Synthetic Biology Parts, Devices and Applications

Table 12.2

DNA scaffolds used to order enzymes of biosynthetic pathways.

Product

Enzyme

DNA‐binding protein and its target site

Chimeric protein

DNA scaffold

a)

Comments

References

Operators

No. units (n)

Spacer (bp)

l‐Threonine

Homoserine dehydrogenase (HDH)

ADB1CAAGCTAGGGAG

HDH‐ADB1 (E1)

[1

:^1

:^ 1]

n

1

8, 18, 28

Reduced production time by more than 50%

[10]

Homoserine kinase (HK)

ADB2GACGAGGGGGTG

HK‐ADB1 (E2)

[1

:^1

:^ 2]

n

[1

:^1

:^ 3]

n

[1

:^1

:^ 4]

n

1

8

Threonine synthase (TS)

ADB3GAAGGGGGGGTA

TS‐ADB1 (E3)

trans

‐

Resveratrol

4‐Coumarate–CoA ligase (4CL)

Zif268GCGTGGGCG

Zif268‐4CL (E1)

[1

:^ 1]

n

4, 16

2, 4, 8

More than threefold improvement in resveratrol production

[11]

[1

:^ 1]

n

4, 16

2, 4, 8

Stilbene synthase (STS)

PBSIIGTGTGGAAA

PBSII‐STS (E2)

1,2‐Propanediol

Methylglyoxal synthase (MgsA)

ZFaGTCGATGCC

MgsA‐ZFa (E1)

[1

:^1

:^ 1]

n

[1

:^2

:^ 1]

n

[1

:^2

:^ 2]

n

[1

:^4

:^ 1]

n

[1

:^4

:^ 2]

n

4, 8, 16

12

More than fourfold improvement in 1,2‐propanediol production

[11]

2,5‐Diketo‐d‐gluconic acid reductase (DkgA)

ZFbGCGGCTGGG

DkgA‐ZFb (E2)

[1

:^1

:^ 1]

n

[1

:^2

:^ 1]

n

[1

:^2

:^ 2]

n

[1

:^4

:^ 1]

n

[1

:^4

:^ 2]

n

1, 2, 4, 8, 16

4

Glycerol dehydrogenase (GldA)

ZFcGAGGACGGC

GldA‐ZFc (E3)

(Continued)