1. Industrial Biotechnology

2. Industrial biotechnology
Application of biotechnology for industrial purposes
Manufacturing
Alternative energy (bioenergy)
Biomaterials
It includes the practice of using cells or components of cells like enzymes to generate industrially useful products.
Division of IB
Industrial
Pharmaceutical biotechnology.
Growing fungus to produce antibiotics, e.g. penicillin from the penicillium fungi.

3. Important Applications
Production of primary metabolites (Acids & Alcohol)
Secondary metabolites (Antibiotic)
Production of whole microbial cells (Food, Vaccine)
Biotransformation reactions (Enzymes, steroids)
Exploitation of metabolism (Leaching and wastes treatment)
Recombinant proteins (Therapeutic proteins, gene delivery vectors, etc.)

4. Cell factories Biofuels Biomaterials Biochemicals Primary metabolite
secondary metabolite
Sugars

5. The IB Value Chain Bulk Biofuels Sugars Feedstocks Biomaterials
Ethanol
Sugars
Feedstocks
Renewable
- Fossil
Biomaterials
Polylactic acid
1,3 propane diol
PHAs
Biochemicals
Food Ingredients
Pharmaceuticals
Fine Chemicals
Bioprocesses
Fine

6. Bioenergy

7. Some definitions
Bioenergy is energy of biological origin, derived from biomass, such as fuelwood, livestock manure, municipal waste, energy crops.
Biofuels are fuels produced from biomass, usually of agricultural origin.
Bioethanol
Biodiesel
Biogas
Energy crops are crops specifically cultivated to provide bioenergy, mainly biofuels but also other forms of energy.
e.g. miscanthus, eucalyptus.

8. Liquid Fuels
Ethanol production by fermentation of carbohydrates is rather expensive and is influenced by the
Yield of ethanol
Ethanol tolerance of fermenting organism
Ethanol is relatively toxic to microbes.. Limited conc. Can accumulate
Ethanol-tolerant strain

9. Main bioenergy feedstocks
Wood
Forest management residues
Fuel timber
Crops
Annual (cereals, oilseed rape, sugar beet)
Perennial (miscanthus, reed canary grass, short rotation coppice)
Wastes
Straw
Animal manure

10. FUEL ETHANOL FROM BIOMASS
Energy can be extracted from biomass
by direct combustion (Common Method) or
by first converting the biomass to another fuel (ethanol, methanol, or methane) and then combusting it.
Cellulose, hemicelluloses, and starches are a vast renewable source of sugars convertible to ethanol by microbial fermentation.
Production of ethanol From the polysaccharides of biomass proceeds in three stages:
Degradation of polysaccharides to fermentable sugars;
fermentation; and
alcohol recovery.
Disruption of the physical structure of lignocellulose makes cellulose and hemicelluloses accessible to enzymatic attack. Disruption is done by
Steam explosion
Acid hydrolysis

11. Production of Alcohol (S. cerevisiae)
Preparation of Medium
Addition of water to molasses to decrease sugar conc to %.
Addition of acid to adjust pH
Addition of yeast
Adjustment of temperature
Thorough mixing of yeast inoculum with molasses
Fermentation
Vigorous fermentation leads to production of CO2, a by product of alcohol industry
Collection of CO2
Separation of ethyl alcohol
Removal of unused substances of molasses
Separation from other impurities
Purification
Purification with the help of rectifying columns

12. Production of Alcohol (Z. mobilis)
Zymomonas mobilis, a bacterium isolated from fermenting sugar-rich plant juices, produces ethanol up to 97% of the theoretical maximum value.
The advantages of Z. mobilis over S. cerevisiae with respect to producing bioethanol:
higher sugar uptake and ethanol yield (up to 2.5 times higher)
lower biomass production
higher ethanol tolerance up to 16% (v/v),
does not require controlled addition of oxygen during the fermentation,
amenability to genetic manipulations.

