Kentucky State University
Prevailing Adenylsuccinate Lyase of Exiguobacterium Acetylicum Strain SN Using Peptide Synthetase Primers
Institution
Kentucky State University
Faculty Advisor/ Mentor
Williard Mazhawidza; Narayanan Rajendran
Abstract
For prokaryotic cells, the ability to biosynthesize essential elements such as peptides and nucleotides for DNA and RNA synthesis is pivotal. The enzyme peptide biosynthetase is critical to cell survival. In an effort to reveal such peptide synthetase biosynthetic mechanisms in a soil isolate Exiguobacterium acetylicum, the genomic DNA was probed in a PCR experiment using peptide synthetase primers. Amplified DNA was resolved by agarose gel electrophoresis and cloned at our molecular microbiology lab, and then sequenced at University of Louisville sequence facility. DNA sequence analysis was performed using MEGA 4.1 software. The Blast search analysis putatively identified our amplicon as Adenylsuccinate Lyase. Further DNA sequencing analysis and multiple sequence alignments revealed the conserved amino acids specific to that family of enzymes. Phylogenetic analysis of related bacteria based on matching sequences revealed close relationships of Exiguobacterium acetylicum strain SN related to some soil microbes. The revelation of similar microbes with regard to the biosynthetic complex, adenylsuccinate lyase suggests a potential common evolutionary trajectory amongst the matching bacteria. Further studies are underway.
Prevailing Adenylsuccinate Lyase of Exiguobacterium Acetylicum Strain SN Using Peptide Synthetase Primers
For prokaryotic cells, the ability to biosynthesize essential elements such as peptides and nucleotides for DNA and RNA synthesis is pivotal. The enzyme peptide biosynthetase is critical to cell survival. In an effort to reveal such peptide synthetase biosynthetic mechanisms in a soil isolate Exiguobacterium acetylicum, the genomic DNA was probed in a PCR experiment using peptide synthetase primers. Amplified DNA was resolved by agarose gel electrophoresis and cloned at our molecular microbiology lab, and then sequenced at University of Louisville sequence facility. DNA sequence analysis was performed using MEGA 4.1 software. The Blast search analysis putatively identified our amplicon as Adenylsuccinate Lyase. Further DNA sequencing analysis and multiple sequence alignments revealed the conserved amino acids specific to that family of enzymes. Phylogenetic analysis of related bacteria based on matching sequences revealed close relationships of Exiguobacterium acetylicum strain SN related to some soil microbes. The revelation of similar microbes with regard to the biosynthetic complex, adenylsuccinate lyase suggests a potential common evolutionary trajectory amongst the matching bacteria. Further studies are underway.