Fermentation- Bacterial, Fungal and Yeast, Biochemistry of fermentation.

Introduction
 Fermentation is one of the ancient food processing technologies.
 Fermentation is defined as a process in which chemical changes occur in an organic
substrate through the action of enzymes produced by microorganisms.
 For example, yeast enzymes convert sugars and starches into alcohol, while proteins
are converted to peptides/amino acids.
 Fermentation takes place in the lack of oxygen that produces ATP (energy).
 It turns NADH and pyruvate produced in the glycolysis step into NAD+ and various
small molecules depending on the type of fermentation.
 The fermenting microorganisms mainly involve L.A.B. like Enterococcus,
Streptococcus, Leuconostoc, Lactobacillus, and Pediococcus and yeasts and molds
like Debaryomyces, Kluyveromyces, Saccharomyces, Geotrichium, Mucor,
Penicillium, and Rhizopus species.

Principle of fermentation
 The main principle of fermentation is to derive energy from carbohydrates in the
absence of oxygen.
 Glucose is first partially oxidized to pyruvate by glycolysis.
 Then pyruvate is converted to alcohol or acid along with regeneration of NAD+
which can take part in glycolysis to produce more ATP.
 Fermentation yields only about 5% of the energy obtained by aerobic respiration.
Different Fermentation pathway of bacteria
1. Alcoholic fermentation
 In this pathway first glucose is converted into Pyruvate by glycolysis. And then
alcohol dehydrogenase reduces the pyruvate into ethanol and CO2.
 Metabolism of pyruvate to produce ethanol occurs in two steps.
 1st step: pyruvate is first decarboxylated into Acetaldehyde and CO2. This reaction is
catalyzed by the enzyme Pyruvate decarboxylase with Thymine pyrophosphate (TPP)
as co-enzyme.
 2nd step: Acetaldehyde is then reduced to ethanol by NADH2. This reaction is
catalyzed by enzyme Alcohol dehydrogenase. NAD+ is regenerated in this step.

Net equation for alcoholic fermentation is

Glucose+ 2ADP +2Pi…………………. 2Ethanol +2CO2+2ATP
 Examples: Sacharomyces cereviseae, Pseudomonas
Application:
 Alcoholic fermentation is important in food and industrial microbiology and is used to
produce beer, wine, distilled sprits etc.
 It is also used in production of fermented food products.

2. Lactic acid fermentation:

 In this pathway pyruvate is reduced to lactic acid. This is a single step reaction carried
out by Lactic acid bacteria (LAB)
 There are two types of lactic acid fermentation.i. Homo lactic fermentationii. Hetero
lactic fermentation
Homolactic fermentation:
 In homolactic fermentation, end product is lactic acid.
 Pyruvate is reduced to lactate or lactic acid by the enzyme lactate dehydrogenase
(Pyruvate reductase).
 Homolactic bacteria: Streptococcus thermophiles, Streptococcus lactis, lactobacillus
lactis, Lactobacillus bulgarius, Pediococcus, Enterococcus
Application:
 Homolactic fermentation is important in dairy industry for souring of milk to produce
various fermented products
 Streptococcus mutant, a bacteria responsible for dental caries is a homolactic
fermenting bacteria.
 Lactobacillus spp in the digestive tract of human helps in digestion of lactose present
in milk.
 Lactobacillus spp are used as probiotic.

Heterolactic fermentation:
 In hetero lactic fermentation, end product is ethanol and CO2 in addition to lactic
acid.
  In this reaction glucose is first metabolized to pyruvate, acetic acid and CO2 by
Pentose phosphate pathway.
 Pyruvate is then reduced to lactic acid whereas acetic acid is reduced to ethanol and
CO2.
 Heterolactic bacteria: Leuconostoc mesenteroides, Lactobacillus bifermentous,
Leconostoc lactis

3. Mixed acid fermentation:

 In mixed acid fermentation, mixture of acids such as lactic acid, acetic acid, ethanol,
formic acid etc are produced as the end product.
 At first pyruvate is cleaved by the enzyme Pyruvate formate lyase to yield formic acid
and Acetyl coA.
 From formic acid various other end products such as acetic acid, lactic acid, succinic
acid, ethanol or CO2 and water are formed according to types of pathway and types of
bacteria. However formic acid is always the intermediate product in this pathway.
  This pathway is followed by member of Enterobacteriaceae family such as E. coli,
Salmonella, Klebsiella etc.
 This fermentative pathway is the basis of Methyl red test.

