Li Zhou, Ying Zhu, Zhongzhe Yuan, Guangqing Liu, Zijin Sun, Shiyu Du, He Liu, Yating Li, Haili Liu, and Zhemin Zhou (2022). Evaluation of Metabolic Engineering Strategies on 2-Ketoisovalerate Production by Escherichia coli. Applied and Environmental Microbiology. 88(17): e00976-22.

Strains

• Escherichia coli B0016-050
• Escherichia coli 050T
• Escherichia coli 050T1
• Escherichia coli 050T2
• Escherichia coli 050T3
• Escherichia coli 050T4
• Escherichia coli 050T4-1
• Escherichia coli 050T4-2
• Escherichia coli 050TY
• Escherichia coli 050T4/pCTSDT
• Escherichia coli 050TY/pCTSDTQ487S-RBS55
• Escherichia coli B0016-050/pCTSDT
• Escherichia coli 050T/pCTSDT
• Escherichia coli BL21(DE3)/pCTSDT
• Escherichia coli 050T3/pCTSDT
• Escherichia coli 050T1/pCTSDT
• Escherichia coli 050T2/pCTSDT
• Escherichia coli 050T3/pCTSDT+pACYC-pntAB
• Escherichia coli 050T4-1/pCTSDT
• Escherichia coli 050T4-2/pCTSDT
• Escherichia coli 050T4/pCTSDTQ487S
• Escherichia coli 050T4/pCTSDTQ487S-RBS55

Plasmids

• pETDuet-1
• pCTSDTQ487S
• pCTSDT
• pACYC-pntAB
• pCTSDTQ487S-RBS42
• pCTSDTQ487S-RBS55

Metabolite production

• isobutanol
  Substrate yield
  • 0.036 mol/mol of substrate (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)
  • 0.284 mol/mol of substrate (Escherichia coli 050T4/pCTSDT in Fed-Batch)
  Titer
  • 13.1 g/L (Escherichia coli 050T4/pCTSDT in Fed-Batch)
  • 0.25 g/L (Escherichia coli 050T4/pCTSDTQ487S in Flask)
  • 0.57 g/L (Escherichia coli 050T4/pCTSDTQ487S-RBS55 in Flask)
• pyruvate
  Substrate yield
  • 0.022 mol/mol of substrate (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)
  • 0.016 mol/mol of substrate (Escherichia coli 050T4/pCTSDT in Fed-Batch)
• succinate
  Substrate yield
  • 0.04 mol/mol of substrate (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)
  • 0.119 mol/mol of substrate (Escherichia coli 050T4/pCTSDT in Fed-Batch)
• acetate
  Substrate yield
  • 0.134 mol/mol of substrate (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)
  • 0.075 mol/mol of substrate (Escherichia coli 050T4/pCTSDT in Fed-Batch)
• 3-methyl-2-oxobutanoate
  Titer
  • 7.39 g/L (Escherichia coli 050T2/pCTSDT in Flask)
  • 7.36 g/L (Escherichia coli 050T1/pCTSDT in Flask)
  • 55.8 g/L (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)
  • 22.0 g/L (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Flask)
  • 46.4 g/L (Escherichia coli 050T4/pCTSDT in Fed-Batch)
  • 15.2 g/L (Escherichia coli 050T4/pCTSDT in Flask)
  • 19.2 g/L (Escherichia coli 050T4/pCTSDT in Flask)
  • 10.83 g/L (Escherichia coli 050T4/pCTSDT in Flask)
  • 14.88 g/L (Escherichia coli 050T4/pCTSDTQ487S in Flask)
  • 19.2 g/L (Escherichia coli 050T4/pCTSDTQ487S-RBS55 in Flask)
  • 7.56 g/L (Escherichia coli 050T4-1/pCTSDT in Flask)
  • 7.69 g/L (Escherichia coli 050T4-2/pCTSDT in Flask)
  • 8.44 g/L (Escherichia coli 050T3/pCTSDT in Flask)
  • 9.35 g/L (Escherichia coli 050T3/pCTSDT+pACYC-pntAB in Flask)
  • 8.01 g/L (Escherichia coli 050T/pCTSDT in Flask)
  • 0.33 g/L (Escherichia coli B0016-050/pCTSDT in Flask)
  • 1.7 g/L (Escherichia coli BL21(DE3)/pCTSDT in Flask)
  Volumetric productivity
  • 2.14 g/L/h (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)
  • 1.78 g/L/h (Escherichia coli 050T4/pCTSDT in Fed-Batch)
  Substrate yield
  • 0.852 mol/mol of substrate (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)
  • 0.94 mol/mol of substrate (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Flask)
  • 0.644 mol/mol of substrate (Escherichia coli 050T4/pCTSDT in Fed-Batch)
  • 0.797 mol/mol of substrate (Escherichia coli 050T4/pCTSDT in Flask)
  Specific production rate
  • 0.112 g/g DCW/h (Escherichia coli 050T4/pCTSDT in Fed-Batch)
  • 0.162 g/g DCW/h (Escherichia coli 050TY/pCTSDTQ487S-RBS55 in Fed-Batch)