Long M, Xu M, Ma Z, Pan X, You J, Hu M, Shao Y, Yang T, Zhang X, Rao Z. Significantly enhancing production of trans-4-hydroxy-l-proline by integrated system engineering in Escherichia coli. Sci Adv. 2020 May 22;6(21):eaba2383.

Strains

• Escherichia coli PM-14
• Escherichia coli M1
• Escherichia coli M2
• Escherichia coli M3
• Escherichia coli M4
• Escherichia coli M5
• Escherichia coli M1/∆putP
• Escherichia coli M1/∆proP
• Escherichia coli M1/∆aceA
• Escherichia coli M6
• Escherichia coli M5/ProB_D107N
• Escherichia coli M5/proB_E143A
• Escherichia coli M6R
• Escherichia coli HYPA
• Escherichia coli M7-H
• Escherichia coli M7-1
• Escherichia coli M7-2
• Escherichia coli M7-3
• Escherichia coli M8-H
• Escherichia coli M8-1
• Escherichia coli M8-2
• Escherichia coli M8-3
• Escherichia coli HYP1
• Escherichia coli HYP2
• Escherichia coli HYP3
• Escherichia coli HYP2M
• Escherichia coli HYP0
• Escherichia coli BL21(DE3)

Plasmids

• pET-28a(+)
• pET-28aA
• pDXW-10
• pDXW-10-Datp4h
• pDXW-10-P_eaS-gfp_dt
• pDXW-10-E2_L170A/P172N

Metabolite production

• L-proline
  Titer
  • 18.1 g/L (Escherichia coli HYPA in Fed-Batch)
  • 10.4 g/L (Escherichia coli HYP1 in Fed-Batch)
  • 4.3 g/L (Escherichia coli HYP2 in Fed-Batch)
  • 8.9 g/L (Escherichia coli HYP3 in Fed-Batch)
  • 7.6 g/L (Escherichia coli HYP0 in Fed-Batch)
  • 0.2 g/L (Escherichia coli HYP2M in Fed-Batch)
  • 0.816 g/L (Escherichia coli PM-14 in Flask)
  • 18.2 g/L (Escherichia coli M1 in Fed-Batch)
  • 26.0 g/L (Escherichia coli M2 in Fed-Batch)
  • 30.4 g/L (Escherichia coli M3 in Fed-Batch)
  • 31.4 g/L (Escherichia coli M4 in Fed-Batch)
  • 32.2 g/L (Escherichia coli M5 in Fed-Batch)
  • 37.5 g/L (Escherichia coli M6 in Fed-Batch)
  • 33.8 g/L (Escherichia coli M5/proB_E143A in Fed-Batch)
  • 20.5 g/L (Escherichia coli M1/∆putP in Fed-Batch)
  • 21.6 g/L (Escherichia coli M1/∆proP in Fed-Batch)
  • 23.6 g/L (Escherichia coli M1/∆aceA in Fed-Batch)
  Volumetric productivity
  • 0.455 g/L/h (Escherichia coli M1 in Fed-Batch)
  • 0.65 g/L/h (Escherichia coli M2 in Fed-Batch)
  • 0.76 g/L/h (Escherichia coli M3 in Fed-Batch)
  • 0.785 g/L/h (Escherichia coli M4 in Fed-Batch)
  • 0.805 g/L/h (Escherichia coli M5 in Fed-Batch)
  • 0.938 g/L/h (Escherichia coli M6 in Fed-Batch)
  • 0.845 g/L/h (Escherichia coli M5/proB_E143A in Fed-Batch)
  • 0.513 g/L/h (Escherichia coli M1/∆putP in Fed-Batch)
  • 0.54 g/L/h (Escherichia coli M1/∆proP in Fed-Batch)
  • 0.59 g/L/h (Escherichia coli M1/∆aceA in Fed-Batch)
• trans-4-hydroxy-L-proline
  Titer
  • 15.7 g/L (Escherichia coli HYPA in Fed-Batch)
  • 8.8 g/L (Escherichia coli HYP0 in Fed-Batch)
  • 27.3 g/L (Escherichia coli HYP1 in Fed-Batch)
  • 43.2 g/L (Escherichia coli HYP2 in Fed-Batch)
  • 54.8 g/L (Escherichia coli HYP2M in Fed-Batch)
  • 15.4 g/L (Escherichia coli HYP3 in Fed-Batch)
  Volumetric productivity
  • 0.3 g/L/h (Escherichia coli HYPA in Fed-Batch)
  • 0.65 g/L/h (Escherichia coli HYP1 in Fed-Batch)
  • 1.03 g/L/h (Escherichia coli HYP2 in Fed-Batch)
  • 0.91 g/L/h (Escherichia coli HYP2M in Fed-Batch)
  • 0.06 g/L/h (Escherichia coli HYP3 in Fed-Batch)
  • 0.21 g/L/h (Escherichia coli HYP0 in Fed-Batch)
  Substrate yield
  • 0.05 g/g of sustrate (Escherichia coli HYPA in Fed-Batch)
  • 0.05 g/g of sustrate (Escherichia coli HYP0 in Fed-Batch)
  • 0.12 g/g of sustrate (Escherichia coli HYP1 in Fed-Batch)
  • 0.18 g/g of sustrate (Escherichia coli HYP2 in Fed-Batch)
  • 0.24 g/g of sustrate (Escherichia coli HYP2M in Fed-Batch)
  • 0.37 g/g of sustrate (Escherichia coli HYP3 in Fed-Batch)