biostudies-literatureNilpa PUniversiti Teknologi MalaysiaTaif Universitye0265969https://www.ebi.ac.uk/biostudies/studies/S-EPMC8975109biostudies-literatureUnknown17(4)The tissue of insects, pests, and fungi has a chitin layer followed by protein in the cell membrane. The complete biodegradation of chitin and protein-present in the waste requires the action of two enzymes, namely chitinase, and protease. Combining chitinase and protease in a single protein/enzyme will serve as a bifunctional enzyme that can efficiently degrade the chitin and protein-rich biomass. The present study was aimed to fuse these two enzymes to produce a single protein and study the kinetics of the recombinant fusion protein. A chitinase and alkaline protease genes were isolated, cloned, and expressed successfully as a fusion product in heterologous host Escherichia coli. The two native genes were successfully fused in E.coli by using flexible glycine-serine (G4S)2 linker (GGGGS,PloS oneFormation of recombinant bifunctional fusion protein: A newer approach to combine the activities of two enzymes in a single protein.PMC8975109RJ130000.7609.4C359 and RJ130000.7609.4C336TURSP-2020/208Sayyed RZAlhazmi AChintan KEl Enshasy HEl Adawi HNilpa PAlmalki AHHaque SfalseFormation of recombinant bifunctional fusion protein: A newer approach to combine the activities of two enzymes in a single protein.The tissue of insects, pests, and fungi has a chitin layer followed by protein in the cell membrane. The complete biodegradation of chitin and protein-present in the waste requires the action of two enzymes, namely chitinase, and protease. Combining chitinase and protease in a single protein/enzyme will serve as a bifunctional enzyme that can efficiently degrade the chitin and protein-rich biomass. The present study was aimed to fuse these two enzymes to produce a single protein and study the kinetics of the recombinant fusion protein. A chitinase and alkaline protease genes were isolated, cloned, and expressed successfully as a fusion product in heterologous host Escherichia coli. The two native genes were successfully fused in E.coli by using flexible glycine-serine (G4S)2 linker (GGGGS,2022-01-01T00:00:00Z20222026-05-31T01:04:19.605Z2025-04-04T11:31:50.777ZS-EPMC89751093536379610.1371/journal.pone.0265969