Dataset Information

Genome Mining, Heterologous Expression, Antibacterial and Antioxidant Activities of Lipoamides and Amicoumacins from Compost-Associated Bacillus subtilis fmb60.

ABSTRACT: Bacillus subtilis fmb60, which has broad-spectrum antimicrobial activities, was isolated from plant straw compost. A hybrid NRPS/PKS cluster was screened from the genome. Sixteen secondary metabolites produced by the gene cluster were isolated and identified using LC-HRMS and NMR. Three lipoamides D-F (1-3) and two amicoumacin derivatives, amicoumacins D, E (4, 5), were identified, and are reported here for the first time. Lipoamides D-F exhibited strong antibacterial activities against harmful foodborne bacteria, with the MIC ranging from 6.25 to 25 µg/mL. Amicoumacin E scavenged 38.8% of ABTS+ radicals at 1 mg/mL. Direct cloning and heterologous expression of the NRPS/PKS and ace gene cluster identified its importance for the biosynthesis of amicoumacins. This study demonstrated that there is a high potential for biocontrol utilization of B. subtilis fmb60, and genome mining for clusters of secondary metabolites of B. subtilis fmb60 has revealed a greater biosynthetic potential for the production of novel natural products than previously anticipated.

SUBMITTER: Yang J

PROVIDER: S-EPMC8036425 | biostudies-literature | 2021 Mar

REPOSITORIES: biostudies-literature

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Genome Mining, Heterologous Expression, Antibacterial and Antioxidant Activities of Lipoamides and Amicoumacins from Compost-Associated Bacillus subtilis fmb60.

Yang Jie J Zhu Qingzheng Q Xu Feng F Yang Ming M Du Hechao H Bian Xiaoying X Lu Zhaoxin Z Lu Yingjian Y Lu Fengxia F

Molecules (Basel, Switzerland) 20210326 7

Bacillus subtilis fmb60, which has broad-spectrum antimicrobial activities, was isolated from plant straw compost. A hybrid NRPS/PKS cluster was screened from the genome. Sixteen secondary metabolites produced by the gene cluster were isolated and identified using LC-HRMS and NMR. Three lipoamides D-F (1-3) and two amicoumacin derivatives, amicoumacins D, E (4, 5), were identified, and are reported here for the first time. Lipoamides D-F exhibited strong antiba ...[more]

PMID: 33810551

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Project description:Grafting generally means stress to a plant and this triggers antioxidant defense systems. An imbalance in reactive oxygen species may negatively affect the grafting success. Several research projects have studied the association with plant growth-promoting rhizobacteria (PGPR) and it has been documented that they enhance nutrient acquisition, regulate hormone levels, and influence the antioxidant response in crops. However, little is known about the strategy of inoculating grafted herbaceous plants with PGPR and its effect on the antioxidant response. The effects of inoculating a strain of Bacillus subtilis on the antioxidant metabolism of grafted tomato were evaluated. In this study, two different rootstocks were used for tomato (Solanum lycopersicum L. var. Rio Grande (RG)): [S. lycopersicum L. var. cerasiforme (Ch)] and eggplant [(Solanum melanogena L. (Ber)] to establish a compatible graft (RGCh) and a semicompatible graft (RGBer). Enzyme activities involved in the antioxidant defense system: superoxide dismutase (SOD), catalase (CAT), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), peroxidase (POD), and total phenols were measured during 4 weeks after grafting. The results show that for RGCh, during the first two weeks after grafting, the tendency was a decrease of the enzyme activity for SOD, CAT, PAL when inoculated with B. subtilis; while in the semicompatible graft RGBer, PPO and PAL decreased their activity after inoculation. For both combinations, the quantity of total phenols varied depending on the day. In both graft combinations, applications of B. subtilis resulted in 86 and 80% callusing compared with the uninoculated control where the percentages were 74 and 70% for RGCh and RGBer, respectively. The highest significant graft success (95%) was recorded 28 days after grafting for inoculated RGBer. These findings imply that B. subtilis induced antioxidant mechanisms in grafted plants and suggest that inoculation with this growth-promoting bacterium can represent a biotechnological approach to improve success in tomato grafting.

Project description:BackgroundSubtilin is a peptide antibiotic (lantibiotic) natively produced by Bacillus subtilis ATCC6633. It is encoded in a gene cluster spaBTCSIFEGRK (spa-locus) consisting of four transcriptional units: spaS (subtilin pre-peptide), spaBTC (modification and export), spaIFEG (immunity) and spaRK (regulation). Despite the pioneer understanding on subtilin biosynthesis, a robust platform to facilitate subtilin research and improve subtilin production is still a poorly explored spot.ResultsIn this work, the intact spa-locus was successfully integrated into the chromosome of Bacillus subtilis W168, which is the by far best-characterized Gram-positive model organism with powerful genetics and many advantages in industrial use. Through systematic analysis of spa-promoter activities in B. subtilis W168 wild type and mutant strains, our work demonstrates that subtilin is basally expressed in B. subtilis W168, and the transition state regulator AbrB strongly represses subtilin biosynthesis in a growth phase-dependent manner. The deletion of AbrB remarkably enhanced subtilin gene expression, resulting in comparable yield of bioactive subtilin production as for B. subtilis ATCC6633. However, while in B. subtilis ATCC6633 AbrB regulates subtilin gene expression via SigH, which in turn activates spaRK, AbrB of B. subtilis W168 controls subtilin gene expression in SigH-independent manner, except for the regulation of spaBTC. Furthermore, the work shows that subtilin biosynthesis in B. subtilis W168 is regulated by the two-component regulatory system SpaRK and strictly relies on subtilin itself as inducer to fulfill the autoregulatory circuit. In addition, by incorporating the subtilin-producing system (spa-locus) and subtilin-reporting system (PpsdA-lux) together, we developed "online" reporter strains to efficiently monitor the dynamics of subtilin biosynthesis.ConclusionsWithin this study, the model organism B. subtilis W168 was successfully established as a novel platform for subtilin biosynthesis and the underlying regulatory mechanism was comprehensively characterized. This work will not only facilitate genetic (engineering) studies on subtilin, but also pave the way for its industrial production. More broadly, this work will shed new light on the heterologous production of other lantibiotics.

Dataset Information

Genome Mining, Heterologous Expression, Antibacterial and Antioxidant Activities of Lipoamides and Amicoumacins from Compost-Associated Bacillus subtilis fmb60.

Publications

Genome Mining, Heterologous Expression, Antibacterial and Antioxidant Activities of Lipoamides and Amicoumacins from Compost-Associated <i>Bacillus subtilis</i> fmb60.

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