[Expression and characterization of a new class IIa bacteriocin].
NB-Cl gene is a potential class IIa bacteriocin gene. To obtain its soluble expression, NB-C1 was fused with the green fluorescent protein (GFP) gene and a recombinant expression vector plVEX 2.4d-GFP-NB-C1 was constructed, which was transformed into Escherichia coli BL21(DE3) pLysS. The expressed fusion protein GFP-NB-CI was purified by Ni-NTA affinity chromatography and the bioactivity was examined using Listeria monocytogenes as the indicator bacteria. The results showed that the expressed fusion protein GFP-NB-C1 was soluble and the final concentration of the purified fusion protein was 36.1 mg/L E. coli culture and had the purity above 95%. The antimicrobial assay of GFP-NB-C1 was analyzed and showed its high activity against Listeria monocytogenes.
School of Food Science and Technology, Jiangnan University, Wuxi 214122, China.
Yan
Y
Xie
Haiqin
H
Chen
Qiuxiang
Q
Zhang
Fengwei
F
Tian
Yongquan
Y
Chen
Hao
H
Zhang
Wei
W
Chen
D000595:Amino Acid Sequence; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics; D002846:Chromatography, Affinity; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D005822:Genetic Vectors / Q000235:genetics; D049452:Green Fluorescent Proteins / Q000096:biosynthesis / Q000235:genetics; D008089:Listeria monocytogenes / Q000187:drug effects*; D008969:Molecular Sequence Data; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics / Q000494:pharmacology*
D000900:Anti-Bacterial Agents; D001430:Bacteriocins; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins