Matriks metalloproteinaz -1 ve -13'ün maküler kornea distrofisi tedavisi için rekombinant üretimi ve karakterizasyonu

Macular corneal dystrophy (MCD) is a rare hereditary disease characterized by the accumulation of insoluble aberrant substances in the corneal stroma. MCD may result in a decrease of visual acuity that can lead to blindness. There is still no cure for the disease, and corneal transplantation is possible but it can lead to likelihood of dystrophy recurrence and graft-related risks. Matrix metalloproteinases -1 and -13, which are necessary for corneal reshaping, have been reported to be decreased in MCD. The goal of this study is to provide a new treatment method for MCD by topical application of MMP-1 and MMP-13 to the cornea. MMP-1 and MMP-13 enzymes were being produced using recombinant DNA technology as a preliminary step in this research. Human MMP-1 and MMP-13 genes were cloned into pET-17b plasmid for E.coli expression. Auto-induction media wass used for bacterial expression (E. coli). Using a novel auto-induction methodology, our data indicated that active MMP-1 and MMP-13 enzymes may be generated in E. coli periplasm and released into production media. Promising protein yields were found, and production parameters were further optimized. Purified MMP-1 and MMP-13 enzymes were characterized by biophysical and biochemical tests. Protein thermal shift assay and enzyme kinetics assay were used to examine the profiles of recombinant enzymes. For MMP-1 and MMP-13 enzymes, this work presents a unique autoinduction recombinant production and characterization approach. These enzymes are expected to be used as a treatment method for MCD after completion of proof-of-concept studies.