Construction of lipopeptide mono-producing Bacillus strains and comparison of their antimicrobial activity

Wu, Guojun; Zhou, Jinghai; Zheng, Jie; Abdalmegeed, Dyaaaldin; Tian, Jingjing; Wang, Mengxi; Sun, Shengwei; Sedjoah, Rita-Cindy Aye-Ayire; Shao, Yuting; Sun, Shuhong; Xin, Zhihong

doi:10.1016/j.fbio.2023.102813

Cited by 7 publications

(4 citation statements)

References 58 publications

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“…Strain HM-Δ sfp exhibited no detectable iturin A production in contrast to the stain HM618 (Figure d,e). Mass spectrometry analysis revealed that iturin A produced by the stain HM618 comprised homologues with m / z values of 1043.48 [M + H] + , 1057.49 [M + H] + , 1071.51 [M + H] + , and 1085.52 [M + H] + , corresponding to C14-iturin A, C15-iturin A, C16-iturin A, and C17-iturin A (Figure S4), consistent with the date in the literature. , Additionally, the stain HM-Δ sfp displayed reduced wrinkling and larger colony diameter compared to the wild-type bacteria (Figure f), suggesting the crucial role of sfp in the morphological development of strain HM618 colonies. Utilizing sfp gene knockout as an example, the results demonstrate the efficacy of the CRISPR/Cas9 system for precise gene editing in B.…”

Section: Resultssupporting

confidence: 83%

Enhanced Iturin A Production of Engineered Bacillus amyloliquefaciens by Knockout of Endogenous Plasmid and Rap Phosphatase Genes

Hou,

Cao,

Gao

et al. 2024

J. Agric. Food Chem.

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Iturin A biosynthesis has garnered considerable interest, yet bottlenecks persist in its low productivity in wild strains and the ability to engineer Bacillus amyloliquefaciens producers. This study reveals that deleting the endogenous plasmid, plas1, from the wild-type B. amyloliquefaciens HM618 notably enhances iturin A synthesis, likely related to the effect of the Rap phosphatase gene within plas1. Furthermore, inactivating Rap phosphatase-related genes (rapC, rapF, and rapH) in the genome of the strain also improved the iturin A level and specific productivity while reducing cell growth. Strategic rap genes and plasmid elimination achieved a synergistic balance between cell growth and iturin A production. Engineered strain HM-DR13 exhibited an increase in iturin A level to 849.9 mg/L within 48 h, significantly shortening the production period. These insights underscore the critical roles of endogenous plasmids and Rap phosphatases in iturin A biosynthesis, presenting a novel engineering strategy to optimize iturin A production in B. amyloliquefaciens.

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Section: Resultssupporting

confidence: 83%

Enhanced Iturin A Production of Engineered Bacillus amyloliquefaciens by Knockout of Endogenous Plasmid and Rap Phosphatase Genes

Hou,

Cao,

Gao

et al. 2024

J. Agric. Food Chem.

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“…It is evident that volatile compounds from biocontrol bacteria represent a good development prospect in the future and provide new ideas for controlling tobacco target spot disease. Lipopeptides are synthesized by non-ribosomal pathway and contain antibacterial active substances such as iturin, surfactin, and plipastatin ( Wu et al 2023 ). The different types of crude lipopeptides secreted by CTXW 7-6-2 can be further studied to improve the productivity of lipopeptides and provide a scientific basis for efficient and green control of tobacco target spot disease.…”

Section: Discussionmentioning

confidence: 99%

“…The sterile fermentation broth, crude lipopeptide, and volatile compounds of CTXW 7-6-2 showed potent inhibitory effects against the growth of R. solani, mainly owing to the antagonistic action of the volatile compounds. Volatile compounds from other Bacillus species are potent against plant pathogens, including Monilinia fructicola in peach fruit (Liu et al 2018), Alternaria solani in potatoes (Zhang et al 2020), Ceratocystis fimbriata in postharvest sweet potatoes (Xu et al 2021) Lipopeptides are synthesized by non-ribosomal pathway and contain antibacterial active substances such as iturin, surfactin, and plipastatin (Wu et al 2023). The different types of crude lipopeptides secreted by CTXW 7-6-2 can be further studied to improve the productivity of lipopeptides and provide a scientific basis for effi-cient and green control of tobacco target spot disease.…”

Section: Discussionmentioning

confidence: 99%

Biocontrol and Growth Promotion Potential of Bacillus subtilis CTXW 7-6-2 against Rhizoctonia solani that Causes Tobacco Target Spot Disease

