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BT2001: Biotechnology > Microbial Biotechnology > Flashcards

Microbial Biotechnology Flashcards

1
Q

What are FOUR common industrial biotechnology strains?

A
  • E. coli
  • B. subtilis
  • S. cereviseae
  • A. niger
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2
Q

What bacterial strain is used in products to treat IBS?

A

Bifidobacterium longum

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3
Q

What yeast strain is used to make Quorn?

A

Fusarium venenatum

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4
Q

What is the Metagenome?

A

The collective genomes of an assemblage of microorganisms

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5
Q

What are the steps in Metagenomic Screening?

A
  1. Isolate combined DNA
  2. Chop it up
  3. Clone fragments into a suitable vector
  4. Transform into a suitable host strain
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6
Q

What are the steps in Sequence-Based Screening?

A
  1. Sequencing of metagenomic DNA
  2. Gene prediction
  3. Gene annotation
  4. Primer design and amplification
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7
Q

What are the steps in Sequence-Based Screening?

A
  1. Sequencing of metagenomic DNA
  2. Gene prediction
  3. Gene annotation
  4. Primer design and amplification
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8
Q

What are the steps of Function-Based Screening?

A
  1. Construction of metagenomic library
  2. Activity detection
  3. Isolation and sequencing of positive clones
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9
Q

How many species of microbes are known?

A

5000

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10
Q

What is the percentage of microbial cells in the body?

A

90%

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11
Q

What is the percentage of microbe from the Earth’s biomass?

A

60%

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12
Q

How many prokaryotic cells are there on Earth?

A

4-6 x 10^30

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13
Q

How many bacterical cells are there in a gram of soil?

A

10^9

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14
Q

How many bacteria are there per ml of ocean water?

A

10^6

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15
Q

What is the total number of bacteria are there in the marine environment?

A

3.67 x 10^30

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16
Q

What is the percentage of microbes in seawater that can be cultured?

A

0.001 - 0.1%

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17
Q

What was the first antibiotic to be discovered and by who?

A

Penicillin discovered by Fleming in 1928

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18
Q

From what fungal strain is Penicillin produced?

A

Penicillium notatum

(Now Penicillium chrysogenum used)

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19
Q

What bacterial strain does Penicillin inhibit?

A

S. aureus

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20
Q

What is a mould?

A

A fungus that grows in the form of multicellular filaments called hyphae

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21
Q

Name FIVE commercially significant secondary metabolites used as antibiotics

A
  1. Streptomycin
  2. Tetracycline
  3. Chloramphenicol
  4. Kanamycin
  5. Neomycin
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22
Q

What antibiotics inhibit cell wall synthesis?

A

Beta lactams (penicillin, cephalosporin), Vancomycin, Bacitracin, and Polymyxin (membrane)

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23
Q

What antibiotics inhibit nucleic acid synthesis?

A
  • Sulfonamides and Trimethoprim (folate synthesis
  • Quinolones (DNA gyrase)
  • Rifampin (RNA polymerase)
24
Q

What antibiotics inhibit protein synthesis?

A
  • Macrolides & Chloramphenicol (50S)
  • Tetracycline & Aminoglycoside (30S)
25
Q

How is antibiotic resistance spread in humans?

A
  • A person develops resistance in the gut and then it spreads to the general community

OR

  • In hospital, resistant microbes spread directly to other patients
26
Q

How is antibiotic resistance spread in animals?

A

Animal develops resistance, drug-resistant bacteria remain on meat, and if not cooked/handled properly it can spread to humans.

27
Q

What is the “Perfect Storm” of antimicrobial resistance?

A

Companies pulling out of antibiotic research and fewer antibiotics are discovered due to AMR

28
Q

Where does Penicillin production take place?

A

In a fermenter / bioreactor

29
Q

What are the THREE typical requirements for producing Penicillin in a bioreactor?

A
  • Aseptic operation
  • Agitation & aeration
  • Measurement & control
30
Q

What are the THREE possible products after yeasts / moulds being in a production fermenter?

A
  • Compressed yeast cakes
  • Active dry yeast
  • Nutritional dry yeast
31
Q

What is a fermenter / bioprocessor / bioreactor?

A

A vessel and ancillaries which has been designed to facilitate the growth and/or activities of microorganisms under controlled and monitored conditions

32
Q

What is the size of a bioreactor for diagnostics enzymes production?

A

1000-20,000 litres

33
Q

What is the size of a bioreactor for antibiotic and some enzyme production?

A

40,000-80,000 litres

34
Q

What is the size of a bioreactor for penicillin production?

A

100,000-150,000 litres

35
Q

What is the size of a bioreactor for amino acid, beer, and wine production?

A

200,000-500,000 litres

36
Q

Does penicillin production need oxygen?

A

Yes

37
Q

What does agitation in the fermenter provide?

A

Uniform, controllable conditions and avoids nutrient depletion and product build up

38
Q

What does aeration in the fermenter ensure?

A

Oxygen supply to all cells

39
Q

What are the FOUR phases of microbial growth?

A
  1. Lag phase
  2. Acceleration/Log phase
  3. Stationary phase
  4. Death phase
40
Q

What is the second stage for penicillin commercial production?

A

Innoculum

41
Q

What are THREE culture storage methods?

A
  1. Refrigeration (2-6°C)
  2. Lyophilisation (freeze dry)
  3. Frozen storage (-18°C to -80°C)
42
Q

What is the kill rate of frozen storage?

A

95%

43
Q

What is the temperature range of glycerol suspensions?

A

-80°C to -196°C

44
Q

Which phase does PRIMARY metabolism occur?

A

Trophophase (Lag & Acceleration phase)

45
Q

Which phase does SECONDARY metabolism occur?

A

Idiophase (Stationary & Death phase)

46
Q

What are Point Mutations?

A

Any mutation where only a single base of the DNA is affected (most common type)

47
Q

What are Frameshift Mutations?

A

Addition or Deletion of one or two base pairs

48
Q

What are the THREE typical effects of frameshift in a transitional region?

A
  1. Destroys encoded product function due to loss of reading frame
  2. Discloses nonsense signals and may have polar effects
  3. May be restored by nearby compensatory frameshift
49
Q

What are Chromosome Mutations?

A

Change in the order or location of genes

50
Q

How do Chromosome Mutations change the order or location of genes?

A
  • Deletion
  • Disruption
  • Duplication
  • Translocation
  • Amplification
  • Inversion
51
Q

What are the essential requirements for bioreactors that also reflect important physiochemical aspects of microbes?

A
  • Maintain sterility
  • Homogeneity
  • Low shear forces
  • Thermal stability
  • Suspend solids
  • Gas exchange (oxygen)
52
Q

What is the advantage of metabolic versatility?

A

Diverse arrays of reactions possible

53
Q

What is the major motivation for industrial strain improvement?

A

Economic

54
Q

What was the output for the original penicillin production strain and the mutated one in g/L?

A

0.01 g/L
After mutation based strain development it becomes >7 g/L

55
Q

What is responsible for penicillin optimisations?

A

Repeated classical mutagenesis and screening

56
Q

Is the spontaneous mutation frequency high or low?

A

10^-8
(Very Low)

57
Q

What increases the rate of occurrence of mutations?

A

Random mutagenesis by interfering with DNA synthesis and DNA repair mechanisms

BT2001: Biotechnology (10 decks)

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