Biomolecules (Mainly Carbohydrates, Proteins, Lipids and Nucleic Acids ) Production form Microorganisms and their Industrial applications were discussed....

1. BIOMOLECULES – PRODUCTION & INDUSTRIAL
APPLICATIONS
G. KANTHARAJAN
ICAR-CIFE

2. Biomolecules
• Biomolecules are organic molecules that occur naturally in living
organisms.
• Macromolecules - proteins, carbohydrates, lipids and nucleic acids.
• Small molecules - primary and secondary metabolites and natural
products.
• Biomolecules consists mainly of carbon and hydrogen with
nitrogen, oxygen, sulphur, and phosphorus.
• Most of the biomolecules are very large and extremely complex –
complex reactions
Living organism Organs  Tissue  cells  Organelles  Biomolecules

4. Characteristics of Biomolecules
1. Most of them are organic compounds.
2) They have specific shapes and dimensions.
3) Functional group determines their chemical properties.
4) Many of them arc asymmetric.
5) Macromolecules are large molecules and are constructed from small
building block molecules.
6) Building block molecules have simple structure.

5. Biomolecules
Micro molecule
Small sized, low mol wt .,
18 - 800 daltons
Found in the acid soluble
pool
Minerals,
Gases,
Water
Sugar,
Amino acids, Nucleotides
Macro molecule
Large sized, high mol wt.
> 10000 daltons
Found in the acid insoluble
pool
Carbohydrate,
Lipid,
Protein,
Nucleic acids
Types of Biomolecules

6. CARBOHYDRATES
The word "carbohydrate" includes polymers and other compounds
synthesized from polyhydroxylated aldehydes and ketones.
Entire carbohydrate family called as saccharides
Composed of carbon, along with hydrogen and oxygen (CH2O) - usually in
the same ratio as that found in water (H2O).
They originate as products of photosynthesis, an endothermic reductive
condensation of carbon dioxide requiring light energy and the pigment
chlorophyll.
Serve as a structural material (cellulose), a component of the energy
transport compound ATP, recognition sites on cell surfaces, and one of three
essential components of DNA and RNA

7. ‘’The unique reaction, which
makes life possible on the Earth,
namely the assimilation of the
green plants, produces sugar,
from which origination of all
other components of living
organisms directly or indirectly’’

8. Types of Carbohydrates
• one sugar (Glucose, Galactose and Fructose)
Monosaccharides
• two sugars (Sucrose, Lactose and Maltose)
Disaccharides
• more than two simple sugars (Raffinose)
Oligosaccharides
Polysaccharides
monomers - consist of 1000 of repeating glucose (Starch, Cellulose)

9. Major
group
Organism Compound Functions Reference
Bacteria Gluconacetobacter xylinus,
Agrobacterium tumefaciens
E. coli
Cellulose
Murein
(Peptidoglycan)
Skin therapy, Artificial blood vessels,
Potential scaffold for tissue
engineering, Wounjd care products,
Tablet modifications etc…
Structural protection
https://www.omicsonli
ne.org/open-
access/bacterial-
cellulose-production-
and-its-industrial-
applications-2155-
9821.1000150.php?aid
=22705
Macro
algae
Gracilaria sp.,
Pyropia, Gelidium
Chondrus, Eucheuma etc..
Phaeophyceae –
Macrocystis
Agar, Agarose
Carrageenan
Alginic acids
Gelling of products, Biotechnological
works etc..
Thickening and stabilizing agents
Binder, Stabilizer, Emulsifier in
Toothpastes, soap, ice cream
Indra Jasmine, 2010.
Fundamentals of
Biochemistry, FC&RI –
TANUVAS.
Fungus Gliocladium virens fructooligosacch
arides (FOS)
Used as functional ingredients to
improve nutritional and technological
properties of foods
http://www.sciencedire
ct.com/science/article/
pii/S134035401200028
9
Carbohydrate biomolecules – Production and Applications

10. PROTEINS
• Majority of biomolecules present in a cell.
• Proteins are responsible for many enzymatic functions in the cell and play an
important structural role .
• Proteins are composed of subunits called amino acids. There are 21 different types
of amino acids (including the less known selenocysteine).
Functions of proteins:
• Structural functions:
Certain proteins perform ‘brick and mortar’ roles and are primarily responsible for
structure and strength of body. These include collagen and elastin found in bone
matrix, vascular system and other organs and a- keratin present in epidermal tissues.
• Dynamic functions:
More diversified in nature- proteins acting as enzymes, hormones, blood clotting
factors, immunoglobulin’s, membrane receptors, storage proteins, besides their
function in genetic control, muscle contraction, respiration etc.

