Michelle M Gerst1, Árpád Somogyi2, Xu Yang3, Ahmed E Yousef1,3. 1. Department of Microbiology, The Ohio State University, Columbus, Ohio, USA. 2. Mass Spectrometry and Proteomics Facility, Campus Chemical Instrument Center, The Ohio State University, Columbus, Ohio, USA. 3. Department of Food Science and Technology, The Ohio State University, Columbus, Ohio, USA.
Abstract
AIM: To detect and characterize novel lantibiotics from a collection of Bacillus spp. using a multifaceted analytical approach. METHODS AND RESULTS: A previously completed microassay identified 45 Bacillus isolates with anti-Listeria activity. The isolates were PCR screened using degenerate primers targeting conserved sequences in lanM-type lantibiotics. B. velezensis GF610 produced a PCR product whose sequence, along with genome mining and bioinformatics, guided the liquid chromatographic analysis of strain's cell-free extracts and the mass spectrometry of purified fractions. Results revealed a new amyloliquecidin variant (designated GF610) produced by the strain. Amyloliquecidin GF610 is a two-component lantibiotic with α and β peptides having monoisotopic masses of 3026 and 2451 Da, and molecular formulae C130 H191 N35 O39 S5 and C110 H158 N26 O30 S4 , respectively. Amyloliquecidin GF610 is active against Listeria monocytogenes, Clostridium sporogenes, Clostridioides difficile, Staphylococcus aureus and Alicyclobacillus acidoterrestris with minimum inhibitory concentrations (MICs) in the range of 0.5-7.0 µmol l-1 . CONCLUSIONS: The proposed multifaceted analytical approach was valuable to provide a deep and proper characterization of a novel bacteriocin, amyloliquecidin GF610, with high antimicrobial activity against Gram-positive bacteria. SIGNIFICANCE AND IMPACT: The discovered Amyloliquecidin GF610 is potentially useful in food, agricultural or medical applications. The analytical approach followed may facilitate future discoveries of two-component lantibiotics, which are challenging compounds to detect and characterize.
AIM: To detect and characterize novel lantibiotics from a collection of Bacillus spp. using a multifaceted analytical approach. METHODS AND RESULTS: A previously completed microassay identified 45 Bacillus isolates with anti-Listeria activity. The isolates were PCR screened using degenerate primers targeting conserved sequences in lanM-type lantibiotics. B. velezensis GF610 produced a PCR product whose sequence, along with genome mining and bioinformatics, guided the liquid chromatographic analysis of strain's cell-free extracts and the mass spectrometry of purified fractions. Results revealed a new amyloliquecidin variant (designated GF610) produced by the strain. Amyloliquecidin GF610 is a two-component lantibiotic with α and β peptides having monoisotopic masses of 3026 and 2451 Da, and molecular formulae C130 H191 N35 O39 S5 and C110 H158 N26 O30 S4 , respectively. Amyloliquecidin GF610 is active against Listeria monocytogenes, Clostridium sporogenes, Clostridioides difficile, Staphylococcus aureus and Alicyclobacillus acidoterrestris with minimum inhibitory concentrations (MICs) in the range of 0.5-7.0 µmol l-1 . CONCLUSIONS: The proposed multifaceted analytical approach was valuable to provide a deep and proper characterization of a novel bacteriocin, amyloliquecidin GF610, with high antimicrobial activity against Gram-positive bacteria. SIGNIFICANCE AND IMPACT: The discovered Amyloliquecidin GF610 is potentially useful in food, agricultural or medical applications. The analytical approach followed may facilitate future discoveries of two-component lantibiotics, which are challenging compounds to detect and characterize.