BIOMOLECULES – PRODUCTION & INDUSTRIAL APPLICATIONS

G. KANTHARAJANICAR-CIFE

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

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. 

Biomolecules

Micro moleculeSmall sized, low mol

wt ., 18 - 800 daltons Found in the acid

soluble pool 

Minerals,Gases,Water

Sugar, Amino acids, Nucleotides

Macro moleculeLarge sized, high mol

wt. > 10000 daltonsFound in the acid

insoluble pool

Carbohydrate,Lipid,

Protein,Nucleic acids

Types of Biomolecules

CARBOHYDRATES

The word "carbohydrate" includes polymers and other compounds synthesized from polyhydroxylated aldehydes and ketones.

Entire carbohydrate family called as saccharidesComposed 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

‘’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’’

Types of Carbohydrates

• one sugar (Glucose, Galactose and Fructose)

Monosaccharides

• two sugars (Sucrose, Lactose and Maltose)

Disaccharides

• more than two simple sugars (Raffinose)

Oligosaccharides

Polysaccharidesmonomers - consist of 1000 of repeating glucose (Starch, Cellulose)

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.omicsonline.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 fructooligosaccharides (FOS)

Used as functional ingredients to improve nutritional and technological properties of foods

http://www.sciencedirect.com/science/article/pii/S1340354012000289

Carbohydrate biomolecules – Production and Applications

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.

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/PMC4557026/

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/bitstream/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/2014/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/PMC3906537/

Protein biomolecules – Production and Applications

 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

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.

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

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.

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).

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.

Photo Bioreactor Production System

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

Species Name Important Lipid ApplicationsAcinetobacter sp. Triglycerides (Wax Ester) lubricants, cosmetics, linoleum and printing inksRhodococcus sp.Streptomyces

E. coliFree fatty acids, Triglycerides, Ethyl ester, Poly ester

Good cholesterol

Raulstonia eutropha Poly ester Clothing and chemical industryPseudomonas Rhamnolipids Biosurfactants - soil remediationCyanobacteria Thylakoid lipids functional integrity of the photosystems

 Bacteria for the production of industrially relevant lipids

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

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.

• 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

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