Bioactivity of Microbacterium barkeri (LMA4) In Vitro and Candidate Gene Annotation In Silico

Appl Biochem Biotechnol. 2023 May 11. doi: 10.1007/s12010-023-04519-5. Online ahead of print.

Abstract

Actinomycetes are considered a never-ending treasure trove of biometabolites, which always fascinated researchers. However, to combat with newly emerging bacterial strains, the search for novel or analogs of existing therapeutic agents is recommended. In this context, this research work was carried out to search for a biopotent Actinomycetal strain grown in untapped soil, near the Hirakud dam. This Gram-positive bacteria was subjected to screening for its bioactivity against the medically important bacteria, isolated from local hospital sample using "co-culture" method, following both qualitative and quantitative assays. Further, the 16 s rRNA sequencing, BLASTn analysis, and GC% calculation were carried out. Based upon its bioactivity, a prediction-based genomics work was pursued, considering the gene sequence deposited in public domain. The reverse translation, elution of protein structural file, and the putative protein were predicted. The strain was identified as Microbacterium barkeri, with 54.1% GC content. From Gene Ontology term annotation, it was predicted that the α/β hydrolase fold of hydrolase protein could have been responsible for antibiotic/biometabolite synthesis, in silico. The in vitro-based sequence (from Whole Genome Sequence data) had inferred that there was elution of alpha/beta hydrolase fold, substantiated with conserved domain analysis, ORF finding more over Gene Ontology (GO) terminology annotations. The GO annotations had suggested that the protein had been produced in response to a bacteria, under the influence of external stimuli more so in stress.

Keywords: Biopotent; High GC%; Novel actinomycetes strain; Persister generator; Putative hydrolase enzyme ontology annotation.