Bioprocess Course Mejda Salhi 2018

Bioprocesses applied to the agri-food industry

COURSE OUTLINE.....................................................................................................................1

1. Définition..........................................................................................................................1

2. Domains of application...................................................................................................1

3. Types................................................................................................................................1

4. Technology of fermentation

5. History........................................................................................................................2

6. Steps development of an industrial bioprocess................................................3

7. Steps of an industrial bioprocess............................................................3

7.1. Formulation from the culture medium............................................................................3

7.2. Stérilisation..............................................................................................................4

7.3. Preparation of the inoculum.......................................................................................4

7.4. Bioreactor of production......................................................................................4

7.5. Separation, Recovery and purification of products..........................................4

7.6. Treatment effluents

8. Applications bioprocesses

8.1. Fermentation...........................................................................................................5

8.2. Culture of higher eukaryotic cells...........................................................6

8.3. Bioprocesses environmental

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Bioprocess Courses Mejda Salhi 2018

1. Definition

A bioprocess is a set of techniques and methods that rely on

cells (animal, plant, or microbial or their components) (enzymes,
proteins, genes..) for the production of goods or services.

Examples: yogurt production, wine production, penicillin production, wastewater treatment...

2. Application areas

Bioprocesses are used in various industries:

- Agri-food industries: beverages, yeasts, animal proteins.

- Chemical industries: production of organic acids, ethanol...
- Pharmaceutical industries: production of antibodies, vaccines, hormones..
- Environmental industry: treatment of liquid, solid and
gaseous..

3. Types

They are thus classified into three main areas:

- The fermentation processes;

- Animal and plant cell cultures;
- Environmental processes.

4. Defermentation technology

The term fermentation, in biochemistry, refers to the process of biological degradation.

in the absence of air.

In industry, fermentation refers to any process that allows the production of a compound
or large-scale biomass by using a microorganism present
or in the absence of air.

Bioreactor:

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Bioprocessing Course Mejda Salhi 2018

Bioreactors or fermenters are vessels or tanks used for the realization

fermentations. Their role is to ensure a controlled environment for cultivation
microbial in order to create a well-defined application. The reactor is made of
system for adjusting physicochemical parameters (pH, temperature, concentration)
substrates, oxygen content..) and helps to avoid any source of contamination.

5. History

Discovered remnants have shown that the use of fermentation dates back to antiquity.
times (7000 BC). The Egyptians as well as other peoples of the Mediterranean
have managed to master the transformation of grape juice into wine and the preservation of
foods by fermentation.

The term fermentation comes from the Latin word "fermentare" used by the Romans and which
derive from the word 'fervere' to describe the bubbling aspect of fermented grape juice.

Au cours de la pré-histoire, le phénomène de la fermentation a été considéré comme

a miracle attributed to the gods (Osiris) who offer the products to humans for
to soothe their bodies and their souls.

In the Middle Ages, this phenomenon was explained by the theory of spontaneous generation.
the scientists who supported this theory postulated for the transformation
spontaneous organic matter.

In the 16th century, microorganisms were discovered thanks to the first microscope.
invented by the Dutch Antonie Van Leeuwenhoek.

This theory was only rejected in 1850 by the French chemist Louis Pasteur who
demonstrated the microbial actions in alcoholic and acetic fermentations.

Currently, an entire field, industrial microbiology, is dedicated to the

mastery of fermentation for the achievement of constant and large-scale productions
quantities. Genetic engineering has allowed us to surpass the limits of systems of
animal and plant production and to produce, through fermentation, molecules
recombinants such as insulin.

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Bioprocess Course Mejda Salhi 2018

6. Stages of development of an industrial bioprocess

The story of a bioprocess begins in a laboratory with mini-reactors of 10

At 100ml of volume, this is the research phase. During the second phase, a
development pilot from 10 to 100 L is designed. We then move on to the industrialization of
this pilot in a process with a capacity of 10 to 100 m3 that is entering production.

7. Steps of an industrial bioprocess

The implementation of a bioprocess is done in 6 steps:

1- Formulation of the culture medium

2- Sterilization of the reactor, its accessories, and the culture medium;
3- Development of an inoculum;
4- Fermentation;
5- Recovery and purification of the product;
6- Elimination of process residues.

7.1. Formulation of the culture medium

The culture medium must provide the nutritional needs of the cultured strain while considering
account of the fermentation objective: carbon source, energy source, sources
nitrogen, sulfur, growth factors… the media synthesized in the laboratory
are more expensive than those obtained from the valorization of by-products of industries
agri-food products such as molasses and whey.

Table 1. The different types of culture media

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Bioprocess Course Mejda Salhi 2018

7.2. Sterilization

Before starting fermentation, we apply sterilization using steam.

under pressure in the culture medium inside the bioreactor. Sterilization allows
to eliminate the contaminants that ferment with the same strain
growth parameters.

