Screening and identification of potential antibiotic-producing Actinobacteria from cemetery soil

A. F. Farihati; E. M. Tyastuti; L. Agustina; N. A. Setiyadi; Y. Sidiq; T. Rahayu

doi:10.15421/012523

A. F. Farihati Universitas Muhammadiyah Surakarta
E. M. Tyastuti Universitas Muhammadiyah Surakarta
L. Agustina Universitas Muhammadiyah Surakarta
N. A. Setiyadi Universitas Muhammadiyah Surakarta
Y. Sidiq Universitas Muhammadiyah Surakarta
T. Rahayu Universitas Muhammadiyah Surakarta

Keywords: Actinobacteria, antibiotic production, cemetery soil, 16S rRNA gene, Streptomyces.

Abstract

Actinobacteria which dominated in a cemetery in Surakarta, Central Java, Indonesia has been discovered from a previous study. A total of 29 isolates of Actinobacteria were sucessfully collected. This study is challenging as the information on antibiotics producing Actinobacteria from cemetery soil is restricted. The aim of this research wa s to identify and characterize Actinobacteria isolated from cemetery soil , also to screen it for its antibiotic producing potential. A total of 29 Actinobacteria isolates collected from cemetery soil were screened for antibiotics using the agar plug diffusion method against the test bacteria Escherichia coli , Staphylococcus aureus , and Bacillus subtilis . Isolates with strong potential were characterized by colony morphology, spore type, and molecular identification based on the 16S rRNA gene. There are two isolates that can inhibit both S. aureus and B. subtilis , namely T5 (19. 3 and 17 .0 mm) and S6 (18. 3 and 10 .0 mm). Four isolates show ed moderate inhibition against E. coli , namely T15, T31, T34, and T42. Two selected isolates, T5 and S6, we re respectively white and yellow (aerial mycelium), while their vegetative mycelium wa s yellowish-brown with closed spiral spore type. Based on the analysis of partial sequencing of the 16S rRNA gene isolate , T5 was identified as Streptomyces sp. VEL17 (99. 9 %) and S6 as Streptomyces sp. strain ADE 004 (83.5%). Genetic di s tance of T5 to the 8 nearest strains based on 16S rRNA show s a close relationship range from 0.001 to 0.005, while by contrast genetic distance of S6 to the 8 nearest strains based on 16S rRNA show s a distant relationship range from 1.057 to 1.074. From this research, it can be concluded that the cemetery soil harbors Actinobacteria that show strong antibiotic potential against Gram-positive bacteria and are very close to Streptomyces sp. yet less potential against Gram-negative bacteria. Two selected isolates are prospective candidates for the biotechnology of potential antibiotic compounds.

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