Aim: The phenotypic and phylogenetic characterization of an alkalophilic bacterium producing two industrially important enzymes – alkaline protease and alkaline amylase, was performed. A Gram-positive, rod-shaped, spore-forming... more
Aim: The phenotypic and phylogenetic characterization of an alkalophilic bacterium producing two industrially important enzymes – alkaline protease and alkaline amylase, was performed. A Gram-positive, rod-shaped, spore-forming alkalophilic bacterium (2b) isolated from alkaline dairy soil of Indore region of Madhya Pradesh, was selected in this work.
Methods
The alkalophile was subjected to a taxonomic investigation, including standard bacteriological characterization and 16S rRNA gene sequencing. Phylogenetic trees were constructed by different treeing algorithms: neighbour joining, maximum parsimony tree, maximum-likelihood and UPGMA method using MEGA5 software. The secondary structures of 16S rRNA gene sequence were predicted using GeneBee package and UNAFOLD, a Linux based software.
Results: Its overall biochemical, physiological and phylogenetic characteristics indicated that strain 2b is an alkaliphilic Bacillus belonging to the species agaradhaerens. The free energy of the secondary structure of rRNA was −171.7 and −265.13 kcal/mol respectively. The structure obtained with a Gibb's free energy, ΔG −265.13 kcal/mol by UNAFOLD seems to be more stable in the present study.
Conclusion: The taxonomic investigation identifies 2b as Bacillus agaradhaerens.
Methods
The alkalophile was subjected to a taxonomic investigation, including standard bacteriological characterization and 16S rRNA gene sequencing. Phylogenetic trees were constructed by different treeing algorithms: neighbour joining, maximum parsimony tree, maximum-likelihood and UPGMA method using MEGA5 software. The secondary structures of 16S rRNA gene sequence were predicted using GeneBee package and UNAFOLD, a Linux based software.
Results: Its overall biochemical, physiological and phylogenetic characteristics indicated that strain 2b is an alkaliphilic Bacillus belonging to the species agaradhaerens. The free energy of the secondary structure of rRNA was −171.7 and −265.13 kcal/mol respectively. The structure obtained with a Gibb's free energy, ΔG −265.13 kcal/mol by UNAFOLD seems to be more stable in the present study.
Conclusion: The taxonomic investigation identifies 2b as Bacillus agaradhaerens.
Probiotic microorganisms are those which exert a positive effect on the growth of the host, when administered as a dietary mixture in an adequate amount. They form the best alternative to the use of antibiotics for controlling enteric... more
Probiotic microorganisms are those which exert a positive effect on the growth of the host, when administered as a dietary mixture in an adequate amount. They form the best alternative to the use of antibiotics for controlling enteric diseases in poultry farm animals, especially in the light of the gruesome problems of development of antibiotic resistance in enteric pathogens and the contamination of poultry products with antibiotics. 16S rDNA sequencing which has gained wide popularity amongst microbiologists for the molecular characterization and identification of newly discovered isolates provides accurate identification of isolates down to the level of sub-species (strain). It's most important advantage over the traditional biochemical characterization methods are that it can provide an accurate identification of strains with atypical phenotypic characters as well. The following work is an application of 16S rRNA gene sequencing approach to identify a novel, alkaline protease producing bacteria, from poultry farm waste. The sample was collected from a local poultry farm in the Guntur district, Andhra Pradesh, India. Subsequently the sample was serially diluted and the aliquots were incubated for a suitable time period following which the suspected colony was subjected to 16S rDNA sequencing. The results showed the isolate to be a novel, high alkaline protease producing bacteria, which was named Bacillus firmus isolate EMBS023, after characterization the sequence of isolate was deposited in GenBank with accession number JN990980.
Research Interests:
Probiotic microorganisms are those which exert a positive effect on the growth of the host, when administered as a dietary mixture in an adequate amount. They form the best alternative to the use of antibiotics for controlling enteric... more
Probiotic microorganisms are those which exert a positive effect on the growth of the host, when
administered as a dietary mixture in an adequate amount. They form the best alternative to the use of antibiotics
for controlling enteric diseases in poultry farm animals, especially in the light of the gruesome problems of development
of antibiotic resistance in enteric pathogens and the contamination of poultry products with antibiotics.
16S rDNA sequencing which has gained wide popularity amongst microbiologists for the molecular characterization
and identification of newly discovered isolates provides accurate identification of isolates down to the level
of sub-species (strain). It’s most important advantage over the traditional biochemical characterization methods
are that it can provide an accurate identification of strains with atypical phenotypic characters as well. The
following work is an application of 16S rRNA gene sequencing approach to identify a novel, alkaline protease
producing bacteria, from poultry farm waste. The sample was collected from a local poultry farm in the Guntur
district, Andhra Pradesh, India. Subsequently the sample was serially diluted and the aliquots were incubated for
a suitable time period following which the suspected colony was subjected to 16S rDNA sequencing. The results
showed the isolate to be a novel, high alkaline protease producing bacteria, which was named Bacillus firmus
isolate EMBS023, after characterization the sequence of isolate was deposited in GenBank with accession number
JN990980.
administered as a dietary mixture in an adequate amount. They form the best alternative to the use of antibiotics
for controlling enteric diseases in poultry farm animals, especially in the light of the gruesome problems of development
of antibiotic resistance in enteric pathogens and the contamination of poultry products with antibiotics.
16S rDNA sequencing which has gained wide popularity amongst microbiologists for the molecular characterization
and identification of newly discovered isolates provides accurate identification of isolates down to the level
of sub-species (strain). It’s most important advantage over the traditional biochemical characterization methods
are that it can provide an accurate identification of strains with atypical phenotypic characters as well. The
following work is an application of 16S rRNA gene sequencing approach to identify a novel, alkaline protease
producing bacteria, from poultry farm waste. The sample was collected from a local poultry farm in the Guntur
district, Andhra Pradesh, India. Subsequently the sample was serially diluted and the aliquots were incubated for
a suitable time period following which the suspected colony was subjected to 16S rDNA sequencing. The results
showed the isolate to be a novel, high alkaline protease producing bacteria, which was named Bacillus firmus
isolate EMBS023, after characterization the sequence of isolate was deposited in GenBank with accession number
JN990980.