4. 2,3-Butanediol fermentation:
 In this pathway 2,3-butanediol is the end product.
 Some Pyruvate produced during glycolysis is metabolized as in mixed acid
fermentation but most of the pyruvate is condensed to form α-acetolactate.
 α-acetolactate undergoes decarboxylation in the presence of enzyme pyruvate
decarboxylase to produce Acetoin (acetyl methylcarbainol) which is reduced by
NADH2 to form 2,3-butanediol.

 This pathway is followed by some member of Enterobacteriaceae famil. Eg Klebsiella

 This fermentative pathway is the basis of VP test.

5. Butanol fermentation:
 In this pathway pyruvate is converted into butanol or butyrate. Other end product such
as Acetone and CO2 or Isopropyl alcohol and CO2 may formed by this pathway.
 This pathway is present in Clostridium spp
 At first Clostridium spp convert pyruvate into AcetylcoA aerobically.
  Two molecule of acetylcoA condenses in the presence of enzyme acetyl-transferase to
from AcetoacetylcoA.
 AcetoacetylCoA is reduced to β-hydroxybutyrylcoA by NADH2 in the presence of
enzyme hydroxybutyrate dehydrogenase.
 β-hydroxybutyrylcoA is reduced by enoylcoA hydratase to form CrotonylcoA and
water.
 CrotonylcoA is further reduced to butyrylcoA by an enzyme NAD-linked
dehydrohenase.
 ButyrylcoA and acetate act together with fatty acid coA transferase to form acetylcoA
and butyrate. Acetyl coA then recycle in the reaction.

6. Propionic acid fermentation:

 In this pathway propionic acid and CO2 is the end product.
 This pathway is carried out by Propionic acid bacteria (PAB).
 These bacteria ferment glucose or lactate to propionic acid under anaerobic condition.
 Pyruvate reacts with methyl malonyl coA to form Propionyl coA and Oxaloacetate.
 Oxaloacetate give rise to malate, fumarate and succinate by reverse TCA cycle.
 Propionyl coA transfer its coA to succinate to form succinylcoA and propionate
 Example: Propioonibacterium
Flowchart: Generalized pathways for the production of some fermentation end products
from glucose by various organisms.
 Fermentation is an anaerobic biochemical process that is used for the production of
energy from the partial oxidation of glucose or other carbon sources.
 The oxidation of the substrate, which occurs through the Embden–Meyerhoff (EMP)
or Entner–Doudoroff(ED) pathways, results in the production of pyruvate, ATP, and
NAD (P) H.
 In the absence of external electron acceptors, the pyruvate undergoes reduction with
the regeneration of NAD+(P).
 This step is essential for the fermentation process to progress and it leads to the
production of products (ethanol and organic acids).
 ATP is the main product of fermentation, and it is generated by phosphorylation at the
substrate level.
 NADH is then re-oxidized, reverting to NAD+ in the second phase of fermentation
that reduces pyruvate to the fermentation product, such as ethanol and lactate.
 For example, in the fermentation of glucose by Streptococcus lactis, the pyruvate is
converted to lactic acid to reform NAD+ coenzymes so two ATP molecules are
produced
 In yeasts like Saccharomyces, when pyruvate is converted to ethyl alcohol (ethanol),
NAD+ is reformed.
Types of fermentation
1. Lactic acid homofermentation
Glucose → Lactic acid
 Homolactic fermentation is carried out by bacteria belonging to the
genera Lactococcus, Enterococcus, Streptococcus, and Pediococcus, and by some
species of the genus Lactobacillus.
 Homofermentative LAB ferment glucose to lactic acid.
 Lactococcus spp. is used in the dairy starter culture.
2. Lactic acid heterofermentation
Glucose → Lactic acid + Acetic acid + Ethyl alcohol + 2CO2 + H2O
 Heterolactic fermentation is carried out by bacteria of the
genera Leuconostoc, Oenococcus, and Weissella, and by
heterofermentative lactobacilli.
 Heterofermentative LAB ferment glucose with lactic acid, ethanol/acetic acid, and
carbon dioxide (CO2 ) as by-products.
3. Propionic acid fermentation
Glucose → Lactic acid + Propionic acid + Acetic acid + CO2 + H2O
 Propionic acid fermentation is carried out by several bacteria that belong to the
genus Propionibacterium and the species Clostridium propionicum.
 During propionic acid fermentation, both sugar and lactate can be used as the initial
substrate.
 When sugar is available, these bacteria use the EMP pathway to produce pyruvate; the
pyruvate is carboxylated to oxaloacetate and then reduced to propionate via malate,
fumarate, and succinate.
 The other end products of propionic fermentation are acetic acid and CO2.
4. Diacetyl and 2,3-butylene glycol fermentation
Diacetyl
↑
Citric acid→ Pyruvic acid + Acetylmethylcarbon
↓
2,3-Butylene glycol
 Butanediol fermentation is carried out by members of the genera Enterobacter,
Erwinia, Hafnia, Klebsiella, and Serratia.
  The reactions that lead to the production of 2,3-butanediol involve a double
decarboxylation step.
5. Alcoholic fermentation
Glucose → Ethyl alcohol
 Alcoholic fermentation is the best known of the fermentation processes.
 It is carried out by yeasts and some other fungi and bacteria.
 The first step of the alcoholic fermentation pathway involves pyruvate, which is
formed by yeast via the EMP pathway, while it is obtained through the ED pathway in
the case of Zymomonas (bacteria).
 The redox balance of alcoholic fermentation is achieved by the regeneration of
NAD+ during the reduction of acetaldehyde to ethanol.