11. Major group Organism Compound Functions References
Macro algae
(Green algae - 9–
26 g protein 100
g−1 dw)
Micro algae
Porphyra tenera,
Undaria pinnatifida,
Aphanizomenon flos-
aquae. Arthrospira sp
Phycobiliproteins
Oxidative Stress and Macrophage
Stimulation, Intestinal Mucosal
Barrier Function
Natural dyes and pharmaceutical
industry
https://www.ncbi.nlm.nih.
gov/pmc/articles/PMC4557
026/
Bacteria E. coli Insulin
(Regulatory
protein)
Blood glucose regulation
Fungi Agaricus bisporus,
Aspergillus niger
Mucor rouxii
Chitin
Chitosan
Chitin treated seeds – Agri,
Packaging technology, Food
processing, Cosmetics etc…
http://nopr.niscair.res.in/bi
tstream/123456789/5397/1/
JSIR%2063(1)%2020-31.pdf
Transgenic Cattle - recombinant
Human serum
albumin
maintenance of oncotic pressure
and the transportation of various
biomolecules and pharmaceuticals
http://inter-
use.com/Journals/JSAB/20
14/Volume%2002%20Issue%
2002/2014/0318/63.html
Mammalian cells,
Microbes
S. cerevisiae, P.
pastoris, and E. coli,
Mono clonal
antibodies
use in diagnostics, potential for
developing bioactive peptides
https://www.ncbi.nlm.nih.
gov/pmc/articles/PMC390
6537/
Protein biomolecules – Production and Applications

12. Cell-free bio-production
 Insulin, antibodies for use in vaccines and cancer medicines, enzymes for the food, cosmetics,
and detergent industries: many such substances can already be produced on a large scale using
biotechnology.
 Currently, demand for biomolecules is often still met by making use of living cells or organisms.
 This involves researchers adding the gene that codes for the target protein to bacteria, yeasts,
or cultures of animal or plant cells.
 These modified organisms are then cultivated en masse in bioreactors before the protein is
finally isolated and purified.
 There is no doubt that the technology is very effective, but it does have disadvantages, as many
of the steps in the process are costly and time-consuming.
 What’s more, the bacteria and other cells themselves consume part of the resources to stay
alive – which reduces the efficiency of the protein synthesis process.
 "Many proteins cannot be produced in cells – or else the results are very poor. Take membrane
proteins, for instance, which play a major role in pharmacological research. Or proteins that
poison a cell when present in high concentrations – making them potentially very useful for
treating cancer."
https://www.fraunhofer.de/en/press/research-news/2013/september/Biomoleculesfortheproductionline.html

13. Producing proteins without cells
• In cell-free techniques, instead of employing intact, living cells they take only
those elements of a cell needed for protein synthesis.
• researchers must break down the cells to obtain a mixture – known as a lysate –
that contains all the necessary elements for protein synthesis.
• Alongside enzymes, this also includes biologically active organelles and
membrane parts that synthesize the proteins according to their genetic coding.
• The desired genes can be added straight to the lysate; there is no longer any
need for them to be implanted laboriously into the cells’ DNA first.
• The first stage was the development of automated cell harvesting and extraction
techniques to produce lysates from bacteria, tobacco, and insect cells.
• A completely automated process supplies these lysates with amino acids and
selected genetic material so that the synthesis of specific proteins can get
underway.

15. Two bioreactor concepts
• One idea consists of small synthesis chambers with a partially permeable
membrane through which fresh stores of ingredients for the reaction can be
fed to the lysate and harmful metabolites removed. A supply and disposal
system of this kind allows protein synthesis to keep going for several days.
• The other idea is for a microfluidic platform on which the reading of the
genes and actual protein synthesis occur in separate places – in much the
same way as it does in animals and plants. This system is particularly suited
to lysates from animal and plant cells

16. Lipids
Lipids are composed of long hydrocarbon chains (-CH2-).
These molecules hold an incredible amount of energy and are therefore
energy storage molecules.
lipids are the major component of cell membranes.
Cholesterol and other sterols are also types of lipids and are necessary
components of cell membranes.

