Skip to main content

Table 1 Expression and recovery conditions of previously reported endolysins at bench scale

From: Evaluation of factors influencing expression and extraction of recombinant bacteriophage endolysins in Escherichia coli

Endolysin

Description

Expression vector/E. coli strain

Induction conditions

Lysis buffer

Purification yields (mg/L of bacterial culture)

LysH5 [18]

Endolysin from phage ΦH5: N-terminal CHAP, a central amidase domain (N-acetylmuramyl-l-alanine amidase), and a C-terminal SH3b CBD

pRSETB (N-ter His-tag)/BL21(DE3)/pLys

1 mM IPTG/18 h—19 °C

20 mM NaH2PO4,500 mM NaCl, 20 mM Imidazole, pH 7.4

2 mg/L

Lys-phiSA012 [27]

endolysin from phage phiSA012: N-terminal CHAP, a central amidase domain (N-acetylmuramyl-l-alanine amidase), and a C-terminal SH3b CBD

pGEX-6P-2 (C-ter GST-tag)/BL21(DE3)

0.1 mM IPTG/O.N.—25 °C

50 mM Tris–HCl, 1 M MgCl2 and 10% NP-40

Not stated

plyGRCS [28]

endolysin from phage GRCS: N-terminal both CHAP and endopeptidase activity and a C-terminal SH3b CBD

pBAD24 (C-His-tag)/BL21 (DE3)

0,25% ara/O.N.—18 °C

Not stated

Not stated

LysGH15 [17]

Endolysin from phage GH15: N-terminal CHAP, a central amidase domain (N-acetylmuramyl-l-alanine amidase), and a C-terminal SH3b CBD

pET15b (N-ter His-tag)/BL21 (DE3) (Codon Plus)

1 mM IPTG/6 h—25 °C

20 mM PBS

Not stated

2638A [29]

Endolysin from phage 2638A: N-terminal M23 peptidase domain, a mid-protein amidase 2 domain, and a C-terminal SH3b_5 (SH3b) CBD

pET21a (C-ter His-tag)/BL21(DE3)

1 mM IPTG/18 h—10 °C

Not stated

Not stated

ClyS [15]

Quimeric endolysin: N-terminal catalytic domain of Twort phage endolysin and C-terminal CBD from phi13 phage endolysin

pJML6/DH5α

2% lactose/O.N.—30 °C

20 mM NaH2PO4, 1 mM DTT

Not stated

CHAP-AMIDASE [30]

Quimeric endolysin: N-terminal CHAP domain and C-terminal amidase-2 domain from Phage K endolysin (LysK). GS-His tag linker

pET-22b (pelB leader)/BL21 (DE3)

1 mM IPTG/4 h—37 °C

Not stated

8–12 mg/L

LysP108 [31]

Endolysin from phage P108: N-terminal amidase and C-terminal SH3b CBD

pET21a (C-ter His-tag)/BL21 (DE3)

0,1 mM IPTG/10 h—23 °C

Not stated

Not stated

LysDW2 [32]

Endolysin from phage DW2: N-terminal CHAP, a central amidase domain (N-acetylmuramyl-l-alanine amidase), and a C-terminal SH3b CBD

pQE-60 (C-ter His-tag)/XL1Blue

1 mM IPTG/5 h—37 °C

BugBuster® protein extraction reagent, Millipore, Merk

Not stated (only CHAP truncated domain was soluble)

SAL-1 [33]

endolysin from phage SAP-1: SAL-1 differs from LysK at three residues (ILE- > VAL 26, GLN- > GLU 114, and HIS- > GLN 486)

pBAD-TOPO (no His-tag)/BL21 (DE3)

0.2 mM ara/10 h—19 °C

50 mM Na2HPO4 (pH 7.5), 10 mM EDTA, 1 mM DTT

80 mg/L

LysCSA13 [26]

endolysin from phage CSA13: N-terminal CHAP and C-terminal SH3-5 CBD

pET28a (N-ter His-tag)/BL21 (DE3)

0,5 mM IPTG/20 h–18 °C

50 mM NaH2PO4, 300 mM NaCl, pH 8.0

10 mg/L

  1. Ara arabinose