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Princ of Biiop > 07 Growth Kinetics > Flashcards

07 Growth Kinetics Flashcards

1
Q

What is the overall goal of bioprocessing?

A

To produce our target biomolecule

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2
Q

Upstream processsing & examples

A 
- growth & production
>stock culture/ shake flasks
>seed fermentor
>sterilize fermentor & media
>production fermentor
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3
Q

downstream processing & examples

A
  • seperation & purification
    >cell seperation
    >product purification
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4
Q

bioprocess analysis

A
  • upstream process

- determines the best means for achieving optimal yield during fermentation phase

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5
Q

Growth Associated Product Formation

A
  • Product expression occurs as a consequence of growth

- Product formation is linked to energy metabolism

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6
Q

Non-Growth Associated Product Formation

A

Product expression occurs as a product of secondary metabolism, and is not linked to energy metabolism.

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7
Q

biomass

A
  • can be an indicator of product formation

>if growth associated

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8
Q

substrate

A
  • cells can only grow/ from bioproduct when sufficient substrate is available
  • limiting it can inhibit growth
  • over addition = less than ideal growth
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9
Q

Substrate uses:

A
  • maintenance
    > limiting substrates: carbon & oxygen
  • cell mass
  • products
    >synthesis of chemical products
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10
Q

cell yield formula

A

lChange in Cell Mass l
Yx/s = l———————————l
lSubstrate Consumed l

       lchngeX  l Yx/s = l-------------l
       lchngeS  l
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11
Q

Product yield formula

A

lchngeP l
Yp/s = l————- l
lchngeS l

P= delta product
S= delta substrate
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12
Q

Growth kinetics formula

A

(lnN2-lnN1)
u = ——————
(t2-t1)

N = # of cells at instance 1 & 2
t = time at instance 1 & 2
** growth rate constant = k, u, [h^-1]

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13
Q

Doubling time formula

A

ln(2)
td = ———–
u

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14
Q

Growth rate & Doubling time formulas (OD)

A

ln(OD at time 2)-ln(OD at time 1)
Uod = ———————————————–
elapsed time 2-elapsed time 1

   ln2 tD =------
     u
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15
Q

Trypan Blue

A
  • dye assay to determine cell viability
  • more accurate than OD600nm
  • Dead/Non viable cells are stained blue
  • Living/Viable remain unaffected
    • helps estimate volume of living and dead cells in culture
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16
Q

Hemocytometer

A
  • thick glass slide with etched grid lines

- count # of cells in specific volume (area) of solution

17
Q

Media nutrient review

A
  1. Carbon
    - Carbs are primary energy source
  2. Nitrogen
    - makes amino acids, nucleic acids, & other molecules
  3. Oxygen
  4. pH, Temp, Secondary factors
18
Q

pH can change due to:

A
  • utilization of substrate
  • NH4 production
  • Cellular by products
  • Cellular lysis/ cell toxins
19
Q 
pH range:
Bacteria
Yeast
Mammalian
Plant
A

> 3-8
3-6
6.5-7.5
5-6

20
Q

dissolved CO2

A
  • 2 degree factor

- toxic at extremely high or low levels

21
Q

Ionic strength

A
  • 2 degree factor
  • too high, inhibitory to normal cell membrane fxn
    > membr transport
    > osmotic balance
22
Q

Additives that can also inhibit growth:

A
  • Glucose > 100g/L
  • EtOH > 10g/L
  • NH4+ > 5g/L
23
Q

dissolved oxygen (DO)

A
  • O2 can be limiting substrate for aerobic ferment.
  • not readily soluble in water
  • too much can be detrimental to cellular production
24
Q

Oxygen Transfer Rate (OTR)

A

Ro2 = Kla(C*-Cl)

  • Ro2 = oxygen transfer rate
  • Kla = volumetric transfer coefficient (constant)
  • C* = saturated conc of oxygen (constant)
  • Cl = actual DO
25
Q

Oxygen Uptake Rate (OUR)

A

chngeO2
qo2X = ————–
chnge t

  • mols O2 consumed/ time
26
Q

The ___ the size of the bubble/globule, the ____ the SA available for mass transfer

A
  • smaller

- larger

27
Q

Impeller (Agitator)

A
  • helps diminish the size of air bubbles
  • Provide a large interaction area for oxygen transfer
  • decrease the diffusion path
  • maintain a uniform envmt
  • shear damage
28
Q

Operational Mode- Batch

A
  • simplest ferment operation
  • sterilization can be performed in reactor
  • all nutrients added before inoculation
  • max levels or C & N limited due to inhibition of cell growth
  • cells harvested when levels start to decline
29
Q

Continuous Flow rates:

A

Dilution > Growth
- culture washes out

Dilution < Growth
- culture overgrows

Dilution = Growth
- steady state culture

Princ of Biiop (10 decks)

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