Formal Expression of Lytic Transglycosylases SLBT and MLTG of Gram-negative Bacteria Pseudomonas aeruginosa

Pseudomonas aeruginosa (p. aeruginosa) is a gram-negative bacterium notorious for causing infections of immunocompetent and immunocompromised cells and for its antibiotic resistance. The bacteria’s cell wall can be severed by lytic transglycosylases (LTs), which are enzymes that cleave the peptidoglycan structures of bacterial cell walls. In p. aeruginosa, the bacteria consist of eleven LTs in its domain which fall under different families. This study focuses on two LTs out of the eleven in the domain for p. aeruginosa (SLTB1 and MLTG), as the expression of the enzymes must proceed successfully in order to conduct further investigation on the effectiveness of breaking down the peptidoglycan in the bacteria. The efficiency of the enzyme can be observed by the success of its inhibition of IVYP1, a protein that inhibits the activity of lysozyme. To measure the inhibition, the LTs endure protein preparation which initiates with a transformation, followed by protein expression and pellet homogenization, and concludes with protein lysis and purification. The resultant was tested to ensure the protein can be applicable for use in p. aeruginosa. Throughout gel-electrophoresis and several muramidase assays, the LTs show observation of success in inhibiting the IVY protein, thus providing the liberty for further examination of the enzymes