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Table 1 Comparison between expression systems for the encapsulation of CYPBM3 inside P22

From: Design of a VLP-nanovehicle for CYP450 enzymatic activity delivery

Expression system (plasmids)

CYP/capsida

Total CYPb

Abs280 (µM)

Active CYP

CO assayc (µM)

% active CYPd

(CO/Abs280)

Co-expression (pETDuet)

156.0 (±0.4)

123.5

9.2

7

Differential expression 1 (pBAD + pRSF)

129.5 (±0.1)

135.3

31.8

23

Differential expression 2 (pBAD + pRSF)

109.7 (±2.8)

123.7

42.9

35

  1. aDetermined by Eq. 1
  2. bTotal CYP determined using an extinction coefficient at 280 nm assuming a molar extinction coefficient ε280 = 44,920 M−1 cm−1 for coat protein and ε280 = 52,830 M−1 cm−1 for CYP-SP (theoretically calculated using ProtParam, Gasteiger, 2005). The concentration of protein was calculated using the Lambert–Beer equation, AbsT = CCP·ɛCPl + CCYP-SP·ɛCYP-SPl as described previously [15]
  3. cActive CYP determined by the formation of CO-CYP complex in reducing medium with a extinction coefficient of ε450 = 0.091 nM−1cm−1 [26]
  4. dActive/total CYP ratio expressed in percentage