Biotechnology

Question 1

White biotech

Answer 1

Use of living organisms or their derivatives to make industrial products

Chemicals

AAs

Vitamins

Enzymes

Question 2

Industrial chemical production

Answer 2

Acetic acid - fermentation of ethanol or methanol by microbes - 200,000 tonnes produced annually

Butanol - From petroleum or fermentation - used in plastics, paint, resins and brake fluid

Lactic acid - Half of lactic acid in Europe is made by microbes - rest is chemical - used as acidifier - preservative in plastics

Question 3

Enzymes

Answer 3

traditionally obtained from microorganisms, plants and animals (amylase/pepsin/rennet/trypsin/lipases/proteases)

Enzymes from fungi - Aspergillus oryzae was grown on straw to obtain amylases and proteases - ‘emersed culture’

Produced in large bioreactors after 1950s - submerged culture - high yields/cheap/continuous

Question 4

Uses of enzymes

Answer 4

Pectinases - break down pectin in manufacture of fruit juice and baby food

Proteases - many uses including in leather tanning

Phytases - added to animal feed to enable digestion of phosphate

Substillsin - detergent

Alkaline proteases, amylases and lipases - detergent

Question 5

Immobilised enzymes

Answer 5

Enzymes that are fixed in a gel or to a membrane

recyclable

Increased stability

Absent from end product

Lower production costs

Question 6

Glucose isomerase

Answer 6

Immobilised enzyme

converts glucose into fructose

isolated from streptomyces

Immobilisation successfully increased fructose yield by 42% and reduced production cost by 40%

Question 7

Red biotechnology

Answer 7

Health related

Biopharmaceuticals

Recombinant proteins

Vaccines

Stem cells

Animal models

Gene therapy

Question 8

Recombinant proteins

Answer 8

Over 100 recombinant proteins are in use:

50 antibodies - $50 billion

Insulin - $16 billion

Blood clotting factors - $16 billion

Others - $25

Main uses:

Replacement for missing or defective proteins

Inhibition of infectious agents

Question 9

Insulin production in E. coli

Answer 9

Type 1 diabetes - lack of insulin

High blood sugar

Gene for insulin production put into plasmid - E. coli take up plasmid and replicate

Question 10

Subunit vaccines

Answer 10

Fragments from the pathogen

Developed before recombinant DNA tech

Pathogens could be grown in liquid culture and secreted proteins were used in the vaccine

Hep B was the first example

Question 11

Inactivated vaccines

Answer 11

Killed pathogen

Hep A, influenza. rabies

Can not be isolated or cultured in vitro or are too expensive to culture

Risk of infection if alive

Question 12

Attenuated vaccines

Answer 12

Live, weakened pathogens - no longer express toxin gene

Can be a natural or GM mutant

Safer to produce but need a lot of research to identify the toxic genes

Risk they can revert to pathogenic strain

Question 13

Sequence genomes of pathogens to develop new vaccines

Answer 13

Sequence pathogen genomes to determine which proteins are responsible for the immune response

Question 14

Use viral genomes to develop new vaccines

Answer 14

clone genes of interest into a plasmid and insert into vaccine genome

Recombinant vaccinia virus as a vaccine against smallpox or influenza e.g.

Question 15

DNA based vaccines

Answer 15

Add the gene encoding the antigen into plasmid

Bind the DNA to a charged particle and inject

DNA will bind with genomic DNA particles and the antigen will be expressed temporarily - triggering a localised immune response

cheaper to make and easier to store

first done in 2005 against Wests Nile virus

Question 16

Edible vaccines

Answer 16

Can express antigens in plants and then eat the plants

GM potatoes containing Hep B vaccines are in trials

Have to eat enough of it and had to be raw

Question 17

Stem cells

Answer 17

Potency:

totipotent cells can differentiate into all cell lineages to regenerate a whole organism - only embryonic stem cells can do this in mammals

Pluripotent cells are capable of forming all the cell lineages within an embryo but not extraembryonic lineages

Mulitpotent cells have the potential to differentiate into many, but not all, cell lineages

Question 18

Hematopoietic stem cells

Answer 18

HSCs

found in bone marrow

multipotent - replenish red and white blood cells

Found near blood vessels within the marrow ‘vascular niche’ and at the interface between the bone and marrow ‘endosteal niche’

Lymphoid and myeloid progenitors - differentiate into two further cell lines - make up the different red and white blood cell types

Question 19

Intestinal epithelial stem cells

Answer 19

ISCs

Found in small intestine

Can differentiate into 4 diff cells:

Absorptive epithelial cells

Goblet cells

Enteroendocrine cells

paneth cells

Question 20

Embryonic stem cells

Answer 20

ESCs

derived from blastocyst

Pluripotent - capable of forming all cell lineages within an embryo

Question 21

Induced pluripotent stem cells

Answer 21

iPSCs

differentiated adult cells into stem cells

changes in gene expression allow this reversion

Not clear if identical to ESCs

Low efficiency of production

Can cause tumours

Question 22

Stem cell therapy

Answer 22

HSCs used with bone marrow transplants

Tissue regeneration

Diabetes

Spinal cord injuries

Leukaemia, lymphoma, sickle cell anaemia

Question 23

Treatment for diabetes

Answer 23

Induce stem cells to differentiate into pancreatic B cells that produce insulin

Treat those that don’t produce insulin

More work needed to prevent immune system attacking B ells

Question 24

Animal cloning

Answer 24

Nuclear transplantation

Remove a nucleus from an egg

Add to a different nucleus

Allow to develop into frog

Shows that a nucleus from a differentiated cell can regenerate an entire organism

Question 25

Forensic biotech

Answer 25

Fingerprints

Blood type

DNA technology

DNA fingerprinting

DNA databases

DNA barcoding

Biometrics

Question 26

Blood typing

Answer 26

Glycolipid antigens A,B and O

Different alleles of the same gene

O has one fewer carbohydrate

A has N-acetyl-galactosamine

B has galactose

Question 27

DNA fingerprinting

Answer 27

Analysis of DNA fragments for ID

DNA extracted, cut with restriction enzyme and run on gel

Southern blot was used - DNA transferred to a nylon filter

DNA probes used and bound to complementary DNA

radiation sensative film placed over blot and DNA bound to the probes appear as bands on film

restriction pattern matched to the victim

Question 28

Phenotypic biometrics

Answer 28

Look at phenotypes to identify individuals

Fingerprints

Retinal scans - pattern of blood vessels in back of eye

Iris recognition systems - pattern of iris

Facial recognition

Question 29

DNA barcoding

Answer 29

Used to identify species

Uses conserved regions of genome:

fungi (ITS)
Plants (chloroplasts)
Animals (COI)

Fish in restaurants

horse meat scandal

ID of insect larvae

Question 30

Bioremediation

Answer 30

Use of living organisms or their products to break down waste and pollutants in the environements

Contaminated drinking water:

trapped in soil and rocks - can’t wash ground water

If oil and gas is mixed with ground water - need a bioreactor containing bacteria to separate the clean water

Question 31

Bioremediation microbes

Answer 31

Petroleum-eating bacteria

E. coli for heavy metals - use metallothioneins to neutralise heavy metals - cadmium and mercury

Question 32

Bioremediation of oil spills

Answer 32

Exxon Valdez - 1989

42 million litres of oil off Alaskan coast

Clear from surface using skimmers and vaccums

Wash rocks with water

Added fertiliser to encourage growth of bacteria to degrade

Oil will remain for hundreds of years

Deepwater horizon - 2010

Explosion released 600 million litres of oil into Gulf of Mexico

various removal methods - Bioremediation degraded 50% of oil released

Question 33

Biorefining of fossil fuels

Answer 33

FFs with high sulphur contnet:

Thiobacillus + Sulfolobus bacteria can be used to convert inorganic FeS2 to sulphates - can be washed away - only accounts for 30% of sulphur in fossil fuels

Rhodacoccus can break down thiophene (where rest of sulphur is found)

Question 34

Other bioremediation EGs

Answer 34

Recovering valuable metals

Degrading radioactive chemicals

White rot fungi to degrade lignin - waste agricultural product

Question 35

Phytoremidiation

Answer 35

Using plants for bioremediation of soil, water and air

Question 36

Biofuel

Answer 36

Fuels produced through biological processes

Issues - expensive - fuel instead of food? - waste products

Question 37

Biogas

Answer 37

Burning biomass for fuel

Can also produce gas from animal dung in small bioreactors instead

Question 38

Bioethanol

Answer 38

Sugarcane or maize - raw materials

Microbes convert sugars into ethanol - production is expensive

14 litres of sugar residue and 100 litres of water needed to make 1 litre of ethanol

Fuel additive

Question 39

Biodiesel

Answer 39

Derived from plant oils + animal fats

Standalone fuel - replacement for diesel

Question 40

Biofuel from algae

Answer 40

algae and cyanobacteria

No competition for land use - oils easily refined into diesel

Easy to genetically manipulate algae to produce ethanol and butanol

Can also make biohydrogen and biomethane

Produce 10x the output of traditional biofuel feedstocks

Needs a lot of fertilizer

Question 41

Green biotechnology

Answer 41

Use of crop plants and agricultural systems

GM crops very succesful:

4 GM crops dominate global production

Soybean - 50%

Maize - 31%

Cotton - 13%

Oilseed rape - 14%

Two main traits of GM crops - Herbicide tolerance and insect resistance

Question 42

Herbicide tolerance

Answer 42

Resistance to glyphosate

Glyphosate blocks AA synthesis

Introduction of a mutant EPSPS gene - glyphosate cannot bind - resistance

Question 43

Insect resistance

Answer 43

Bt toxin - toxic protein that binds to inset epithelial cells in digestive tract and creates holes