TYPES OF

FERMENTATIONS
SUBMITTED BY-PRACHI PANWAR
ROLL NO-19001554014
MSC BIOTECHNOLOGY,4TH SEMESTER
SUBMITTED TO-DR.MANJEET K.SEHWAL
INTRODUCTION TO FERMENTATION

• Fermentation is a metabolic process that produces chemical changes in organic substrates through the action of enzymes.
In biochemistry, it is narrowly defined as the extraction of energy from carbohydrates in the absence of oxygen. In food
production, it may more broadly refer to any process in which the activity of microorganisms brings about a desirable
change to a foodstuff or beverage. The science of fermentation is known as zymology.
• In microorganisms, fermentation is the primary means of producing adenosine triphosphate (ATP) by the degradation of
organic nutrients anaerobically. Humans have used fermentation to produce foodstuffs and beverages since
the Neolithic age. For example, fermentation is used for preservation in a process that produces lactic acid found in such
sour foods as pickled cucumbers, kombucha, kimchi, and yogurt, as well as for producing alcoholic beverages such as
wine and beer. Fermentation also occurs within the gastrointestinal tracts of all animals, including humans.
• Fermentation has been widely used for the production of a wide variety of substances that are highly beneficial to
individuals and industry.
• Over the years, fermentation techniques have gained immense importance due to their economic and Environmental
advantages.
• Ancient techniques have been further modified and refined to maximize productivity. This has also involved the
development of new machinery and processes.
• Two broad fermentation techniques have emerged as a result of this rapid development: Solid State Fermentation (SSF).
Submerged Fermentation (SmF).
• At the research level, both SSF and SmF have been used;however, some techniques yielded better results than others.
SOLID STATE FERMENTATION
• Solid-state fermentation (SSF) is a common technique for the production of microbial metabolites .
• The process of SSF is performed on a solid substrate with a low moisture content, with the advantages of a high product
concentration but only a relatively low energy being required .
• The required water content in SSF is absorbed by the substrate in a solid matrix and offers more advantages for the growth
of microorganism for the transfer of oxygen.
• Several agricultural wastes, such as rice straw, sugarcane bagasse, wheat straw, rice hulls, and corn cobs, are being used as
substrate for SSF.
• Industrially, metabolites production is mostly carried out under submerged fermentation (SmF) conditions due to the
simpler downstreaming process of SmF compared to SSF .
• In the case of SmF, there is a possibility of strong and simple control over the various parameters, whereas there is a
knowledge gap in the SSF process. Although SmF is an easier process in terms of operating parameters than SSF, the
microbial mycelial morphology is better suited for microbial secondary metabolites production in SSF.
• Thus SSF exhibits several advantages, like easy gaseous transportation, less water consumption, use of cellulosic waste,
pH control, and smaller fermenters are required for SSF and thus less effort is needed for downstream processing.
• However, SSF technology is needed to be explored in order to improve production of metabolites at a large scale .
• During the last 18 years, much research have been devoted either in the form of research/review articles or books on SSF
and the recovery of microbial metabolites, due to the fact that this is a well-suited process of microbial metabolism for the
production of various value-added metabolites 
• Apart from this, pollution causing solid wastes, which are rich source of cellulose, protein may be utilized as substrates
for SSF. Specifically, Japan is frequently using SSF for the production of various products such as
enzymes, amylase, protease, and soya sauce.
• Further, much SSF-related equipment has been designed and adapted well in Japan for the production of various
industrially important microbial metabolites .
• Additionally, in India Biocon Company started the SSF process for enzyme production more than 20 years ago.
• Though SSF is an easy process to conduct and has several advantages, the process still suffers from certain drawbacks
which need to be overcome for large-scale processing. It has been noted that SSF is slower than SmF with difficult
downstreaming properties.
• Moreover, the SSF process also has difficulties with heat dissipation, which can be limited by inter- and intraparticle
conflicts and hence are uncontrolled because of the absence of satisfactory sensors and supportive solid handling
techniques for continuous operations .
• In addition, various issues such as an efficient process to improve SSF, bioreactor design, and sustainability are some
critical limiting factors for SSF which need to be tailored based on the existing advancements in SSF technique. Fig.
23.2 explains the bioethonal production process via SSF route.
Selection of Micro-organism
• This is one of the key factor for improved yields of the product. Bacteria, Yeast and Filamentous Fungi can be used.
• Filamentous Fungi has shown better results growing in the solid substrate fermentation.

Substrate
• Substrate also plays important role in determining the growth of micro- organisms, there by increasing the product yield.
• Substrate is chosen such a way that it should provide physical support as well as nutrients to the growing culture.
• There are many biotechnological processes that involve the growth of organisms on solid substrates in the absence or near
absence of free water.
