDH). The main anaplerotic reactions include
the synthesis of OAA from pyruvate catalyzed by PC during
production from glucose and the glyoxylate cycle during
growth on n-alkanes. Accumulation of isocitrate (10–50% of
the citrate produced) in excess with respect to the predicted
equilibrium of aconitase probably is due to the high perme-
ability of yeast mitochondria to isocitrate compared with
citrate. Low cytoplasmic levels of citrate may be responsible for
reduced feedback inhibition of glycolysis. Improvement of
yeast for citric acid production is directed to obtain strains with
reduced isocitrate dehydrogenase and aconitase activities.
Methods of Manufacture
Citric acid production is mainly accomplished by the
submerged fermentation process, probably because of the
smaller contribution of investment and labor costs to its overall
production costs. The surface fermentation process currently
accounts for only 5–10% of the world supply. In Europe, all
surface fermentation plants have been shut down during the
past decade. Smaller amounts of citric acid (<1%) are reported
to be extracted from citrus fruits in Mexico and South America
and to be produced by the solid-state process in Japan.
806 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)
 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic) 807

Both submerged- and surface-culture fermentation pro- Table 2 Composition for the production media used in the
cesses use beet molasses or glucose syrup as the main raw laboratory- and industrial-scale production of citric acid by A. niger
material and use A. niger as the fermenting organism.
Range of
Submerged fermentation is carried out either in 150–200 m3
Component concentrations Typical values Unit
stirred-tank reactors or in 300–500 m3 (up to 1000 m3 as claimed
by some manufacturers) bubble-column reactors. The main Sucrose or glucose 125–225 180 kg m3
advantages of these techniques are improved asepsis during NH4NO3 (or other 0.5–3.5 1.5 kg m3
fermentation, automatic control of inoculation and fermentation NHþ 4 salt)
procedures, shorter fermentation times, and greater product KH2PO4 0.5–2 0.5 kg m3
yields. MgSO47H2O 0.1–2.0 0.25 kg m3
Feþ2 2–1300 <200 mg m3
In spite of the old and renewed interest in citric acid
Znþ2 0–2900 200–1500a mg m3
production by yeast grown submerged in sugar- or hydro-
Cuþ2 1–10 200 200–1500a mg m3
carbon-based media to overcome the main disadvantages of Mnþ2 0–46 <2 mg m3
traditional mold fermentation (i.e., high sensitivity to trace Initial pH 2.5–6.5 2.2 –
metals and low production rates), no yeast-based process is
a
currently known to be operating worldwide. To overcome the detrimental effects of iron and manganese on mycelium structure
and restore proper morphology.

Production Media
Citric acid is produced from media containing high concen-
trations of simple sugars (molasses or glucose syrup) (see
Fermentation (Industrial): Media for Industrial Fermenta-
tions), in which mycelium growth is restrained by nutrient
(phosphorous, manganese, iron or zinc) limitation. Their range
of composition is given in Table 2.
Nitrogen is usually added as ammonium nitrate or sulfate.
Metals are removed by pretreatments of raw materials, espe-
cially molasses, with cation-exchange resins or the addition of
potassium hexacyanoferrate (HCF). The optimal iron concen-
tration seems to depend on the fungal strain, but iron levels of
200 mg m3 were found to inhibit citrate production. The
inhibitory effect of Feþþ can be counterbalanced by the addi-
tion of copper and zinc salts during the inoculum development
or during early mycelium growth in the production medium.
Manganese concentration has to be kept as low as possible
(<10 mg m3).
Some ingredients (methanol, 3–6% w/v; corn, peanut, and
olive oils, 0.1–0.5% w/v; starch, 0.025–0.5% w/v) have been
claimed to enhance the citric acid yield.
Media sterilization is carried out batch wise at 121
C for
15–30 min at the laboratory or pilot scale or continuously
using a plate–heat exchanger unit at the industrial scale.
Fermentation Process
Inoculation is generally carried out by transferring aseptically
the stock culture maintained on agar slants on other working
slants. After w24 h incubation at 30
C, the conidia crop is
inoculated in starch-rich seed-production media to yield up to
1011 spores cm3. This culture may be directly transferred into
10–20 m3 seed fermenters to obtain a pellet-type inoculum
consisting of 1–5  105 pellets dm3 (0.1–0.2 mm in diam-
eter), which in turn is used as inoculum (5–10% v/v) for the
industrial-scale production medium.
The fungus will develop different morphological forms
(Figure 2).
The formation of a loose mycelium with long, unbranched
hyphae is to be avoided because this results in an enormous
increase in the apparent viscosity of the culture broth, thus
limiting the effective oxygen transfer rate with little or no citric

