SEQUENCING OF A LACCASE GENE FROM POLYETHYLENE- DEGRADING BACTERIUM

Background and Aim:Environmental pollution is one of the major and most important problems of the modern world. Polyethylene is widely used for various applications that lead to a large quantity of plastic waste causing serious environmental problems. Laccase is involved in biodegradation of polyethylene. This enzyme can help in the oxidation of the hydro-carbon backbone of polyethylene.Methods:In this study, sequencing of a laccase gene from a bacterium, isolated from PE waste depot was carried out. The isolate was cultured in a medium with PE as the sole carbon source. Before this step the molecular identification of the isolate had been done and the phylogenic tree determined that Acidovorax ebreus was the closest species. The FASTA sequences of laccase genes of 52 acidovorax species was received from NCBI database, then was aligned with MEGA7 software. After analyzing of conserved sequences 3 forward and 3 reverse primers were designed with these data. The primers were synthesized by sinaclon company. Bacterium extracted genome were amplified by the primers Pairwise. PCR optimization was carried out. To cover the entire gene sequence another primer design and PCR was done.Results:The agarose gel depicted the size of the PCR product was about 900bp.Conclusion:There are only a few reports on enzymes degrading synthetic plastics. Cloning of the genes encoding PE-degrading enzyme are under study.