Exchange of single amino acids at different positions of a recombinant protein affects metabolic burden in Escherichia coli

Table 1 Applied clones and abbreviations

Clone	Abbreviation
E. coli BL21(DE3):pET22b(+)–His₆-LOV	His-LOV
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-wt	Wild type enzyme (His-LOV-BSLA)
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Ala1Trp	A1W
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-His10Asp	H10D
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Ile12Cys	I12C
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Phe17Pro	F17P
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Lys23stop	K23stop
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Ser56Pro	S56P
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Asp91Arg	D91R
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Gly93Tyr	G93Y
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Val99Lys	V99K
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Leu102Trp	L102W
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Ser167Pro	S167P
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Lys170Glu	K170E
E. coli BL21(DE3):pET22b(+)–His₆-LOV-BSLA-Gly175Phe	G175F

The investigated E. coli BL21(DE3) clones contain plasmid pET22b(+) harboring the gene encoding fusion tags, wild type BSLA or BSLA variants. Each variant contains a single amino acid exchange at a different position chosen randomly and distributed over the entire enzyme sequence of 181 amino acids.

ISSN: 1475-2859