Molecular Cloning and Characterization of Fengycin Synthetase Gene
            <i>fenB</i>
            from
            <i>Bacillus subtilis</i>

Lin, Guang-Huey; Chen, Chyi‐Liang; Tschen, Johannes Scheng-Ming; Tsay, San-San; Chang, Yu-Sun; Liu, Shih‐Tung

doi:10.1128/jb.180.5.1338-1341.1998

Cited by 30 publications

(24 citation statements)

References 25 publications

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“…Recently, fenB, which is a fengycin synthetase gene and a homolog of ppsE, was cloned and sequenced. FenB overexpression revealed that FenB is responsible for isoleucine activation (20). This result agrees with our prediction of activation of isoleucine by PpsE.…”

Section: Discussionsupporting

confidence: 92%

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The Genes degQ , pps , and lpa-8 ( sfp ) Are Responsible for Conversion of Bacillus subtilis 168 to Plipastatin Production

Tsuge

Ano

Hirai

et al. 1999

Antimicrob Agents Chemother

121

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Bacillus subtilis YB8 produces the lipopeptide antibiotic plipastatin. B. subtilis MI113, which is a derivative of strain 168, was converted into a new plipastatin producer, strain 406, by competence transformation with the chromosomal DNA of YB8. Transposon mini-Tn10 insertional mutagenesis was applied to strain 406, which revealed that lpa-8(sfp) (encoding 4′-phosphopantetheinyl transferase) and thepps operon (located between 167 and 171°) are essential for plipastatin production. The pps operon was previously suggested to encode putative peptide synthetases (A. Tognoni, E. Franchi, C. Magistrelli, E. Colombo, P. Cosmina, and G. Grandi, Microbiology 141:645–648, 1995) and was thought to be the fengycin operon (V. Tosato, A. M. Albertini, M. Zotti, S. Sonda, and C. V. Bruschi, Microbiology 143:3443–3450, 1997). We claim that the pps operon is the pli operon, encoding plipastatin synthetase. By using a new high-performance liquid chromatography system, we revealed that strain 168 expressing onlylpa-8 can also produce plipastatin, although the yield is very low. However, the introduction of the pleiotropic regulatordegQ of strain YB8 into strain 168 expressinglpa-8 resulted in a 10-fold increase in the production of plipastatin.

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Section: Discussionsupporting

confidence: 92%

“…However, ambiguity still exists regarding the structure of fengycin. The fengycin operon had been cloned and sequenced from B. subtilis F29-3 (2,20) and has significant homology with the putative peptide synthetase operon in strain 168 (20,39).…”

Section: Discussionmentioning

confidence: 99%

The Genes degQ , pps , and lpa-8 ( sfp ) Are Responsible for Conversion of Bacillus subtilis 168 to Plipastatin Production

Tsuge

Ano

Hirai

et al. 1999

Antimicrob Agents Chemother

121

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“…The final module of a peptide synthetase, except for the one involved in the termination process, typically contains an epimerization domain (12) which converts an L-amino acid to a D-amino acid. The module in-volved in the termination of peptide synthesis contains a thioesterase domain in the C-terminal region (11,24,30,32,33).…”

mentioning

confidence: 99%

Functional and Transcriptional Analyses of a Fengycin Synthetase Gene, fenC, from Bacillus subtilis

Lin¹,

Chen

Chang

et al. 1999

J. Bacteriol.

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A 37-kb DNA fragment containing five fengycin synthetase genes, including fenC, fenD, fenE,fenA, and fenB, was cloned and sequenced. Among these genes, fenC encodes a fengycin synthetase 2,560 amino acids long with an estimated molecular mass of 287 kDa. This protein contains two amino acid activation modules, FenC1 and FenC2, which activate l-glutamic acid and l-ornithine, respectively. Primer extension, using mRNA isolated from the log-phase cells, identified a transcription start site located 86 nucleotides upstream from the initiation codon of fenC, implying that a promoter is located upstream from the start site. Primer extension using total RNA isolated from stationary-phase cells also identified a transcription start site located 61 nucleotides upstream from the initiation codon of fenC. Gene fusion studies demonstrated that in nHA medium, the cells transcribe the fengycin synthetase genes at two different stages of cell growth. The promoter is active during the log phase, and the activity reaches the highest level during the late log phase. The activity decreases sharply but is maintained at a low level for approximately 24 h after cells enter the early stationary phase. The results of this investigation also suggest that the transcription of fenC is positively regulated during the late log phase. Results presented herein provide further insight into fengycin synthesis by B. subtilis F29-3.

