Whole-genome sequencing combined with mass spectrometry to identify bacteriocin and mine silent genes
2022
Li, Hongbiao | Guo, Lidan | Zhang, Xiaoyan | Mu, Haoqiang | Sha, Shenfei | Lin, Yi | Hou, Hongwei | Wang, Liping
Lactobacillus plantarum WUH3 and Lactobacillus plantarum M4L1 have high homology and contain genes encoding Plantaricin EF. The bacteriocin-WUH3 and bacteriocin-M4L1, using ethyl acetate, anion exchange, gel chromatography, and RP-HPLC, were purified for identifying the bacteriocins and mining silent genes. The molecular weight of bacteriocin-WUH3 measured by MALDI-TOF-MS was 4299.801 Da, highly matched with the molecular weight of Plantaricin E analyzed by whole-genome sequencing. Q-TOF-MS/MS analysis predicted the amino acid sequence of novel bacteriocin-M4L1 and molecular weight as 3748.9643 Da. The whole-genome sequencing results were compared with Bagel4 to determine the bacteriocin encoded by Lactobacillus plantarum, and then the genes encoding bacteriocin were mined through uniport. The results of whole-genome sequencing and mass spectrometry showed that the expression levels of Enterocin _ X _ chain _ beta, Pediocin, Plantaricin K, and Plantaricin EF were very low, so the genes encoding these bacteriocins were silent. Mining silenced genes can provide functional gene donors for subsequent expression, regulate bacteriocin synthesis and improve production.
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