Novel Methods for Genetic Transformation of Natural
            <i>Bacillus subtilis</i>
            Isolates Used To Study the Regulation of the Mycosubtilin and Surfactin Synthetases

Duitman, Erwin H.; Wyczawski, Dobek; Boven, Ludolf G.; Venema, Gerard; Kuipers, Oscar P.; Hamoen, Leendert W.

doi:10.1128/aem.02751-06

Cited by 58 publications

(56 citation statements)

References 52 publications

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“…In B. subtilis, overexpression of comK allows for genetic access of non-competent wild isolates (Duitman et al 2007;Nijland et al 2010). Artificially induced competence was successfully applied for close relatives of B. subtilis as well: in B. licheniformis MW3.1, overexpression of the homologous comK yielded up to 2.97×10 5 transformants per microgram of chromosomal DNA-an efficiency that allows for direct gene knock outs and even conditional knock outs of essential genes (Hoffmann et al 2010).…”

Section: Artificial Induction Of Genetic Competencementioning

confidence: 99%

What renders Bacilli genetically competent? A gaze beyond the model organism

Jakobs

Meinhardt

2014

Appl Microbiol Biotechnol

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Natural genetic competence enables bacteria to take in and establish exogenously supplied DNA and thus constitutes a valuable tool for strain improvement. Extensively studied in the Gram-positive model organism Bacillus subtilis genetic competence has indeed proven successful for genetic manipulation aiming at enhancement of handling, yield, and biosafety. The majority of Bacilli, particularly those relevant for industrial application, do not or only poorly develop genetic competence, although rather homologous DNA-uptake machineries are routinely encoded. Establishing the competent state solely due to high cell densities (quorum sensing dependency) appears to be restricted to the model organism, in which the small signalling peptide ComS initiates the regulatory pathway that ultimately leads to the expression of all genes necessary for reaching the competent state. Agreeing with the lack of a functional ComS peptide, competence-mediated transformation of other Bacilli depends on nutrient exhaustion rather than cell density. Genetically, competent strains of the model organism B. subtilis, cultivated for a long time and selected for laboratory purposes, display probably not least to such selection a point mutation in the promoter of a regulatory gene that favors competence development whereas the wild-type progenitor only poorly displays genetic competence. Consistent with competence being a matter of deregulation, all strains of Bacillus licheniformis displaying efficient DNA uptake were found to carry mutations in regulator genes, which are responsible for their genetic competence. Thus, strain-specific genetic equipment and regulation as well as the proven role of domestication for the well-established laboratory strains ought to be considered when attempting to broaden the applicability of competence as a genetic tool for strains other than the model organism.

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Section: Artificial Induction Of Genetic Competencementioning

confidence: 99%

What renders Bacilli genetically competent? A gaze beyond the model organism

Jakobs

Meinhardt

2014

Appl Microbiol Biotechnol

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“…Its expression is highest at the end of logarithmic phase (Tsuge et al 2001b). Duitman et al (2007) have shown recently that mycosubtilin synthetase expression is under the influence of a regulatory cascade depending on AbrB, one of the main transition state regulators in B. Fig.…”

Section: Quorum Sensing and Surfactin Effluxmentioning

confidence: 99%

“…Performed on surfactin synthetase, the same promoter exchange did not allow the improvement of the surfactin production (Coutte et al 2010a). The native promoter P srf of Bacillus subtilis 168 is considered very efficient, frequently allowing the concentration of surfactin in the culture supernatant to be higher than 1 g l À1 (Duitman et al 2007). The horizontal transfer of iturin operon in Bacillus subtilis 168 together with a functional sfp gene and the pleiotropic regulator degQ allowed conversion of this strain to an efficient iturin producer (Tsuge et al 2005).…”

Section: Directed Biosynthesis and Molecular Optimisationmentioning

confidence: 99%

Surfactin and Other Lipopeptides from Bacillus spp.