17. Lipids perform several important functions
1. They are the concentrated fuel reserve of the body (triacylglycerol’s).
2. Lipids are the constituents of membrane structure and regulate the
membrane permeability (phospholipids and cholesterol).
3. They serve as a source of fat soluble vitamins (A, D, E and K).
4. Lipids are important as cellular metabolic regulators (steroid hormones
and prostaglandins).

18. Marine Micro algae - Polyunsaturated fatty acids
• PUFAs include docosahexaenoic acid, eicosapentaenoic acid, arachidonic acid,
γ-linolenic acid and have been widely recognized as beneficial towards human
health.
• Μarine microalgae have significantly higher DHA contents compared to fresh
water microalgae.
• DHA is the characteristic PUFA of the marine dinoflagellates. Crypthecodinium
cohnii is a non-photosynthetic, marine dinoflagellate producing DHA
predominantly.
• Nannochloropsis sp. has been proposed as a source of PUFAs due to its high
contents of EPA.

19. Photo Bioreactor Production System

20. Microbes
• Wax esters (WE), composed of long chain alcohol and fatty acid
• Usually, they are harvested from plants, such as jojoba or carnauba wax, which
require rather time-consuming and expensive cultivation.
• Certain bacteria can offer an interesting alternative source of WE with a similar
composition compared to jojoba oil.
• The ability to accumulate significant amounts of transesterification of
triacylglycerols as intracellular storage deposits is predominantly found in bacteria
belonging to the Gram-positive Actinomycetales,
• e.g. Arthrobacter, Dietzia, Gordonia, Nocardia, Rhodococcus or Streptomyces species
• Gram-negative isolate, Aeromonas sp. 3010, achieved a total lipid content of approx.
12%, whereof up to 30% is eicosapentaenoic acid [20:5 n-3, EPA], a PUFA with great
relevance for pharmaceutical or food industries

21. Species Name Important Lipid Applications
Acinetobacter sp. Triglycerides (Wax Ester) lubricants, cosmetics, linoleum and printing inks
Rhodococcus sp.
Streptomyces
E. coli
Free fatty acids, Triglycerides,
Ethyl ester, Poly ester
Good cholesterol
Raulstonia eutropha Poly ester Clothing and chemical industry
Pseudomonas Rhamnolipids Biosurfactants - soil remediation
Cyanobacteria Thylakoid lipids functional integrity of the photosystems
Bacteria for the production of industrially relevant lipids

22. Lipids from FUNGI-BACTERIAL SYNERGISTIC
Filamentous fungi, Aspergillus fumigatus can efficiently flocculate the
unicellular cyanobacteria Synechocystis PCC 6803 and its genetically
modified derivatives that have been altered to enable secretion of free fatty
acids into growth media. Secreted free fatty acids are potentially used by
fungal cells as a carbon source for growth and ex-novo production of lipids.
For most of genetically modified strains the total lipid yields extracted from
the fungal-cyanobacterial pellets were found to be higher than additive
yields of lipids and total free fatty acids produced by fungal
and Synechocystis components when grown in mono-cultures.
The synergistic effect observed in fungal-Synechocystis associations was also
found in bioremediation rates when animal husbandry wastewater was used
an alternative source of nitrogen and phosphorus.
33 fungal strains isolated from wastewater sludge for their lipid content and
flocculation efficiency against 15 photosynthetic microalgae
https://biotechnologyforbiofuels.biomedcentral.com/articles/10.1186/s13068-015-0364-2

23. NUCLEIC ACIDS
• These molecules are responsible for all of our genetic information – DNA
and RNA.
• Nucleic acids are formed from subunits called nucleotides.
• There are 5 different types of nucleotides in the cell; Adenine, Thymine,
Guanine, Cytosine and Uracil.
• Each nucleotide is composed of a nitrogenous base, a 5-carbon sugar, and
3 phosphate groups.
• The bonds that form between nucleotides are called phosphodiester
bonds.
• Nucleotides are responsible for more than just composing DNA and RNA,
as ATP is a nucleotide and is also the energy currency of the cell.

24. • Brevibacterium ammoniagenes ATCC 6872 accumulates 5′-GDP and -GTP,
or 5′-ADP and -ATP together with GMP or AMP in nucleotide fermentation
by salvage synthesis.
• Nucleic acid techniques-Food industry monitoring

25. Thank u