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4b - Biological Method

The document discusses three biological methods for producing citric acid through fermentation: solid-state fermentation (SSF), liquid surface fermentation (LSF), and submerged culture fermentation (SCF). SSF uses agricultural waste on a solid substrate, while LSF grows microorganisms on the surface of liquid. SCF is the most common industrial method, using a liquid medium in a bioreactor. SCF offers the best control and is typically preferred for large-scale production.

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43 views4 pages

4b - Biological Method

The document discusses three biological methods for producing citric acid through fermentation: solid-state fermentation (SSF), liquid surface fermentation (LSF), and submerged culture fermentation (SCF). SSF uses agricultural waste on a solid substrate, while LSF grows microorganisms on the surface of liquid. SCF is the most common industrial method, using a liquid medium in a bioreactor. SCF offers the best control and is typically preferred for large-scale production.

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Nizar Mousa
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© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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2- Biological Method

It involves the fermentation of citric acid using microorganisms. The following


three methods can carry out biological fermentation of citric acid :

1-Koji Process
It is called “Solid-State Fermentation (SSF) ”, It is related to the use of agro-industrial residues
for citric acid production. In the Koji process, raw materials like apple pomace, sugar cane, beet
molasses etc. are generally used. The Aspergillus Niger utilizes raw materials.
SSF involves growing microorganisms on a solid substrate with minimal water content. It's
typically used for producing citric acid from agricultural waste products .
SSF can be more challenging to control and optimize compared to other processes, as
maintaining uniform conditions and mixing can be difficult in solid-state systems .
However, SSF can be cost-effective and environmentally friendly for certain feedstocks and is
suitable for certain strains of microorganisms.
SFF has several advantages, such as lower water usage, reduced contamination risks, and the
ability to use low-cost substrates .
Here's a general overview of the steps involved in solid-state fermentation for citric acid
production:
1- Selection of Microorganism: Typically, the fungus Aspergillus niger is used .
2- Substrate Selection: Common substrates include wheat bran, rice bran, sugarcane bagasse .
3- Inoculation: The selected microorganism (Aspergillus niger) is inoculated onto the solid
substrate.
4- Fermentation Conditions:
- Temperature: Typically maintained between 28°C to 30°C.
- pH: Initially, the pH is maintained in the acidic range (pH 2.0-3.5) to encourage citric acid
production. It may be adjusted during the fermentation process.
-Moisture Content: The moisture content of the substrate is maintained at an optimal level to
support fungal growth and citric acid production .
5- Aeration and Mixing
6- Harvesting
7- Purification and Recovery

2- Surface Culture Process


It is also called “liquid surface fermentation ( LSF )”, it involves growing microorganisms
(usually Aspergillus Niger) is grown on the surface of a liquid medium . LSF can be used in
laboratory settings and for small-scale production.
LSF process steps are topically same as SSF , the difference between them is just in Surface
culture , conditions , medium state .
LSF offers advantages in terms of easier monitoring and control compared to solid-state
fermentation. However, it requires the use of liquid medium and aeration systems, which can
lead to higher energy and operational costs.
The design and calculations for these processes may be somewhat simpler than SSF, but still
more complex than submerged culture.

3- Submerged Culture Process


It is also called “Submerged culture fermentation ( SCF ) ”. About 80% of the citric acid
production is carried out through SCF. SCF makes the use of black Aspergillus, i.e. A. japonicus.
It is performed in a bioreactor made of stainless steel compiled with proper aeration, cooling
system, impellers etc.

SFC is one of the most common and widely used methods for both industrial-scale citric acid
production and laboratory research scale .

Microorganisms are grown in a liquid medium, and conditions such as temperature, pH,
aeration, and nutrient supply can be more easily controlled and optimized in this method.
This method is more efficient and cost-effective compared to SSF AND LSF . Here's an overview
of the submerged culture process for citric acid production:
1- Selection of Microorganism: Aspergillus niger is the most commonly used microorganism due
to its ability to produce high levels of citric acid efficiently.
2- Inoculum Preparation: A small culture of the chosen microorganism is grown separately to
create a sufficient amount of biomass, known as the inoculum. This inoculum is then used to
seed the main fermentation vessel.
3- Fermentation Medium: A fermentation medium is prepared, which typically contains a
carbon source (usually a sugar, like glucose or sucrose), nitrogen sources (such as ammonium
salts), and various salts and trace elements required for growth and citric acid production. The
medium is sterilized to eliminate any competing microorganisms.
4- Inoculation: The prepared inoculum is introduced into the sterilized fermentation medium to
start the fermentation process.
5- Fermentation: The fermentation process takes place in a submerged culture, where the
microorganism grows and produces citric acid. It typically occurs under controlled conditions,
including temperature (30 degrees Celsius ), pH ( 2.5 to 4.0) , and aeration.

6- Control Parameters: Several key parameters are closely monitored and controlled during
fermentation, including temperature (usually around 28-30°C), pH (usually around 2.0-3.0), and
aeration. The pH is maintained by the addition of acids or bases as needed.
7- Nutrient Feeding: To sustain the growth of microorganisms and maximize citric acid
production, additional nutrients, such as carbon and nitrogen sources, may be fed into the
culture at specific intervals.
8- Citric Acid Production: As the microorganism grows, it consumes the carbon source and
produces citric acid as a metabolic byproduct ( Kreb’s cycle) . This citric acid accumulates in the
fermentation broth over time.

9- Harvesting and Recovery: When the citric acid concentration reaches its peak, the
fermentation process is stopped, and the citric acid is separated from the fermentation broth.
the separation and purification including filtration and precipitation, are used to recover the
citric acid.
10- Post-processing: The recovered citric acid may undergo further purification and
crystallization to obtain a high-purity product, suitable for various industrial applications.

In summary, the choice of the easiest and most suitable method for citric acid
production depends on factors such as the scale of production, the specific microorganisms
used, the substrate, and the available resources. For large-scale, industrial production, the
SCF is often preferred due to its ease of control and scalability. However, for certain
applications or when using specific waste materials, solid-state fermentation or surface
culture processes may be viable options.

Furthermore , if we interest for an easier and more predictable process in terms of


calculations and design for citric acid production, the submerged culture process is typically
the preferred choice. It offers better control, well-established methodologies, and a wealth
of available data and resources for designing and optimizing the production process .

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