Ariza, AntonioAlsaytari, Abdulmalik Ibrahim2023-11-142023-11-142023-11-14https://hdl.handle.net/20.500.14154/69682Plastics have become an essential part of modern life due to their versatility, low cost and multiple uses. However, most types of synthetic plastic have a downside: they tend to last for hundreds of years before they break down and degrade, leaving behind harmful substances known as microplastics. The methods currently used to dispose of and recycle plastics have not proven to be the most ideal solutions. The lack of a highly efficient tool to solve the problem of plastic pollution and its accumulation in the ecosystem has been worrying scientists for years. The search for biodegrading organisms has emerged as a promising alternative to conventional methods of dealing with plastic waste. The discovery of the PETase enzyme system, consisting of two novel enzymes: IsPETase and IsMHETase, in the bacterium Ideonella sakaiensis, which can use PET (polyethylene terephthalate), one of the most widely produced and distributed types of plastic, as an energy source, has opened up the possibility of developing biotechnological tools that could use microorganisms to degrade and detoxify the plastic waste that has been choking the environment for years. The search for improving the functional and catalytic activities of these newly discovered classes of enzymes, as well as the search for other organisms that may have developed a mechanism for degrading plastic, is now more necessary than ever. The novel enzyme KsPETase, which holds structural similarities to IsPETase, was investigated in this study as a potential enzyme to be studied for its structural and functional properties that are expected to be resemble those of IsPETase. It was also intended for the investigation of whether it performs the catalytic functions more efficiently.32enPlasticBiotechnologyMolecular BiologyPlasticsTransformationBacterial TransformationPlasmid extractionCompetent cellsProtein expressionSDS-PAGEProtein PurificationPlastic wastePlastic degradationPlastic clean-upThesis