=
Figure 1 Metabolic pathways for citric acid overproduction in Aspergillus niger. Only relevant enzyme activities, substrate, products, and effectors
(, inhibitor; þ, activator) are shown. Enzymes and transport systems: INV, membrane bound invertase; GC, low-affinity glucose carrier; FC: Fructose
carrier; PP, proton pump; CC, putative citrate carrier; HK, Hexokinase; PGI, phosphoglucose isomerase; PFK1, phosphofructokinase; PFK2,
6-phosphofructo-2-kinase; ALD, aldolase; PK, pyruvate kinase; PC, pyruvate carboxylase; MDH, malate dehydrogenase; PT, pyruvate transport system;
TCC, citrate transport system; PDH, pyruvate dehydrogenase; CS, citrate synthase; ACT, aconitase; IDH, isocitrate dehydrogenase; KDH, a-ketoglutarate
dehydrogenase; AOX, alternative oxidase system. Substrates and products: glu, glucose; glu6P, glucose-6-phosphate; fru6P, fructose-6-phosphate;
fru1,6 dP, fructose-1,6-bisphosphate; fru2,6 dP, fructose-2,6-bisphosphate; gly, glycerol; dhp, dihydroxiaceton phosphate; pep, phosphoenolpyruvate;
pyr, pyruvate; oaa, oxaloacetate; mal, malate; cit, citrate; acCoA, acetyl-coenzyme A; aco, cis-aconitate; ica, isocitrate; a-kg, a-ketoglutarate; sucCoA,
succinyl-coenzyme A; tre, trehalose.
The most important steps in controlling glycolytic flux are glucose transport (simple diffusion is the main mechanism at high sugar concentrations,
although A. niger has both low-affinity and high-affinity carriers for glucose) and hexokinase (HK) activity, which initially is inhibited by trehalose-6-
phosphate. PFK1 is feedback inhibited by citrate, but the inhibition is counteracted by the presence of high levels of NHþ4 and by fructose-2,6-biphosphate
(FBP). A phosphorylated fragment of PFK1, which is insensitive to citrate inhibition, may be responsible for acidogenesis. PFK2 activity is increased at high
substrate concentration: its product, FBP lowers the Michaelis–Menten constant (Km) of PFK1, counteracts citrate inhibition, and inhibits gluconeogenesis,
thus increasing carbon flux through glycolysis during acidogenesis. CS activity in A. niger is regulated by the level of OAA, which is produced in the
anaplerotic reaction catalyzed by PC. Malate is produced from OAA by cytosolic MDH and acts as a counterion for citrate efflux from the mitochondrion,
where it is oxidized back to OAA. Low activity of NADPþ-specific IDH and KDH are a consequence of the effective removal of citrate, which has a higher
affinity than ACT.
A salicylhydroxamic acid–sensitive, alternate oxidase system is used during acidogenesis to reoxidize the NADH produced during glycolysis.
Malfunction of the normal respiratory chain is due to diminished activity of NADH ubiquinone reductase and other respiratory chain enzymes.
808 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)

Figure 2 Pellet morphology of A. niger NRRL 2270 during citric production in a laboratory 2 dm3 stirred fermenter: (a) young pellet (100); (b) stubby,
bulbous hyphae with frequent branching, which are characteristic of citric acid production (400); and (c) degenerating pellet with pointed unbranched
hyphae protruding from the pellet core (100).

