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Isolation and Identification of Lipopeptide Antibiotics from Paenibacillus elgii B69 with Inhibitory Activity Against Methicillin-Resistant Staphylococcus aureus
Rui Ding , Xue-Chang Wu , Chao-Dong Qian , Yi Teng , Ou Li , Zha-Jun Zhan , Yu-Hua Zhao
J. Microbiol. 2011;49(6):942-949.   Published online December 28, 2011
DOI: https://doi.org/10.1007/s12275-011-1153-7
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  • 39 Crossref
AbstractAbstract
Two lipopeptide antibiotics, pelgipeptins C and D, were isolated from Paenibacillus elgii B69 strain. The molecular masses of the two compounds were both determined to be 1,086 Da. Mass-spectrometry, amino acid analysis and NMR spectroscopy indicated that pelgipeptin C was the same compound as BMY-28160, while pelgipeptin D was identified as a new antibiotic of the polypeptin family. These two peptides were active against all the tested microorganisms, including antibiotic-resistant pathogenic bacterial strains such as methicillin-resistant Staphylococcus aureus (MRSA). Time-kill assays demonstrated that pelgipeptin D exhibited rapid and effective bactericidal action against MRSA at 4×MIC. Based on acute toxicity test, the intraperitoneal LD50 value of pelgipeptin D was slightly higher than that of the structurally related antimicrobial agent polymyxin B. Pelgipeptins are highly potent antibacterial and antifungal agents, particularly against MRSA, and warrant further investigation as possible therapeutic agents for bacteria infections resistant to currently available antibiotics.

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Journal Article
Cyclic Lipopeptide Profile of Three Bacillus subtilis Strains; Antagonists of Fusarium Head Blight
Christopher A. Dunlap , David A. Schisler , Neil P. Price , Steven F. Vaughn
J. Microbiol. 2011;49(4):603-609.   Published online September 2, 2011
DOI: https://doi.org/10.1007/s12275-011-1044-y
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  • 59 Crossref
AbstractAbstract
The objective of the study was to identify the lipopetides associated with three Bacillus subtilis strains. The strains are antagonists of Gibberella zeae, and have been shown to be effective in reducing Fusarium head blight in wheat. The lipopeptide profile of three B. subtilis strains (AS43.3, AS43.4, and OH131.1) was determined using mass spectroscopy. Strains AS43.3 and AS43.4 produced the anti-fungal lipopeptides from the iturin and fengycin family during the stationary growth phase. All three strains produced the lipopeptide surfactin at different growth times. Strain OH131.1 only produced surfactin under these conditions. The antifungal activity of the culture supernatant and individual lipopeptides was determined by the inhibition of G. zeae. Cell-free supernatant from strains AS43.3 and AS43.4 demonstrated strong antibiosis of G. zeae, while strain OH131.1 had no antibiosis activity. These results suggest a different mechanism of antagonism for strain OH131.1, relative to AS43.3 and AS43.4.

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Research Support, Non-U.S. Gov'ts
Antifungal Activity and Mechanism of Fengycin in the Presence and Absence of Commercial Surfactin Against Rhizopus stolonifer
Yang Tao , Xiao-mei Bie , Feng-xia Lv , Hai-zhen Zhao , Zhao-xin Lu
J. Microbiol. 2011;49(1):146-150.   Published online March 3, 2011
DOI: https://doi.org/10.1007/s12275-011-0171-9
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AbstractAbstract
The antifungal activity and mechanism of fengycin in the presence and absence of commercial surfactin against Rhizopus stolonifer were investigated. The MIC (minimal inhibitory concentration) of fengycin without commercial surfactin added was 0.4 mg/ml while the MIC of fengycin with commercial surfactin added was 2.0 mg/ml. Fengycin acted on cell membrane and cellular organs and inhibited DNA synthesis. The antifungal effect of fengycin was reduced after commercial surfactin was added. All these results suggest that the fungal cell membrane may be the primary target of fengycin action and commercial surfactin may reduce the antifungal activity of fengycin.
Isolation and Characterization of Antifungal Peptides Produced by Bacillus amyloliquefaciens LBM5006
Lisianne Brittes Benitez , Renata Voltolini Velho , Marcia Pagno Lisboa , Luis Fernando da Costa Medina , Adriano Brandelli
J. Microbiol. 2010;48(6):791-797.   Published online January 9, 2011
DOI: https://doi.org/10.1007/s12275-010-0164-0
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AbstractAbstract
Bacillus amyloliquefaciens LBM 5006 produces antagonistic activity against pathogenic bacteria and phytopathogenic fungi, including Aspergillus spp., Fusarium spp., and Bipolaris sorokiniana. PCR analysis revealed the presence of ituD, but not sfp genes, coding for iturin and surfactin, respectively. The antimicrobial substance produced by this strain was isolated by ammonium sulfate precipitation, gel filtration chromatography and 1-butanol extraction. The ultraviolet spectrum was typical of a polypeptide and the infrared spectrum indicates the presence of peptide bonds and acyl group(s). The antimicrobial substance was resistant to proteolytic enzymes and heat treatment, and was reactive with ninhydrin. Mass spectroscopy analysis indicated that B. amyloliquefaciens LBM 5006 produces two antimicrobial peptides, with main peaks at m/z 1,058 Da and 1,464 Da, corresponding to iturin-like and fengycin-like peptides, respectively. B. amyloliquefaciens LBM 5006 showed significant activity against phytopatogenic fungi, showing potential for use as a biocontrol agent or production of antifungal preparations.

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Natarajan Velmurugan , Mi Sook Choi , Sang-Sub Han , Yang-Soo Lee
J. Microbiol. 2009;47(4):385-392.   Published online September 9, 2009
DOI: https://doi.org/10.1007/s12275-009-0018-9
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AbstractAbstract
The antifungal activity of bacterial strains Bacillus subtilis EF 617317 and B. licheniformis EF 617325 was demonstrated against sapstaining fungal cultures Ophiostoma flexuosum, O. tetropii, O. polonicum, and O. ips in both in vitro and in vivo conditions. The crude active supernatant fractions of 7 days old B. subtilis and B. licheniformis cultures inhibited the growth of sapstaining fungi in laboratory experiments. Thermostability and pH stability of crude supernatants were determined by series of experiments. FT-IR analysis was performed to confirm the surface structural groups of lipoproteins present in the crude active supernatant. Partial purification of lipopeptides present in the crude supernatant was done by using Cellulose anion exchange chromatography and followed by Sephadex gel filtration chromatography. Partially purified compounds significantly inhibited the sapstaining fungal growth by in vitro analysis. The lipopeptides responsible for antifungal activity were identified by electrospray ionization mass spectrometry after partial purification by ion exchange and gel filtration chromatography. Four major ion peaks were identified as m/z 1023, 1038, 1060, and 1081 in B. licheniformis and 3 major ion peaks were identified as m/z 1036, 1058, and 1090 in B. subtilis. In conclusion, the partially purified lipopeptides may belong to surfactin and iturin family. In vivo analysis for antifungal activity of lipopeptides on wood was conducted in laboratory. In addition, the potential of extracts for fungal inhibition on surface and internal part of wood samples were analyzed by scanning electron microscopy.

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