Abstract
The inhibitory properties of novel antimicrobial proteins against food-borne pathogens such as Listeria monocytogenes offer extensive benefits to the food and medical industries. In this study, we have identified antimicrobial proteins from a milk curd–derived bacterial isolate that exhibits antilisterial activity using genome mining and mass spectrometry analysis. The analysis of the draft genome sequence identified the isolate as Paenibacillus polymyxa Kp10, and predicted the presence of antimicrobial paenibacillin, paenilan, paeninodin, sactipeptides, thiazole-oxazole modified microcin, and histone-like DNA binding protein HU encoded in its genome. Interestingly, nanoLC-MS/MS analysis identified two histone-like DNA binding proteins HU as predicted in silico earlier, exhibiting antilisterial activity. Additionally, translation initiation factor IF-1 and 50S ribosomal protein L29 were also discovered by the mass spectrometry in the active fractions. The antilisterial activity of the four proteins was verified through heterologous protein expression and antimicrobial activity assay in vitro. This study has identified structural regulatory proteins from Paenibacillus possessing antilisterial activity with potential future application in the food and medical industries.
Highlights
Food spoilage caused by pathogenic microbes such as Listeria monocytogenes (L. monocytogenes) is a common problem, imposing serious detrimental economic effects
The current study aims to identify potential antimicrobial proteins from a local milk curd isolate through genome mining and mass spectrometry
The cell-free culture supernatant (CFCS) of an unidentified pure bacterial culture isolated from milk curd has preliminarily been found to confer high antilisterial activity at 1051.84 ± 11.84 AU (Figure S1)
Summary
Food spoilage caused by pathogenic microbes such as Listeria monocytogenes (L. monocytogenes) is a common problem, imposing serious detrimental economic effects. Its ability to form a biofilm on the surface of food processing equipment and its development of resistance to the standard sterilizing procedure have imposed further burdens on the food industry (Skowron et al, 2018). Common food preservatives to eradicate pathogen contamination include acetic acid, potassium acetate, Antilisterial Proteins From Paenibacillus polymyxa Kp10 benzoic acid, benzoates, and nitrites and nitrates. The cytotoxic and mutagenic activity of sodium benzoate, a common preservative in dairy products, has been found in lymphocytes (Pongsavee, 2015). The use of antimicrobial proteins in food and medical industries has increased in the past decades due to their inhibitory activity against food-borne pathogens and multidrug-resistant bacteria
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