acid production. On the contrary, small spherical, dense
pellets (Figure 2(a)) with short stubby hyphae (Figure 2(b))
are generally regarded as the best morphological form
for optimal citrate yields. Frequent observation of pellet
morphology during this early stage of the fermentation using
a microscope allows hyphae proliferation to be controlled by
the addition, in case of adverse development (Figure 2(c)), of
appropriate amounts of inhibiting compounds, such as HCF
or zinc and copper sulfates. Mycelial clumps (whose structure
is less compact than pellets) also may develop under high
agitation speed.
Fermentation is exothermic and temperature has to be kept
in the range 28–35
C. Assuming that the overall heat transfer
coefficient and effective temperature difference between the
fermenting medium and cooling water are of the order of
500 kJ m2 h1 and 5
C, respectively, the heat transfer surface
required to keep the fermentation temperature constant would
be w3.2 m2 per m3 of fermentation medium.
Low pH and high DO concentration are essential for citric
acid production. Initial decrease of pH is due to ammonium
uptake. Extreme pH values (<1.6) limit productivity, and the
addition of alkali (NH3) is used to control pH at 2.2–2.6 once
production of citric acid has started. The typical industrial-scale
productivities of 1–1.5 kg m3 h1 result in microbial oxygen
demand rates of 0.3–0.5 kg O2 m3 h1, that are met by
sparging 0.1–0.4 volumes of air per medium volume per
minute (vvm) at pressures at the sparger section of the
fermenter not less than 0.3–0.4 MPa and at the tank top,
ranging from 0.25–0.35 MPa to 0.12–0.15 MPa, depending on
the (stirred or air-lift) fermenter type used. Foaming is
controlled by adding food-grade antifoam agents.
Temporary interruption to the air supply during fermenta-
tion does not seem to affect the performance of the culture on
the condition that the DO level is greater than 20% of the
saturation value. DO values of about 0 for as long as 85 min,
followed by restoration of the air supply, do not inhibit
permanently mycelial growth and citrate production, but they
do reduce the product yield coefficient up to 20%.
Figure 3 shows the evolution of a typical batch citric acid
fermentation in glucose-based media by A. niger NRRL 2270 in
2-dm3 stirred fermenter and by a mutant strain of A. niger in
400 m3 bubble-column fermenter.
Two distinct phases are evident: during the primary growth
phase (trophophase), no acid production occurs; during the
second growth phase (idiophase), acid production by almost
nongrowing cells is observed.
 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)
Figure 3 Time course of a typical batch citric acid fermentation from
glucose-based media by A. niger NRRL 2270 in a 2 dm3 stirred fermenter
(closed symbols) and by an industrial strain in a 400 m3 bubble-column
fermenter (open symbols): Concentrations of mycelial biomass (X: A, >),
glucose (S: l, B), citric acid (P: n, ,), and ammoniac nitrogen (N: D)
versus time (t). The industrial-scale trial was gently provided by Dr A.
Trunfio c/o Palcitric SpA, Calitri, Italy, and the laboratory-scale trial was
performed by the authors at the University of Basilicata (Potenza, Italy).
Table 3 shows the simplified overall stoichiometric reac-
tions occurring during the trophophase and idiophase of the
fermentation examined. In particular, it was assumed that
during the trophophase the microorganism (represented by
a raw formula based on elemental analysis: CHnOpNq) repli-
cates itself at the expanses of a generic carbon source in the
presence of ammonia as the only nitrogen source; during the
idiophase, it undergoes further growth while decreasing
progressively its intracellular nitrogen content and excreting
citric acid in a medium practically devoid of any nitrogen
source (Figure 3).
Assuming that no carbon atom of sugar is converted into
biomass, carbon dioxide, or other by-products as shown by the
reaction in Eqn [2] (i.e., when y and d are equal to 0), the
theoretical molar yield (z) of citric acid would be one or two if
glucose or sucrose is used. This would be equivalent to 1.17 (or
1.23) kg of citric acid monohydrate per kilogram of glucose
(or sucrose) consumed. In practice, the industrial yields range
from 57 to 81% of this theoretical value, with the smaller
figure generally being associated with the surface fermentation
technique.
The citric acid fermentation may be mathematically
described by means of the set of kinetic equations shown
in Table 3.
In accordance with the Herbert–Pirt maintenance concept,
both product formation (rP) and substrate consumption (rS)
rates may be linearly related to cell growth rate (rx) and cell
concentration (X). In this way, the well-known Luedeking–
Piret kinetics for product formation has to be regarded as
a special case: In fact, the first term in Eqn [10] may be
described as the product formation rate in association with the
mycelial growth rate, whereas the second term may be regarded
as the nongrowth-associated product formation rate. In both of
the fermentation trials shown in Figure 3, citric acid fermen-
tation may be classified to be of the mixed-growth-associated
product formation type.
809
A microscopic description of this fermentation using
A. niger pellets also has to account for oxygen diffusion
phenomena from the bulk of the fermenting medium to the
pellet surface and through the porous structure of the pellet
itself. The low effective diffusivity of oxygen within the pellet
limits mycelial activity to a peripherical spherical shell only,
with the oxygen penetration depth ranging from 110 to 300 mm
in 2 mm pellets.
Recovery and Purification Processes
Citric acid may be recovered from the broths resulting from
either the surface- or submerged-culture fermentations, using
almost the same three methods – namely, direct crystallization
upon concentration of the filtered liquor, precipitation as
calcium citrate tetrahydrate, or liquid extraction (see Fermenta-
tion (Industrial): Recovery of Metabolites). Direct crystalliza-
tion cannot be applied unless refined raw materials, such as
sucrose syrups or crystals, are used. Liquid extraction is used by
Tate & Lyle Co. (formerly Haarmann & Reimer Co., a subsidiary
of Bayer Co.) in the Dayton (OH, USA) and Elkhart (IN, USA)
plants. The precipitation process is used by the great majority of
world citric acid manufacturers, including ADM in the United
States.
A simplified process flowsheet of this method is shown
in Figure 4.
Mycelia and suspended particles are separated by
continuous belt filters under vacuum. Citric acid is precipi-
tated as calcium citrate by the addition of lime to the filtrate.
Liming temperature is critical. Amorphous tricalcium citrate
tetrahydrate generally is obtained at w70
C, while crystal-
line dicalcium acid citrate is obtained at 90
C. No removal
of oxalic acid is needed if the submerged-culture fermenta-
tion is used.
The residual citrate in the filtrate is precipitated as tricalcium
citrate by further addition of lime to set the pH to 5.8. The
crystals are recovered using another continuous belt filter and
then recycled back to the liming step, while the filtrate has to be
disposed.
Precipitation of dicalcium acid citrate results in one-third
less consumption of lime and consequently of sulfuric acid for
the subsequent regeneration of citric acid, in greater filterability
and washability because of its crystalline structure, but 10–25%
of the expected product yield is needed as seed. The precipitate
is washed to remove the impurities adsorbed onto it (i.e.,
residual sugars and contaminants from the raw carbon source
and soluble proteins from the autolysis of the fungus). The
washed crystals and 98% w/w sulfuric acid are simultaneously,
but separately, fed to a mixer containing a 40% citric acid
solution at pH 0.5–0.6, to free the citric acid with the formation
of a precipitate of calcium sulfate dihydrate (gypsum). Final
refining of the filtrate is performed by decolorization on acti-
vated carbon and removal of residual calcium sulfate and iron
and nickel salts on strong cation exchange and weak anion
exchange (demineralization step). The resulting solution
(250–280 kg m3 of citric acid anhydrous) is concentrated
using multiple-effect evaporators to about 700 kg m3, before
feeding a vacuum crystallizer operating at temperatures below
(35
C) or above (62
C) the transition temperature (36.6
C)
between the monohydrate and anhydrous forms, depending
810 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)

Table 3 Citric acid fermentation: overall stoichiometric reactions, kinetic equations, and instantaneous concentrations of
mycelia, product, sugar, and nitrogen sources

