Negative Interplay between Biofilm Formation and Competence in the Environmental Strains of
            <i>Bacillus subtilis</i>

She, Qianxuan; Hunter, Evan; Qin, Yuxuan; Nicolau, Samantha E.; Zalis, Eliza A.; Wang, Hongkai; Yun, Chen; Chai, Yunrong

doi:10.1128/msystems.00539-20

Cited by 13 publications

(10 citation statements)

References 66 publications

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“…In the wild type strain GP2630, about 40% of the cells expressed GFP, similar to previously published results (She et al, 2020), and slightly higher numbers were obtained for the control strain lacking ComEC, which is not impaired in comK and subsequent comG expression. In contrast, the control strain lacking DegU showed decreased amount of GFP expressing cells as compared to the wild type (Figure 1), which reflects the role of DegU on the activation of comK expression.…”

Section: Comk-dependent and -Independent Functions Of Proteins Are Required For The Development Of Genetic Competencesupporting

confidence: 89%

Influence of the ABC Transporter YtrBCDEF of Bacillus subtilis on Competence, Biofilm Formation and Cell Wall Thickness

et al. 2021

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Bacillus subtilis develops genetic competence for the uptake of foreign DNA when cells enter stationary phase and a high cell density is reached. These signals are integrated by the competence transcription factor ComK, which is subject to transcriptional, post-transcriptional and post-translational regulation. Many proteins are involved in the development of competence, both to control ComK activity and to mediate DNA uptake. However, for many proteins, the precise function they play in competence development is unknown. In this study, we assessed whether proteins required for genetic transformation play a role in the activation of ComK or rather act downstream of competence gene expression. While these possibilities could be distinguished for most of the tested factors, we assume that two proteins, PNPase and the transcription factor YtrA, are required both for full ComK activity and for the downstream processes of DNA uptake and integration. Further analyses of the role of the transcription factor YtrA for the competence development revealed that the overexpression of the YtrBCDEF ABC transporter in the ytrA mutant causes the loss of genetic competence. Moreover, overexpression of this ABC transporter also affects biofilm formation. Since the ytrGABCDEF operon is naturally induced by cell wall-targeting antibiotics, we tested the cell wall properties upon overexpression of the ABC transporter and observed an increased thickness of the cell wall. The composition and properties of the cell wall are important for competence development and biofilm formation, suggesting that the observed phenotypes are the result of the increased cell wall thickness as an outcome of YtrBCDEF overexpression.

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Section: Comk-dependent and -Independent Functions Of Proteins Are Required For The Development Of Genetic Competencesupporting

confidence: 89%

Influence of the ABC Transporter YtrBCDEF of Bacillus subtilis on Competence, Biofilm Formation and Cell Wall Thickness

et al. 2021

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“…Biofilm is considered as a suppressor of lipopeptides (e.g., surfactin) synthesis, so disruption of biofilm formation is favorable for improving lipopeptides production ( Wu et al, 2019 ). Here, we knocked out several genes related to biofilm formation such as comK ( She et al, 2020 ), sigD ( Fan et al, 2016 ), dhbF (a gene of dhb operon for biosynthesis of siderophore) and fur (a repressor of dhb ) ( Oliveira et al, 2017 ; Pi and Helmann 2017 ), abrB and sinR (regulators to suppress biofilm formation) ( Newman et al, 2013 ; Klausmann et al, 2021 ), and sinI (a regulator to promote biofilm formation) ( López et al, 2009 ). Deletion of respective genes resulted in an obvious change of colony morphology and float pellicle, but only deletion of dhbF led to a significant increase of antifungal activity.…”

Section: Discussionmentioning

confidence: 99%

Systemically engineering Bacillus amyloliquefaciens for increasing its antifungal activity and green antifungal lipopeptides production

Wang

Wang²,

Zhao

et al. 2022

Front. Bioeng. Biotechnol.

