Professional Documents
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The Organism
The Organism
The Organism
Lecture 2
The
Outline
Properties of a Useful
Industrial Microorganism
Properties of a Useful
Industrial Microorganism
Genome known.
Gene transfer systems available.
Genetically stable.
Safe.Bacillus anthricis?
Well known industrially.
Culture collections.
Biodiscovery
Screening
Screening
More complex studies.
Medium/process
optimisation, genetic
stability etc.
Decreasing No. of
Isolates
Simple/High
throughput assays
Strain Improvement
Note
Organisms, medium and process will be
discussed separately during this course, but
they must always be considered TOGETHER
when developing or improving an industrial
process.
Titre
Year
Induction
Enzyme
Immediate or final
product
Inhibition/Repression
stops or reduces enzyme activity
Induction
Substrate
Induction
Enzyme
Immediate or
subsequent
product
Inhibition/Repression
Constitutive Mutants
The Chemostat
sequential cultures of
Escherichia coli alternating lactose and
glucose will enrich for mutants constitutive
for beta galactosidase.
Inhibition/Repression
Substrate
Enzyme
Induction
Inhibition/Repression
Immediate or
subsequent
product
Our strategy:
pathways to avoid
inhibitor/repressor build-up.
Simple
L,L DAP
Meso DAP
L-lysine + CO2
Feedback
Control
In
Cultures
Strain A
Glycerol
Cannot
L,L DAP
Meso DAP
L-lysine + CO2
Strain B
The
mixture contains:
Large
Control Point
Aspartate semi-aldehyde
Lysine
Homoserine
Methionine
CONCERTED
CONTROL
Threonine
Isoleucine
NOTE
No
Lysine
Homoserine
Methionine
Threonine
Isoleucine
Use
Catabolite repression
When
Catabolite Repression
(Glucose Effect)
galactosidase
- glucose
Glucose
added
+ glucose
Time (hr)
+ lactose
Your Strains
How
Uncontaminated.
True to their known
characteristics, both
qualitative and
quantitative.
Starters must be
provided in a suitable
and active form.
Lyophilisation (freeze
dried stocks)
Glycerol suspensions at
80oc to -196oc
Freeze onto cryobeads
(The Protect system)
Agar slope cultures
overlaid with mineral oil
and stored at 20oc