Equation or reaction

Trophophase reaction C6 H12 O6 þ a NH3 þ b O2 / y CHn Op Nq þ d CO2 þ e H2 O ½1

glucose mycelium

Idiophase reaction C6 H12 O6 þ b O2 / y CHn Op Nq þ z C6 H8 O7 þ d CO2 þ e H2 O ½2

glucose mycelium citric acid

Kinetic equations dX
rX ¼ ¼ mX X ½3
dt


dN dX
rN ¼ ¼ YN=X for N  Nlim ½4
dt dt

dN
rN ¼ ¼ 0 for N < Nlim ½5
dt

m ¼ 0 for t  to ½6

m ¼ mM for t  tlim ½7


X
m ¼ mM 1  for t > tlim ½8
XM

dP
rP ¼ ¼ 0 for t  tlim ½9
dt


dP dX
rP ¼ ¼ YP=X þ mP X for t > tlim ½10
dt dt


dS dX
rS ¼  ¼ YS=X þ mS X ½11
dt dt

Integral solutions of the X ¼ X0 for t  to ½12

differential kinetic equations
ðt t0 Þ
X ¼ X0 e mM for t  tlim ½13

XM
X ¼
 for t > tlim ½14
XM
1þ  1 e mM ðt tlim Þ
X0

N ¼ N0 for t < t0 ½15

N ¼ N0  YN=X ðX  X0 Þ for t  tlim ½16

N ¼ Nlim for t > tlim ½17

P ¼ P0 þ mP Aðt Þ þ YP=X ðX  X0 Þ ½18

S ¼ S0  ½mS Aðt Þ þ YS=X ðX  X0 Þ ½19

Aðt Þ ¼ 0 for t  tlim ½20

 i
XM X h
Aðt Þ ¼ ln 1  M 1  e mM ðt tlim Þ
for t > tlim ½21
mM mM

Nomenclature: A(t), cumulative nongrowth contribution to product formation; b, y, z, d, and e, stoichiometric coefficients; mP (mS), specific rate of
product formation (or substrate consumption) at zero cell growth rate; Nlim, critical concentration of nitrogen at the onset of citric acid production; P,
citrate concentration; ri, conversion rate of any reagent or product; tlim, overall duration of the citrate lag phase; to, overall duration of the cell lag phase;
S, substrate concentration; X, mycelium concentration; XM, maximum mycelium concentration; m, specific cell growth rate; mM, maximum specific cell
growth rate; YN/x, YP/x, and YS/x, yield factors for ammoniac nitrogen, citrate, and substrate on unit cell biomass. Subscripts: lim, referred to limiting
concentration of the nitrogen source; N, nitrogen; P, citric acid; S, glucose; X, mycelium; 0, referred to the initial value.
 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)
Figure 4 Process flowsheet of a typical citric acid fermentation from glucose-based media by A. niger. Equipment and utility identification items: AE, anion exchanger; AF, antifoam agent; AL, alkaline reagent; BD,
fluidized-bed drier; BF, vacuum belt filter; C, centrifuge; c, Condensate; CA, activated carbon adsorber; CE, cation exchanger; CR, vacuum crystallizer; cw, cooling water; CY, cyclone; D, holding tank; dcc, dicalcium
citrate; DW, demineralized water; E, heat exchanger; EA, exhausted air; EV, evaporator; F, production-bubble fermenter; FI, sterile pressure filter; GR, grinder; HT, holding tube; LS, lime slurry; HA, hot air;

