. 2022 Jun 10;9(6):251. doi: 10.3390/bioengineering9060251

Table 1.

Nisin variants produced by bioengineering in different hosts.

Host	Methods	Prenisin	Activation of Prenisin	Nisin Sensitive Indicator	Variants	Variant Features	References
L. lactis NZ9800 ¹	site-directed mutagenesis	No	N/A	M. flavus, S. thermophilus	ΔN20, ΔM21	inactive in the pore formation assay	[22]
L. lactis NZ9800 ¹	site-directed mutagenesis	No	N/A	M. flavus S. faecalis, L. monocytogenes, S. cerevisiae, G. candidum.	N20K, M21K	higher solubility, displayed antimicrobial activity against some G− strains	[49]
L. lactis NZ9800 ¹	random mutagenesis, saturation mutagenesis	No	N/A	L. lactis ssp. cremoris HP, E. faecium, S. agalactiae, S. aureus, L. monocytogenes.	N20P, M21V, K22S	enhanced antimicrobial activity	[50]
L. lactis NZ9800 ¹	random mutagenesis, saturation mutagenesis, site-directed mutagenesis	No	N/A	A series of G+ positive and G− bacteria ⁵	S29A/D/E/Q	enhanced antimicrobial activity against both G+ and G− bacteria	[56]
L. lactis NZ9800 ¹	saturation mutagenesis	No	N/A	Streptococcus mitis, L. lactis UCC90, L. lactis HP, L. monocytogenes, S. agalactiae	SVA, NAK	enhanced antimicrobial activity against L. monocytogenes in complex matrices	[76]
L. lactis NZ9800 (L. lactis NZ9700ΔnisA)	saturation mutagenesis, site-directed mutagenesis	No	N/A	Six Gram-positive bacteria	K12A	enhanced antimicrobial activity	[44]
L. lactis NZ9000	site-directed mutagenesis	Yes	NisP (expressed by indicator strain)	L. lactis NZ9000 -pNZnisPT	variants of precursor nisin with negatively charged residues ¹¹	severe decrease in antimicrobial activity	[10]
L. lactis NZ9800 ¹	saturation mutagenesis	No	N/A	S. agalactiae, L. lactis HP M. smegmatis	AAK, NAI, SLS	enhanced antimicrobial activity	[52]
L. lactis NZ9000	site-directed mutagenesis	Yes	trypsin	A series of Gram-positive bacteria ⁶	20NK21, 20NLMK23, 20NVMK23, 20NIMK23 20NIVMK24	enhanced antimicrobial activity against specific strains at certain temperatures	[48]
L. lactis NZ9800 ¹	saturation mutagenesis	No	N/A	S. pseudintermedius, S. intermedius, S. aureus, L. lactis HP, S. uberis, B. cereus	I4V	enhanced antimicrobial activity and anti-biofilm activity against S. pseudintermedius and S. intermedius	[46]
E. coli	ASM ⁴	Yes	trypsin	L. lactis NZ9000 NlacZ	L16A, L16H, L16V, M21A, M21D, M21N	increased induction activity and antimicrobial activity	[77]
L. lactis NZ9800 ¹	site-directed mutagenesis	No	N/A	S. aureus SA113, S. pseudintermedius DSM21284	M21V, I4V	The activity of the nisin derivative and antibiotic combination was higher than that of the nisin and antibiotic combination	[29]
Trp-auxotrophic Lactococcus lactis NZ9000	SPI ²	Yes	NisP	L. lactis MG1363	four different positions of nisin Trp and Trp analogue variants ¹²	Nisin variants containing tryptophan analogues	[64]
E. coli	SCS ³	Yes	trypsin	M. flavus	Ser3TAG	Nisin variant with novel macrocyclic topologies	[65]
Pro-auxotrophic E. coli strain	SPI ²	Yes	NisP (expressed by indicator strain)	L. lactis NZ9000	P9X ⁷	Nisin variant with 6 proline analogues	[63]
Lactococcus lactis NZ9000, E. coli	SCS ³	Yes	NisP (expressed by indicator strain)	L. lactis NZ9000	I4BocK ⁸, K12BocK ⁸	enhanced antimicrobial activity	[78]
E. coli, C321.ΔprfA-T7RNAPΔrneΔompTΔlon	site-directed mutagenesis	Yes	trypsin	L. lactis HP	Ser5m-BrPhe	Nisin variant with Phe analogues	[67]
L. lactis NZ9000	site-directed mutagenesis	Yes	NisP	L. lactis NZ9000Cm/NisI/NisFEG	₂₀NMKIV₂₄	decreased recognition of immunity protein	[54]
L. lactis NZ9000	saturation mutagenesis	Yes	NisP	L. lactis NZ9000-Cm/NisI/NisFEG/SaNSR /SaNsrFP	I1X ⁹	I1X variants influenced antimicrobial activity and the efficiency of the immunity and resistance proteins.	[47]
L. lactis NZ9800 ¹	saturation mutagenesis	No	N/A	L. lactis subsp. diacetylactis DRC3 (expressing the nisin resistance protein (NSR))	S29P	The variant exhibited a 20-fold increase in specific activity against a strain expressing the nisin resistance protein.	[60]
L. lactis NZ9000	site-directed mutagenesis	Yes	NisP	L. lactis NZ9000 pNZ-SV-Erm/SaNSR/SaNSRS236A	C28P	3 times more efficient against SaNSR-expressing L. lactis cells	[59]
L. lactis NZ9800 ¹	site-directed mutagenesis	No	N/A	Lb. plantarum UCC16, Lb. brevis SA-C12, L. lactis ssp. cremoris HP	P9T, P9S	The variants retain induction capacity, while most of the antimicrobial activity is abolished.	[79]
Met-auxotrophic Lactococcus lactis NZ9000	SPI ²	Yes	NisP	L. lactis and six Gram-positive pathogenic strains ¹⁰	M21V-M17Aha + M21VM17Hpg	The variant is the most active dimeric nisin construct	[68]
L. lactis NZ9800 ¹	site-directed mutagenesis	No	N/A	L. innocua FH1836lux	M17Q + N20P, M17Q + S29E	The combinations of nisin derivative exhibited enhanced anti-listerial activity when used together compared to when used alone	[80]
L. lactis NZ9000	site-saturation mutagenesis	Yes	Not mentioned	S. aureus RF122, S. aureus NCDO1499, S. agalactiae ATCC13813, L. lactis HP, L. lactis MG1363	M17Q, T2L, HTK	improved specific activity against some Staphylococci but unchanged or reduced activity against dairy Lactococci	[81]
L. lactis NZ9800 ¹	site-directed mutagenesis	No	N/A	T. thermophilus HB27, T. scotoductus Se-1	M17Q, M21F	enhanced specific activity against Thermus strains	[55]

