Details
| Original language | English |
|---|---|
| Pages (from-to) | 10820-10830 |
| Number of pages | 11 |
| Journal | ACS Sustainable Chemistry and Engineering |
| Volume | 12 |
| Issue number | 29 |
| Publication status | Published - 22 Jul 2024 |
Abstract
Baeyer-Villiger monooxygenases (BVMOs) show great potential for the selective oxidation of a wide scope of ketones with exceptional regio-, chemo-, or stereoselectivity to produce diverse value-added (cyclic) esters. However, the technical application of BVMOs remains challenging due to their oxygen- and cofactor-dependency, instability, and susceptibility to substrate/product inhibition. The use of nonaqueous media is considered a straightforward solution but is limited due to enzyme instability and the unavailability of specific amounts of water required for enzyme activation and cofactor regeneration. Fusion approaches provide enzymes with the possibility of recycling cofactors even under low-water conditions by shortening the diffusion distance between active sites, whereas the stability and reusability of enzymes can be achieved by immobilization techniques. On all bases, a fusion enzyme of cyclohexanone monooxygenases (CHMOs) and alcohol dehydrogenases (ADHs) was immobilized on a newly reported resin, Chromalite MIDA (Methacrylate IminoDiAcetic) loaded with a variety of metal ions, via a one-pot metal affinity approach of simultaneous purification and immobilization. The immobilization process was rationally optimized in terms of activity and immobilization yields for both enzymes by the design of experiments (DoE). The catalytic performance of the immobilized fused enzyme was examined for the linear cascade to synthesize ϵ-caprolactone as a polymer precursor in cyclopentyl methyl ether (CPME). The immobilized fused enzyme showed an improved productivity of ϵ-caprolactone in 99.5 vol.% CPME than the buffer system (50 vs 30 mM), as well as a higher productivity and operational stability in comparison with the free enzyme (49 vs 12 mM) with prospective reusability of seven times. The upscale of the cascade in a rotating bed reactor at 125 mL in 99 vol.% CPME was established by obtaining 37 mM ϵ-caprolactone, demonstrating the feasibility of using ADH-CHMO fusion on a technical scale with further improvements.
Keywords
- Baeyer−Villiger monooxygenases, biocatalytic cascades, design of experiments, metal affinity immobilization, rotating bed reactor
ASJC Scopus subject areas
- Chemistry(all)
- Environmental Science(all)
- Environmental Chemistry
- Chemical Engineering(all)
- Energy(all)
- Renewable Energy, Sustainability and the Environment
Sustainable Development Goals
Cite this
- Standard
- Harvard
- Apa
- Vancouver
- BibTeX
- RIS
In: ACS Sustainable Chemistry and Engineering, Vol. 12, No. 29, 22.07.2024, p. 10820-10830.
Research output: Contribution to journal › Article › Research › peer review
}
TY - JOUR
T1 - Metal Affinity Fusion Enzyme Immobilization
T2 - Batch Process Showcase for Cascading Alcohol Oxidation and Bayer-Villiger Oxidation in Microaqueous Media
AU - Vernet, Guillem
AU - Ma, Yu
AU - Serban, Simona
AU - Basso, Alessandra
AU - Zhang, Ningning
AU - Kara, Selin
N1 - Publisher Copyright: © 2024 American Chemical Society.
PY - 2024/7/22
Y1 - 2024/7/22
N2 - Baeyer-Villiger monooxygenases (BVMOs) show great potential for the selective oxidation of a wide scope of ketones with exceptional regio-, chemo-, or stereoselectivity to produce diverse value-added (cyclic) esters. However, the technical application of BVMOs remains challenging due to their oxygen- and cofactor-dependency, instability, and susceptibility to substrate/product inhibition. The use of nonaqueous media is considered a straightforward solution but is limited due to enzyme instability and the unavailability of specific amounts of water required for enzyme activation and cofactor regeneration. Fusion approaches provide enzymes with the possibility of recycling cofactors even under low-water conditions by shortening the diffusion distance between active sites, whereas the stability and reusability of enzymes can be achieved by immobilization techniques. On all bases, a fusion enzyme of cyclohexanone monooxygenases (CHMOs) and alcohol dehydrogenases (ADHs) was immobilized on a newly reported resin, Chromalite MIDA (Methacrylate IminoDiAcetic) loaded with a variety of metal ions, via a one-pot metal affinity approach of simultaneous purification and immobilization. The immobilization process was rationally optimized in terms of activity and immobilization yields for both enzymes by the design of experiments (DoE). The catalytic performance of the immobilized fused enzyme was examined for the linear cascade to synthesize ϵ-caprolactone as a polymer precursor in cyclopentyl methyl ether (CPME). The immobilized fused enzyme showed an improved productivity of ϵ-caprolactone in 99.5 vol.% CPME than the buffer system (50 vs 30 mM), as well as a higher productivity and operational stability in comparison with the free enzyme (49 vs 12 mM) with prospective reusability of seven times. The upscale of the cascade in a rotating bed reactor at 125 mL in 99 vol.% CPME was established by obtaining 37 mM ϵ-caprolactone, demonstrating the feasibility of using ADH-CHMO fusion on a technical scale with further improvements.
AB - Baeyer-Villiger monooxygenases (BVMOs) show great potential for the selective oxidation of a wide scope of ketones with exceptional regio-, chemo-, or stereoselectivity to produce diverse value-added (cyclic) esters. However, the technical application of BVMOs remains challenging due to their oxygen- and cofactor-dependency, instability, and susceptibility to substrate/product inhibition. The use of nonaqueous media is considered a straightforward solution but is limited due to enzyme instability and the unavailability of specific amounts of water required for enzyme activation and cofactor regeneration. Fusion approaches provide enzymes with the possibility of recycling cofactors even under low-water conditions by shortening the diffusion distance between active sites, whereas the stability and reusability of enzymes can be achieved by immobilization techniques. On all bases, a fusion enzyme of cyclohexanone monooxygenases (CHMOs) and alcohol dehydrogenases (ADHs) was immobilized on a newly reported resin, Chromalite MIDA (Methacrylate IminoDiAcetic) loaded with a variety of metal ions, via a one-pot metal affinity approach of simultaneous purification and immobilization. The immobilization process was rationally optimized in terms of activity and immobilization yields for both enzymes by the design of experiments (DoE). The catalytic performance of the immobilized fused enzyme was examined for the linear cascade to synthesize ϵ-caprolactone as a polymer precursor in cyclopentyl methyl ether (CPME). The immobilized fused enzyme showed an improved productivity of ϵ-caprolactone in 99.5 vol.% CPME than the buffer system (50 vs 30 mM), as well as a higher productivity and operational stability in comparison with the free enzyme (49 vs 12 mM) with prospective reusability of seven times. The upscale of the cascade in a rotating bed reactor at 125 mL in 99 vol.% CPME was established by obtaining 37 mM ϵ-caprolactone, demonstrating the feasibility of using ADH-CHMO fusion on a technical scale with further improvements.
KW - Baeyer−Villiger monooxygenases
KW - biocatalytic cascades
KW - design of experiments
KW - metal affinity immobilization
KW - rotating bed reactor
UR - http://www.scopus.com/inward/record.url?scp=85198049943&partnerID=8YFLogxK
U2 - 10.1021/acssuschemeng.4c02236
DO - 10.1021/acssuschemeng.4c02236
M3 - Article
AN - SCOPUS:85198049943
VL - 12
SP - 10820
EP - 10830
JO - ACS Sustainable Chemistry and Engineering
JF - ACS Sustainable Chemistry and Engineering
SN - 2168-0485
IS - 29
ER -