Figure: Alcohol Fermentation. Created with biorender.com.

6. Butyric acid fermentation
Glucose → Acetic acid + Butyric acid
 Butyric acid fermentation is characteristic of several obligate anaerobic bacteria that
mainly belong to the genus Clostridium.
 Pyruvate is in turn oxidized to acetyl-CoA, with the production of CO2 and H2.
 Part of the acetyl-CoA is converted into acetic acid, with ATP production.
  Some bacteria, such as Clostridium acetobutylicum, produce fewer acids and more
neutral products, thus carrying out acetone butanol fermentation.
Applications of fermentation

Figure: Application of fermentation.

Application in medicine
 Production of antibiotics
 Production of insulin
 Production of growth hormones
 Production of vaccines
 Production of interferon
Application in the food industry
 Production of fermented foods as cheese, wine, beer, and bread to high-value products
 Food grade bio preservatives
  Functional foods/Neutraceuticals
 Production of single-cell protein
Other Applications
 It is also used for waste management such as biofuels production (biodiesels,
bioethanol, butanol, biohydrogen, etc).
 It is also used to produce bio-surfactant, polymers production such as bacterial
cellulose production.
 Development of bioremediation processes (involving microbes or their isolated
enzymes) for soils and wastewater treatments.

Fermentation in yeast
Yeasts
Yeasts are eukaryotic microorganisms that live in a wide variety of ecological niches, mainly
in water, soil, air and on plant and fruit surfaces. Perhaps the most interesting habitat at this
point is the latter, since they directly intervene in the decomposition of ripe fruit and
participate in the fermentation process. In this natural environment, yeasts can carry out their
metabolism and fermentation activity satisfactorily as they have the necessary nutrients and
substrates The production and tolerance to ethanol, organic acids and SO2 are also important
tools to differentiate among species. The reproduction of yeasts is mainly by budding, which
results in a new and genetically identical cell. Budding is the most common type of asexual
reproduction, although cell fission is a characteristic of yeasts belonging to the
genus Schizosaccharomyces. According to technological convenience, yeasts are divided into
two large groups namely Saccharomyces and non-Saccharomyces.
Morphologically, Saccharomyces yeasts can be round or ellipsoidal in shape depending on
the growth phase and cultivation conditions. S. cerevisiae is the most studied species and the
most utilized in the fermentation of wines and beers due to its satisfactory fermentative
capacity, rapid growth and easy adaptation. They tolerate concentrations of SO2 that
normally most non-Saccharomyces yeasts do not survive. However, despite these advantages,
it is possible to find in the nature representatives of S. cerevisiae that do not necessarily have
these characteristics.