7.3. Preparation of the inoculum

The inoculum consists of the biomass of the strain used for seeding the
fermenter. This biomass can consist of bacteria, mycelium, spores,
of cells. The inoculum must be homogeneous and free of contaminants.

The transition from the industrial strain in its lyophilized or frozen form to a
inoculum is done on different scales:

1- Stirred culture in an Erlenmeyer flask;

2- Preparation of the pre-culture or "starter culture" in a bioreactor
propagation having a volume of 5 to 10% of the fermenter;
3- Inoculation of the fermenter.

7.4. Production Bioreactor

There is a wide variety of fermenter technologies depending on the production.

realize and the requirements of the strain to be cultivated.

7.5. Separation, recovery, and purification of products

The product can be either biomass, growth metabolites, or compounds

Intracellular. The biomass is recovered by filtration, sedimentation, centrifugation.
It is then washed, dried, and packaged. In the case where the product to be recovered is
Intracellularly, the biomass undergoes lysis, the product is recovered, purified, and then concentrated.

For the metabolites, they are obtained by chemical extraction using organic solvent.
culture medium filter.

7.6. Treatment of effluents

The residual wastewater from bioprocesses is rich in organic matter, in addition

large quantities of acids and bases used for reactor cleaning. The

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Bioprocessing Course Mejda Salhi 2018

the treatment of these wastewaters will be subject to environmental processes such as the

methanization, composting...

8. Applications of bioprocesses

8.1. Fermentation

Biomass production

We distinguish two types of processes: the production of viable biomass and the production
of dead biomass. Viable biomass, in the form of ferment, is used to
the sowing and the production of food or pharmaceutical products,
industrial or domestic.

Baker's yeast, lactic ferments..

Viable biomass also includes probiotics (Bifidus regularis) and agents of

biological control (Bacillus thuringiensis).

Dead biomass is used as a protein-rich dietary supplement.

called Unicellular Organism Proteins (UOP).

Example: Candida utilis containing 50% protein by dry biomass.

Production of metabolites

Microorganisms, through the diversity of their metabolic pathways, allow for the
production of very complex molecules to be produced by the chemical industry.

We distinguish:

- Primary metabolites, produced during the growth phase

exponential through common metabolic pathways. They are the residues of
catabolism. They are alcohols, solvents, and organic acids exploited in
IAA and in Chemical Industries.
- Secondary metabolites, produced during the stationary phase (or of
decline) through specific metabolic pathways providing a competitive advantage to
certain species of bacteria and molds. They are derived from anabolism,
enzyme inhibitors, growth factors, pigments..

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Bioprocess Course Mejda Salhi 2018

Production of microbial enzymes

They are produced in larger quantities and at a lower cost compared to

animal and plant enzymes. Enzymes, primary metabolites, are exploited in
IAA. Enzymes, secondary metabolites, are exploited in the pharmaceutical industry.

Production of Recombinant Proteins

These proteins are obtained from animal or plant genes, inserted into the genome.
of a microorganism. The main recombined microorganisms are: E. coli
Saccharomyces cerevisiae. Recombinant proteins are for pharmaceutical use:
insulin, Growth hormone, Factor VIII...

Recombinant plasmids

Plasmids are small circular molecules of extrachromosomal DNA coding

for secondary traits such as resistance to an antibiotic, toxins, ..They
are distinguished by their autonomous replication. They are used as gene carriers.
recombinants coding for defective or non-existent proteins for purposes
therapeutics.

Bioconversion

It is the transformation of a molecule into a high value-added product.

The basic biochemical reactions are: dehydrogenation, oxidation, deamination,
decarboxylation, isomerization. These enzymatic reactions are advantageous because
regarding the physicochemical conditions that require drastic conditions
as well as the use of chemical catalysts.

8.2. Culture of higher eukaryotic cells

Animal and plant cells have developed enzymatic machinery. They are
used for the production of therapeutic products that cannot be
synthesized by GM microorganisms.

- Animal cells derived from hamsters, rats, or mice are used for the
manufacturing of vaccines, therapeutic proteins or diagnostic such as
monoclonal antibodies; hormones...

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Bioprocessing Course Mejda Salhi 2018

- Plant cells are used for the production of secondary metabolites.

therapeutics: caffeine, nicotine, antioxidants, anti-cancer..

8.3. Environmental Bioprocesses

They rely on the use of microorganisms for protection.

the environment.

- Wastewater treatment by aerobic or anaerobic digestion (methanation) of

waste.
- Solid waste treatment through composting, methanization…
- Bioremediation of contaminated soils
- Biolixiviation of metals
- air decontamination by biofilters