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Bringing functions together with fusion enzymes—from nature’s inventions to biotechnological applications
Publikationstyp
Journal Article
Date Issued
2014-12-24
Sprache
English
Author(s)
Institut
Volume
99
Issue
4
Start Page
1545
End Page
1556
Citation
Applied Microbiology and Biotechnology 99 (4): 1545-1556 (2015-02-01)
Publisher DOI
Scopus ID
PubMed ID
25535094
Publisher
Springer
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.
Subjects
Bifunctionality
Biomass degradation
End-to-end fusion
Multifunctionality
Synergism
DDC Class
570: Biowissenschaften, Biologie
600: Technik