86 Antonie van Leeuwenhoek 49 (1983)

Fermentative production of lactic acid

G. L. NANNINGA

C.V. Chemic Combinatie Amsterdam, P.O. Box 21,

4200 AA Gorinchem, The Netherlands

Lactic acid has been produced in The Netherlands since 1936. C.C.A. together with its Brazilian
subsidiary produces 65~ of the world's supply of fermentative lactic acid.

Characteristics of the fermentation process

Sucrose is fermented to lactic acid by a culture of lactic acid bacteria. The fermentation is carried
out batchwise at a temperature above 50 ~ Strict sterile conditions are not necessary. In contrast
to earlier described industrial processes, in this process the pH is kept constant by the addition
of lime. The yield of lactic acid is more than 90%, with an L(+)-isomer content of about 97%.

Purification
At the end of the fermentation, lime is added and the bacteria are killed by heat. After sedimenta-
tion and filtration, lactic acid is liberated by addition of sulfuric acid. The gypsum formed is filtered
and the lactic acid solution obtained is purified by ion-exchange resins after active carbon treatment.
Concentration leads to "edible-grade" lactic acid. Further purification by esterification, distillation
and hydrolysis gives a reagent-grade product.

Application and other products

Most of the lactic acid is used in the food industry. Some of the numerous applications are:
acidulation of confectionery products and beverages, pH adjustment of beer, wine, cider, addition
to brines for green olives and gherkins, in pickles and dairy products and as a moisturizer in hand
skin lotions. Calcium lactate is used as calcium source in pharmaceutical preparations, in infant
food, pet food, fish feed and as strengthening agent in fruit canning.
Sodium lactate can partially replace sodium chloride in sausage production. Ferrous lactate is
a readily absorbable iron source. Ethyl-lactate is an excellent solvent for lacquers.
A recent development is the production of a lactic acid powder with 60~ free lactic acid for
those applications in which a'dry or powdered acid is necessary.

Microbial production of citric acid

F. SMEETS

Citrique Beige, B-3300 Tienen, Belgium

Citric acid is a natural constituent of numerous plants and animals. The presence of one hydroxyl
and three carboxylic acid groups makes it the most versatile and widely used acidulant of food
and pharmaceutical products. It has been generally recognized as safe (GRAS).
Antonie van Leeuwenhoek 49 (1983) 87

History
Citric acid was first isolated and crystallized by Schule (1784). As raw material was used the
juice of lemons, which may contain up to 8~ of the acid. Many years later (1913) a process for
recovering citric acid from citrus fruits has been developed by Italians on a commercial scale. Later
on, fermentation processes were developed to find an alternative way of making citric acid.
A forerunner in this development was Wehmer, who used a Penicillium strain for an industrial
fermentation in the "Fabriques chimiques de Thann et de Mulhouse" - Alsace (1894). The produc-
tion, however, was abandoned after two years.
The first to succeed (1919) were"Les Produits Organiques de Tirlemont", Tienen, Belgium. After
some ten years of an intense struggle against the lemon-based citric acid, an agreement was finally
reached with the Italians, and a joint-venture put up under the name of "Citric Belge". From 1923
on, several newcomers entered the field in a number of countries (U.S.A., Czechoslovakia, Germany
etc.). World production has since grown steadily and is actually estimated at about 350000 tons/year.

Raw materials and microorganisms

Nowadays, commercial citric acid is produced almost exclusively by strains of the mould Aspergil-
lus niger. Occasionally other microorganisms have been tried but without success. For some time,
fermentation of n-alkanes by yeasts (e.g. Candida lipolytica) looked promising. However, it failed,
because of a number of reasons, the increasing price of the raw material probably being the most
important one.
Sugar, refined or raw, supplemented with nutrients, has been the main carbon source until World
War II. Since then, sugar molasses, especially those from beets, gradually replaced the more expen-
sive pure material.
Because of the ready availability in regions where corn is grown, hydrolysates of starch are also
used by some manufacturers.

Techniques offermentation
Citric acid is produced by two different processes. The oldest one, the surface process, comprises
the inoculation of shallow trays, containing diluted molasses with fungal spores. A mycelial mat
develops on the surface of the liquid, whereafter the acid is produced. Fans supply air, which also
control the temperature by evaporative cooling. Yields depend on a number of parameters, such
as type of strain, composition of the medium and physical conditions.
The more recent, submerged process, comprises the inoculation of large tanks. Small spherical
beads, so-called pellets, develop from clusters of spores. Compressed air is bubbled through the
broth and dispersed by mechanical agitation. Temperature is controlled by water cooling. Basically,
yields are related to the same parameters in both methods. Additional monitoring of some factors
is possible and necessary for obtaining optimal results with the submerged fermentation.
The last two decades have seen an increasing percentage of the total citric acid output performed
by the submerged process. However, the surface technique will not necessarily become obsolete.
The submerged system requires less manpower, but requires more energy.
Acid production in tanks or towers is faster, but the level of the usable concentration of raw
materials is lower than in trays.
Yields from a sugar-based fermentation are more or less comparable, exceeding 2/3 of the theoreti-
cal value of 122% (w/w) expressed as citric acid monohydrate.
Summarising one could say that both microbial techniques have a fair chance of survival; so
far, they look superior to all known chemical routes.
Replacement of citric acid by other organic acids seems unlikely in the near future. Due to its
mildly sour taste and its high complexing and buffering capacity it will probably continue to remain
in demand for a large number of applications.