Details
| Original language | English |
|---|---|
| Article number | e202200794 |
| Journal | CHEMBIOCHEM |
| Volume | 24 |
| Issue number | 8 |
| Early online date | 7 Feb 2023 |
| Publication status | Published - 17 Apr 2023 |
Abstract
Baeyer-Villiger monooxygenases (BVMOs) are attractive for selectively oxidizing various ketones using oxygen into valuable esters and lactones. However, the application of BVMOs is restrained by cofactor dependency and enzyme instability combined with water-related downsides such as low substrate loading, low oxygen capacity, and water-induced side reactions. Herein, we described a redox-neutral linear cascade with in-situ cofactor regeneration catalyzed by fused alcohol dehydrogenase and cyclohexanone monooxygenase in aqueous and microaqueous organic media. The cascade conditions have been optimized regarding substrate concentrations as well as the amounts of enzymes and cofactors with the Design of Experiments (DoE). The carrier-free immobilization technique, crosslinked enzyme aggregates (CLEAs), was applied to fusion enzymes. The resultant fusion CLEAs were proven to function in microaqueous organic systems, in which the enzyme ratios, water contents (0.5–5 vol. %), and stability have been systematically studied. The fusion CLEAs showed promising operational (up to 5 cycles) and storage stability.
Keywords
- biotransformations, crosslinked enzyme aggregates, cyclohexanone monooxygenase, enzyme immobilization, fusion enzymes
ASJC Scopus subject areas
- Biochemistry, Genetics and Molecular Biology(all)
- Biochemistry
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Medicine
- Biochemistry, Genetics and Molecular Biology(all)
- Molecular Biology
- Chemistry(all)
- Organic Chemistry
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In: CHEMBIOCHEM, Vol. 24, No. 8, e202200794, 17.04.2023.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Crosslinked Aggregates of Fusion Enzymes in Microaqueous Organic Media
AU - Vernet, Guillem
AU - Ma, Yu
AU - Zhang, Ningning
AU - Kara, Selin
N1 - Funding Information: The Deutsche Forschungsgemeinschaft (DFG; grant no. KA 4399/3‐2) and China Scholarship Council (CSC; grant no. 202108610066) are acknowledged for their kind financial support.
PY - 2023/4/17
Y1 - 2023/4/17
N2 - Baeyer-Villiger monooxygenases (BVMOs) are attractive for selectively oxidizing various ketones using oxygen into valuable esters and lactones. However, the application of BVMOs is restrained by cofactor dependency and enzyme instability combined with water-related downsides such as low substrate loading, low oxygen capacity, and water-induced side reactions. Herein, we described a redox-neutral linear cascade with in-situ cofactor regeneration catalyzed by fused alcohol dehydrogenase and cyclohexanone monooxygenase in aqueous and microaqueous organic media. The cascade conditions have been optimized regarding substrate concentrations as well as the amounts of enzymes and cofactors with the Design of Experiments (DoE). The carrier-free immobilization technique, crosslinked enzyme aggregates (CLEAs), was applied to fusion enzymes. The resultant fusion CLEAs were proven to function in microaqueous organic systems, in which the enzyme ratios, water contents (0.5–5 vol. %), and stability have been systematically studied. The fusion CLEAs showed promising operational (up to 5 cycles) and storage stability.
AB - Baeyer-Villiger monooxygenases (BVMOs) are attractive for selectively oxidizing various ketones using oxygen into valuable esters and lactones. However, the application of BVMOs is restrained by cofactor dependency and enzyme instability combined with water-related downsides such as low substrate loading, low oxygen capacity, and water-induced side reactions. Herein, we described a redox-neutral linear cascade with in-situ cofactor regeneration catalyzed by fused alcohol dehydrogenase and cyclohexanone monooxygenase in aqueous and microaqueous organic media. The cascade conditions have been optimized regarding substrate concentrations as well as the amounts of enzymes and cofactors with the Design of Experiments (DoE). The carrier-free immobilization technique, crosslinked enzyme aggregates (CLEAs), was applied to fusion enzymes. The resultant fusion CLEAs were proven to function in microaqueous organic systems, in which the enzyme ratios, water contents (0.5–5 vol. %), and stability have been systematically studied. The fusion CLEAs showed promising operational (up to 5 cycles) and storage stability.
KW - biotransformations
KW - crosslinked enzyme aggregates
KW - cyclohexanone monooxygenase
KW - enzyme immobilization
KW - fusion enzymes
UR - http://www.scopus.com/inward/record.url?scp=85150595676&partnerID=8YFLogxK
U2 - 10.1002/cbic.202200794
DO - 10.1002/cbic.202200794
M3 - Article
VL - 24
JO - CHEMBIOCHEM
JF - CHEMBIOCHEM
SN - 1439-4227
IS - 8
M1 - e202200794
ER -