• Solid state fermentation (SSF) deals with substrates that are solid and contain low moisture levels.
• The most regularly used solid substrates are cereal grains (rice, wheat, barley, and corn), legume seeds, wheat bran,
lignocellulose materials such as straws, sawdust or wood shavings, and a wide range of plant and animal materials.
• Most of these compounds are polymeric molecules – insoluble or sparingly soluble in water – but most are cheap and
easily obtainable and represent a concentrated source of nutrients for microbial growth.
Figure: Schematic of solid state fermenter for conversion of lignocellulosic
biomass to enzymes. Source: Amit K. Jaiswal.
Organisms Used in Solid State Fermentation (SSF)
• The microbiological components of SSF can occur as single pure cultures, mixed identifiable cultures or totally mixed
indigenous microorganisms.
• Some SSF processes e.g., tempeh and ontjom production, requires selective growth of organisms such as molds that need
low moisture levels to carry out fermentation with the help of extracellular enzymes secreted by fermenting
microorganisms.
• However, bacteria and yeasts, which require higher moisture content for efficient fermentation, can also be used for SSF,
but with a lower yield.
Steps in Solid State Fermentation (SSF)
SSF is normally multistep processes involving the following steps:
1. Pre-treatment of substrate raw materials either by mechanical, chemical or biochemical processing to enhance the
availability of the bound nutrients and also to reduce the size of the components, e.g., pulverizing straw and shredding
vegetable materials to optimize the physical aspects of the process. However, the cost of pre-treatment must be balanced with
the eventual product value.
2. Hydrolysis of primarily polymeric substrates, e.g., polysaccharides and proteins.
3. Utilization (fermentation) of hydrolysis products.
4. Separation and purification of end products.
The low moisture content of SSF enables a smaller reactor volume per substrate mass than LSF and also simplifies product
recovery.
However, serious problems arise with respect to mixing, heat exchange, oxygen transfer, moisture control and gradients of
pH, nutrient and product as a consequence of the heterogeneity of the culture.
The latter characteristic of SSF renders the measurement and control of the above-mentioned parameters difficult, laborious
and often inaccurate, thereby limiting the industrial potential of this technology. Due to these problems, the micro-organisms
that have been selected for SSF are more tolerant to a wide range of cultivation conditions.
Applications of Solid State Fermentation (SSF)
• Solid-state fermentation has emerged as a potential technology for the production of microbial products such as feed, fuel,
food, industrial chemicals, and pharmaceutical products.
• It is widely applied to producing several enzymes, organic acids, flavoring compounds etc., which must be extracted and
purified and then used in different products.
• Its application in bioprocesses such as bioleaching, bio-beneficiation, bioremediation, bio-pulping, etc. has offered several
advantages.
Advantages of Solid State Fermentation (SSF)
•The main advantage of such methods is that it produces a minimum amount of waste and liquid effluent thus not very
damaging to the environment.
•Solid substrate fermentation employs simple natural solids as the media.
•Low technology, low energy expenditure and requires less capital investment.
•No need for sterilization, less microbial contamination, and easy downstream processing.
•Utilization of agro-industrial residues as substrates in SSF processes provides an alternative avenue and value-addition to
these otherwise under- or non-utilized residues. 
•The yield of the products is reasonably high.
•Bioreactor design, aeration process, and effluent treatment are quite simple.
•Many domestic, industrial and agricultural wastes can be fruitfully used in SSF.
Limitations of Solid State Fermentation (SSF)
• The microorganisms that tolerate only low moisture content can be used.
• Precise monitoring of SSF (e.g., O2 and CO2 levels, moisture content) is not possible.
• The organisms grow slowly and consequently, there is a limitation in product formation.
• Heat production creates problems, and it is very difficult to regulate the growth environment.  
Submerged Liquid Fermentations
• Submerged production began in the 1930’s and is the main method used today.
• Submerged liquid fermentations are traditionally used for the production of microbially derived enzymes.
• In the submerged process, the substrate used for fermentation is always in liquid state which contains the nutrients
needed for growth.
• The fermentor which contains the substrate is operated and the product biomass is harvested from the fermenter by using
different techniques then the product is filtered or centrifuged and then dried.
• Submerged fermentation is a method of manufacturing biomolecules in which enzymes and other reactive compounds
are submerged in a liquid such as alcohol, oil or a nutrient broth
• The process is used for a variety of purposes, mostly in industrial manufacturing
• The process can be used to make products such as citric acid, glycerol or lactic acid.
• Submerged culture fermentation has been widely used for the production of enzyme because in submerged fermentation
unwanted metabolites are not produced and purification of enzymes takes place in an easy way
• Submerged fermentation involves submersion of the microorganism in an aqueous solution containing all the nutrients
needed for growth.
