Department of Microbiology, The Ohio State University, Columbus, Ohio, USA.
Mass Spectrometry and Proteomics Facility, Campus Chemical Instrument Center, The Ohio State University, Columbus, Ohio, USA.
J Appl Microbiol. 2022 Feb;132(2):994-1007. doi: 10.1111/jam.15290. Epub 2021 Sep 15.
To detect and characterize novel lantibiotics from a collection of Bacillus spp. using a multifaceted analytical approach.
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 C H N O S and C H N O S , 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 .
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.
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.
使用多方面的分析方法从芽孢杆菌属中发现和鉴定新型类细菌素。
先前完成的微量分析鉴定出 45 株具有抗李斯特菌活性的芽孢杆菌分离株。使用针对 lanM 型类细菌素保守序列的简并引物对分离株进行 PCR 筛选。B. velezensis GF610 产生了一个 PCR 产物，其序列以及基因组挖掘和生物信息学指导了对菌株无细胞提取物的液相色谱分析和纯化馏分的质谱分析。结果显示，该菌株产生了一种新的短小芽孢杆菌素变体（命名为 GF610）。短小芽孢杆菌素 GF610 是一种二组分类细菌素，其 α 和 β 肽的分子量分别为 3026 和 2451 Da，分子式分别为 C H N O S 和 C H N O S 。短小芽孢杆菌素 GF610 对李斯特菌、梭状芽孢杆菌、艰难梭菌、金黄色葡萄球菌和嗜酸耐热节杆菌具有活性，最小抑菌浓度（MIC）在 0.5-7.0 µmol l 范围内。
所提出的多方面分析方法对于深入和正确表征新型细菌素短小芽孢杆菌素 GF610 非常有价值，该细菌素有很高的抗革兰氏阳性菌活性。
发现的短小芽孢杆菌素 GF610 在食品、农业或医学应用中具有潜在用途。所采用的分析方法可能有助于未来发现二组分类细菌素，这是一种具有挑战性的化合物，难以检测和表征。