Non-Saccharomyces Yeasts
Non-Saccharomyces yeasts are a group of microorganisms used in numerous fermentation
processes, since their high metabolic differences allow the synthesis of different final
products. Generally, many of these yeasts capable of modifying the sensory quality of wines
are considered as contaminants, so eliminating them or keeping them at low levels was a
basic objective in the past.In order to eliminate their activity in wine fermentation, it is usual
to disinfect the tanks and fermentation containers using sulfite. This perception has been
modified year after year, gaining relevance the action of these yeasts in the spontaneous
fermentation, since they contribute positively in the final sensory quality of the wine. These
yeasts are the majority in the initial phase of spontaneous fermentation to the point where the
concentration of ethanol reaches 4 and 5% v/v. At that point, between alcohol and the
exhaustion of dissolved oxygen, their growth is inhibited. When the process is
completed, Saccharomyces yeasts, the most resistant to ethanol, predominate and complete
the fermentation. It has been reported that some non-Saccharomyces yeasts are able to
survive toward the end of the spontaneous fermentation and exert their metabolic activity,
thus contributing positively to the sensory quality of wines. Based on this evidence, in recent
years, many researchers have focused their studies in understanding the nature and
fermentative activity of the non-Saccharomyces yeasts.The findings demonstrated the
enormous potential of these yeasts for use in the fermentation of traditional and
nontraditional beverages. Despite the fact that most non-Saccharomyces yeasts show some
technological disadvantages compared to S. cerevisiae such as lower fermentative power and
production of ethanol, non-Saccharomyces yeasts possess characteristics that in S.
cerevisiae are absent, for instance, production of high levels of aromatic compounds such as
esters, higher alcohols and fatty acids.
Yeast Fermentation Processes
2.1. Alcoholic Fermentations
The production of alcoholic beverages from fermentable carbon sources by yeast is the oldest
and most economically important of all biotechnologies. Yeast plays a vital role in the
production of all alcoholic beverages. Yeast plays a vital role in the production of all
alcoholic beverages and the selection of suitable yeast strains is essential not only to
maximise alcohol yield, but also to maintain beverage sensory quality
Wine Fermentation
In wine fermentation, strains with specific characteristics are needed, for instance, highly
producers of ethanol to reach values of 11–13% v/v, typically found in this beverage. On the
other hand, beers and ciders contain less amounts of ethanol with a balanced and distinctive
sensory profile characteristic of each one. In recent years, new consuming trends and
requirements for new and innovative products have emerged. This situation led to rethink
about the existing fermented beverages and to meet the demands of consumers. Yeasts are
largely responsible for the complexity and sensory quality of fermented beverages. Based on
this, current studies are mainly focused on the search of new type of yeasts with technological
application. Non-Saccharomyces yeasts have always been considered contaminants in the
manufacture of wine and beer. Therefore, procedures for eliminating them are routinely
utilized such as must pasteurization, addition of sulfite and sanitization of equipment and
processing halls. In recent years, the negative perception about non-Saccharomyces yeasts
has been changing due to the fact that several studies have shown that during spontaneous
fermentations of wine, these yeasts play an important role in the definition of the sensory
quality of the final product. Based on this evidence, the fermentative behavior of some non-
Saccharomyces yeasts is being studied in deep with the purpose of finding the most adequate
conditions and the most suitable strain to be utilized in the production of fermented
beverages.
Beer Fermentation
Beer is the most consumed alcoholic beverage worldwide. It is traditionally made from four
key ingredients: malted cereals (barley or other), water, hops, and yeast. Each of these
ingredients contributes to the final taste and aroma of beer. During fermentation, yeast cells
convert cereal-derived sugars into ethanol and CO2. At the same time, hundreds of secondary
metabolites that influence the aroma and taste of beer are produced. Variation in these
metabolites across different yeast strains is what allows yeast to so uniquely influence beer
flavor
Although most breweries use pure yeast cultures for fermentation, spontaneous or mixed
fermentation is nowadays used for some specialty beers. These fermentation procedures
involve a mix of different yeast species (and bacteria as well) that contribute to the final
product sequentially, giving the beer a high degree of complexity. Commonly, breweries have
their own stock of selected yeasts for their specific beers. As it is well-known, two types of
yeast are used in brewing: S. cerevisiae as the top-fermenting yeast to make ales while S.
pastorianus is a bottom-fermenting yeast used in lager brewing processes.
Cider Fermentation
Cider is another alcoholic beverage derived from the apple fruit industry, very popular in
different countries in the world, mainly Europe, North America, and Australia . Although
traditional ciders are produced from spontaneous fermentation of juice carried out by
autochthonous yeasts, selected S. cerevisiae strains are also commonly used to carry out
alcoholic fermentation. This ensures a consistent quality of the finished products . Some other
non-Saccharomyces yeast species are involved in spontaneous fermentation of apple juice for