811
HS, sulfuric acid; M, mixer; NA, nutrients and additives; PC, centrifugal pump; PE, plate–heat exchanger; S, low-pressure steam; SA, sterile compressed air; Se, dicalcium citrate seed; SF, seed-bubble fermenter;
tcc, tricalcium citrate; WE, water evaporated.
812 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)
on the form manufactured. Crystals are separated by centrifu-
gation and dehydrated in a two-stage fluidized-bed dryer, the
first one using hot air at 90
C and the second one using air
conditioned at 20
C and relative humidity (RH) of 30–40%
because of crystal hygroscopicity. The mother liquor is partly
(w20%) diluted with equipment-cleansing waters, decolor-
ized, and fed back to liming; the remainder is in sequence
decolorized and demineralized before being recycled to the
crystallization unit. In this way, citric acid crystals do not need
additional purification steps to meet specifications for U.S.
Pharmacopeia or Food Chemical Codex material.
In the liquid extraction process, citric acid may be extracted
from the fermentation broth using a highly selective, low-price,
and nontoxic food-grade solvent (i.e., water-insoluble amines,
namely trilaurylamine, n-octanol, C10 or C11 isoparaffin, tri-n-
butyl phosphate, alkysulphoxides). The extract is then heated
and washed countercurrently with water, resulting in about
90% recovery yield and an aqueous citric acid concentrated
solution, which is passed through a granular activated–carbon
column before undergoing the aforementioned evaporation
and crystallization steps.
Future Developments
Production of citric acid has not been much in the focus of
modern molecular biology presumably because it is considered
a mature area. Any improvement of strains of A. niger usually
were carried out by mutagenesis and selection, but metabolic
and genetic engineering are likely to improve acidogenesis.
The replacement of the current batch fermentation with
semicontinuous processes, to increase volumetric productivity
and reduce specific production costs, presently is hampered in
fungal processes by the deterioration of mycelial structure, the
mechanism of which still is unknown. Although the effect of
nitrogen deficiency on citric acid accumulation by A. niger is
well known, a low level of ammonium ions (i.e., 30 g m3,
equivalent to 2 mmol of intracellular NHþ 4 per gram of dry cell)
was found to inhibit the morphological degeneration of pellets
and postpone sporulation. NHþ 4 ions simply are not deposited
into the cell to form the so-called ammonium pool, but they
enter the cell to combine with glucose and form glucosamine,
that is straight away released in the medium. The effective
relationship between the different compounds of the TCA cycle
is to be studied further and controlled before the present batch-
production technology may be converted effectively into
a prolonged fed-batch or continuous production process.
Similarly, the possibility of maintaining microbial cells
active and controlling their growth and production processes
for several weeks or months by immobilization within organic
or inorganic matrices represents a further challenge to the
technological modernization of this sector.
The traditional recovery technology results in several
problems because of disposal of liquid effluents (their chem-
ical oxygen demand being about 20 kg m3) and solid
by-products (i.e., about 0.15 kg of dried mycelium and 2 kg of
gypsum per kilogram of citric acid anhydrous). Several process
alternatives have been suggested thus far to minimize the
overall environmental impact of this process. The replacement
of molasses with raw or hydrolyzed starch- or raw sucrose-
based materials would simplify only the downstream
processing. On the contrary, the recovery of tricalcium (or
trisodium) citrate from clarified, decolorized fermentation
broths by electrodialysis, as well the adsorption of citric acid
onto weakly basic anionic-exchange resins or zeolites using the
simulated-moving bed chromatographic technology (Citrex
Process, UOP, Des Plaines, IL, USA) followed by desorption
with water or dilute acidic solutions, or the use of liquid
membranes, would allow the citric acid to be separated in
a single step and to be recovered without the formation of solid
wastes for disposal.
The environmental aspects of citric acid production have
been assessed. Despite the fact that most raw materials are of
biological origin, many ingredients, such as ammonium
nitrate, lime, and sulfuric acid, are hazardous chemicals. For
instance, it was found that the environmental impact of citric
acid production using whey was smaller than that using corn
starch.
Gluconic Acid
D-Gluconic acid (2,3,4,5,6-pentahydroxy pentane-1-carboxylic
acid: C6H12O7) is an oxidation product of D-glucose, which, in
an aqueous solution, leads to a complex equilibrium between
gluconic acid and its two lactones: 1,5-lactone (D-glucono-
d-lactone) and 1,4-lactone (D-glucono-g-lactone).
D-Gluconic acid is commercially available as 50% aqueous
solution (density of 1230 kg m3 at 20
C and pH 1.82). This
acid and its derivatives are used in the pharmaceutical, food,
feed, and chemical industry because of their low toxicity and
their ability to form water-soluble complexes with metallic
ions (e.g., Caþ2, Feþ3), especially in the presence of 5–10% of
sodium hydroxide. Sodium gluconate is the main industrial
product and it is used as a sequestering agent (e.g., bottle
washing, metal surface cleaning, and rust removal) and to
plasticize and retard the curing process of cement mixes. The
calcium and iron gluconates are used in medicine to treat
diseases of calcium and iron deficiency (such as osteoporosis
and anemia). D-Glucono-d-lactone is used as a latent acid in
baking powders for use in dry cake mixes, meat processing, and
instant chemically leavened bread mixes, whereas D-glucono-
g-lactone is made only in small quantities as a specialty
chemical.
The conversion of glucose to gluconic acid is a simple
oxidation process and may be carried out by a variety of
processes – namely, microbial fermentation, chemical, elec-
trochemical, or enzymatic catalysis. Currently, these processes
appear to be either more expensive, unstable, or less efficient
than the fermentation process, which presently is the only
method of choice.
After the first isolation of calcium gluconate (1880) from
glucose fermentation in the presence of CaCO3 by a strain of
Mycoderma aceti, the Chas. Pfizer & Co., Inc. (New York, USA)
started industrial-scale production of gluconic acid in 1923.
Further research at the U.S. Department of Agriculture in
cooperation with the Iowa State College led to the semi-
continuous production of sodium gluconate from glucose
using A. niger NRRL 67.
Several filamentous fungi of the genera Penicillium and
Aspergillus, the yeastlike fungus Aureobasidium pullulans, and
 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic) 813

