Purification, De Novo Characterization and Antibacterial Properties of a Novel, Narrow-Spectrum Bacteriostatic Tripeptid
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RESEARCH ARTICLE-BIOLOGICAL SCIENCES
Purification, De Novo Characterization and Antibacterial Properties of a Novel, Narrow-Spectrum Bacteriostatic Tripeptide from Geotrichum candidum OMON-1 Sunday Osaizua Omeike1 · Sarafadeen Olateju Kareem2 · Hemraj Nandanwar3 · Adebayo Aliyu Lasisi4 · Flora Oluwafemi2 · Manoj Jangra3 Received: 29 June 2020 / Accepted: 4 October 2020 © King Fahd University of Petroleum & Minerals 2020
Abstract Antibiotic resistance and need for novel antibiotics have seen focus on antimicrobial peptides (AMPs) increase in recent years. The study investigated purification and de novo characterization of an AMP compound produced by bioactive Geotrichum candidum OMON-1. Antimicrobial extract of G. candidum OMON-1 was purified via reverse-phase high-performance liquid chromatography (RP-HPLC). Purified compound (GP-2B) was characterized on liquid chromatography–mass spectrometry (LC–MS) and MALDI-TOF tandem mass spectrometry (MS/MS), and spectra were interpreted using de novo peptide sequencing protocols. Minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of GP-2B were determined in vitro against Gram-positive clinical isolates and time-kill kinetics against Staphylococcus aureus investigated, while antimicrobial activity was also observed on scanning electron microscope (SEM). GP-2B was successfully purified on RP-HPLC with trifluoroacetic acid-buffered acetonitrile. LC–MS confirmed GP-2B as pure and positively charged with molecular mass 409.23 amu, while characterization of MS/MS spectrum via de novo peptide sequencing described it a tripeptide with carboxymethylcystyl–asparagyl–aspartate amino acid sequence. GP-2B had MIC 8 and 32 µg/mL against antibiotic-sensitive and -resistant S. aureus, respectively, and 64 µg/mL for Enterococci strains. MBC was fourfold MIC, while GP-2B reduced S. aureus by 21.8% and 30% at 5× and 10× MIC, respectively, in time-kill kinetics. SEM images also confirmed bacteriostasis of S. aureus cells. This study successfully purified and characterized via de novo sequencing, a novel bacteriostatic AMP from G. candidum OMON-1. Keywords Antimicrobial peptide · Geotrichum candidum OMON-1 · De novo peptide sequencing · Carboxymethylcysteine · Aspartic acid · Asparagine
1 Introduction Electronic supplementary material The online version of this article (https://doi.org/10.1007/s13369-020-05024-1) contains supplementary material, which is available to authorized users.
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Sunday Osaizua Omeike [email protected]
1
Department of Biological Sciences, McPherson University, PMB 2094, Seriki Sotayo, Abeokuta, Ogun State, Nigeria
2
Department of Microbiology, Federal University of Agriculture, Abeokuta, Ogun State, Nigeria
3
Clinical Microbiology and Bioactive Screening Laboratory, CSIR-Institute of Microbial Technology, Chandigarh, India
4
Department of Chemistry, Federal University of Agriculture, Abeokuta, Ogun State, Nigeria
Antimicrobial resistance and limited antibiotics with improved and diverse activity spectrum in clinical use are
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