Huang,

Jin,

Wen

et al. 2024

Polish Journal of Microbiology

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Fungal diseases form perforated disease spots in tobacco plants, resulting in a decline in tobacco yield and quality. The present study investigated the antagonistic effect of Bacillus subtilis CTXW 7-6-2 against Rhizoctonia solani, its ability to promote the growth of tobacco seedlings, and the expression of disease resistance-related genes for efficient and eco-friendly plant disease control. Our results showed that CTXW 7-6-2 had the most vigorous growth after being cultured for 96 h, and its rate of inhibition of R. solani growth in vitro was 94.02%. The volatile compounds produced by CTXW 7-6-2 inhibited the growth of R. solani significantly (by 96.62%). The fungal growthinhibition rate of the B. subtilis CTXW 7-6-2 broth obtained after high-temperature and no-high-temperature sterile fermentation was low, at 50.88% and 54.63%, respectively. The lipopeptides extracted from the B. subtilis CTXW 7-6-2 fermentation broth showed a 74.88% fungal growth inhibition rate at a concentration of 100 mg/l. Scanning and transmission electron microscopy showed some organelle structural abnormalities, collapse, shrinkage, blurring, and dissolution in the R. solani mycelia. In addition, CTXW 7-6-2 increased tobacco seedling growth and improved leaf and root weight compared to the control. After CTXW 7-6-2 inoculation, tobacco leaves showed the upregulation of the PDF1.2, PPO, and PAL genes, which are closely related to target spot disease resistance. In conclusion, B. subtilis CTXW 7-6-2 may be an efficient biological control agent in tobacco agriculture and enhance plant growth potential.

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“…The following supporting information can be downloaded at: [ 52 , 53 , 54 ], Figures S1–S8: An integrated pipeline and overexpression of a novel efflux transporter, YoeA, significantly increase plipastatin production in Bacillus subtilis . Table S1, The strains used in this study; Table S2, The plasmids used in this study; Table S3, The primer sequences used in this study; Table S4, Experimental results of Box-Behnken designs for product yield; Table S5, Factor levels for response surface methodology; Table S6, Effects of carbon source on the diameter of the inhibition zone of M-2435Δ abrb ; Table S7, Effects of glucose concentration on the diameter of the inhibition zone of M-2435Δ abrb ; Table S8, Effects of amino acids on the diameter of the inhibition zone of M-2435Δ abrb ; Table S9, Effects of glu concentration on the diameter of the inhibition zone of M-2435Δ abrb ; Table S10, Effects of Inorganic Nitrogen Source on the Diameter of the Inhibition Zone of M-2435Δ abrb .…”

mentioning

confidence: 99%

An Integrated Pipeline and Overexpression of a Novel Efflux Transporter, YoeA, Significantly Increases Plipastatin Production in Bacillus subtilis

Wang,

Zheng,

Sun

et al. 2024

Foods

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Plipastatin, an antimicrobial peptide produced by Bacillus subtilis, exhibits remarkable antimicrobial activity against a diverse range of pathogenic bacteria and fungi. However, the practical application of plipastatin has been significantly hampered by its low yield in wild Bacillus species. Here, the native promoters of both the plipastatin operon and the sfp gene in the mono-producing strain M-24 were replaced by the constitutive promoter P43, resulting in plipastatin titers being increased by 27% (607 mg/mL) and 50% (717 mg/mL), respectively. Overexpression of long chain fatty acid coenzyme A ligase (LCFA) increased the yield of plipastatin by 105% (980 mg/mL). A new efflux transporter, YoeA, was identified as a MATE (multidrug and toxic compound extrusion) family member, overexpression of yoeA enhanced plipastatin production to 1233 mg/mL, an increase of 157%, and knockout of yoeA decreased plipastatin production by 70%; in contrast, overexpression or knockout of yoeA in mono-producing surfactin and iturin engineered strains only slightly affected their production, demonstrating that YoeA acts as the major exporter for plipastatin. Co-overexpression of lcfA and yoeA improved plipastatin production to 1890 mg/mL, which was further elevated to 2060 mg/mL after abrB gene deletion. Lastly, the use of optimized culture medium achieved 2514 mg/mL plipastatin production, which was 5.26-fold higher than that of the initial strain. These results suggest that multiple strain engineering is an effective strategy for increasing lipopeptide production, and identification of the novel transport efflux protein YoeA provides new insights into the regulation and industrial application of plipastatin.

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Construction of lipopeptide mono-producing Bacillus strains and comparison of their antimicrobial activity

Cited by 7 publications

References 58 publications

Enhanced Iturin A Production of Engineered Bacillus amyloliquefaciens by Knockout of Endogenous Plasmid and Rap Phosphatase Genes

Enhanced Iturin A Production of Engineered Bacillus amyloliquefaciens by Knockout of Endogenous Plasmid and Rap Phosphatase Genes

Biocontrol and Growth Promotion Potential of Bacillus subtilis CTXW 7-6-2 against Rhizoctonia solani that Causes Tobacco Target Spot Disease

An Integrated Pipeline and Overexpression of a Novel Efflux Transporter, YoeA, Significantly Increases Plipastatin Production in Bacillus subtilis

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