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. 2016 Dec;14(6):5155-5163.
doi: 10.3892/mmr.2016.5900. Epub 2016 Oct 27.

Extracts containing CLPs of Bacillus amyloliquefaciens JN68 isolated from chicken intestines exert antimicrobial effects, particularly on methicillin-resistant Staphylococcus aureus and Listeria monocytogenes

Jen-Ni Chen  1 , Chyou-Wei Wei  1 , Hsiao-Chun Liu  1 , Shu-Ying Chen  2 , Chinshuh Chen  3 , Yu-Min Juang  4 , Chien-Chen Lai  4 , Giou-Teng Yiang  5
Affiliations

Extracts containing CLPs of Bacillus amyloliquefaciens JN68 isolated from chicken intestines exert antimicrobial effects, particularly on methicillin-resistant Staphylococcus aureus and Listeria monocytogenes

Jen-Ni Chen et al. Mol Med Rep. 2016 Dec.

Abstract

Bacillus amyloliquefaciens JN68, which has been discussed with regards to its antimicrobial activities, was successfully isolated from healthy chicken intestines in the present study. Using the spot-on-the-lawn antagonism method, the preliminary study indicated that a suspension culture of the B. amyloliquefaciens JN68 strain can inhibit the growth of Aspergillus niger and Penicillium pinophilum. Furthermore, the cyclic lipopeptides (CLPs) produced by the B. amyloliquefaciens JN68 strain were further purified through acid precipitation and Bond Elut®C18 chromatography, and their structures were identified using the liquid chromatography‑electrospray ionization‑mass spectrometry (MS)/MS method. Purified CLPs exerted broad spectrum antimicrobial activities on various pathogenic and foodborne bacteria and fungi, as determined using the agar well diffusion method. Listeria monocytogenes can induce listeriosis, which is associated with a high mortality rate. Methicillin‑resistant Staphylococcus aureus (MRSA) is a major pathogenic bacteria that causes nosocomial infections. Therefore, L. monocytogenes and MRSA are currently of great concern. The present study aimed to determine whether B. amyloliquefaciens JN68 extracts could inhibit L. monocytogenes and MRSA. The results indicated that extracts of B. amyloliquefaciens JN68 have CLP components, and can successfully inhibit the growth of L. monocytogenes and MRSA.

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Figures

Figure 1.
Figure 1.
Screening of potential antimicrobial strains against (A) Aspergillus niger and (B) Penicillium pinophilum, as demonstrated from the zones of inhibition produced using the spot-on-the-lawn method.
Figure 2.
Figure 2.
LC-ESI-MS/MS analysis of (A and B) surfactin standard and (C and D) biosurfactant lipopeptide extract. (A and C) Exact ion current chromatogram of surfactin (m/z 1,036). (B and D) Product ion spectra of the protonated molecules [M+H]+ of surfactin at m/z 1,036. LC-ESI-MS/MS, liquid chromatography-electrospray ionization-mass spectrometry/mass spectrometry.
Figure 3.
Figure 3.
LC-ESI-MS/MS analysis of (A and B) iturin A standard and (C and D) biosurfactant lipopeptide extract. (A and C) Exact ion current chromatogram of iturin A (m/z 1,043). (B and D) Product ion spectra of the protonated molecules [M+H]+ of iturin A at m/z 1,043. LC-ESI-MS/MS, liquid chromatography-electrospray ionization-mass spectrometry/mass spectrometry.
Figure 4.
Figure 4.
LC-ESI-MS/MS analysis of biosurfactant lipopeptide extract. (A) Exact ion current chromatogram of fengycin A (m/z 1,464). (B) Product ion spectra of the protonated molecules [M+H]+ of fengycin A at m/z 1,464. Significant fragment ions of fengycin A were marked with an asterisk. LC-ESI-MS/MS, liquid chromatography-electrospray ionization-mass spectrometry/mass spectrometry.
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