Table 1 Characteristics of CatIBs/FIBs All constructs were characterized regarding CatIB/FIB formation efficiency, quantified as the relative activity of the insoluble CatIB/FIB-containing pellet fraction compared to the crude cell extract (set to 100%), the initial rate activity (kcat; µmolProduct s−1, per subunit) of the lyophilized CatIB preparation, activity retention compared to the soluble enzyme, the relative protein and lipid content based on the initial weight of the lyophilizate and the yield of CatIBs obtained from 100 g wet E. coli cells
From: Tailoring the properties of (catalytically)-active inclusion bodies
Construct | CatIB/FIB formation efficiency [%] | Activity kcat [s−1] | Residual activity [%]d | Rel. protein content lyophilizate [%] | Yield \(\frac{{g_{lyophilizate} }}{{100 g_{cells} }}\) g | Lipid content [%] |
|---|---|---|---|---|---|---|
Constructs showing robust CatIB/FIB formation efficiency | ||||||
 TDoT fusions | ||||||
  TDoT-L-YFP | 53.8 ± 7.4 (6)b | na | na | 70.0 ± 5.3 (4) | 4.9 ± 0.6 (3) | nd |
  TDoT-YFP | 65.4 ± 4.9 (3) | na | na | 69.2 ± 6.8 (2) | 5.5 (1) | nd |
  TDoT-mCherry | 31.8 ± 8.2 (4)b | na | na | 85.7 ± 8.3 (2) | 3.2 (1) | nd |
  TDoT-L-BsLAa | 114.1 ± 3.1 (1) | nd | nd | 79 (1) | 8.6 (1) | nd |
  TDoT-L-AtHNLa | 76.4 ± 3.5 (1) | 4.3 ± 0.2 (1) | 11.1 | 85 (1) | 7.3 (1) | nd |
  TDoT-L-EcMenDa | 90.3 ± 0.2 (1) | nd | nd | 93 (1) | 12.2 (1) | nd |
  TDoT-L-RADH | 87.5 ± 3.2 (4)b | 0.054 ± 0.008 (3)b | 2.0b | 84.6 ± 3.9 (3)b | 9.7 ± 1.7 (4)b | 14.3 ± 0.3 (1) |
  TDoT-L-PfBAL | 87.7 ± 6.8 (4)b | 0.77 ± 0.12 (4)b,c | 1.0b,c | 71.9 ± 4.5 (4)b,c | 8.8 ± 1.0 (8)b | 16.4 ± 1.0 (1)c |
  TDoT-EcLDC | nd | 6.2*10−7 (1) | nd | nd | nd | nd |
  EcLDC-L-TDoT | nd | 0.71 (1)e | nd | 67.9 ± 5.9 (3)e | 12.4 ± 3.0 (3) | 12.9 ± 3.2 (1) |
 3HAMP fusions | ||||||
  3HAMP-L-RADH | 75.4 ± 3.7 (4) | 0.33 ± 0.02 (3) | 12.0 | 50.9 ± 7.6 (3) | 3.8 ± 0.5 (3) | 30.6 ± 8.3 (1) |
  3HAMP-L-PfBAL | 75.8 ± 8.0 (5) | 13.9 ± 2.9 (3)c | 18.1c | 33.8 ± 5.2 (3)c | 3.3 ± 0.5 (4) | 30.1 ± 4.7 (1)c |
  3HAMP-L-LbADH | 67.0 ± 21.7 (3) | 0.60 ± 0.20 (3) | 1.0 | 54.6 ± 8.0 (3) | 8.1 ± 1.3 (3) | 34.7 ± 1.36 (1) |
  3HAMP-L-PpBFD | 61.3 ± 35.4 (3) | 23.4 ± 6.1 (4) | 10.3 | 35.5 ± 6.7 (4) | 6.6 ± 1.4 (3) | 27.9 ± 3.7 (1) |
  EcLDC-L-3HAMP | nd | 0.80 (1) | nd | 56.5 ± 6.5 (2) | 7.5 ± 6.5 (4) | 17.7 ± 0.6 (1) |
Constructs showing low CatIB/FIB formation efficiency | ||||||
 TDoT-L-mCherry | 3.5 ± 1.9 (3) | na | na | 15.8 ± 0.5 (1) | 2.8 (1) | nd |
 TDoT-L-LbADH | 5.4 ± 5.9 (3) | 3.63 ± 0.90 (3) | 5.8 | 43.4 ± 5.5 (3) | 2.5 ± 0.4 (3) | 25.2 ± 0.73 (1) |
 TDoT-L-PpBFD | 1.2 ± 0.6 (3) | 9.2 ± 4.7 (4) | 4.1 | 26.9 ± 4.1 (4) | 1.6 ± 0.7 (3) | 19.1 ± 0.8 (1) |
 3HAMP-L-YFP | 6.3 ± 3.2 (4) | na | na | 49.0 ± 5.7 (3) | 5.4 ± 1.0 (3) | nd |
 3HAMP-L-mCherry | 5.5 ± 0.2 (5) | na | na | 36.4 ± 4.1 (4) | 3.0 ± 0.9 (4) | nd |