13. Disadvantages
In spite of these attractive advantages, several factors prevent the commercial usage of Z. mobilis in cellulosic ethanol production.
Substrate Limitation: Utilize only glucose, fructose and sucrose.
Wild-type Z. mobilis cannot ferment C5 sugars like xylose and arabinose which are important components of lignocellulosic hydrolysates.
Unlike E. coli and yeast, Z. mobilis cannot tolerate toxic inhibitors present in lignocellulosic hydrolysates such as acetic acid and various phenolic compounds.
Concentration of acetic acid in lignocellulosic hydrolysates can be as high as 1.5% (w/v), which is well above the tolerance threshold of Z. mobilis.

14. Bioenergy crops

15. Overview of Biofuel Production Technologies First Generation of Biofuels
Biofuel type
Specific name
Feedstock
Conversion Technologies
Pure vegetable oil
Pure plant oil (PPO),
Straight vegetable oil (SVO)
Oil crops (e.g. rapeseed, oil palm, soy, canola, jatropha, castor, …)
Cold pressing extraction
Biodiesel
Biodiesel from energy crops: methyl and ethyl esters of fatty acids
Biodiesel from waste
Waste cooking/frying oil
Cold and warm pressing extraction, purification, and transesterification
Hydrogenation
Bioethanol
Conventional bio-ethanol
Sugar beet, sugar cane, grain
Hydrolysis and fermentation
Biogas
Upgraded biogas
Biomass (wet)
Anaerobic digestion

16. Conversion Technologies
Overview of Biofuel Production Technologies Second/Third* Generation Biofuels
Biofuel type
Specific name
Feedstock
Conversion Technologies
Bioethanol
Cellulosic bioethanol
Lignocellulosic biomass and biowaste
Advanced hydrolysis & fermentaion
Biogas
SNG (Synthetic Natural Gas)
Lignocellulosic biomass
and residues
Pyrolysis/Gasification
Biodiesel
Biomass to Liquid (BTL), Fischer-Tropsch (FT) diesel, synthetic (bio)diesel
Lignocellulosic biomass and residues
Pyrolysis/Gasification & synthesis
Other biofuels
Biomethanol, heavier (mixed) alcohols, biodimethylether (Bio-DME)
Gasification & synthesis
Biohydrogen
Gasification & synthesis or biological process
*Use GMO as a feedstock to facilitate hydrolysis / technologies for hydrogen production

17. Biofuel transformation processes
First generation
ETBE: Ethyl tetra butyl Ether
Fatty Acid Methyl Ester
Second generation
Fatty Acid Ethyl Ester

18. Biofuel uses Bioethanol Biodiesel
Used as neat ethanol (E95, blend of 95% ethanol and 5% water)
Used as E85 (85% volume ethanol with petrol) in flex-fuel vehicles
Used as blend smaller than 5% volume (E5) in ordinary petrol or as its derivative ETBE
Biodiesel
Current maximum 5% in diesel blends, otherwise can only be used in modified diesel engines

19. Manufacturing factories

20. Secondary metabolites
Secondary metabolites have no function in the growth of the producing cultures (although, in nature, they are essential for the survival of the producing organism), functioning as:
(1) sex hormones;
(2) Antibiotics
(3) ionophores;
(4) competitive weapons against other bacteria, fungi, amoebae, insects and plants;
(5) agents of symbiosis etc.
Microbially produced secondary metabolites are extremely important for health and nutrition.
Antibiotics
Other medicinals
Toxins
Biopesticides
Animal and plant growth factors

21. Antibiotics
The best-known group of the secondary metabolites are the antibiotics.
Their targets include
DNA replication (Actinomycin)
Transcription (Rifamycin)
Translation (Chloramphenicol, tetracycline, erythromycin and streptomycin)
Cell wall synthesis (cycloserine, bacitracin, penicillin, cephalosporin and vancomycin)

22. Enzyme production
The production of enzymes by fermentation was an established business before modern microbial biotechnology.
However, recombinant DNA methodology was so perfectly suited to the improvement of enzyme production technology that it was almost immediately used by companies involved in manufacturing enzymes.
Important enzymes are proteases, lipases, carbohydrases, recombinant chymosin for cheese manufacture and recombinant lipase for use in detergents.
Recombinant therapeutic enzymes already have a market value of over US$2 billion, being used for thromboses, gastrointestinal and rheumatic disorders, metabolic diseases and cancer.
They include tissue plasminogen activator, human DNAase and Cerozyme.

