Course on Industrial Biotechnology

Prof. Debabrata Das

Department of Biotechnology
Indian Institute of Technology Kharagpur
Mod01 Lecture01
Introduction to Industrial Biotechnology

Course that I am going to offer that is industrial biotechnology. It has two components, one is
biotechnology another is a industrial. Biotechnology again it has the two parts, one is bio
another is technology as you that technology basically it is combination of science and
engineering. Science tells why it is happening? How it is happening and engineering is the
application of science. Basically whatever we develop in the day to day life and when we go
for the application the both the combination of science and engineering is required that (())
(0:58) is call technology and bio stands for biological component, if the technology comes
through with the help of some biological component we call in biotechnology. An industrial
biotechnology basically it is what is in practice in the industry. There is industrial
biotechnology.

(Refer Slide Time: 1:31)

Now if you look at the definition of industrial biotechnology, is the set of practices, because
what are the different practices we have in the industry to get a purified products as, for
example, suppose we want to here what we used the living cells, bacteria, yeast different
components, not only the living cells we can use some non-living components also, which is
derive from the living of organism as for example, enzymes which is derive from the both
plant system, microbial system, animal system which is use for the production of different
products.

(Refer Slide Time: 2:02)

Now industrial biotechnology, it has the it comprises of different components, one is medium
preparation and then inoculum preparation using industrial strain. medium preparation plays
very important role as we know that media comprises of not only carbon not only a single
component, it comprises of carbon source, nitrogen source, minerals and vitamin for the
living cells, but for the non-living cells, suppose if we use enzyme we required it particular
state, particular pH and temperature to have a particular reaction to get the definite products.

The inoculum preparation usually the industry we use the industrial strain, it is not the this is
the specially prepared for the industry and it is it is we consider as a robust strain(())(2:57).
Why it is robust strain (())(2:59), because it can has high productivity and I shall discuss
more in details in subsequence slides, then it comes to the fermentation process.

Fermentation process is the actually that is when here organism that you know function it has
the metabolic pathways (())(3:17) and it produces the (())(3:20) we through the metabolic
pathway (())(3:21) the different products and the science of fermentation basically it also
known as zymology and then when you get the desired amount desired food and then we go
for some physico-chemical separation process to get the purified product when purified
product whatever product we marketed that should be in the purified form, then after getting
this purified product then it undergo packaging where and then for the this is packaging.
Now with this packaging system has been proved (())(3:54) extent for marketing purpose and
after that you know that the during the product formation we also generate lot of organic
waste that is to be treated. So you know what we call effluent treatment process that we have.

(Refer Slide Time: 4:11)

Now I want to give typical example of the brewing industry, because throughout the world
that you know different brewing processes are in practice, we know that we have (())(4:23)
we have (())(4:24) in India we produce that allibear (())(4:27) and it comprises the different
stage. So as for example molting, then milling then massing then lautering then you have this
boiling then you have then whill pooling then cooling fermentation, maturing, filtration then
packaging.

So what I want to mean that if you look at the this is the fermentation process, this is the what
we you call brewery process and this is before material comes to the brewery process. There
are so many units involved we have the molting process, we have milling process, we have
massing process, we have lautering process, we have boiling process, we have whill pool the
pooling that is process (())(5:14) then we have cooling then it comes to the fermentation.
After fermentation it goes to the different separation process (())(5:22) this is call maturation
process then we have filtering then packaging can be done in different ways either in the can,
in the bottle, cellular barrels (())(5:33) and then it is marketing with the help of some kind of
transport arrangements.
(Refer Slide Time: 5:45)

Now this the course I divided into different parts. Now initially I shall restricted myself on
microbe the microbes and enzymes for the industrial importance as I have give you some
detail information on that, then I shall discuss about different types of bioreactors and
bioreactor design and this would be followed by tutorial 1 and stoichiometry is very
important part of any kind of fermentation process, because, because the stoichiometry give
the material analogy analysis of the system then microbial growths, substrate degradation and
product formation kinetics and this will be followed by instrumentation and sterilization of
air, media and reactor. Then we I shall discuss the upstream and down-stream processing.

