Benítez-Mateos, Ana I; Schneider, Andreas; Hegarty, Eimear; Hauer, Bernhard; Paradisi, Francesca (2022). Spheroplasts preparation boosts the catalytic potential of a squalene-hopene cyclase. Nature Communications, 13(1), p. 6269. Springer Nature 10.1038/s41467-022-34030-0
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Squalene-hopene cyclases (SHCs) are a highly valuable and attractive class of membrane-bound enzymes as sustainable biotechnological tools to produce aromas and bioactive compounds at industrial scale. However, their application as whole-cell biocatalysts suffer from the outer cell membrane acting as a diffusion barrier for the highly hydrophobic substrate/product, while the use of purified enzymes leads to dramatic loss of stability. Here we present an unexplored strategy for biocatalysis: the application of SHC spheroplasts. By removing the outer cell membrane, we produced stable and substrate-accessible biocatalysts. SHC spheroplasts exhibited up to 100-fold higher activity than their whole-cell counterparts for the biotransformations of squalene, geranyl acetone, farnesol, and farnesyl acetone. Their catalytic ability was also higher than the purified enzyme for all high molecular weight terpenes. In addition, we introduce a new concept for the carrier-free immobilization of spheroplasts via crosslinking, CLS (crosslinked spheroplasts). The CLS maintained the same catalytic activity of the spheroplasts, offering additional advantages such as recycling and reuse. These timely solutions contribute not only to harness the catalytic potential of the SHCs, but also to make biocatalytic processes even greener and more cost-efficient.
Item Type: |
Journal Article (Original Article) |
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Division/Institute: |
08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP) |
UniBE Contributor: |
Benitez Mateos, Ana Isabel, Hegarty, Eimear, Paradisi, Francesca |
Subjects: |
500 Science > 540 Chemistry |
ISSN: |
2041-1723 |
Publisher: |
Springer Nature |
Funders: |
[4] Swiss National Science Foundation |
Projects: |
[UNSPECIFIED] Artificial Enzymatic Factories (200021_192274) |
Language: |
English |
Submitter: |
Francesca Paradisi |
Date Deposited: |
09 Nov 2022 14:44 |
Last Modified: |
05 Dec 2022 16:26 |
Publisher DOI: |
10.1038/s41467-022-34030-0 |
PubMed ID: |
36271006 |
BORIS DOI: |
10.48350/173992 |
URI: |
https://boris.unibe.ch/id/eprint/173992 |
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