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Identification, Purification, Characterization and Biopreservation Potential of Antimicrobial Peptide of Pediococcus acidilactici NCDC 252

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Abstract

Lactic acid bacteria (LAB) are economically important because of their fermentative aptitude and they also confer health and nutritional benefits. Pediococcus acidilactici NCDC 252 (NCDC 252) is potential probiotic with novel genome that exhibited all in-vitro probiotic attributes, colonization in intestine and survival in conditions that mimick gastric, intestinal and vaginal fluid. Genome was screened for its ability to produce antimicrobial peptide (AMP). AMP from NCDC 252 was isolated and purified against indicator strain (Escherichia coli). AMP production was optimized in MRS broth at 32 °C, pH 6 and upto 36 h of incubation under both aerobic and anaerobic conditions. AMP was purified 2.2 fold with yield of 7.2% and specific activity of 64.5 U mg/mL using ammonium sulphate precipitation (upto 70%) and gel filtration chromatography. Molecular weight of purified AMP was calculated to be 2.5 KDa. Tricine SDS-PAGE and LC/MS studies confirmed it to be a low molecular weight peptide. AMP was stable over pH range of 3 to 9 and temperature range of 40 °C to 100 °C. Enzymatic treatment inactivated the AMP whereas, detergent treatment enhanced its activity. Biopreservation potential of purified AMP and whole NCDC 252 cells was studied by incubating them with some dairy products, apple juice, apricot pulp and pre-pasteurized wine over 72 h. Bacterial growth in AMP treated food products reduced significantly after 72 h. Viable cell count decreased to approximately half in both purified AMP treated and NCDC 252 treated food products. Purified AMP demonstrated exceptional resistance to heat treatment over a wide pH range, enabling it to establish itself as a biopreservative. Studies on biopreservation potential revealed NCDC 252 derived AMP to be effective even at low concentrations and it can be utilised to extend the shelf life of food goods.

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Acknowledgements

Authors are thankful to Kurukshetra University, Kurukshetra for providing necessary facilities. Authors also acknowledge Professor Brajesh Sawhney, Department of English and Dean, Faculty of Languages for editing manuscript for English language.

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  1. Department of Biochemistry, Kurukshetra University, Kurukshetra, Haryana, 136119, India

    Shweta Dhanda, Praveen Kumar, Jasbir Singh & Suman Dhanda

  2. Department of Biotechnology, Maharishi Markandeshwar University, Mullana Ambala, Haryana, 133207, India

    Poonam Bansal

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  1. Shweta Dhanda
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SD, PB and PK: methodology and writing, SD: data curation, visualization, conceptualization, writing-reviewing and editing of original draft preparation. JS: editing in revised manuscript.

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Dhanda, S., Kumar, P., Bansal, P. et al. Identification, Purification, Characterization and Biopreservation Potential of Antimicrobial Peptide of Pediococcus acidilactici NCDC 252. Int J Pept Res Ther 29, 15 (2023). https://doi.org/10.1007/s10989-022-10485-z

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