Introduction to Biotechnology

• Biotechnology is the use of living organisms to create products.
• It includes the production of food, medicine, and other industrial products.
• Karl Ereky coined the term "biotechnology" in 1919.
• Biotechnology has enhanced fermentation, flavor, texture, and nutritional value in food production
• Genetically modified organisms, proteins, hormones, enzymes, and food additives are parts of biotechnology

Definition of Biotechnology

• Biotechnology combines 'bios' (life) and 'technology' (application of knowledge).
• It applies living organisms or biological systems in industrial processes and waste treatment.
• The UN Convention on Biological Diversity defines biotechnology as using biological systems to modify products.
• The European Federation of Biotechnology defines it as integrating biochemistry, microbiology, and engineering for technological applications.
• The OECD defines it as applying scientific and engineering principles to process materials with biological agents.
• The FDA defines it as applying biological systems to technical and industrial processes.
• Germany defines it as introducing biological processes into technical and industrial production, involving microbiology, biochemistry, and technical chemistry.
• The Biotechnology Industry Organization defines it as using cellular and molecular processes to solve problems or make products.

Traditional Food Fermentation

• Fermentation is an old biotechnology for producing foods with extended shelf-life and desirable properties.
• It improves microbial stability/safety.
• It can increase nutritional value/digestibility of raw materials.
• It enhances palatability for consumers

Roles of Fermentation

• Preservation of food through inhibitory metabolites is a key role of fermentation
• Improving food safety via pathogen inhibition or toxin removal is also important
• Enhancing nutritional value and organoleptic qualities of food are additional benefits
• Microorganisms in food fermentations include bacteria, yeasts, and molds.
• Lactic acid bacteria (Lactobacillaceae) are most important to produce carbohydrates
• Acetobacter produces acetic acid
• Bacillus ferments legumes
• Saccharomyces (especially S. cerevisiae) are beneficial yeasts to produce enzymes that create desirable biochemical reactions for wine, beer, ethanol, and bread.
• Lactic acid bacteria (LAB) are commonly found in fermented foods with functions that contribute to acceptance as safe for consumption
• Bacillus species bring about alkaline fermentation through the hydrolysis of protein to amino acids and peptides and releasing ammonia

Background of Fermentation

• Louis Pasteur first studied fermentation in the mid-19th century, showing that living cells (like yeast) could demonstrate yeast fermentation.
• Eduard Buchner won a Nobel Prize in 1907 for demonstrating fermentation in yeast cells.
• Arthur Harden and Hans Euler-Chelpin won a Nobel Prize in 1929 for their work on fermentation enzymes.
• Antibiotics by fermentation were being produced by the 1940s.
• Chain Weizmann developed fermenters for acetone production in WW1, but large-scale fermenters for yeast were created in 1944, and industrial fermenters with aseptic systems were founded in India in 1950

Types of Fermentation

• Solid State Fermentation (SSF) grows microorganisms on a moist solid substrate with little free water.

Uses of SSF

• enzyme production (pectinase, cellulase, protease, amylase, phytase).
• organic acids (gallic, and lactic acid).
• secondary metabolites (gibberellic acid and antibiotics).
• unsaturated fatty acids and biocontrol agents.
• SSF has advantages like high product titer, little water use/waste, no foaming, low expenditure, and low energy requirements.
• Disadvantages include moisture control, oxygen availability, pH control, and limited species usage.

Submerged Fermentation (SmF)

• Submerged Fermentation cultivates microorganisms in enriched liquid broth
• It is used for industrial production to give high oxygen
• In the medium, the substrate composition is set to optimize the product or target molecule
• SmF consists of batch, fed-batch, and continuous modes
• Batch mode is simple - involves adding all prerequisites at the beginning and running it until nutrients deplete or the target molecule maximizes
• Fed-batch mode adds substrates/nutrients/inducers as needed to increases the productive phase of microorganism

Advantage of Fed-Batch mode

• High cell density is obtained from from fed batch mode
• metabolite production
• controlling organism growth/oxygen requirements
• Continuous mode feeds fresh nutrients and removes spent medium/cells to maintain constant conditions.

