Influence of PepF peptidase and sporulation on microcin J25 production in Bacillus subtilis

• Published in: Microbiology spectrum Vol. 12; no. 7; p. e0374823
• Main Authors: Zhang, Guangwen, Feng, Saixiang, Qin, Miaomiao, Sun, Juan, Liu, Yutong, Luo, Changqi, Lin, Min, Xu, Siqi, Liao, Ming, Fan, Huiying, Liang, Zhaoping
• Format: Journal Article
• Language: English
• Published: United States American Society for Microbiology 02-07-2024
• Subjects: Antimicrobial Chemotherapy; Bacillus subtilis; microcin J25; PepF; peptidase; Research Article; sporulation; Bacillus subtilis; peptidase; PepF; microcin J25; sporulation
• ISSN: 2165-0497; 2165-0497
• DOI: 10.1128/spectrum.03748-23

The lasso peptide microcin J25 (MccJ25) possesses strong antibacterial properties and is considered a potential effective component of bacterial disease treatment drugs and safe food preservatives. Although MccJ25 can be heterologously expressed in
as we have previously reported, its regulation and accumulation are yet to be understood. Here, we investigated the expression level and stability of MccJ25 in
strains with disruption in peptidase genes
,
and
. Oligoendopeptidase F (PepF) was found to be involved in reduction of the production of MccJ25 by degradation of its precursor peptide. In the
mutant, the MccJ25 reached a concentration of 1.68 µM after a cultivation time exceeding 60 hours, while the wild-type strain exhibited a concentration of only 0.14 µM. Moreover, the production of MccJ25 in
downregulated the genes associated with sporulation, and this may contribute to its accumulation. Finally, this study provides a strategy to improve the stability and production of MccJ25 in
.
MccJ25 displays significant antibacterial activity, a well-defined mode of action, exceptional safety, and remarkable stability. Hence, it presents itself as a compelling candidate for an optimal antibacterial or anti-endotoxin medication. The successful establishment of exogenous production of MccJ25 in
provides a strategy for reducing its production cost and diversifying its utilization. In this study, we have provided evidence indicating that both peptidase PepF and sporulation are significant factors that limit the expression of MccJ25 in B. subtilis. The Δ
and Δ
mutants of
express MccJ25 with higher production yield and enhanced stability. To sum up, this study developed several better engineered strains of
, which greatly reduced the consumption of MccJ25 during the nutrient depletion stage of the host strain, improved its production, and elucidated factors that may be involved in reducing MccJ25 accumulation in
.