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The biosynthesis of antifungal lipopeptides iturin and fengycin has attracted broad interest; however, there is a bottleneck in its low yield in wild strains. Because the key metabolic mechanisms in the lipopeptides synthesis pathway remain unclear, genetic engineering approaches are all ending up with a single or a few gene modifications. The aim of this study is to develop a systematic engineering approach to improve the antifungal activity and biosynthesis of iturin and fengycin in Bacillus amyloliquefaciens. First, blocking the carbon overflow metabolic pathway to increase precursor supply of the branched-chain amino acids by knockout of bdh, disrupting sporulation to extend the stage for producing antifungal lipopeptides by deletion of kinA, blocking of siderophore synthesis to enhance the availability of amino acids and fatty acids by deletion of dhbF, and increasing Spo0A∼P by deletion of rapA, could improve the antifungal activity by 24%, 10%, 13% and 18%, respectively. Second, the double knockout strain ΔbdhΔkinA, triple knockout strain ΔbdhΔkinAΔdhbF and quadruple knockout strain ΔkinAΔbdhΔdhbFΔrapA could improve the antifungal activity by 38%, 44% and 53%, respectively. Finally, overexpression of sfp in ΔkinAΔbdhΔdhbFΔrapA further increased the antifungal activity by 65%. After purifying iturin and fengycin as standards for quantitative analysis of lipopeptides, we found the iturin titer was 17.0 mg/L in the final engineered strain, which was 3.2-fold of the original strain. After fermentation optimization, the titer of iturin and fengycin reached 31.1 mg/L and 175.3 mg/L in flask, and 123.5 mg/L and 1200.8 mg/L in bioreactor. Compared to the original strain, the iturin and fengycin titer in bioreactor increased by 22.8-fold and 15.9-fold in the final engineered strain, respectively. This study may pave the way for the commercial production of green antifungal lipopeptides, and is also favorable for understanding the regulatory and biosynthetic mechanism of iturin and fengycin.

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“…This study showed that natural competence could not transfer plasmids to undomesticated B. subtilis strains tested ( Figure 1 ). The result may be due to a lack of natural competence-related genes or the negative interplay between biofilm formation and competence ( She et al, 2020 ). Some strains may have competence inhibitor genes ( Konkol et al, 2013 ).…”

Section: Discussionmentioning

confidence: 99%

Cell Factory Engineering of Undomesticated Bacillus Strains Using a Modified Integrative and Conjugative Element for Efficient Plasmid Delivery

Jeong

Kim

et al. 2022

Front. Microbiol.

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A large number of Bacillus strains have been isolated from various environments and many of them have great potential as cell factories. However, they have been rarely developed as cell factories due to their poor transformation efficiency. In this study, we developed a highly efficient plasmid delivery system for undomesticated Bacillus strains using a modified integrative and conjugative element (MICE), which was designed to be activated by an inducer, prevent self-transfer, and deliver desired plasmids to the recipient cells. The MICE system was demonstrated to successfully introduce a gfp-containing plasmid into all 41 undomesticated Bacillus subtilis strains tested and eight other Bacillus species. The MICE was used to deliver a cytosine base editor (CBE)-based multiplex genome-editing tool for the cell factory engineering of the Bacillus species. The introduced CBE enabled one-step inactivation of the major extracellular protease genes of the tested strains. The engineered strains were used as hosts for heterologous expression of nattokinase, which resulted in various enzyme expression levels. The results suggested that the MICE and CBE systems can be powerful tools for genetic engineering of undomesticated Bacillus strains, and greatly contribute to the expansion of the Bacillus cell factory.

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Negative Interplay between Biofilm Formation and Competence in the Environmental Strains of Bacillus subtilis

Cited by 13 publications

References 66 publications

Influence of the ABC Transporter YtrBCDEF of Bacillus subtilis on Competence, Biofilm Formation and Cell Wall Thickness

Influence of the ABC Transporter YtrBCDEF of Bacillus subtilis on Competence, Biofilm Formation and Cell Wall Thickness

Systemically engineering Bacillus amyloliquefaciens for increasing its antifungal activity and green antifungal lipopeptides production

Cell Factory Engineering of Undomesticated Bacillus Strains Using a Modified Integrative and Conjugative Element for Efficient Plasmid Delivery

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