SELECTIVE BREEDING
 Artificial selection
 Process by which humans breed plants and animals
of a particular characteristic
 Farming: hunter-gatherer  farmer
 Started with breeding wild crops into food crops (rice
from streams and wetlands to paddy fields for larger
regular grains)
 Vs. natural selection
 Proposed by Charles Darwin as the way species
evolve
 Same gene can encode for varying traits
 Varying traits are favorable or non-favorable
 “Superior” trait persists
 Nature decides which species is more fit
 Steps
Decide which characteristics are important
Choose parents that show these
Select the best offspring to breed for next gen
Repeat the process continuously
 Applications
 Utility
 Insect resistance in food and crops
 Animals that produce more meat or milk
 Domestication
 Dogs with gentle nature
 Przewalki’s horse  modern horse
 Athletic horse & horse for carriages
 South Asian wolf  domestic dog
 Hypoallergenic cat (short hair)
 Munchkin cat (short legs)
 Aesthetics
 Large or unusual flowers
 Garden flowers for bright colors and nice smell
 Dogs for cute traits
 Livestock
 Size of chicken has increased
 Auroch  domestic cattle
 Friesan cow: dairy breed
 Aberdeen Angus bull: beef
 Plants
 Quality of crop increases
 Disease-resistant wheat
 Teosinte  modern corn (larger cobs with
more kernels that are softer and more edible)
 Wild mustard plant: broccoli (flower
development), cabbage (intermode length),
kale (leaves enlargement), cauliflower (sterility
of flowers), kohlrabi (enhancement of lateral
meristems)
 Problems
 Species are selected for pleasing traits not favorable
for survival (corgi, bull terrier, English bulldog,
boxer)
 Limitations
 Production of desirable trait requires generations for
breeding
 “Unnatural selection”, weak traits propagated
 Reduced genetic variability which may obliterate an
entire population when there is high environmental
stress
 Human intervention results in unintentional artificial
selection (fish population in fishing, small fish
instead of both small and large)
 Eugenics
 Selective breeding in humans
 Francis Galton
 Improve intellectual, economic, and social level of
humankind
 Support in late 19th and early 20th century
 Legislation passed favoring forces sterilization of
criminals, those with mental disorders, and specific
ethnic groups
 Ignored the fact that human traits are influenced by
genetic and environmental factors (mental
disorders)
 Hereditarianism: misconception that human traits
are determined solely by genetic inheritance
 Used to pass restrictive immigration laws in the
United States (immigrants from south and eastern
EU were said to be genetically inferior)
 Followed by the rise of the Nazi movement (1933 law
for the protection against genetically defensive
offspring in Germany, allowed killing of the
“defective” individuals)
 Nazis imposed forced sterilization/infanticide
 Eugentics was used to justify the holocaust
PROTEINS
 Mendel’s study established that physical trait is due to
presence of a gene
 Protein ties gene to trait
 Given protein assumes a specific function (structural,
mechanical, enzymes, hormones, antibodies, fluid
balance, acid-base balance, channels and pumps,
transport)
 Gene for yellow peas codes for a protein that breaks
down chlorophyll (PAO)
 Gene for tall pea plants codes for protein that makes
gibberellin (growth hormone)
 Gene for purple flower codes for a protein that makes
anthocyanin (purple pigment)
 Foods high in protein: meat, fish, cheese, eggs, beans,
bread, hummus, nuts and seeds
 Proteins are source of amino acids
 Made up of amino acids
 Body breaks down the protein we eat into amino
acids
 Body takes carbs to break it down to glucose (source
of energy)
  Body takes protein to break down to amino acids
(needed to make more proteins that the body needs;
tissue, plasma, ammonium, etc.)
