DISCOVERY OF POLYGALACTURONASE PRODUCING BACILLUS TEQUILENSIS STRAIN ARMATI USING 16S rRNA GENE SEQUENCING
Objective: To identify a novel strain of polygalacturonase producing Bacillus tequilensis from poultry farm.
Methods: Poultry feces sample was serially diluted, and the pure isolate was subjected to morphological tests, biochemical tests, genomic DNA
isolation, polymerase chain reaction amplification, amplicon purification and 16S rRNA gene sequencing. The strain was screened for polygalacturonase
production using plate assay method. The nucleotide sequences obtained from the isolate were subjected to BLAST tool for the pairwise alignment.
RNA secondary structure was predicted through RNAStructure Web Server. Multiple antibiotic resistance (MAR) index was determined against six
antibiotics through disc diffusion method. Phylogenetic trees were inferred using the neighbor-joining algorithm in Molecular Evolution Genetic
Analysis software version 4.0.
Results: B. tequilensis strain ARMATI showed the production of polygalacturonase. MAR index of this novel strain was found to be zero. RNA
secondary structure with a minimum free energy of âˆ’281.40 kcal/mol was obtained. Phylogenetic tree of Bacillus 16S rRNA genes separate each
Bacillus according to their taxonomic positions and were supported statistically.
Conclusion: The present investigation clearly indicates the isolation, molecular characterization and sequence analysis of B. tequilensis strain ARMATI
from poultry farm, and industrial application of this novel strain for the production of the enzyme.
Keywords: Bacillus tequilensis, Multiple antibiotic resistance index, Polygalacturonase, Phylogenetic tree, RNA secondary structure, RNAStructure
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