I come (())(6:36) for biochemical block fermentation processes like tax and non-tax alcohol,
then brewing industry, wine making industry, vinegars, citric-acid production, penicillin,
streptomycin, high fructose corn syrup then we have cheese making then single cell protein,
vaccine production and metal leaching. We also try to cover this bioenergy, because
bioenergy is very important part. Nowadays in because replacing the conventional fossil fuels
bioenergical is very important also different bioenergy that is this produce where you in
practice nowadays, who are one use the bio methane and another is bioethanol, there largely
used biodiesel also used in (())(7:22) industry, I shall discussed about that how it produced
and finally I shall discuss about the aerobic and un-aerobic waste water treatment process.

(Refer Slide Time: 7:33)

Now how question comes that what are the bio product? Bio production of commercially
useful product means from the use of biological as for example microbes and enzymes or
renewable materials like biomass is derived from the agricultural residues or food processing.
So the different types of microbes and enzyme catalyze the various reaction what about the
generation of bio products. So we have bacteria, we have algae, we have fungi, we have yeast
that can involve to give produce different type of bio products then we have the enzyme that
catalyze (())(8:11) is enzyme(())(8:11) that the that acts on the hydrogen peroxide produce the
water and hydrogen molecules.
(Refer Slide Time: 8:19)

And this is the examples of the bio products that we have I just the talk about the antibiotics
that is we have the lot of antibiotics in the nowadays available in the market to cure lot of
diseases we have different oxy-chemicals that can be produces the biological means (())(8:37)
what is ethanol then acetic acid, butenol and the citric acid. Different enzymes the enzymes
can be used not only for the pharmaceutical industry. It can be used for the industrial
processes like high fructose corn syrup formation. So we have the amylases we have
proteases, xynalases and rennin, then we have vaccine that is nowadays, it is very much
important to in the to protect our disease, because different type of vaccines that is usually
apply to the babies so that you know they can be free from certain diseases. Hormones that is
used particular in the hor (())(9:19) agricultural sector to improve the agricultural goal to a
great extent. Then single cell protein like the baker’s yeast largely used by the baker
industries.
(Refer Slide Time: 9:32)

Now why bio products, the reason is that it is sustainable, because we can use these bio
products for longer period of time and second is the less carbon and water footprint that is
very much essential for this and it creates rural employment opportunity, because not only is
this bio process can be can be applied for the both in the small scale and bigger scales and
less emission of the to the emission of pollutant to the environment that is another important.
Biodegradability and recyclability. This is very important factors then we have high
productivity and lastly we use the raw materials for the from the local sources. So this is
different advantages of using bio products while (())(10:16) this is in practice.

(Refer Slide Time: 10:20)

Now initially we should compare that how this bio process, they compatible with a chemical
processes. Now chemical processes one thing I want to tell you in the chemical process that if
you want to produce the any kind of chemicals you any kind of product you your if you
change the product your raw material will be changes in the chemical process, your product is
complicated to your raw material will be complicated, but in the bio process is something
different, because the raw material same carbon source can produce the different type of
compound, thus (())(10:53) as for example I can give the example of glucose, glucose can be
produce ethanol, it can produce citric acid. It can produce lactic acid. It can produce lactic (())
(11:03) acid.

So different type of product formation can be taking place with the help of this this particular
bio process and another advantage of the bio process is that it can be operated as the ambient
temperature and atmospheric pressure and in the chemical process if you look at is usually
operated at high high temperature and high pressure for that we we this quite anything to (())
(11:27) and it is easily operated one advantages is that one it is usually operated under (())
(11:34), but biological processes we allow our desire organism to grow in a environment, so
your involvement should be sterilized and for the sterility purpose our (())(11:47) that is it is
some energy is required to maintain the sterility of the system that is major drawback of this
system and it also in the chemical process we required very costly say catalyst like platinum,
for biological process we required enzymes which is not costly, it is very chip and produce
the limited verities of product we can have in the chemical process, but in the biochemical
process we can have wide varieties of product and we have greater toxicity in the chemical
process. So it mean itself (())(12:19) is very complicated. In a bioprocess whatever there the
they have the effluent generesis (())(12:25) that usually bio degradable.
(Refer Slide Time: 12:28)

Now the bio products they can be available in three different forms, one is here one is call
low value high volume product, another is medium value and medium value and medium
volume product and high value low volume products.