bacteria (Acetobacter suboxidans, Pseudomonas ovalis, Glucono- adjusted to 6–6.5 with sodium hydroxide, and a 2–5% v/v
bacter spp.) produce gluconic acid from glucose-based media, inoculum generally is used. For inoculum development, con-
but industrial processes have been developed only for the idia are recovered from stock agar slants and are inoculated into
production gluconic acid from glucose syrups with A. niger and vegetative seed–culture media (106 conidia cm3); pellet-like
Gluconobacter oxydans. Industrial strains are not freely available, mycelia is obtained after incubation at 30
C for 15–24 h and is
but a few gluconic acid–producing strains (A. niger NRRL 3, used to inoculate seed fermenters at a density of 20–50
NRRL 67) can be obtained from international culture collec- pellets cm3.
tions. Penicillium spp. generally produce less gluconate than The fermentation is carried out under continuous automatic
Aspergillus, but they have the advantage of excreting the glucose control of sterile air sparging (1.0–1.5 vvm), temperature
oxidase (an important by-product) into the medium, which (33
C), pressure on the tank top (2–3 bar), pH (5.5–6.5 by
makes its recovery easier. addition of 30–50% NaOH solution to neutralize the gluconic
The formation of gluconic acid by A. niger is controlled by acid formed), and foam level. It is completed within w30 h
the enzyme glucose oxidase, an omodimer containing two with yield factors of 0.97–1 kg of gluconic acid per kilogram of
flavin adenine dinucleotide (FAD) moieties. Such enzyme glucose consumed (against a theoretical yield of 1.09 kg kg1)
abstracts two hydrogen atoms from glucose, thus yielding the and gluconate productivities of 9–13 kg m3 h1.
glucono-d-lactone, which to some extent hydrolyzes to glu- In the fed-batch operation, the mycelium may be reused up
conic acid. The FADH2 reacts with oxygen to form hydrogen to five times without any loss in gluconate productivity
peroxide, which is converted into oxygen and water by the provided that the levels of glucose oxidase activity and other
enzyme catalase. Both glucose oxidase and catalase are microelements (i.e., iron and manganese) are kept under
constitutive endoenzymes in A. niger. control. Stepwise addition of glucose may be used to increase
A highly productive process of gluconic acid using free- gluconate concentration to 580 kg m3.
growing cells of A. pullulans DSM 7085 recently has been At the end of fermentation, the mycelium is removed using
developed. Its high conversion yields (90–98%) and rates aseptic centrifugation, under vacuum-belt filtration or cross-
(13–19 kg m3 h1) resulted in as high gluconate concentrations flow microfiltration and may be used as a source of glucose
as 504 or 230–433 kg m3 in fed-batch or chemostat trials. oxidase or may be disposed off via incineration. The clarified
Although this novel fermentation process offers a new opportu- broth, generally containing w300 kg m3 of sodium gluconate,
nity for commercial gluconic acid production, as well as many is filtered, decolorized using a granular activated–carbon
advantages over the traditional microbial fermentation pro- column, concentrated under vacuum to 45–50% total solids,
cesses, it is still confined to laboratory-scale applications. Thus, neutralized to pH 7.5 with NaOH, and then spray or drum
only the sodium process by batch-submerged fermentation from dried. If 50% gluconic acid is required, the concentrated liquor
glucose syrups using A. niger will be described in the following may be passed through a cation exchanger to remove Naþ ions.
paragraphs. Further crystallization at 30–70
C or at more than 70
C
Glucose syrups of 70
Brix strength are generally used as allows crystals of the d-lactone or g-lactone to be precipitated,
carbon source in the preparation of the fermentation medium. respectively.
Table 4 lists the typical composition of the seed and
production media used in laboratory- and industrial-scale
trials. Lactic Acid
After the pH is adjusted at 4.5 with sulfuric acid, the
medium is sterilized at 121
C for 15–30 min, cooled at 33
C, Lactic acid (2-hydroxypropionic acid: C3H6O3) may be
and then transferred into the fermentation vessel. The pH is produced by chemical synthesis or fermentation. Of the two
enantiomers, L-(þ) and D-() lactic acid, only the L-(þ) isomer
is used by human metabolism and, because of the slight
Table 4 Composition for the production media used in the
toxicity of the D-() isomer, it is preferred for food uses.
laboratory- and industrial-scale production of gluconic acid by A. niger Because the chemical route yields a mixture of L-lactic acid and
D-lactic acid and relies on costly raw materials, all lactic acid
Vegetative seed Gluconic acid manufacturing industries have switched to fermentation-based
Component –culture media production media Unit technologies (Table 1). The free acid is used as an acidulant/
preservative in several food products (cheese, meat, jellies,
Glucose 40 120–350 kg m3
NH4NO3 (or other 2.4 0.4–0.5 kg m3 beer) (see Preservatives: Traditional Preservatives – Organic
NHþ 4 salt)
Acids); sodium lactate is used for carcass decontamination;
KH2PO4 1.5 0.1–0.3 kg m3 ammonium lactate is used as a source of nonprotein nitrogen
MgSO4$7H2O 3.57 0.1–0.3 kg m3 in feeds; sodium and calcium stearoyl lactylates are used as
Agar 1 0 kg m3 emulsifiers and dough conditioners. The large increase in lactic
Yeast extract 1 0 kg m3 acid production is due to its use in the synthesis of polylactic
Corn-steep liquor 0 0.2–0.4 kg m3 acid (PLA), a polyester used for biodegradable plastics for food
ZnSO4$7H2O 100 0 mg m3 packaging, compost, and garbage bags, and disposable table-