 9 Essential Amino Acids
 Human body cannot synthesize these amino acids
 We get them from the proteins we eat
 Histidine, Isoleucine, Leucine, lysine, Methionine,
Phenylalanine, Threonine, Tryptophan, Valine
 Protein Molecule  Digestion  Amino Acid
 Proteins are workhorses of the cell
 Insulin (sugar metabolism)
 Cytochrome C (cell respiration)
 Growth Hormone (anti-aging)
 Hemoglobin (oxygen transport)
 Hexokinase (glycolysis)
 Gamma Globulin (immune system in blood)
 Myosin (muscle action)
 50% of cell is made of proteins
 4 macromolecules: carbs, lipids, proteins, nucleic
acids
 Proteins have complex structures that allow them to
have various functions
 Proteins fold in such a way that hydrophilic is
exposed and hydrophobic is hidden from the water
environment
 Primary, secondary, tertiary, quaternary
 Amino acids have charges that make them hydrophilic
(charge), hydrophobic (no charge), or neutral
 Microtubules are made up of tubulin proteins
 Histones are round, allowing DNA to wound around
them
 Aquaporins are channels through which water enters the
cell
 Different uses of protein in the body
 Transport: hemoglobin (carry oxygen and circulate it
around the body, RBC)
 Hormone: insulin (tells body to store glucose)
 Neurotransmitters: serotonin
 Enzyme: amylase
PROTEINS IN BIOTECH
 Proteins are important to all organisms
 Microorganisms produce proteins that can be utilized by
humans
 Penicillin
 Produced in great amounts by letting mold grow in
tanks
 Binds to a protein in the cell wall and prevents cell
wall synthesis
 Without cell wall, bacterial cell dies (only gram
positive bacteria thick cell wall)
 Treatment of bacterial infections (Pharyngitis by
Streptococcus, Pneumonia by Pnemococcus, Skin
infections by Staphy)
 Not effective against gram negative bacteria like E.
Coli and Salmonella
 Waksman studied soil microorganisms to screen for
naturally occurring antibiotic compounds (discovered
antibiotics actinomycin and steptomysin, coined
“antibiotic”)
 Antibiotics are antimicrobial drugs used to treat bacterial
infections
 Actinomycin, from soil bacteria steptomyces
antibioticus, is a protein that can inactivate DNA
 Effective against a wide range of bacteria
 Structure-function at work
 Actinomycin fits right in DNA groove
 When DNA s inactivated, the cell cannot perform its
functions properly, such as cell division
 Why would a bacterium be producing an antibiotic
protein?
 Ward off competition
 Steptomyces lives in soil and competes for resources
 Actinomycin was developed to become an anti-cancer
drug (cannot replicate)
 Streptomycin: binds to bacterial ribosomes to prevent
protein production
 Bacteria harnessed to secrete bioluminescent proteins
(Luciferin protein is needed)
 Luciferase catalyzes
 Allows oxygen to combine with luciferin
 Reaction produces light
 Squid lives in symbiosis with a fluorescence-producing
bacteria (Vibrio fischeri)
 Squid uses the bioluminescent bacteria to emit light
in its body parts and ward off predators, attract prey,
or find mates
 Special organ were bacteria can live
 Bioluminescent proteins: bacteria from squid are isolated
and cultured as a marker for successful genetic
modification of proteins
 Proteins by animals also used in biotech
 Insulin
 Tells body to store glucose
 Diabetes: insulin production is defective
 Pigs and cows were source of insulin in traditional
biotech
 Pic/cow pancreas are run through grinder; insulin
proteins are filtered out then packaged
 Antivenom
 Horse as source of antibody
 Antibodies: defensive proteins against foreign
materials
 Antibodies are shaped to fit their target
 Bacteria, viruses, snake venom look different
 Venom extraction, animal immunization, blood
collection, Antivenom
 Antivenom antibody “IgG”
 Treatment has to be immediate, so antivenom
antibodies are supplied externally in drug form
 Plants can also be source of protein-based meds
 Cortisone is steroid hormone and can be synthesized
from wild yam root
 Cortisone relieves pain in arthritis
 37 steps to synthesize it before and expensive
  Fungus to cut down synthesis to 11 steps, allowed for
microbial hydroxylation
 Starting substance: diosgenin to synthesize
cortisone
 Wild yam root & Rhizopus fungus
ENZYMES
 Every chemical synthesis needs a starting reactant to
jump-start the process
 Bovine bile acid (starting material of reaction)
 Proteins are useful in research
 Marine organism houses bioluminescent bacteria to
ward off predators, attract prey, and find mates
 Bacteria use bioluminescence for communication:
Quorum Sensing
 Cellular processes that need more bacteria to
become efficient are achieved by this (antibiotic
production, conjugation, virulence factors, biofilm
formation, bioluminescence)
 Need many cells for a process to exert a significant
function (bacteria are small)
 BLB use it to monitor the environment for the
presence of other BLB (reach a quorum by sending
out a signal)
 Should we luminesce?