(Refer Slide Time: 12:49)

The examples there in the low volume products are those, which cost is around 6 pound per
kg as for example, citric acid, xanthum gum, this is this is low value product and the high
medium value and high medium volume products the cost is the about 60 pounds per kg the
examples are different antibiotics like penicillin, streptomycin kanamyne kanamycin usually
more costly as compared to penicillin. So this is consider as medium value and medium
volume products, but the high value low volume products are human insulin that is largely
used nowadays and interferon that is that is in practice this is produce very low volume but
the cost is very high.
(Refer Slide Time: 13:38)

Now the bio that bio products can be divided into three different types; one is call bioenergy,
biochemicals and biomaterials. Bio I told you that bioenergy we have three different data
types one is call solid fuel, another is the liquid fuel and the gaseous fuel. Solid fuel we have
the biomass and with the liquid fuel we have bioethanol, biobutenol and gaseous fuel we have
bio hydrogen bio methane. So different type of gases we can produce. Biochemicals we have
biopharmaceutical, like vaccine can be produce. We have industrial chemicals like we have
citric acid, then enzymes then bio cosmetic, like body creams can be produce through cells
(())(14:25) and biomaterials we have bioplatic, biocomposit bioplatic nowadays quite highly
demanded, because plastic causes the environmental pollution we know it. It is very difficult
to biodegrade. So bio plastic has lot of demand in the market, biocomposit that is also largely
used and then we have biofoam and bio rubber that can be also produced through this.
(Refer Slide Time: 14:51)

Now if you look at in the biotechnology industrial seen arrow, the India is the among the top
12 biotechnology destinations in the world and ranks second in Asia after china where china
is the first then the second is the India in Asia and the last year the growth of the
Biotechnology industry about 57.14 percent and is estimated to reach about 11.1 billion dollar
by 2017. The bio pharmaceutical segments accounted for the latest revenue. Now here we can
see that you know in bio pharmaceutical how they are shearing the line part of these bio
products.

(Refer Slide Time: 15:38)

Now this are the different made market values of different products that we have that we
have, we have ethanol, we have ethylene glycol, we acetic acid, acetic acid the different type
of chemicals how it is produced through the biological process and their market value is
given here.

(Refer Slide Time: 15:57)

And this is the examples how this bio products is usually produced, I have given the example
of the ethanol, because the alcohol is produced to a great extent, because nowadays because
alcohol not only use for the human consumption, but also used in the pharmaceutical sector,
also as uses the power alcohol and also it is use as a chemicals install (())(16:23) by the
chemical industries.

Now if you look at the how it is produce how the ethanol is produce by the fermentation
industry, the by we can form if you take the example of simple compound like glucose or
sucrose, sucrose when hydrolyze sugar when hydrolyze it produce fructose and glucose and
glucose and fructose, they will undergo (())(16:47) which produce the pyruvic acid and
pyruvic acid then it converted to acetaldehyde then to ethanol. So this is the biochemical
pathways how it produce the ethanol through the fermentation process.
(Refer Slide Time: 17:03)

Now the range of fermentation process 4 major groups of commercial important fermentation
exist, the one is the production of the microbial cells as the product; examples include the
baker’s yeast, of for single cell protein and probiotics. This is the this is one types and second
is the production of microbial enzymes, the enzymes as I told you that enzymes not only use
for the pharmaceutical sectors but also it is for the industrial purpose to for the detergents we
use this and also for the production of high fructose corn syrup we produce that.

Also, it is used for the here the bio sensor formation of the bioreactors sensors. Production of
microbial metabolites, we have several I just I giving the example of ethanol beside that we
have citric acid, vitamins, acetone, butenol and glutamic acid, lysine etcetera. The
modification of a compound which is added to the fermentation with the transformation
process that can be used to a great extent as the as for example the steroid, antibiotics and
prostaglandin that we used.
(Refer Slide Time: 18:25)

Now there is very interesting seen arrow that how this bio product development has been
develop in the chronological order if you look at pre 1900 that the alcohol with the vinegar
this is usually produce through the batch fermentation process, then in between 1900 and
1940 the baker’s yeast, glycerol, citric acid, lactic acid, acetone, butenol, that is usually
produce through the fed batch fermentation process, then 1940 on the to up to date that we
have several antibiotics in the market like penicillin, streptomycin other antibiotics we have
gibberellin, this kind of (())(19:08) growth hormone then we have amino acid, nucleotides,
enzymes and transformation takes place.

Then 1960 whereas onwards we have single cell protein that continuous medium cycle
genetic engineering of productions (())(19:25) strains. Then 1979 to onwards, we have
following compounds not normally produce microbial cells as for example insulin, interferon,
genetic engineering to introduce the foreign gene into microbial host. This is largely in
practice. The present development of industrial fermentation products is the microbial cell of
probiotic in the that is use in the form of capsule, in the form of drinks, beverage, amylase
and glucose glucose isomerase for high fructose corn syrup production that is mostly use in
the western country.