Jacques

2010

Microbiology Monographs

162

167

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“…The chemical principles for the biosynthesis of iturin-like compounds are also largely characterized (12,44). In contrast, little is known about the regulatory mechanisms that control the expression of the iturin-like lipopeptides (13).…”

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confidence: 99%

DegU and YczE Positively Regulate the Synthesis of Bacillomycin D byBacillus amyloliquefaciensStrain FZB42

Koumoutsi

Chen

Vater

et al. 2007

Appl Environ Microbiol

108

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Environmental strain Bacillus amyloliquefaciens FZB42 differs from the domesticated model organism of the same genus, Bacillus subtilis 168, in its ability to promote plant growth and suppress plant-pathogenic organisms present in the rhizosphere. This behavior is exerted mainly through the production of several nonribosomal cyclic lipopeptides and polyketides, which exhibit a broad range of action against phytopathogenic bacteria, fungi, and nematodes. Here, we provide evidence that the synthesis of the main antifungal agent of B. amyloliquefaciens Plant growth-promoting Bacillus amyloliquefaciens strain FZB42 is applied as a biocontrol agent, as it suppresses soilborne plant-pathogenic organisms. Its antagonistic function is attributed mainly to its ability to produce several secondary metabolites with antibacterial and antifungal activities (5). We have previously reported that FZB42 is a producer of the antibacterial polyketides bacillaene, difficidin/oxydifficidin (6), and a third polyketide which has been identified recently as macrolactin (5). Three cyclic lipopeptides are also produced by B. amyloliquefaciens FZB42 (referred to hereafter as FZB42): surfactin, fengycin, and bacillomycin D. Bacillomycin D has been shown to exert the main antifungal activity (20).Bacillomycin D belongs to the iturin family of lipopeptides, with members such as iturin A and mycosubtilin. It is a cyclic heptapeptide with a ␤-amino fatty acid moiety and is synthesized nonribosomally according to the multicarrier thiotemplate mechanism (40). The bmy operon (37.2 kb) directs the biosynthesis of bacillomycin D and consists of four genes (bmyD, bmyA, bmyB, and bmyC) without orthologues in Bacillus subtilis 168. Several studies have elucidated the physicochemical and biological properties of peptides that belong to the iturin group (3, 25, 34). The chemical principles for the biosynthesis of iturin-like compounds are also largely characterized (12, 44). In contrast, little is known about the regulatory mechanisms that control the expression of the iturin-like lipopeptides (13).Nonribosomal peptide synthetases require posttranslational modification to be functionally active (14). In bacilli, this is achieved by the sfp gene, which encodes a 4Ј-phosphopantetheinyl transferase and posttranslationally converts inactive apoenzyme peptide synthetases to their active holoenzyme forms (14, 18). Strains containing intact synthetases but a dysfunctional sfp gene, such as B. subtilis 168, which contains intact srf and fen operons but has a frameshift mutation in the sfp gene (27), are unable to produce the antibiotics. However, when B. subtilis 168 is complemented with a functional 4Ј-phosphopantetheinyl transferase, its antibiotic production is restored (30,45).It has also been reported that increased expression of the pleiotropic regulator DegQ in B. subtilis 168 (1) enhances the antibiotic production (45, 46). Interestingly, most of the natural Bacillus isolates that express peptide antibiotics show significantly elevated degQ expression c...

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Novel Methods for Genetic Transformation of Natural Bacillus subtilis Isolates Used To Study the Regulation of the Mycosubtilin and Surfactin Synthetases

Cited by 58 publications

References 52 publications

What renders Bacilli genetically competent? A gaze beyond the model organism

What renders Bacilli genetically competent? A gaze beyond the model organism

Surfactin and Other Lipopeptides from Bacillus spp.

DegU and YczE Positively Regulate the Synthesis of Bacillomycin D byBacillus amyloliquefaciensStrain FZB42

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