CuSO4$5H2O 20 0 mg m3
ware, as well as several medical applications, such as reab-
FeCl3$6H2O 300 0 mg m3
sorbable sutures, orthopedic implants, and controlled drug
MnSO4$7H2O 0 30 mg m3
Initial pH 6.5 6.0 – release. The current economically viable industrial process for
PLA production is via the dehydrated cyclic dilactate ester
814 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)
(lactide) formation. In brief, lactic acid is first polymerized into
oligomers of PLA consisting of 30–70 lactyl units (–CHCH3–
CO–O–). These oligomers are then depolymerized by
increasing the polycondensation temperature and lowering the
pressure in the presence of transition metal–based catalysts
(i.e., stannous octoate at 0.05%) to distil the lactide. Finally, by
opening its ring, it is possible to obtain high molar mass
polymers (100–300 kDa) with appropriate optical and
mechanical properties (i.e., tensile strength >50 MPa). Other
comonomers, such as caprolactone, hydroxybenzoic acid, and
others, can be incorporated to provide environmentally safe
materials. PLA appears to be a sustainable alternative to
petroleum-based plastics, because lactide is produced from the
fermentation of renewable resources, such as corn starch (as in
the industrial plant of Nature Works LLC, Blair, Nebraska, USA:
140 000 metric tons per year). Nevertheless, to minimize
competition for land and food, research studies have started to
develop second-generation PLA products from lignocellulosic
hydrolysates (e.g., crushed corncobs).
Organisms and Metabolic Pathways Involved
Lactic acid can be produced using homofermentative lactic
acid bacteria (LAB), facultatively anaerobic Bacillus species
(B. coagulans), and molds (Rhizopus microsporus, Rhizopus
oryzae). Recently, lactic acid producing genetically modified
strains of Escherichia coli and Saccharomyces cerevisiae have
been developed. The choice of the species depends on several
considerations, including the ability to use the type of sugars
available in the substrate, growth temperature, nutritional
needs, acid tolerance, and type of lactic acid isomer
produced.
Thermophilic lactobacilli (Lactobacillus delbrueckii subsp.
delbrueckii, L. delbrueckii subsp. bulgaricus, Lactobacillus helveticus)
tolerate higher concentrations of lactate and higher temperatures
(48–52
C), thus involving higher productivity and yields
and reduced contamination risks. They produce D-() or DL-lactic
acid (some industrial strains that have been claimed to be
L. delbrueckii produce L-(þ) lactic acid) and may be less suitable
for food, feed, or biomedical applications. Lactococci, mesophilic
lactobacilli (Lactobacillus casei subsp. casei, Lactobacillus amylophi-
lus), and thermophilic streptococci (Streptococcus thermophilus)
have lower temperature optima or reduced acid tolerances, but
they may be desirable for other reasons (e.g., production of pure
L-(þ) lactic acid, hydrolysis of starch). Recently, genetic engi-
neering has been used to produce L. helveticus and Lactobacillus
plantarum strains, which produce optically pure L-(þ) or D-()
lactic acid (see Lactobacillus: Introduction; Lactococcus: Introduc-
tion; and Streptococcus thermophilus).
Homofermentative LAB ferment hexoses via the glycolytic
pathway (see Metabolic Pathways: Release of Energy (Anaer-
obic)). Pyruvate is reduced to lactate by stereospecific lactate
dehydrogenase(s) (L-LDH or D-LDH). LDH is allosteric (acti-
vators: fructose-1,6-bisphosphate and Mnþ2) in lactococci and
nonallosteric in homofermentative lactobacilli. Undissociated
lactic acid acts as a noncompetitive inhibitor for growth and
lactic acid production by diffusing through the membrane and
decreasing intracellular pH: pH control during fermentation
reduces the inhibition, but the maximum lactic acid concen-
tration achievable is usually lower than 150 kg m3.
Concomitant substrate and product inhibition has been
reported for several species.
LAB are fastidious microorganisms and require supple-
mentation of fermentation media with peptides and growth
factors, usually in the form of yeast extract. Because this may
account for 30–35% of substrate costs, they may be replaced by
less demanding B. coagulans and R. oryzae, both being
L(þ)-lactate producers, even if smaller yields (as low as 70%
for R. oryzae because of concomitant fumaric acid and ethanol
production) and acid tolerance may offset the advantage of
using lower amounts of supplements.
Substrate Production and Recovery
Lactic acid can be produced from a variety of raw substrates
(whey and whey permeate, beet and cane molasses, starch and
corn starch hydrolysates, wood hydrolysates; see Fermentation
(Industrial): Media for Industrial Fermentations). Some species
(Lb. amylophilus) can hydrolyze starch, but the pseudoplastic
behavior of starchy substrates makes the pH control difficult.
When whey permeate is used, supplementation with milk
protein hydrolysates (5–10 kg m3) and yeast extract (up to
20 kg m3) is required. Lactic acid production is usually
a growth-associated production process, but nongrowth-
associated production becomes significant when growth is
limited by a lack of nutrients or high undissociated acid
concentration. The pH is controlled at 5–6.5 by the automatic
addition of NaOH, Na2CO3, or NH4OH or by the addition of
CaCO3. Fermentation is carried out under anaerobic or micro-
aerophilic conditions and lactic acid yield is usually between 85
and 98% with isomer purity as high as 99%. Batch fermenta-
tions result in high product concentration (120–150 kg m3)
but in low productivity (2 kg m3 h1). Conversely, continuous
fermentations with cell-recycle or immobilized cells give rise to