• Fermentation takes place in large vessels (fermenter) with volumes of up to 1,000 cubic meters.
• The fermentation media sterilizes nutrients based on renewable raw materials like maize, sugars, and soya.
• Most industrial enzymes are secreted by microorganisms into the fermentation medium in order to break down the
carbon and nitrogen sources.
• Batch-fed and continuous fermentation processes are common.
• In the batch-fed process, sterilized nutrients are added to the fermenter during the growth of the biomass.
• In the continuous process, sterilized liquid nutrients are fed into the fermenter at the same flow rate as the fermentation
broth leaving the system.
• Parameters like temperature, pH, oxygen consumption and carbon dioxide formation are measured and controlled to
optimize the fermentation process.
• Next in harvesting enzymes from the fermentation medium one must remove insoluble products, e.g. microbial cells.
This is normally done by centrifugation.
• As most industrial enzymes are extracellular (secreted by cells into the external environment), they remain in the
fermented broth after the biomass has been removed.
• The enzymes in the remaining broth are then concentrated by evaporation, membrane filtration or crystallization
depending on their intended application.
• If pure enzyme preparations are required, they are usually isolated by gel or ion exchange chromatography.
• Several types of submerged type of fermentors are known and they may be grouped in several ways: shape or
configuration, whether aerated or anaerobic and whether they are batch or continuous.
• The most commonly used type of fermentor is the Aerated Stirred Tank Batch Fermentor.
Aerated stirred tank batch fermentor
• A typical fermentor of this type is an upright closed cylindrical tank fitted with one or more baffles attached to the side of
the wall, a water jacket or coil for heating and/ or cooling, a device for forcible aeration (known as sparger), a
mechanical agitator usually carrying a pair or more impellers, means of introducing organisms and nutrients and of
taking samples, and outlets for exhaust gases.
• Modern fermentors are highly automated and usually have means of continuously monitoring, controlling or recording
pH, oxidation-reduction potential, dissolved oxygen, effluent O2, and CO2, and chemical components.
SUBMERGED FERMANTATOR
Principle of Submerged Fermentation
• Submerged fermentation involves the growth of the microorganism as a suspension in a liquid medium in which various
nutrients are either dissolved or suspended as particulate solids in many commercial media.
• Submerged fermentation is a process involving the development of microorganisms in a liquid broth.
• This liquid broth contains nutrients and it results in the production of industrial enzymes, antibiotics or other products.
• The process involves taking a specific microorganism such as fungi and placing it in a small closed flask containing the
rich nutrient broth.
• A high volume of oxygen is also required for the process. The production of enzymes then occurs when the
microorganisms interact with the nutrients on the broth resulting in them being broken down. 
• The bioactive compounds are secreted into the fermentation broth.
Methods of Carrying Out Submerged Fermentation
• There are two common methods by which submerged fermentation takes place; they are batch-fed fermentation and
continuous fermentation.
• In batch-fed fermentation sterilized growth nutrients are added to the culture. It is most common in bio-industries as it
occurs during the growth of biomass in the fermenter. It helps raise the cell density in the bioreactor and it is typically
highly concentrated to stop dilution. The rate of growth in the culture is maintained by adding nutrients, this also reduces
the risk of overflow metabolism.
• An open system is constructed for continuous fermentation. Then sterilized liquid nutrients are slowly and continuously
added to the bioreactor at the same rate at which the converted nutrient solution is being recovered from the system. This
results in a steady-rate production of the fermentation broth.
• In order to maintain a successful fermentation, certain variables must be monitored, for example, temperature, pH, as well
as oxygen and carbon dioxide levels.
Submerged fermentation substrate
• Some common substrates used in submerged fermentation are soluble sugars, molasses, liquid media, fruit and vegetable
juices, and sewage/wastewater.
Applications of Submerged Fermentation
• SmF is primarily used in the extraction of secondary metabolites that need to be used in liquid form.
• Submerged liquid fermentations are traditionally used for the production of microbially derived enzymes.
Advantages of Submerged Fermentation
• Measure of process parameters is easier than with solid-state fermentation.
• Bacterial and yeast cells are evenly distributed throughout the medium.
• There is a high water content which is ideal for bacteria.
• Inoculum ration is usually small
• Lower total investment costs
• Improved process control
• Reduced fermentation time
• Reduced floor space requirements
• Purification of products is easier
• Lower labor costs
• Simpler operations
• Easier maintenance of aseptic conditions on an industrial scale .
Disadvantages of submerged fermentation:
• High costs due to the expensive media.
• Expenses for equipment are higher
• Consumption of electrical energy is higher
• The process is very sensitive
• Agitation is often essential
• Chance of contamination