8 | metabolic engineering strategies I Flashcards

1
Q

What is the motivation behind coupling product formation to growth?

A
  • force cell to produce desired compound as by-product of growth
  • β†’ ensures production is evolutionarily stable.
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2
Q

What is strong coupling in metabolic engineering?

A

KO strategy: product formation strongly coupled to growth (biomass production)
ensure any growth-supporting flux dist also produces product > threshold.

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3
Q

What is weak coupling?

A

Product is only formed at the optimal growth flux, not necessarily under all conditions.

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4
Q

What is the exhaustive KO strategy covered in lecture 8 on metabolic engineering?

A
  • Try all possible subsets of knock-outs from the list of genes.
  • Evaluate product yield or other objectives, solution = maximum of all maxima
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5
Q

Write the LP for an exhaustive knock out enumeration.

A

For every subset 𝑇 βŠ† 𝐿:
max π‘£π‘β„Žπ‘’π‘šπ‘–π‘π‘Žπ‘™
s.t.
𝑁𝑣 =0
βˆ€π‘— ∈ 𝑇, 𝑣𝑗= 0,
βˆ€π‘–, 𝑖 βˆ‰ 𝑇,π‘£π‘–π‘šπ‘–π‘› ≀ 𝑣𝑖 ≀ π‘£π‘–π‘šπ‘Žπ‘₯
π‘£π‘π‘–π‘œ β‰₯ π›Όπ‘§βˆ—
If we want sufficient high product yield to be at some level of biomass yield:
where π‘§βˆ— is the optimal growth.

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6
Q

What additional constraint ensures viability in exhaustive knock-out tests?

A

vbio β‰₯ Ξ± Β· z, where z is wild-type biomass flux

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7
Q

What are the limitations of exhaustive knock-out search?

A
  • Combinatorial explosion: infeasible for large gene lists
  • Cannot scale to full genome-wide searches
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8
Q

What is a bi-level optimization problem?

A
  • One optimization problem (outer) depends on the solution of another (inner)
  • I.e two nested optimizations
  • The outer objective uses the result of the inner optimization
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9
Q

Why do bi-level problems appear in metabolic engineering?

A
  • The outer problem wants to maximize product formation (engineer)
  • The inner problem responds by optimizing its own growth (cell)
  • The engineering strategy must account for the internal optimization
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10
Q

In OptKnock, what are the outer and inner problems?

A
  • Outer: maximize vtarget (e.g. product formation) by choosing KOs
  • Inner: maximize vbio (growth) given knock-outs
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11
Q

How are knock-outs encoded in OptKnock?

A
  • Binary variable yi ∈ {0, 1}
  • yi = 1 β†’ reaction is active
  • yi = 0 β†’ reaction is knocked out β†’ vi = 0
  • Use bounds: yi * vimin ≀ vi ≀ yi * vimax
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12
Q

What does the inner problem in a bi-level formulation depend on?

A
  • The outer decision variables (e.g. knock-outs y)
  • These change the feasible region for the inner problem
  • The inner LP is parameterized by y
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13
Q

What makes OptKnock a mixed-integer bi-level program?

A
  • Outer layer includes binary variables yi
  • Inner problem is a linear program over flux variables v
  • The feasible set for v depends on y
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14
Q

What is OptKnock?

A
  • A bi-level MILP to find knock-outs that couple growth and production
  • Automates the search for knock-out designs
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15
Q

How is the number of knock-outs restricted in OptKnock?

A

Add a constraint for K, the max allowed deletions: βˆ‘(1βˆ’yi) ≀ K

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16
Q

What is the full program for OptKnock (Burgard et al. (2003) Biotech. & Bioeng.)

A

max over y: vchemical
st:
max vbio
st:
N v = 0
for all i ≀ 1 ≀ m, yi Β· vimin ≀ vi ≀ yi Β· vimax
yi ∈ {0, 1}
βˆ‘i=1m(1βˆ’yi) ≀ K

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17
Q

What does the inner problem in OptKnock represent?

