Engineering of effective redox enzymes by exploring enzyme fusions

Biocatalysis has become a potent and widely embraced methodology, gaining prominence in both academic research and biotechnological applications. Biocatalysts, represented by enzymes, offer numerous benefits compared to chemocatalysts, such as high selectivity, high reaction rate, and biodegradability [1,2]. The utilization of multiple enzymes to catalyze cascade reactions has become a focus of research in the field of biocatalysis nowadays. Cascade reactions enable the performance of multiple chemical reactions in a single reaction vessel without isolating intermediates and eliminating the need for intermediate purification steps. Cascade reactions are highly attractive for enabling efficient cofactor regeneration, especially considering that cofactors are easily degraded under certain conditions and can be expensive to obtain. Efficient methods to regenerate and reuse cofactors is becoming a top priority in the development of an economically sustainable process. Several approaches have been introduced to facilitate cascade reactions for cofactor regeneration, ranging from genetic fusions of enzymes couples to immobilization or encapsulation techniques Except for development of enzyme complexes that efficiently regenerate a cofactor (chapter or utilize a byproduct for performing selective oxidations, also a new biocatalyst was studied in more detail concerning its ability to perform oxidation without the need for a cofactor. Below, the results of the performed research is summarized.