caa a22 4500 605503524 CHVBK 20210128100611.0 cr unu---uuuuu 210128e20150201xx s 000 0 eng 10.1007/s00253-014-6315-1 doi (NATIONALLICENCE)springer-10.1007/s00253-014-6315-1 Elleuche Skander Institute of Technical Microbiology, Hamburg University of Technology (TUHH), Kasernenstr. 12, 21073, Hamburg, Germany aut Bringing functions together with fusion enzymes—from nature's inventions to biotechnological applications [Elektronische Daten] [Skander Elleuche] It is a mammoth task to develop a modular protein toolbox enabling the production of posttranslational organized multifunctional enzymes that catalyze reactions in complex pathways. However, nature has always guided scientists to mimic evolutionary inventions in the laboratory and, nowadays, versatile methods have been established to experimentally connect enzymatic activities with multiple advantages. Among the oldest known natural examples is the linkage of two or more juxtaposed proteins catalyzing consecutive, non-consecutive, or opposing reactions by a native peptide bond. There are multiple reasons for the artificial construction of such fusion enzymes including improved catalytic activities, enabled substrate channelling by proximity of biocatalysts, higher stabilities, and cheaper production processes. To produce fused proteins, it is either possible to genetically fuse coding open reading frames or to connect proteins in a posttranslational process. Molecular biology techniques that have been established for the production of end-to-end or insertional fusions include overlap extension polymerase chain reaction, cloning, and recombination approaches. Depending on their flexibility and applicability, these methods offer various advantages to produce fusion genes in high throughput, different orientations, and including linker sequences to maximize the flexibility and performance of fusion partners. In this review, practical techniques to fuse genes are highlighted, enzymatic parameters to choose adequate enzymes for fusion approaches are summarized, and examples with biotechnological relevance are presented including a focus on plant biomass-degrading glycosyl hydrolases. Springer-Verlag Berlin Heidelberg, 2014 Bifunctionality nationallicence Biomass degradation nationallicence End-to-end fusion nationallicence Multifunctionality nationallicence Synergism nationallicence Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1545-1556 0175-7598 99:4<1545 2015 99 253 https://doi.org/10.1007/s00253-014-6315-1 text/html Onlinezugriff via DOI BK010053 XK010053 XK010000 Metadata rights reserved Springer special CC-BY-NC licence nationallicence 1 review-article jats NATIONALLICENCE NATIONALLICENCE NL-springer NATIONALLICENCE 856 40 https://doi.org/10.1007/s00253-014-6315-1 text/html Onlinezugriff via DOI NATIONALLICENCE 100 1- Elleuche Skander Institute of Technical Microbiology, Hamburg University of Technology (TUHH), Kasernenstr. 12, 21073, Hamburg, Germany aut NATIONALLICENCE 773 0- Applied Microbiology and Biotechnology Springer Berlin Heidelberg 99/4(2015-02-01), 1545-1556 0175-7598 99:4<1545 2015 99 253