[Site-directed mutagenesis of NisinZ and properties of NisinZ mutants].
According to the knowledge gained from engineering of nisinZ, using plasmid pHJ201 DNA as template, NisinZ was mutated by site-directed mutagenesis, NisinZ mutant T8S contains Serine at position 8 instead of Threonine, NisinZ mutant N27K/H31K contains Lysine at position 27 and 31, respectively, instead of Asparagine and Histidine and NisinZ mutant T2S/ H31K contains dehydrobutyrine and Lysine at position 2 and 31 instead of dehydroalanine and Histidine. They are cloned into pMG36e and expressed in L. Lactis NZ9800, the expression products of these mutants purified by Sephadex CM-25 and Sephadex G-25 chromatography, some properties of NisinZ mutants (T8S, T2S/H31K and N27K/H31K) were studied. The results showed that the spectrum of antimicrobial activity and solubility of these mutants had not been changed, their antimicrobial activities were found to be slightly lower than that of the wild-type NisinZ. but mutants T8S and T2S/H31K showed higher stability, which were significantly more stable than wild-type NisinZ at 55 approximately 100 degrees C and pH7 approximately 9.
Yuan
Jing
J
Molecular Microbiology Research Center and 2 State Key Laboratory of Microbial Resource 2, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100080, China.
Zhang
Zhen-Zhong
ZZ
Yang
Wei
W
Chen
Xiu-Zhu
XZ
Huan
Lian-Dong
LD
D000900:Anti-Bacterial Agents / Q000378:metabolism / Q000494:pharmacology; D001419:Bacteria / Q000187:drug effects; D004591:Electrophoresis, Polyacrylamide Gel; D016297:Mutagenesis, Site-Directed / Q000379:methods*; D009561:Nisin / Q000031:analogs & derivatives* / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology; D055550:Protein Stability; D012995:Solubility; D013696:Temperature
D000900:Anti-Bacterial Agents; D009561:Nisin; C071364:nisin Z