Types of Fermenter

• There are six fermenters or bioreactors of varying configurations
• Continous stirred-tank
• Air-lift
• Bubble column
• Fluidized bed
• Packed bed
• Photo bioreactor

Application of Fermentation in Biotechnology

• Fermentation can be used to produce cells or bionass in large quantities

• Microorganisms inoculum grown in production medium achieves maximum growth rate.

• The harvested biomass can be processed to extract the target product.

• Fermentation can also make both primary and derived metabolites from microorganisms that include ethanol and organic acids

• Secondary metabolites are made while during microbes stationary phase like penicillin.

• Cultivation or molecular methods change metabolic pathways

• Fermentation produces recombinant proteins, vaccines, and hormones for pharmaceuticals.

• Fermentation occurs when organisms turn sugar or starch into acids or alcohols

• The science of fermentation is zymology

Process of Fermentation

• The process is anaerobically
• Glycolysis creates 2 ATP molecules with pyruvic acid formed
• Pyruvate then changes into lactic acid or ethanol
• NAD+ is used in glycolysis

Types of Fermentation

• Homofermentation produces one product type
• Heterofermentation produces more than one product
• Lactic acid fermentation: pyruvate generated from glucose
• NAD+ generates from NADH
• Lactic acid bacteria use this process to produce milk curd or cheese

Alcohol Fermentation

• Commonly used to produce biofuels, beer and wine
• Creates alcohol and CO2
• Pyruvic acid converts to CO2 and acetaldehyde
• Acetylaldehyde then creates ethanol that utilizes glycolysis
• Bacteria and yeast carry this out

Acetic Acid Fermentation

• Creates vinegar in a two-step process
• The first step of acid fermentation is forming an ethyl alcohol from anaerobic sugar
• The second step forms acetic acid using acetobacter from the oxidation of acids
• The step is an aerobic process with acetic acetobacter bacteria

Butyric Acid Fermentation

• This is the anaerobic bacteria, that takes place during fiber production
• Is used for the formation of energy for colorectal epithelium
• Sugar oxidizes which then turns into pyruvate, then oxidixed again into CO2 utilizing oxidoreductase enzyme
• This then makes Butyric acid after being reduced

Advantages of Fermentation

• Fermentation suits all environments and can be prokayotes or eukaryotes for making products
• Used to make beer, wine, bread etc

Postives

• Foods containing lactic acid
• Contains certain antibiotics and vitamins
• Helps with intestinal bacteria function
• Prevents cancer and increases the immune system while reducing intolerance of lactose
• Other than the food indstry, fermentation makes methane to treat water from sewage

DownStream Processing

• Needed to restore the process and materials during any processing
• A important step that takes place to manufacture different product
• Such as making products for medicine, and in addition to food
• The molecule is commonly mixed with others that must be separated

Steps

• Separation
• Disruption of cells
• Isolation
• Extraction
• Drying
• Purification

Separation of Particles

• The first step and normally removes substances in liquid form

Process

1. Surface filtration
2. Depth filtration
3. Centrifugal filtration
4. Rotational Filtration
5. Used to decrease aggregates of inorganic salts on the microbial surface
6. This step would not be effective if the bubbles did not completely surround the cells

Extaction

• Recovery of products, from cells using solvent phase
• Achieved by both separation and concentrations together

Forms of extraction

1. Aqueous
2. Organic solvents

Isolation

• Used to obtain the purest product
• By evaporation, evaporation, membrane filtrations, ion exchange.

Downstream of SSF

• Produces low waste and effluent
• Natural solids can subsitute media
• This helps reduces investment since tech is less expensive

Socail concerns

• To maintain security and safety that keeps in alignment with the growth of the industry
• Ethical, and maintains high quality of goods

Ethical Concerns

• To avoid unsafe manipulation practices
• Avoids unethical practices that exploit or damage safety or human interest

Food Safety

• No GM products have been confirmed to cause harm
• No foods can be 100% proven to never cause allergic reactions
• GM salmon has been documented as a cause for higher underwater pollution and a result for loss of aquatics spicies
• Many individuals fear the introduction of new pestisides
• There is a fear that gene transforamtion can lead to potential wiping out of an organism