cider production. However, these yeasts contribute at a lesser extent than Saccharomyces and
can be producers of off-flavours.
Non-Alcoholic Fermentations
Moreover, yeast can act in the fermentation of global non-alcoholic products (bread,
chocolate or coffee, beverages such as kefir, sodas, lemonades, and vinegar or even biofuels
and other chemicals.
2.2.1. Bread Fermentation
The fermentation of the dough made by the yeasts is the most critical phase in the making of
bread. The fermentative yield of yeast cells during this fermentation is crucial and determines
the final quality of the bread. Yeasts not only produce CO2 and other metabolites that
influence the final appearance of the dough, volume, and texture, and of course, the taste of
the bread. The yeast strain, pregrowth conditions, its activity during the dough fermentation
process, the fermentation conditions, as well as the dough ingredients are basic to control the
process. The fermentation rate is also conditioned by the ingredients of the dough, including
the amounts of sugar and salt used in its preparation. Commercial bread producers currently
produce various types of dough such as lean, sweet or frozen dough. Depending on the type
of dough, and to obtain optimal fermentation rates, it is recommended to use suitable yeast
strains with specific phenotypic traits
2.2.2. Coffee Fermentation
Yeasts play an important role in coffee production, in the post-harvest phase. Its performance
can be done in two phases. On the one hand, aerobically, in which the berries just collected
are deposited in a tank and the yeasts are allowed to act. This process is carried out under
control of basic parameters, such as time and temperature. Alternatively, coffee berries are
deposited in a container mixed with water and microorganisms are allowed to act
anaerobically (in the absence of oxygen). This second process is more homogeneous and easy
to control than the aerobic. Sometimes, coffee beans are even fermented in a mixed process,
first in an aerobic and finally anaerobic manner . To develop these processes in a satisfactory
manner, and to preserve/improve the organoleptic properties of coffee, refine its sweetness,
control acidity, give them body or add sensory notes (chocolate, caramel, fruits) mucilage
should be removed. The process is naturally carried out by the yeasts present in the mixture,
although the process can be improved by the addition of appropriate enzymes
(polygalacturonase, pectin lyase, pectin methylesterase)
Chocolate Fermentation
Raw cacao beans have a bitter and astringent taste, because of high phenolic content.
Anthocyanins are one group of these polyphenols, and it both contributes to astringency and
provide the reddish-purple color. Fermentation allows the enzymatic breakdown of proteins
and carbohydrates inside the bean, creating flavor development. This is aided by microbial
fermentation, which create the perfect environment through the fermentation of the cacao
pulp surrounding the beans. This processing step enables the extraction of flavor from cacao
and contributes to the final acidity of the final product. Yeasts (and also bacteria) ferment the
juicy pulp among the cacao beans by different methods, generally following a an anaerobic
phase and an aerobic phase. During the anaerobic phase, the sugars of the pulp (sucrose,
glucose, fructose) are consumed by yeasts using anaerobic respiration to yield carbon
dioxide, ethanol, and low amounts of energy.
Role and Applications of Fungi in Food and Fermentation Technology
Fungi are single- or multicelled eukaryotic organisms such as yeasts, moulds and mushrooms
that play a major role in food and fermentation industry. Various fungi are used as an
important component in the preparation of a range of comestible products such as leavened
and unleavened breads, beverages, probiotics, cheese, tempeh, miso, amino acids, food
pigments, enzymes, organic acids, etc. Fungi, namely, Saccharomyces spp., Penicillium spp.
and Aspergillus spp., are the integral part of industrial production of beverages and bakery
products, organic acids and soy-based fermented foods, respectively. Yeast-mediated
alcoholic fermentation is one of the oldest known biological processes that has been known
for ages. However, there is an apparent paradigm shift in the earlier approach, and more
attention is on the production, proliferation and secretion of extracellular enzymes, secondary
metabolites, peptides of novel functions and antibiotics in alleviating the infectious diseases
from fungi. Several fungal probiotics exert nutraceutical effects by folate biofortification and
biodegradation and absorption of mycotoxins. This chapter presents how fungi are helpful in
producing various food and feed products with a special focus on the recent studies related to
fungal applications in food preservation, sensory changes and current knowledge about
mechanism of action in food industry.
 Traditional vs Modern Biotechnology
Traditional biotechnology refers to the utilisation of live creatures to generate new
things or transform foods and other objects for human use. DNA Fragments, structural
and functional genomes, DNA diagnostic probes, and other genetic altering procedures
are employed in contemporary biotechnology.Traditional biotechnology refers to a
variety of old methods for creating new products or modifying current ones by utilizing
live organisms.
Modern biotechnology encompasses a variety of procedures that entail the deliberate
and controlled manipulation of genes, cells, and live tissue to induce changes in an
organism’s genetic makeup or develop new tissues.
Modern biotechnology includes DNA profiling, genome analysis, transgenesis, DNA
cloning, and the manipulation of genes and living tissues in a controlled environment.
Comparison Table

