Expression and purification of the recombinant murine REG3α protein in Pichia pastoris and characterization of its antimicrobial and antitumour efficacy

Figures & data

Figure 1. SDS-PAGE analysis of the first step purification of recombinant REG3α from the culture supernatant. Note: First purification fractions (lanes 1–8).

Figure 2. SDS-PAGE analysis of the second step purification of recombinant REG3α.

Figure 3. SDS-PAGE and western blot analysis of the recombinant murine REG3α final product.

Table 1. Antimicrobial activity of the recombinant murine REG3α.

Micro-organism	CMCC No.^a	MIC (b – c μmol/L)	MBC (d – e μmol/L)
Escherichia coli	44102	5.08–10.15	10.15–20.3
Salmonella paratyphi A	(B) 50001	5.08–10.15	10.15–20.3
Pseudomonas aeruginosa	(B) 10104	NT	NT
Staphylococcus aureus	26003	2.54–5.08	2.54–5.08
Staphylococcus epidermidis	(B) 26069	2.54–5.08	2.54–5.08
Bacillus subtilis	63501	2.54–5.08	2.54–5.08
Bacillus pumilus	(B) 63202	5.08–10.15	10.15–20.3
Micrococcus luteus	(B) 28001	5.08–10.15	10.15–20.3

Note: a–National centre for medical culture collections (CMCC).

MIC values are expressed as the interval of concentration b–c μmol/L, where b is the highest concentration tested, at which a microbial growth can be observed, and c is the lowest concentration yielding no detectable microbial growth. (The assay was performed in triplicate.)

MBC values are expressed as the interval of concentration d–e μmol/L, where d is the same mean as b and e is the lowest concentration tested that inhibited micro-organism growth or that killed more than 99.9% of the micro-organisms. (The assay was performed in triplicate.)

Not tested (NT).

Figure 4. Killing kinetics of E. coli and S. aureus by recombinant murine REG3α.

Figure 5. MTT assay of the SMMC-7721 tumour cells, incubated with different concentrations of REG3α for 24, 48 and 72 h.

Figure 6. The haemolytic activity of the recombinant murine REG3α towards human erythrocytes.