higher productivities (20–80 kg m3 h1) and lower lactate
concentrations (<50 kg m3). End-product inhibition may be
circumvented by using integrated fermentation processes, in
which lactic acid is removed from the culture broth by several
techniques, including electrodialysis, ion-exchange resins, or
nanofiltration.
Recovery of lactate is made complicated by the high solu-
bility of its salts. The traditional process involves precipitation
of calcium lactate and regeneration of lactic acid by the addi-
tion of sulfuric acid followed by further purification steps (ion
exchange and decolorization). Alternative processes include the
extraction by liquid membranes, electrodialysis, and ion
exchange. In particular, the recent industrial use of electrodi-
alysis with bipolar membranes in France resulted in the virtual
elimination of gypsum waste production. Conventional
recovery by the precipitation method seems to be the most
economical route.
Other Organic Acids Produced by Fermentation
Propionic Acid
Propionic acid (C3H6O2) and its salts are used as mold inhib-
itors in bakery products, although other nonfood uses are
important (see Permitted Preservatives – Propionic Acid). It may
be produced by fermentation by members of the genera
 FERMENTATION (INDUSTRIAL) j Production of Some Organic Acids (Citric, Gluconic, Lactic, and Propionic)
Propionibacterium (P. freudenrichii, thoenii, acidipropionici), Veillo-
nella, Clostridium, and Selenomonas, but currently it is produced
by chemical synthesis because of the shortcomings of the
fermentation route (low productivities, <1 kg m3 h1 in batch
processes; low product concentrations, <50 kg m3; difficulty in
product separation from acetic acid, which invariably is
produced from sugars and lactate). The high microbial
productivities (2–14 kg m3 h1) obtained in continuous
fermentations using immobilized cells or membrane-recycle
reactors, as well the possibility of obtaining pure propionic acid
from alternative low-cost substrates, like crude glycerol from the
biodiesel industry, might refocus industrial manufacturers
toward the fermentation route. For instance, use of a metaboli-
cally engineered strain of P. acidipropionici (ACK-Tet) resulted in
a propionic acid concentration of 106 kg m3 with a product
yield of 0.54–0.71 g per g of glycerol consumed and a propionic
acid-to-acetic acid ratio of 22.4.
See also: Arthrobacter; Aspergillus; Bacillus: Introduction; Bread:
Bread from Wheat Flour; Yarrowia lipolytica (Candida Lipolytica);
Escherichia coli: Escherichia coli; Fermentation (Industrial):
Basic Considerations; Fermentation (Industrial): Media for
Industrial Fermentations; Fermentation (Industrial): Control of
Fermentation Conditions; Fermentation (Industrial): Recovery of
Metabolites; Fermented Foods: Fermentations of East and
Southeast Asia; Fungi: The Fungal Hypha; Fungi: Classification
of the Hemiascomycetes; Fungi: Classification of the
Deuteromycetes; Genetic Engineering; Gluconobacter;
Lactobacillus: Introduction; Metabolic Pathways: Release of
Energy (Aerobic); Metabolic Pathways: Release of Energy
(Anaerobic); Preservatives: Traditional Preservatives – Organic
Acids; Permitted Preservatives – Propionic Acid;
Propionibacterium; Streptococcus thermophilus; Vinegar; Yeasts:
Production and Commercial Uses.
815
Further Reading
Anastassiadis, S., Aivasidis, A., Wandrey, C., 2003. Continuous gluconic acid
production by isolated yeast-like mould strains of Aureobasidium pullulans. Applied
Microbiology and Biotechnology 61, 110–117.
Berovic, M., Legisa, M., 2007. Citric acid production. Biotechnology Annual Review 13,
303–343.
Hofvendahl, K., Hahn–Hägerda, B., 2000. Factors affecting the fermentative lactic acid
production from renewable resources. Enzyme and Microbial Technology 26,
87–107.
Joglekar, H.G., Rahman, I., Babu, S., Kulkarni, B.D., Joshi, A., 2006. Comparative
assessment of downstream processing options for lactic acid. Separation and
Purification Technology 52, 1–17.
John, R.P., Nampoothiri, K.M., Pandey, A., 2007. Fermentative production of lactic
acid from biomass: an overview on process developments and future perspectives.
Applied and Microbiology and Biotechnology 74, 524–534.
Legisa, M., Mattey, M., 2007. Changes in primary metabolism leading to citric acid
overflow in Aspergillus niger. Biotechnology Letters 29, 181–190.
Magnuson, J.K., Lasure, L.L., 2004. Organic acid production by filamentous fungi.
Chapter 12. In: Tkacz, J.S., Lange, L. (Eds.), Advances in Fungal Biotechnology for
Industry, Agriculture, and Medicine. Kluwer Academic/Plenum Publishers, New
York, pp. 307–340.
Okano, K., Tanaka, T., Ogino, C., Fukuda, H., Kondo, A., 2010. Biotechnological production
of enantiomeric pure lactic acid from renewable resources: recent achievements,
perspectives, and limits. Applied Microbiology and Biotechnology 85, 413–423.
Papagianni, M., 2007. Advances in citric acid fermentation by Aspergillus niger: biochemical
aspects, membrane transport and modeling. Biotechnology Advances 25, 244–263.
Sauer, M., Porro, D., Mattanovich, D., Branduardi, P., 2007. Microbial production of
organic acids: expanding the markets. Trends in Biotechnology 26 (2), 100–108.
Singh, O.V., Kumar, R., 2007. Biotechnological production of gluconic acid: future
implications. Applied Microbiology and Biotechnology 75, 713–722.
Yoo, D.-K., Kim, D., 2009. Production of optically pure poly(lactic acid) from lactic acid.
Polymer Bulletin 63, 637–651.
Zhang, A., Yang, S.-T., 2009. Propionic acid production from glycerol by meta-
bolically engineered Propionibacterium acidipropionici. Process Biochemistry 44,
1346–1351.