Parameter of
Traditional Biotechnology Modern Biotechnology
Comparison

Modern biotechnology encompasses a variety of

Traditional biotechnology refers to a
procedures that entail the deliberate and
variety of old methods for creating new
Definition controlled manipulation of genes, cells, and live
products or modifying current ones by
tissue to induce changes in an organism’s
utilizing live organisms.
genetic make-up or develop new tissues.

Traditional biotechnology includes tissue Modern biotechnology includes DNA profiling,

Examples
culture, mutagenesis genome analysis, transgenesis, DNA cloning

Cheese, yoghurt, bread, beer, and wine

Modern biotechnology is used in a variety of
are all made through breeding animals
industries, including food, agriculture, forestry,
Uses and crops. To generate enzymes,
wellness, ecology, minerals, and industrial
good bacteria are used. Enzymes are
activities.
then used in washing powder.

Genetic engineering is the process of

using natural organisms to produce new Genes and live tissues are manipulated in a lab
Involves
foods or improve existing ones for setting.
human consumption.

It was by chance that humans discovered In 1953, scientists discovered the structure of
hundreds of years ago how to use DNA and the transfer of genetic knowledge
Discovery
natural processes that occur constantly from one person to the next, ushering in the
within living cells for their benefit. modern world of biotechnology
What is Traditional Biotechnology?
People have been making beverages, curd, cheese, cider, wine, and bread without recognising
that microorganisms have been involved in the process since the ancient period.
Yeasts, moulds, and lactic acid bacteria have been used to preserve milk, fruits, and
vegetables for ages, and the products have improved people’s lives.
After the First World War, microorganisms were employed for the first time to generate
organic molecules such as citric acid.
Through the years, people discovered how to use natural processes that occur constantly in all
living organisms, including humans.
Even though they didn’t have a scientific explanation for the processes, they used what they
learned in the workplace in their personal life.
For example, they determined that when food matures, it changes in flavour and content
while also becoming less tasteless.
Organic fermentation may be utilised to create bread from wheat flour, wine from grape juice,
and cheese from milk stored in camel belly bags.
Traditional biotechnology can also include cell culture products, segments and sub, or
different disease-eradication tactics, but current biotechnology has a special focus on industry
use of rDNA, cell fusion, and new bioprocessing processes.
For the most critical agronomic characteristics, traditional biotechnology is still the best
option. It is due to its easy way and fruitful results with tons of research already done.
What is Modern Biotechnology?
Biotechnology, in its contemporary definition, refers to the application of newly gained
abilities in microbiological and biochemical technology to applied biology, i.e., the utilization
of biological systems and processes for our benefit.
biotechnology also refers to industrial procedures that involve the utilization of live cells or
their derivatives.
Numerous early and ongoing biotechnological applications employ microbes as agents in
certain processes—some natural, some induced—or as sources of valuable products, such as
fermentation technology for the manufacture of wine and liquors.
These breakthroughs paved the way for the evolution of biotechnology from the conventional
to the contemporary.
In comparison to previous biotechnology approaches, they allow for the regulated and faster
production of desired alterations in an organism by direct manipulation of its genes.
These breakthroughs, along with developments in technology and science (such as
biochemistry and physiology), opened the door to novel biotechnology applications
previously unimaginable.
Soil, water, and air pollution can be cleaned up using organisms or organism components. In
New Zealand, bioremediation has been advocated as a viable option for eliminating the
pesticide DDT from the groundwater.
The modern field of biotechnology is built on recombinant DNA technology. Modern
biotechnology makes use of GE methods, including DNA diagnostics tools, gene Cloning,
and structural and functional biometrics for genetic manipulation.
