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Molecular Biotechnology > Lecture 15 -Bioremediation > Flashcards

Lecture 15 -Bioremediation Flashcards

1
Q

Define bioremediation, what is it based on and what does it involve

A

Current and potential use of microorganisms and plants to treat pollution in the environment
Based on natural ability of living organisms to reduce or eliminate pollution
Involves ENHANCING removal rates of pollutants from an ecosystem

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2
Q

Define pollution

A

the presence in or introduction into environment of harmful or poisonous substances, or excessive levels of light, noise, organic waste etc. Especially as result of human activity

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3
Q

Give 3 broad examples of different types of pollution

A

Oil spills e.g. Exxon valdez spill. 260000-750,000 crude oil barrels spilled
Atmospheric pollution - anything discharged from chimneys
Contaminated land

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4
Q

What are the 4 classifications of pollutants

A

Solid
Liquid
Gas
Ionising radiations and electromagnetic waves

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5
Q

What environments are affected by pollution

A 
ALL
Marine
terrestrial
fresh water
atmosphere
extraterrestrial
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6
Q

What has been the UK’s biggest ‘serious pollution incident’ other than ‘other’ on the pie chart since 2008

A

sewage and waste

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7
Q

What are the challenges faced when quanitfying pollution in an area?

A

There’s no true record
325000 sites have had KNOWN use that COULD have led to contamination
Historically still being effected - e.g. lead poisoning of ground by romans

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8
Q

What are the 2 main reasons/arguments for treating pollution

A

Ecological - destruction of habitat by pollution can lead to losses in species diversity and extinction
Human - pollution can have deleterious effects on both human health and welfare (contaminated land worth less)

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9
Q

What are the 3 broad ways of treating pollution

A

Physical - use mechanical process e.g. containment
Chemical - e.g. neutralising acid
Often combined to a physiochemical treatment
Biological

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10
Q

Define biological treatment

A

Any process where a pollutant is converted into an alternative (usually less harmful) form by the action of a biological system.

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11
Q

What aspect of biological agents does bio treatment exploit?

A

Their metabolic abilities to remove/treat pollutants naturally

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12
Q

Give an historic example of bio treatment

A

manure on fields as fertiliser

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13
Q

What kind of organisms are used in bio treatment

A

Animals, plants and microorganisms

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14
Q

Are animals well suited to treat pollution? Why?

Give an example where they have been used

A 
No
Large
slow growing
ethical issues
chicken feathers to mop up oil
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15
Q

What is phytoremediation

A

The use of plants to treat pollution

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16
Q

List the 6 different phytoremediation processes that can be exploited, giving a small description

A

Phytoextraction-plants translocate pollutant from soil to harvestable body
Phytodegradation-metabolic abilities of plants and associated microorganisms to degrade organic pollutants
Rhizofiltration-plant roots absorb/adsorb pollutants, mainly metals, from water. Then keep them in place
Phytostabilisation-plants reduce bioavailability of pollutants in the environment , by storing them in their plant system
Phytovolatillisation - volatile products taken up with H2O then evaporated, so pollutants are volatised
Rhizodegradation - plant acts as host fro pollutant degrading organism

17
Q

Give a detailed example of phytoremediation using arabidopsis

A

Novel cytochrom P450 in a bacteria (R. rhodochrous) that degrades RDX-a high explosive
Transfered gene to arabidopsis, they then express.
Grew plants on contaminated soil - found that it removes and degrades the explosive from soil

18
Q

What do majority of bio treatments use

A

microbes - bacteria, fungi, protozoa and algae

NOT viruses

19
Q

What are the characteristics that make microorganisms suitable for use in waste and pollution treatment

A
  • Ability to survive hostile envir.
  • rapid growth rates
  • ability to grow on range of C and energy sources
  • genetic plasticity - easy for them or us to transfer genetic info
  • easy to control and study
  • no ethical considerations
20
Q

What are the 2 basic bioremediation strategies, and define them

A

Biostimulation - modify conditions to stimulate INDIGINOUS microbe activity to break down pollutant. Can be done both in and ex situ
Bioaugmentation - introduce large quantities bacteria into contaminated environment to remove pollution.

21
Q

Define in situ and ex situ

A

ex situ - remove contaminat to specialised treatment facility and treat it there
in situ - treat at site

22
Q

How is biostimulation done ex situ

A

In suspended growth reactor

23
Q

How is biostimulation done in situ

A

Liquid delivery - add nutirents and/or o2 to site, set up circulatory system so product can be removed

24
Q

Why does biostimulation not always work

A

pollutant killed indiginous popl.

25
What are the 2 options for providing microbes for use in bioaugmentation
grow up indiginous or naturally occuring and inoculate site | develop genetically modified bacteria
26
What does gene introduction exploit
plasmids, mating of donor and recipient microbes
27
Can gene introduction occur naturally as well as artificially
yes.
28
What's an alternative to natural gene introduction
genes cloned into broad host range plasmid , then optimised via: transcription, promotor, terminator manipulation increase copies of plasmid present improve stability
29
What is the main porblem with gene stability and how might it be solved
- risk of transfer to other bacteria | - incorporate gene into host genome
30
Give an example of gene introduction
Phenyl hydroxylase genes from Cornamonas testoseroni R5 (a bacteria with good phenol degrading ability, but not good at surviving outside the lab) Genes put in rN7 cornamonas that was dominant in sewage sludge Resulting bacteria were able to survive and degrade pheno 3xfaster than before
31
How is gene alteration done
Gene cloned in lab bacteria so easier to use position accordingly with transcription pormotor and terinator regions increase the number of copies of genes in host improve stability of cloned gene proteins
32
Give example of gene alteration
Pseudomonas putida contains pWWO plasmid, whcih allows it to degrade toleune and xylene, byt not ethylbenzoate. DESPITE the presence of all the functional genes needed to degrade it (because EB doesn't induce the pathway) Cloned xylS gene (controls pathway) into ecoli and generated mutant that responds to EB mutant gene transfered to Ps.putida - success
33
What problems might arise in the real environment even after successful gene manipulation
fail to adapt to contaminated environement conditions Insufficent substrate competition with indiginous popl use other oragnic substrates in preferences to pollutants predation, usually by protozoa
34
How could you get around the 'real environment' problems
Horizontal gene transfer
35
Define horizontal gene transfer
introduction of remediation genes into indigious microbes alrewady adapted to survive to that environement
36
In HGT how is the host strain got rid of once it's inserted the genes to the indiginous microbe
insert conditional suicide gene to donors | -if the conatminent is not present host dies
37
Give an example of a conditional suicide system
In presence of inducer contaminent, 3-methylbenzoate, production of lacI repressor blocks Gef protein When contaminent completely degraded, Gef protein is produced which kills cells by producing porins that destabilise cell membrane
38
List molecular techniques used to measure what microbes are doing
``` 16s rDNA PCR Reverse transcription PCR Real time PCR Fluorescent in situ hybridization Microarrays reporter genes ```

Molecular Biotechnology (14 decks)

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