Metabolic Engineering: Redirecting Flux Through a Production Host · seed 1 · A4, ink-friendly. The answer key prints on its own page for grown-ups.

Steering carbon toward your product

Science · Microbiology · ages 23-24
Name ______________________   Date ____________
  1. A native branch and a target branch both use intermediate I. Which process is a competing drain of I away from the target branch?

    • I is converted into a native byproduct
    • I is supplied more slowly from upstream
    • I is converted into the desired target product
  2. A carbon-only model converts one 6-carbon substrate into one 4-carbon product. Only CO2 carries the remaining carbon out, and no carbon accumulates. Which CO2 output completes the balance?

    • One CO2 molecule per substrate molecule
    • Three CO2 molecules per substrate molecule
    • Two CO2 molecules per substrate molecule
  3. Inducing a production pathway after biomass growth, by itself, makes product formation necessary for growth.

    Circle one:   True   False

  4. A host produces 30 mmol ATP/hour. Growth and maintenance use 18 mmol ATP/hour. A model pathway uses 3 mol ATP per mole of product. With no stored ATP available, what upper bound does this ATP budget set?

    • 10 mmol product/hour
    • 12 mmol product/hour
    • 4 mmol product/hour
  5. A branch competing for your target intermediate also supplies an essential growth precursor. Which change is a reasonable first test for redirecting flux?

    • Delete the branch and remove its growth precursor supply
    • Reduce branch activity while checking growth and flux
    • Keep branch activity fixed and assume target flux must rise
  6. A model pathway uses 3 mol NADPH per mole of product. Production has 15 mmol NADPH/hour available after other cellular needs, with no stored reserve. What maximum product flux does this NADPH budget allow?

    • 5 mmol product/hour
    • 15 mmol product/hour
    • 45 mmol product/hour
  7. In a model, growth is impossible at zero product flux. What does this growth coupling establish?

    • Every faster-growing cell has the highest product yield
    • Growth requires some positive product flux
    • Product is made only after growth has completely stopped
  8. After many generations, mutants lacking a costly production pathway become common. They grow faster than producers in this culture. Which process explains their increasing frequency?

    • Pathway loss guarantees a higher product yield per cell
    • Slower growth makes the producing cells divide more often
    • Selection favors the faster-growing mutants
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Answer key

For grown-ups. Fold this page away before handing over the rest.

Steering carbon toward your product W1-mt_fjz3EA-dDa-s1

  1. I is converted into a native byproduct · The native branch consumes the same intermediate needed by the target branch. Slower upstream supply is a different constraint.
  2. Two CO2 molecules per substrate molecule · Six carbons enter and four leave in product. The remaining two carbons leave in two CO2 molecules, with one carbon in each.
  3. False · Induction controls when the pathway is expressed. Making growth require product formation is a separate design condition.
  4. 4 mmol product/hour · Production has 30 - 18 = 12 mmol ATP/hour available. Dividing by the cost of 3 gives an upper bound of 4 mmol product/hour.
  5. Reduce branch activity while checking growth and flux · A partial reduction can test whether less competing consumption helps production while retaining enough precursor supply for growth. A gain is not guaranteed.
  6. 5 mmol product/hour · Each mole of product needs three moles of NADPH. Divide the available NADPH rate by three: 15 / 3 = 5 mmol product/hour.
  7. Growth requires some positive product flux · The model excludes growth without product formation. It does not specify how product yield ranks across cells or strains.
  8. Selection favors the faster-growing mutants · The mutants have a stated growth advantage in this culture. Selection can increase their share of the population while production falls.
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