Industrial Biotechnology Flashcards
Define sustainability
Creating and maintaining the conditions under which humans and nature can exist harmoniously that permits the fullfilling of social, economic and other requirements of the present and the future.
Rationalise the argument of oil vs. bio-feedstock
- thinking about what the supply chain of the future will look like - will it be bio or chemical based
- important to consider competition of land for food vs. land for chemical feedstocks and fuels
- most likely both i.e a mixed oil/bio economy
- biotechnology requires new industrial manufacturing methods and
biocatalyts - use of food based feedstock for e.g. bioplastics increases the pressure on food prices but release pressure on oil prices
What are some current sustainability concerns?
- mounting waste
- scarcity of quality water
- difficulty associated with extracting oil and coal reserves
- diminishing metal / material resources
What are some examples of industrial biotechnology all products from everyday life?
- insulin
- biological washing powders
- stone wash jeans
- cheese
Describe the production of insulin
- originally made from pigs, whose insulin is similar to human insulin
- the gene that makes insulin was found and copies of it made and inserted into E.coli
- this allowed them to group colonies/ batches of E.coli that produce human insulin
What is the role of insulin in the body?
- a hormone that helps take up sugar from the blood
- diabetics cannot make insulin and so have to have regular injections of insulin
Describe the industrial biotechnological process of making stonewashed jeans
- biological washing powders contain a variety of enzymes that break down and remove stains
- enzymes can be effective at temperatures
List 3 enzymes commonly found in detergents and what they down
- proteases - break down proteins e.g. Blood, egg, gravy
- lipases - break down fats and grease
- amylases - break down starches
Describe the industrial biotechnological process of stonewashed jeans and the associated advantages
- produced by washing jeans with the enzyme cellulase produced naturally by a bacteria and fungi
- jeans need to be rinsed few times using less water energy
- jeans are also not damaged as much thereby increasing their durability
What is the original process of stonewashed jeans and the disadvantages associated with it?
- using pumice stone
- stones can damage material making it less durable
- stones can also damage washing machine
- labour intensive because the stones need to be removed from the pockets of the jeans by hand
- jeans need to be rinsed several times which uses more water and adds cost
Describe the biotechnology case study of cheese
- traditionally made by taking milk then adding bacteria and enzymes from the stomached of calves (rennet)
- for vegetarian cheese cheese-makers sometimes use enzymes from plants i.e. fig trees, nettles, and thistles
- the enzymes do not work as well and those from the calf and the cheese doesn’t taste as good
- so the calf’s enzyme gene was inserted into a microorganism to produce recombinant enzyme
What are the three basic facts of natural selection?
- organisms produce more offspring than can survive
- individual characteristics vary
- many characteristics are inherited by offspring
Describe the process of sugar cane to tyres.
- the manufacturing of isoprene from sugar cane, corn, corn cobs, switch grass or other biomass that involve:
- microbial strain development
- large scale fermentation
- recovery and purification
- using bioisoprene reduces the seven gallons of petroleum feedstock per tyre required to close to zero
Describe the IB of rubbish to bio ethanol
- the conversion of cheap biomass and wastes to clean fuel and energy
- delivers a step change in green house gas emissions
- can be achieved safely, reliably, in a cost effective and sustainable way
- process: feedstock –> gasification –> renewable power –> fermentation –> bio ethanol which can be used as fuel
What are some examples of biocatalyts?
- isolated enzymes
- library of protein variants
- microbial culture collection
What are the screening methods associated with developing an active enzyme?
1- whole cells -genetic selection, colorimetric tests
2- microlitre plates -enzyme coupled assays, labelled substrates
3- analytical instruments -GC, HPLC, TLC, NMR, IR, MS
Briefly describe enzymes.
- proteins whose MW is 20-300KD
- usually unstable and low activity outside pH 5-9 therefore often need to be used in buffers
- commercially typically supplied as lyophilised powders and maybe crude preparations
- activity defined in units (U) per mg
Define the unit used to describe enzyme activity
U/mg - 1 Unit is the amount of enzyme that catalyses the conversion of 1μmol of substrate per min understand standard defined conditions of temperature and pH
Describe the process of the production of whole cells
- a sample of the desired microbial strain is obtained- either as an ampoule from a culture collection or glycerol stock stored in a -80 degree freezer
- microbial strain is cultivated on solid phase in a static incubator
- the microorganisms are maintained on agar plates or on slopes
- the microbial strain is grown in liquid phase in a shaking incubator
- enzymes of interest are produced within microorganism
- the microorganism cell containing the enzyme is harvest and centrifuged
- whole cells can be used for biotrnasformations
- cells can be lysed and the enzyme isolated
What is recombinant DNA technology?
- allows a foreign gene to be put into a host organism
- simplest host organisms are bacteria e.g. E.coli
- the organism can be programmed into producing the corresponding protein
- plasmids are used to shuttle DNA into host organism
What are the steps of gene cloning that lead to recombinant protein expression?
- organism is selected
- DNA is isolated and the gene identified
- the gene cloned
- plasmid vector containing the gene is inserted to the microorganism
- bacteria is cultivated into colonies on agar plates
- the protein is then purified from the bacteria cultures
What are plasmids and describe their role.
- circular double stained DNA that contain antibiotic genes
- they contain restriction sites that can be used for the insertion of foreign DNA
- easier to manipulate than chromosomal DNA
- many different plasmids are commercial available