A

The cell’s behavior: maximize biomass production given knock-outs

18
Q

What does the outer problem in OptKnock represent?

A

The engineer’s goal: maximize product formation by choosing which knock-outs to apply

19
Q

What assumptions does OptKnock make about the cell?

A
  • The cell optimizes growth
  • Product formation is only induced indirectly via knock-outs
20
Q

Why are bi-level problems difficult to solve?

A
  • The inner problem changes with each outer decision
  • You can’t just solve them separately
  • to nesting, discrete nature of int vars β†’ problem non-convex, combinatorial
21
Q

What does β€œcombinatorial” mean in the context of bi-level optimization?

A
  • refers to having to choose among many discrete combinations, eg KO sets
  • Each combination may lead to a different inner solution
  • possible combinations can grow very large - β€˜combinatorial explosion’
  • Can make problem hard to solve efficiently
22
Q

Why can’t bi-level problems be solved directly?

A

Inner LP depends on outer variables β†’ standard MILP solvers unable to handle it directly.

23
Q

How can bi-level problems be solved in OptKnock?

A

Two approaches:
* By replacing the inner LP with its dual problem
* Apply Karush-Kuhn-Tucker (KKT) conditions (optimality) to connect primal, dual

24
Q

What are the KKT conditions used to solve bi-level problems?

(not so relevant ! )

A
  • Stationarity
  • Primal feasibility
  • Dual feasibility
  • Complementary slackness
25
Q

What is complementary slackness?

A

For each constraint:
* If it’s active, dual variable may be positive
* If it’s inactive, dual variable is zero

26
Q

What are mixed-integer linear programs (MILPs)?

A

LPs with some variables required to be integers (e.g. binary knock-out indicators)

27
Q

What is LP duality?

A
  • Every LP has a dual problem
  • The dual variables represent the shadow prices of the constraints
28
Q

What is a shadow price in LP duality?

A

Each constraint in the primal has a shadow price.
* = The value of a dual variable corresponding to a primal constraint
* = how much objective function would improve if that constraint relaxed by 1 unit
* AKA marginal value of a resource

29
Q

If you have n variables and m constraints in an LP, how many constraints will you have in its dual?

A
  • m variables
  • n constraints
30
Q

What is the optimality property in LP duality?

A

Let π‘₯ be a feasible solution to a primal LP, and πœ‡ be a feasible solution to the dual LP.
If 𝑐𝑇π‘₯ = π‘π‘‡πœ‡ then π‘₯ and πœ‡ are optimal solutions to the primal and dual LPs, respectively.

31
Q

What is the strong duality property in LP duality?

A
  • If primal has finite optimal solution, then so does its dual, and these values are equal.
  • ie maxπ‘₯ 𝑐𝑇π‘₯ = minπœ‡ π‘π‘‡πœ‡
    That means:
  • Solve the primal β†’ get a value; Solve the dual β†’ get a value. βœ… Both values are equal
    This only holds if:
  • The primal or dual has a feasible solution (satisfies all constraints)
  • The problem is bounded (no infinite solutions)
32
Q

How do you convert a primal LP into its dual (without Lagrangian)?

A
  • Each constraint in the primal becomes a variable in the dual
  • Each variable in the primal becomes a constraint in the dual
  • The primal objective coefficients become RHS values in the dual constraints
  • The primal constraint matrix A is transposed in the dual
33
Q

In LP duality, what happens to β‰₯ constraints in the primal?

A
  • They become ≀ constraints in the dual
  • Their corresponding dual variables must be β‰₯ 0
34
Q

What is the dual of this primal LP?
min cT x
s.t. Aineq x β‰₯ bineq
x β‰₯ 0
Aeq x = beq

A

max bineqT y1 + beqT y2
s.t. AineqT y1 + AeqT
y2 ≀ c
y1 β‰₯ 0

35
Q

Why is LP duality useful in OptKnock?

