TY  - JOUR
AU  - Ölçücü, Gizem
AU  - Baumer, Benedikt
AU  - Küsters, Kira
AU  - Möllenhoff, Kathrin
AU  - Oldiges, Marco
AU  - Pietruszka, Jörg
AU  - Jaeger, Karl-Erich
AU  - Krauss, Ulrich
TI  - Catalytically Active Inclusion Bodies─Benchmarking and Application in Flow Chemistry
JO  - ACS synthetic biology
VL  - 11
IS  - 5
SN  - 2161-5063
CY  - Washington, DC
PB  - ACS
M1  - FZJ-2022-02276
SP  - 1881 - 1896
PY  - 2022
AB  - In industries, enzymes are often immobilized to obtain stable preparations that can be utilized in batch and flow processes. In contrast to traditional immobilization methods that rely on carrier binding, various immobilization strategies have been recently presented that enable the simultaneous production and in vivo immobilization of enzymes. Catalytically active inclusion bodies (CatIBs) are a promising example for such in vivo enzyme immobilizates. CatIB formation is commonly induced by fusion of aggregation-inducing tags, and numerous tags, ranging from small synthetic peptides to protein domains or whole proteins, have been successfully used. However, since these systems have been characterized by different groups employing different methods, a direct comparison remains difficult, which prompted us to benchmark different CatIB-formation-inducing tags and fusion strategies. Our study highlights that important CatIB properties like yield, activity, and stability are strongly influenced by tag selection and fusion strategy. Optimization enabled us to obtain alcohol dehydrogenase CatIBs with superior activity and stability, which were subsequently applied for the first time in a flow synthesis approach. Our study highlights the potential of CatIB-based immobilizates, while at the same time demonstrating the robust use of CatIBs in flow chemistry.
LB  - PUB:(DE-HGF)16
C6  - pmid:35500299
UR  - <Go to ISI:>//WOS:000805370400018
DO  - DOI:10.1021/acssynbio.2c00035
UR  - https://juser.fz-juelich.de/record/907898
ER  -