 Sends our chemical signal “AHL” aka
autoinducer
 AHL signal in environment: not enough
OR
 AHL signal in environment: threshold
concentration reached
 Autoinducers use Lux genes to produce
luciferase
 Luciferase is an enzyme and allows
bioluminescence chemical reaction to proceed
 BLB need to exist in a large number to luminesce
(live in marine organism)
 Luciferase induces bioluminescence chemical reaction
(protein AND ezyme)
 Enzymes are proteins
 Unassisted, chemical reactions need a high amount of
energy to proceed
 Enzyme lowers the energy hill because it provides the
optimal environment for reaction
 Catalyzes the reaction: faster
 Luciferase is an enzyme
 Substrate: luciferin
 Product: Oxyluciferin + light
 Examples of Enzymes
 Amylase
 Saliva
 Digests starch into simpler sugars
 Aldehyde dehydrogenase
 Liver
 Breaks down alcohol metabolite
 Alcohol flush reaction if lacking enzyme
 Pepsin
 Stomach
 Breaks down proteins
 Rennin
 Stomach
 Curdle milk and make it easier to help the
young digest the milk
 Cheese fermentation
 Diastase
 First plant enzyme to be purified and studied
 1800’s “starch-liquefying principle from
germinated barley”
 Isolated an active compound from barley in
powder form
 Can turn starch into liquid
 Ability lost when heated
 When dissolved in water, gained back ability
= Active components can be isolated and can
perform function
 Does fermentation need living organisms?
 Wilhelm Friedrich Kuhne coined the word enzyme
for this type of ferment
 Edward Buchner experiment proved that alcoholic
fermentation can occur in absence of yeast cells
 Zymase: enzyme in yeast that induced alcoholic
fermentation
 Proteins are workhorses of cell; they enact the
instruction encoded in DNA
 Enzyme was behind alcoholic fermentation
 For the first time, it was possible to observe
fermentation in absence of living yeast
 Enzyme actually responsible for the chemical
conversions, not the microorganism
ENZYMES IN BIOTECH
 Industrial biotech ethos = IMRPOVE PROCESS
 Become more efficient
 Fermentation by accident  Large-scale
fermentation in bioreactors  Improved large scale
fermentation in bioreactors
 Isolate enzymes to make certain processes easier
 Why are microorganisms still used in industrial scale
fermentation?
 Enzymes are produced by the microorganisms
 Easier and cheaper to use microorganisms
 Isolating and purifying enzymes has a corresponding
cost
 Microorganisms reproduce on their own
 Enzymes used in the industry
 Amylase for industrial applications (textiles,
brewing, dextrose syrup)
 Amylase used to convert starchy paper into a
smoother and stronger product
 Sucrose vs Glucose vs. Fructose
 Sucrose (sugar cane/beet)
 Glucose not as sweet (corn, potato)
 Fructose 2x sweeter than sucrose
 Starchy materials  syrup
 Amylase used to convert starch to glucose
 Glucose isomerase converts glucose to fructose
 Rennins in dairy industry
 Stomach of calves contain rennin
 Coagulates milk and makes cheese
 Rennin is enzyme that breaks down milk proteins
(curdles milk during fermentation)
 Amylase, glucose, and proteases in beer fermentation
 Enzymes used to break down starch into simpler
sugar, protein into amino acids
 Products are used by yeast for fermentation
 Cellulase in the biofuel industry
 Biofuel used plant material, which is rich in cellulose
 Has to be broke down before it can be used
 Proteases in detergent industry
 Help with the removal of protein stains from clothes
 Protease producing bacteria are cultured in large
flasks
 Protease is later purified and mixed with detergent
product