A
  • Allows removing inner LP, replacing with constraints involving dual variables
  • Converts the bi-level problem to a single-level MILP
36
Q

What are two approaches to flattening bi-level optimization problems?

A
  • LP duality: replace the inner LP with its dual and enforce strong duality
  • KKT conditions: use stationarity, feasibility, and complementary slackness via the Lagrangian
37
Q

Do LP duality and KKT methods give the same solution for bi-level LPs?

A
  • Yes, they both produce the same optimal result
  • They enforce optimality of the inner problem in different ways
38
Q

Do LP duality and KKT produce the same flattened single-level program?

A

No, the formulations differ
* LP duality leads to a more compact and structured MILP
* KKT-based reformulations are often larger and more complex

39
Q

What does it mean to β€œnot use the Lagrangian approach” for duality?

A
  • You should derive the dual using the standard LP rulebook, not calculus or KKT
  • Don’t introduce Lagrange multipliers
  • Just work structurally from constraint-to-variable transformation
40
Q

For a primal minimization problem:

min cTx
s.t.
A x β‰₯ b
x β‰₯ 0

What would be the key steps to formulate the dual?

A

Follow the standard duality principles in optimization.

  1. Identify Primal Components:
  2. Assign Dual Variables:
  3. Formulate Dual Objective:
  4. Formulate Dual Constraints:
41
Q

For a primal minimization problem:

min cTx
s.t.
A x β‰₯ b
x β‰₯ 0

What is the dual?

A

maxπœ‡ 4πœ‡
s.t
πœ‡ = 0

Perplexity explanation:

Original (primal) problem:
Minimize y
st
y β‰₯ 0,
x + y β‰₯ 4.

1. Identify Primal Components:
Variables:
yβ‰₯0,
x (unrestricted).
Objective: Minimize y
Constraints: One explicit constraint
x+yβ‰₯4.

Assign Dual Variables:
Each primal constraint becomes a dual variable. Here, the constraint
x+yβ‰₯4 corresponds to a dual variable
Ξ»β‰₯0.

Formulate Dual Objective:
The dual objective is derived from the right-hand side of the primal constraint multiplied by Ξ»:
4Ξ».

Formulate Dual Constraints:
For each primal variable, create a dual constraint based on its sign:
For x (unrestricted): The coefficient of x in the primal constraint (1) forms an equality constraint in the dual: Ξ»=0.
For y (non-negative): The coefficient of y in the primal constraint (1) forms an inequality constraint: λ≀1.
Combine Constraints:
The dual constraints are Ξ»=0, λ≀1, and Ξ»β‰₯0.

Dual Problem:
Maximize 4Ξ»
subject to
Ξ»=0,
Ξ»β‰₯0,
λ≀1.

Interpretation:
The only feasible solution for the dual is Ξ»=0, yielding an optimal value of 0 This matches the primal optimal value y=0 (achieved when xβ‰₯4), demonstrating strong duality.

Summary:
The dual LP is trivial in this case because the unrestricted variable x forces
Ξ»=0. This ensures the primal and dual optimal values align, satisfying the duality theorems in linear programming.

42
Q

What is the simplified OptKnock reformulation/approximation from the exercise, often used computationally, which:

  • Turns the bilevel OptKnock problem into a single-level MILP
  • Focuses on minimizing the number of knockouts
  • Adds a small constant Ξ΅ to soften knockouts

?

A

min βˆ‘ yi

s.t.

N v = 0
vbio β‰₯ Ξ± Β· z*
vtarget β‰₯ Ξ² Β· wtarget
v β‰₯ 0
βˆ€ i:
 (1 βˆ’ yi) Β· Ξ΅ ≀ vi
 vi ≀ (1 βˆ’ yi) Β· vimax + yi Β· Ξ΅
y ∈ {0,1}