Table 2 Kinetic parameters of candidate pathway enzymes used for in vivo studies

From: Construction of a synthetic metabolic pathway for biosynthesis of 2,4-dihydroxybutyric acid from ethylene glycol

Enzyme

Function

Vmax (U mg−1)

Km (mM)

Vmax/Km (U mg−1 M−1)

Reference

Ec.FucO

EG dehydrogenase

5.7

7

814

Sridhara et al.59

Go.Adh

EG dehydrogenase

4.82 (±0.23)a

2.40 (±0.19)

2008

Zhang et al.57

  

7.06 (±0.28)b

964 (±84)

0.08

Scheffen et al.58

Ec.FsaATA c

D-threose aldolase

5.3 (±0.3)

Donor: 0.094 (±0.008)

2524

Szekrenyi et al.28

   

Acceptor: 2.1 (±0.3)

  

Pc.TadH

D-threose dehydrogenase

3.50 (±0.1)e

–

70

This work

Tt.Lac11

D-threono-1,4-lactonase

1.76 (±0.16)

2.92 (±0.13)

603

This work

Hh.AraD

D-threonate dehydratase

0.19 (±0.04)

1.21 (±0.36)

157

This work

Aa.AraD

D-threonate dehydratase

0.12 (±0.06)

1.95 (±0.53)

62

This work

Ec.Mdh5Q d

OHB reductase

110.31 (±0.19)

1.6 (±0.01)

68,944

Frazão et al.29

  1. aParameters estimated at pH 8.5 (50 mM Tris-HCl) and 0.5 mM NAD+. Range of tested EG concentrations was 0.5–20 mM.
  2. bParameters estimated at pH 7.8 (100 mM MOPS-KOH) and 2.0 mM NAD+. Range of tested EG concentrations was 10–2000 mM.
  3. cCorresponds to double mutant Ec.FsaA L107Y:A129G
  4. dCorresponds to quintuple mutant Ec.Mdh I12V:R81A:M85Q:D86S:G179D.
  5. eVmax values refer to the specific activity obtained at 50 mM substrate concentration, since no saturation was observed under tested conditions.
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