Broad Spectrum Antimicrobial Activity of Forest-Derived Soil Actinomycete, Nocardia sp. PB-52.

Priyanka Sharma,Mohan C Kalita,Debajit Thakur

doi:10.3389/fmicb.2016.00347

Abstract

A mesophilic actinomycete strain designated as PB-52 was isolated from soil samples of Pobitora Wildlife Sanctuary of Assam, India. Based on phenotypic and molecular characteristics, the strain was identified as Nocardia sp. which shares 99.7% sequence similarity with Nocardia niigatensis IFM 0330 (NR_112195). The strain is a Gram-positive filamentous bacterium with rugose spore surface which exhibited a wide range of antimicrobial activity against Gram-positive bacteria including methicillin-resistant Staphylococcus aureus (MRSA), Gram-negative bacteria, and yeasts. Optimization for the growth and antimicrobial activity of the strain PB-52 was carried out in batch culture under shaking condition. The optimum growth and antimicrobial potential of the strain were recorded in GLM medium at 28°C, initial pH 7.4 of the medium and incubation period of 8 days. Based on polyketide synthases (PKS) and non-ribosomal peptide synthetases (NRPS) gene-targeted PCR amplification, the occurrence of both of these biosynthetic pathways was detected which might be involved in the production of antimicrobial compounds in PB-52. Extract of the fermented broth culture of PB-52 was prepared with organic solvent extraction method using ethyl acetate. The ethyl acetate extract of PB-52 (EA-PB-52) showed lowest minimum inhibitory concentration (MIC) against S. aureus MTCC 96 (0.975 μg/mL) whereas highest was recorded against Klebsiella pneumoniae ATCC 13883 (62.5 μg/mL). Scanning electron microscopy (SEM) revealed that treatment of the test microorganisms with EA-PB-52 destroyed the targeted cells with prominent loss of cell shape and integrity. In order to determine the constituents responsible for its antimicrobial activity, EA-PB-52 was subjected to chemical analysis using gas chromatography-mass spectrometry (GC-MS). GC-MS analysis showed the presence of twelve different chemical constituents in the extract, some of which are reported to possess diverse biological activity. These results confirmed that the presence of bioactive constituents in EA-PB-52 could be a promising source for the development of potent antimicrobial agents effective against wide range of microbial pathogens including MRSA.

Highlights

The discovery of antibiotics like penicillin and other antimicrobial agents to treat infectious diseases has revolutionized the field of medicine in the mid-twentieth century
We aimed to investigate the antimicrobial biosynthetic potential of PB-52 strain against a wide range of microbial pathogens
During the exploration of rare actinomycetes prevailing in forest-derived soil samples of Pobitora Wildlife Sanctuary of Assam, India, Nocardia sp

Summary

Introduction

The discovery of antibiotics like penicillin and other antimicrobial agents to treat infectious diseases has revolutionized the field of medicine in the mid-twentieth century. These discoveries have led to the development of improved antibiotics with a hope to serve humanity well (Walsh, 2003). The genus Nocardia, Saccharopolyspora, Micromonospora, Streptosporangium, Streptoverticillium, Actinoplanes, and Actinomadura are considered as the rare group of actinomycetes. It is because these microbes are difficult to isolate and maintain under conventional conditions (Berdy, 2005). Numerous interesting biologically active compounds have been reported from the genus Nocardia such as nargenicin (Celmer et al, 1980), transvalencin (Hoshino et al, 2004), nocardithiocin (Mukai et al, 2009) etc

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