¹L. lactis NZ9800: L. lactis NZ9700∆nisA. ² SPI = selection pressure incorporation. ³ SCS = stop codon suppression. ⁴ ASM = alanine scanning mutagenesis. ⁵ Targets included the antibiotic resistant S. aureus strains ST 528 (MRSA), ST 530 (MRSA), hVISA 32679, as well as S. aureus RF122, Streptococcus mitis, L. lactis HP and MG1363, Bacillus cereus DPC 6088/6089, Enterococcus durans and L. monocytogenes strains 10403S and LO28. ⁶ Enterococcus faecalis VE14089, Listeria monocytogenes, Bacillus cereus 4147, Bacillus cereus 4153, Lactococcus lactis MG1363, Bacillus cereus (L’29) 16, Micrococcus luteus, Streptococcus pneumoniae R6, Bacillus sporothermodurans lC4, S. aureus. ⁷ X represents six proline analogues: (4R)-fluoroproline, (4R)-hydroxyproline, (4R)-methanoproline, (4S)-fluoroproline, (4S)-hydroxyproline, (4S)-methanoproline. ⁸ BocK = Nε-Boc-L-lysine charged amino acids. ⁹ X includes four classes depending on the amino acid property: (1) aliphatic amino acids (L, A, V, G); (2) aromatic amino acids (W, F, Y); (3) C, T, S; (4) charged amino acids (K, R, H, Q, E, N, D). ¹⁰ The tested strains included two Staphylococci, two Enterococci, Bacillus cereus and Listeria monocytogenes. ¹¹ NisA-H6 T2D P9D with two negatively charged residues, NisA-H6 T2D P9D K12D N20E with four negatively charged residues, and NisA-H6 T2D P9D K12D N20E H27D K34E with six negatively charged residues. ¹² Tryptophan and tryptophan analogue variants at four positions of nisin: I1W/5FW/5HW/5MeW, I4W/5FW/5HW/5MeW, M17W/5FW/5HW/5MeW, V32W/5FW/5HW/5MeW.