Main differences between Traditional Biotechnology and Modern Biotechnology
1. Traditional biotechnology refers to a variety of old methods for creating new products
or modifying current ones by utilizing live organisms, whereas Modern biotechnology
encompasses a variety of procedures that entail the deliberate and controlled
manipulation of genes, cells, and live tissue to induce changes in an organism’s
genetic make-up or develop new tissue.
2. Traditional biotechnology includes tissue culture, mutagenesis whereas Modern
biotechnology includes DNA profiling, genome analysis, transgenesis, DNA cloning
3. Traditional biotechnology makes use of biological ecosystems to generate or change
food or other valuable items for human consumption, whereas Modern Biotechnology
manipulates genes and live tissues in a sterile environment.
4. While in traditional biotechnology, people discovered centuries ago how to exploit the
organic processes that constantly occur within living tissue by chance, the structure of
deoxyribonucleic acid (DNA) and the transmission of genetic information from one
generation to the next were discovered in 1953, and it was the beginning of modern
biotechnology.
5. Traditional biotechnology involves breeding animals and crops to make cheese,
yoghurt, wheat, lager, and wine, but modern biotechnology applies current
biotechnology methods to food, farming, forestry, medicine, ecology, minerals, and
industrial processes, as well as food.
Process flow sheeting – block diagrams, pictorial representation in fermentation
biotechnology
In fermentation biotechnology, process flow sheeting visually represents the steps involved in
the fermentation process. A concise overview using both block diagrams and pictorial
representations.
Block Diagram
1. Inoculum Preparation: Culturing microorganisms for fermentation.
2. Fermentation: Microorganisms convert substrates into products in a bioreactor.
3. Harvesting: Separating cells from the fermentation broth.
4. Product Recovery: Extracting and purifying the desired product.
5. Waste Treatment: Treating and disposing of waste materials.
Inoculum Preparation → Fermentation → Harvesting → Product Recovery → Waste
Treatment
Pictorial Representation
1. Inoculum Preparation: Illustrated with a bioreactor where microorganisms are
cultured.
2. Fermentation: Shown as a larger bioreactor where substrate and microorganisms
grow.
3. Harvesting: Depicted with centrifuge or filtration equipment for cell separation.
4. Product Recovery: Shown with distillation or chromatography equipment for
purification.
5. Waste Treatment: Illustrated with treatment tanks for waste management.
Example:
In a biopharmaceutical process, yeast cells are cultured (Inoculum Preparation) in a
bioreactor. The fermentation process involves converting sugars into ethanol and other by-
products. Cells are harvested using centrifugation. Ethanol is recovered and purified via
distillation, while waste is treated and disposed of responsibly.
I hope this gives you a clear snapshot of the fermentation biotechnology process! If you need
more details, feel free to ask.
Process flow sheeting – block diagrams, pictorial representation in fermentation
biotechnology with example in 1000 words with sample diagram
Certainly! Let's dive deeper into the process flow sheeting in fermentation biotechnology
with detailed block diagrams and pictorial representations, including an example with a
sample diagram.
Process Flow Sheeting in Fermentation Biotechnology
Overview
Fermentation biotechnology involves the use of microorganisms to convert substrates into
valuable products. This process is commonly used in the production of pharmaceuticals,
biofuels, food products, and industrial chemicals. A process flow sheet visually represents the
various stages of the fermentation process, helping to understand and optimize the flow of
materials, energy, and information.
Block Diagrams
Block diagrams are simplified representations of the process flow, showing major steps and
the connections between them. Each block represents a specific process step, while arrows
indicate the flow of materials or energy.
Key Stages in Fermentation
1. Inoculum Preparation
 2. Fermentation
3. Harvesting
4. Product Recovery
5. Waste Treatment
Block Diagram Example
Here’s a detailed block diagram for a typical fermentation process:
Inoculum Preparation → Fermentation → Harvesting → Product Recovery → Waste
Treatment
 Inoculum Preparation: Culturing the microorganisms to be used in fermentation.
