The Discovery of New Antilisterial Proteins From Paenibacillus polymyxa Kp10 via Genome Mining and Mass Spectrometry

Nur Fadhilah Khairil Mokhtar; Amalia Mohd Hashim; Irwan Hanish; Aisyah Zulkarnain; Raja Mohd Hafidz Raja Nhari; Asmahani Azira Abdul Sani; Sahar Abbasiliasi; Arbakariya Ariff; Shuhaimi Mustafa; Raha Abdul Rahim

doi:10.3389/fmicb.2020.00960

The Discovery of New Antilisterial Proteins From Paenibacillus polymyxa Kp10 via Genome Mining and Mass Spectrometry

Front Microbiol. 2020 Jul 2:11:960. doi: 10.3389/fmicb.2020.00960. eCollection 2020.

Authors

Affiliations

¹ Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
² Halal Products Research Institute, Putra Infoport, Universiti Putra Malaysia, Selangor, Malaysia.
³ Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
⁴ Mass Spectrometry Technology Section, Malaysia Genome Institute, National Institute of Biotechnology Malaysia, Kajang, Malaysia.
⁵ Bioprocessing and Biomanufacturing Research Centre, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Selangor, Malaysia.
⁶ Chancellory, Universiti Teknikal Malaysia Melaka, Malacca, Malaysia.

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.

Keywords: Listeria monocytogenes; Paenibacillus polymyxa Kp10; antilisterial protein; genome mining; heterologous protein expression; mass spectrometry.