Lecture 6: Industrial Microbiology and Biotechnology
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- Name 5 industrial applications microbiology has.
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1) Food industry
-organic acids, supplements, enzymes
2) Pharmaceutical applications
-Antibiotics
3) Agricultural applications
- Bioinsecticides
4) Environmental biology
- Bioremediation (remove pollutants from environment using microorganisms)
5) Vaccine production - What are the main products of fermentation?
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acid
alcohol - What are some products of the TCA Cycle when the TCA cycle is stopped at citric acid?
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citric acid is used to make:
1) inks and dyes
2) soft drinks
3) chemistry
4) anticoagulants (heart disease therapy)
5) foods - In genetic engineering and biotechnology, products are achieved through ____ and ____. Or through ______.
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mutation and screening of natural gene pool
recombinant DNA techniques - What are the results of recombinant DNA techniques?
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1) redesign of a gene (and the protein it encodes)
2) Modification of gene expression
3) Transfer genetic information between organisms - What are the three tools of Recombinant DNA?
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1) site-directed mutagenesis
2) random mutagenesis using PCR
3) Restriction and modifying enzymes
* site-directed mutagenesis and random mutagenesis are opposites - What is site-directed mutagenesis?
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Use oligonucleotides, DNA primers with designed mutation incorporated.
Pick a site to change on a gene and mutate it. - What is Random mutagenesis using PCR?
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-Use MANGANESE as cofactor instead of magnesium for polymerase chain rxn
-Causes polymerase to be more error prone
A gene goes through PCR, which randomly mutates a gene - Human genes can be cloned in microorganisms
- look on page 252
- Mutation and Screening of Available Gene Pool. Say that your desired product is PENICILLIN. It is the first antibiotic to be mass-produced. Originially, it came from _____, which yielded 5mg/L.
- Penicillium notatum
- What is the other, better, source of penicillin? This source is even further improved by ____ and subsequent screening. Its yield is ___mg/L.
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P. chrysogenum
UV mutation
60,000mg/L - Redesign of Gene using Recombinant DNA. What is Hirudin?
- an anticogulant polypeptide (takes out blood clots)
- Hirudin's effects are similar to ___ or ___.
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warfarin or heparin
Hirudin has 65 amino acids - In redesigning hirudin using Recombinant DNA, use _____ mutagenesis to change ASN47 to ___ or ___
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site-directed
LYS or ARG
* single amino acid change results in improved anticoagulant activity - Control regions of genes can be altered to change _____ levels.
- expression
- In modification of Gene Expression, engineer a _______ to increase expression of desired gene
- promoter construct
- Give an example of a promoter construct that increases an expression of a desired gene.
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AOX1 promoter of the methylotrophic yeast Pichia pastoris.
- Used to express over 500 heterologous proteins including angiostatin and endostatin - Transfer of Genetic Information is achieved by _____
- genetic engineering
- What is the world's toughest bacterium?
- Deinococcus radiodurans- tolerates high levels of radiation
- What bacterium tolerates high levels of radiation?
- Deinococcus radiodurans
- E. coli have genes that degrade ionic _____ compounds
- mercury
- Transfer ____ genes to Deinococcus to create recombinant bacteria that can _____
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E.coli
clean up radioactive toxic waste sites - Primary metabolites are made during _____
- trophophase
- Are primary metabolites associated with growth phase?
- yes
- Give some examples of primary metabolites
- amino acids, fermentation end products, enzymes
- Seconday metabolites are made during _____.
- idiophase
- Do Secondary metabolites have a direct relationship to growth?
- NO
- Give 3 examples of secondary metabolites
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antibiotics
antimicrobials
mycotoxins - Give examples of products of industrial microbiology
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1) Antibiotics
2) Microbes that make bacterial cement
3) Biosurfactants
4) Bioinsecticides
5) Biosensors
6) Bioremediation
7) White Rot Fungus - Give three examples of antibiotics
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- Neomycin (Streptomyces fradiae)
- Polymyxin B (Bacillus polymyxa)
- Griseofulvin (Penicillium griseofulvum) - What digests urea to make ammonia?
- Bacillus pasteurii, sporosarcina ureae
- Ammonia and water create____
- ammonium hydroxide
- Ammonium hydroxide converts calcium chloride to __________ which crystallizes as limestone.
- calcium carbonate
- Give an overview of microbes that make bacterial cement
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- Bacillus pasteurii, sporosarcina ureae digest urea to make ammonia
- Ammonia and water create ammonium hydroxide
- NH2OH converts calcium chloride to calcium carbonate which crystallizes as limestone - Biosurfactants are used for?
- emulsification, solubilization, and phase dispersion
- ______ can make a biosurfactant which is a glycolipid emulsifier - improves oil recovery
- Pseudomonas aeruginosa
- What is used for emulsfication, solubilization, and phase dispersion?
- biosurfactants
- What toxin kills caterpillars?
- Bacillus thuringensis
- Bacillus thuringensis is a toxin that kills caterpillars by ___
- forming pores in caterpillar guts
- What toxin kills mosquitoes
- Bacillus sphaericus
- What toxin kills Japanese beetle larvae?
- Bacillus popilliae
- What genes do you use to create transgenic insect resistant crops?
- Bacillus genes
- Use Bacillus genes to create _____
- transgenic insect-resistant crops
- What are biosensors used for?
- Use microbes to 'sense' whether toxic products are around
- E.coli containing lux genes (from Vibrio or Photobacteirum) emit ____
- light
- E.coli containing lux genes that emit light are used as a warning system to detect failures in _____
- waste-water treatement plants
- What does it mean if bacteria fail to emit light?
- toxic pollutants are still in the water
- What is bioremediation?
- Removing pollutants from environment using microorganisms
- What is PCBs?
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polychlorinated biphenyls
it is a bad pollutant! - Naturally occurring microbes can partially dehalogenate PCBs under ____ conditions
- anaerobic
- You would _____mud to get aerobic bacteria to complete degradation.
- aerate
- Give an example of white rot fungus
- Phanerochate chysosporium
- White Rot FUngus, Phanerochaete chysosporium, degrades ___.
- lignin
- What is lignin and where is it found?
- phenylpropane-based polymeric compound found in wood
- Besides lignin, name 6 other items White Rot Fungus can degrade
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-benzene
-toulene
-agent orange
-TNT
-Xylenes
-Chlorinated compounds such as pesticide and herbicides - What is the problem with sending engineered microbes into the environment?
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- engineered microbes can't actually contact target
- Engineered microbes fail to survive environment and compete with indigenous microbes, so they die or get eaten