 Fermentation: The main process where microorganisms convert substrates into
desired products.
 Harvesting: Separating the microbial cells from the fermentation broth.
 Product Recovery: Extracting and purifying the desired product from the
fermentation broth.
 Waste Treatment: Treating and disposing of waste materials generated during the
process.
Pictorial Representation
Pictorial representation provides a more detailed and realistic view of the process, including
images of equipment and detailed flow paths.
Components of Pictorial Representation
1. Inoculum Preparation: Illustration of a bioreactor where microorganisms are
cultured.
2. Fermentation: A larger bioreactor where substrates and microorganisms interact.
3. Harvesting: Centrifuge or filtration equipment for separating cells from the broth.
4. Product Recovery: Distillation or chromatography equipment for purification.
5. Waste Treatment: Treatment tanks or systems for handling waste.
Example: Ethanol Production from Yeast Fermentation
Let’s take the example of ethanol production using yeast fermentation to create a process
flow sheet.
Inoculum Preparation
In this stage, yeast cells are cultured in a bioreactor with a nutrient-rich medium. The cultured
yeast serves as the inoculum for the main fermentation process.
Fermentation
The inoculum is transferred to a larger bioreactor containing the substrate (such as sugarcane
or corn). Yeast cells convert the sugars into ethanol and carbon dioxide under controlled
conditions.
Harvesting
After fermentation, the microbial cells (yeast) are separated from the fermentation broth
using centrifugation or filtration. This step ensures that only the fermentation broth,
containing ethanol, proceeds to the next stage.
Product Recovery
The fermentation broth, now separated from the yeast cells, is subjected to distillation.
Distillation helps in purifying ethanol from the broth. Additional purification steps like
molecular sieve or rectification columns may be used to achieve the desired ethanol purity.
Waste Treatment
The residual biomass and waste products from the fermentation and recovery processes are
treated in waste treatment systems. This may include anaerobic digestion, aerobic treatment,
or other waste management techniques to ensure environmentally responsible disposal.
Sample Diagram
Below is a sample diagram representing the ethanol production process through yeast
fermentation.
Detailed Description of Each Stage
1. Inoculum Preparation
o Objective: To cultivate a high concentration of active yeast cells.
o Equipment: Small bioreactors, shaker flasks.
o Process: Sterilized growth medium is inoculated with yeast and incubated
under optimal conditions (temperature, pH, aeration).
2. Fermentation
o Objective: To convert sugars into ethanol and carbon dioxide.
o Equipment: Large bioreactors with temperature, pH, and agitation control.
o Process: The inoculum is added to the substrate-containing bioreactor.
Fermentation conditions are maintained to maximize ethanol production.
3. Harvesting
o Objective: To separate yeast cells from the fermentation broth.
o Equipment: Centrifuges, filtration units.
o Process: The fermentation broth is passed through centrifugation or filtration
systems to remove yeast cells, leaving behind the ethanol-rich broth.
 4. Product Recovery
o Objective: To purify ethanol from the fermentation broth.
o Equipment: Distillation columns, molecular sieves, rectification columns.
o Process: The broth is distilled to separate ethanol from water and other
impurities. Further purification is done if required to achieve high-purity
ethanol.
5. Waste Treatment
o Objective: To treat and dispose of waste materials responsibly.
o Equipment: Anaerobic digesters, aerobic treatment systems.
o Process: Waste materials, including residual biomass and by-products, are
treated to reduce environmental impact. Treated waste is then safely disposed
of or repurposed.
Conclusion
Process flow sheeting in fermentation biotechnology is a critical tool for visualizing and
optimizing the production process. Both block diagrams and pictorial representations offer
valuable insights into the flow of materials, energy, and information. By breaking down each
stage and using detailed illustrations, we can ensure a clear and comprehensive understanding
of the fermentation process, ultimately leading to improved efficiency and productivity.
Sample pictorial representation
Sample block diagram