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“…The fengycin biosynthetic gene cluster in the strain consists of ve genes (38 kb) that encode the synthetases FenCDEAB, of which FenC recognizes and carries glutamate and ornithine, FenD recognizes and carries tyrosine and threonine, FenE recognizes and carries glutamate and valine, FenA recognizes and carries proline, glutamine, and tyrosine, and FenB recognizes and carries isoleucine. FenCDEAB recognizes 10 amino acids and carries them to the β-OH FA chain to form fengycin [52][53][54]. However, NCD-2 only had fenEAB, lacking fenC and fenD, compared with the typical cluster structure of fenCDEAB in the FZB42 strain and 10 other Bacillus strains ( Fig.…”

Section: Discussionmentioning

confidence: 94%

Genome mining and UHPLC–QTOF–MS/MS illuminate the potential antimicrobial active compounds and specificity of biosynthetic gene clusters in Bacillus subtilis NCD-2

Su¹,

Chen²,

Liu³

et al. 2020

Preprint

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Background Bacillus subtilis strain NCD-2 is an excellent biocontrol agent against plant soil-borne diseases and shows broad-spectrum antifungal activities. This study aimed to explore some of the secondary metabolite biosynthetic gene clusters and related bioactive compounds in strain NCD-2. An integrative approach, which combined genome mining with structural identification technologies using ultra-high-performance liquid chromatography coupled to quadrupole time-of-flight tandem mass spectrometry (UHPLC-MS/MS), was conducted to interpret the chemical origins of the significant biological activities in strain NCD-2. Results Genome mining revealed that strain NCD-2 contained nine gene clusters having predicted functions involving secondary metabolites with bioactive abilities. They encoded six known products including fengycin, surfactin, bacillaene, subtilosin, bacillibactin, bacilysin and three unknown products. Fengycin, surfactin, bacillaene and bacillibactin were successfully detected from the fermentation broth of strain NCD-2 by UHPLC-QTOF-MS/MS. Bacillaene, subtilosin, bacillibactin, and bacilysin related biosynthetic gene clusters showed 100% amino acid sequence similarity with B. velezensis strain FZB42，however, the biosynthetic gene clusters for surfactin and fengycin showed 83% and 92%, respectively. Further comparison of gene clusters encoding fengycin and surfactin revealed that strain NCD-2 had lost the fenC and fenD genes in the fengycin biosynthetic operon. Moreover, biosynthetic enzyme-related gene srfAB for surfactin had divided into two parts. Bioinformatics analysis predicted that FenE function in strain NCD-2 was same to that of FenE and FenC in strain FZB42, and FenA function in strain NCD-2 was same to that of FenA and FenD in strain FZB42. Five kinds of fengycin, with 26 homologs, and surfactin, with 4 homologs, were detected from strain NCD-2. To the best of our knowledge, this is the first report of a non-typical and unique gene cluster related to fengycin synthesis. Conclusions It was found that there were many gene clusters encoding antimicrobial compounds in the genome of strain NCD-2, and the fengycin synthetic gene cluster might be unique by using genome mining and UHPLC–QTOF–MS/MS. The production of fengycin, surfactin, bacillaene and bacillibactin might explain the biological activities of strain NCD-2.

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Molecular Cloning and Characterization of Fengycin Synthetase Gene fenB from Bacillus subtilis

Cited by 30 publications

References 25 publications

The Genes degQ , pps , and lpa-8 ( sfp ) Are Responsible for Conversion of Bacillus subtilis 168 to Plipastatin Production

The Genes degQ , pps , and lpa-8 ( sfp ) Are Responsible for Conversion of Bacillus subtilis 168 to Plipastatin Production

Functional and Transcriptional Analyses of a Fengycin Synthetase Gene, fenC, from Bacillus subtilis

Genome mining and UHPLC–QTOF–MS/MS illuminate the potential antimicrobial active compounds and specificity of biosynthetic gene clusters in Bacillus subtilis NCD-2

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