23. Sources of Enzymes
Biologically active enzymes may be extracted from any living organism:
Of the hundred enzymes being used industrially,
- over a half are from fungi
- over a third are from bacteria with the remainder divided between animal (8%) and plant (4%) sources .

24. Enzyme Production

25. Sources f Enzymes
Microbes are preferred to plants and animals as sources of enzymes because:
They are generally cheaper to produce.
Their enzyme contents are more predictable and controllable.
- Plant and animal tissues contain more potentially harmful materials than microbes, including phenolic compounds (from plants).

26. E: extracellular enzyme; I: intracellular enzyme
Fungal Enzymes
Enzyme
Sources
Application
a-Amylase
Aspergillus E
Baking
Catalase I
Food
Cellulase
Trichoderma
Waste
Glucose oxidase
Lactase
Dairy
Lipase
Rhizopus
Rennet
Mucor miehei
Cheese
Pectinase
Drinks
Protease
Catalase:catalyzes the decomposition of hydrogen peroxide to water and oxygen.
E: extracellular enzyme; I: intracellular enzyme

27. Bacterial Enzymes Enzyme Sources Application a-Amylase Bacillus E
Starch
b-Amylase
Asparaginase
Escherichia coli I
Health
Glucose isomerase
Fructose syrup
Penicillin amidase
Pharmaceutical
Protease
Detergent
Asparaginase:(EC ) is an enzyme that catalyzes the hydrolysis of asparagine to aspartic acid.
Penicillin amidase: Sakaguchi and Murao1 reported on the presence of an enzyme in the mycelium of Penicillium chrysogenum and Aspergillus oryzae which would split penicillin G (I) into phenylacetic acid (II) and 'penicin' (III) :

28. Therapeutic Proteins
Recombinant protein plays a big role in the creation of therapeutic agents that could modify and repair genetic errors, destroy cancer cells, treat immune system disorders, etc.
For instance, Erythropoietin, a protein hormone produced by recombinant technology can be utilized in treating patients with erythrocyte deficiency, which is a common cause of kidney complications.

29. Categorization of FDA approved PTs based on mechanism of action
PTs replacing a protein that is deficient/abnormal
PTs augmenting an existing pathway
PTs providing a novel function
PTs that interfere with a molecule/organism
PTs that deliver other compounds/proteins
Protein vaccines
Protein diagnostics

31. New Generation of Vaccines:
Recombinant DNA technology is being used to produce a new generation of vaccines.
Virulence genes are deleted and organism is still able to stimulate an immune response.
Live nonpathogenic strains can carry antigenic determinants from pathogenic strains.
If the agent cannot be maintained in culture, genes of proteins for antigenic determinants can be cloned and expressed in an alternative host e.g. E. coli.

32. DNA Vaccines
DNA vaccines are possibly the most hopeful and powerful alternative to traditional vaccines.
A genetically engineered vaccine is already widely used against the liver infection hepatitis B.

33. Production of Vitamin C
Humans, as well as other primates, guinea pigs, the Indian fruit bat, several species of fish, and a number of insects, all lack a key enzyme that is required to convert a sugar, glucose, into vitamin C.
No single bacterial genus or species is known that will carry out all of the reactions needed to synthesize vitamin C.
Two species (Erwinia species and Corynebacterium genus) can perform all but one of the required steps.
In 1985 a gene from one of these genus (Corynebacterium) was introduced into the second organism (Erwinia herbicola), resulting in a new bacterial form.
This engineered organism can be used to produce a precursor to vitamin C that is converted via one chemical reaction into this essential vitamin.