Coloring agent from microorganism for the textile what we considered as natural colored then
we have biodiesel as the energy source to replace the petroleum then bio insecticides we
know nowadays we have lot of problem with the pesticides and pesticides is such a
chemicals, which I keep on accumulating (())(20:25) system (())(20:26) never goes out of
your system and has (())(20:29) property. So if we use the bio insecticides this is the basically
the that the protein molecules that can be easily biodegraded. So we did not cause any kind of
pollution problem, then we have microbial plastic, bio plastics then isoflavon soybean lipase
for detergent.

(Refer Slide Time: 20:55)

Then a list of industrial importance microbes which is involve for the production of different
type of products is listed in this table and we find that in the baker industry that we use the
saccharomyces cerevisiae. This use the saccharomyces cerevisiae also use for the production
of beer and wine, then we have lactic acid bacteria, it produce the yoghurt and we have kefir
and probiotics. We have then Asepergillus oryzae is used Soy Sauce formation, tempeh deep
frying fermented soya beans, the rhizopus oryzae. So different type of product that has been
listed here that is then penicillin chrysogenum that is used for penicillin production process.
(Refer Slide Time: 21:43)

Now we have list of industrial important enzymes and their applications as for example
proteases, the lipase, cellulases, isomerase, xynalases, ligase and nucleases, rennin, pectinase,
beta glucanse, trypsin and tannase, because this is lot of enzymes that is used and I have
given you the protease use in the food processing industry and the desire also in the detergent
industry particular for removing the kind of strain that has been attached to the (())(22:20)
that can be easily removed with the help of protease enzyme as for example, the blood can be
degraded with the help of protease enzyme and different health care, it can be used.

Health care as for example that is used protease enzyme is used. Lipases it has lot of use in
the dairy and food processing industries. Cellulose, nowadays for used in the biofuel industry
for the breakdown of cellulose. Isomerase used for the conversion of glucose to fructose as
we know that fructose is the 10 times sweeter than glucose. So that is why that is used in the
(())(23:01) industry to a great extent, we have lipase ligase and nuclease that is used in the
molecular biology. The rennin is used for the cheese making industry. Pectinase in the food
processing industry, trypsin is used in the pharmaceutical industries.

(Refer Slide Time: 23:18)

Now finally I want to tell you that if you look at the industrial Biotechnology processes, it
broadly it can be divided into 2 parts, one is we call it upstream processing another is the
downstream processing. Upstream processes comprises of medium preparation, inoculum
preparation and the fermentation process. So the different things that is involve as for
example medium preparation medium I told you media comprises of carbon source, nitrogen
source, minerals and vitamin that we mix in a tank then we have we shall have to sterilize it
that is call medium sterilization. Then most of the fermentation process, they are operated
aerobically and most of them so they are mostly operated aerobically, so we required air and
air should be sterilized then inoculum preparation that is also very important that inoculum is
usually prepared with the help of industrial enzyme, then this whole unit where we have
including the fermentation process we call it upstream processing.

In the downstream processing that is nothing but the product purification, because I told you
whenever we market any kind of product, that should be in the purified form, we cannot
market the product, because we know that if you look at the chemical catalog, we have two
type of products that is available, one is call that analytical grade chemicals or that is
commercial grade chemicals. Analytical grade chemicals usually 99.9 percent purity and the
commercial grade chemicals it might be 94, 90 percent 94 percent like this and if you look at
the cost difference of this particular chemical is used may be your analytical chemical, it is 4
times costly at then the commercial grade chemicals.

So with the purity of the chemicals is very important I can give the example of the penicillin
production industry. Penicillin is the marketed in the 2 different forms in the form of capsule,
in the form of injection fluid. Now injection fluid, it should be 100 percent free from
contaminance, we could use directly injected to our blood. In a blood pH is 7. So any kind of
contamination is there, the immunity your body your see our system will be affected, so it
should be 100 percent free from contaminance, but when we take the antibiotics in the form
of capsule, it goes through your stomach and when it goes through your stomach (())(25:45)
here the stomach we know the our pH is 2 even some contamination is there your stomach
pH will take care the contamination problem. So this is the things we are we are going to
cover in this course I hope it will be very helpful to you all, thank you.