Conversion of Unsaturated Short- to Medium-Chain Fatty Acids by Unspecific Peroxygenases (UPOs)
Abstract
:1. Introduction
2. Materials and Methods
2.1. Enzyme Preparations
2.2. Conversion of Medium Chained Fatty Acids (MCFAs)
2.3. Preparation of Epoxides via the Prilezhaev Reaction
2.4. Analyses
2.5. Chemicals
3. Results and Discussion
3.1. Terminally Unsaturated FAs
3.2. Unsaturated FAs with the Double Bond at the β,γ-Position
3.3. Unsaturated FAs with the Double Bond at α,β-Position
- UPOs mainly forming the epoxide between C2 and C3 (α,β-position) with yields of about 80% and producing minor hydroxylation products at different positions (C2, C3, C7 and C8). This group included wild-type and recombinant UPOs deriving from dark-spored agaric Basidiomycota (genera Candolleomyces, Psathyrellaceae), i.e., CeuMQ V, CabMQ II and rCab II.
- One UPO forming the epoxide with yields around 25% but favoring hydroxylation at the C4 position (about 45%); furthermore, the enzyme formed a so far unidentified oxygenation product (C?) and a keto-product (4-oxo-trans-2-nonenoic acid, which was probably deriving from a second hydroxylation at the C4 position (second hydroxylation at the C4 position gives a gem-diol that is in equilibrium with the chemically favored oxo-FA, 14%). CglUPO was the only enzyme in this group.
- UPOs favoring subterminal hydroxylation at C8 (26–36%), C7 (25–29%), and C6 (14%) and additionally, forming the epoxide (5–18%) and other hydroxylation products at C2/3 (about 20% and 3%, respectively). This group included UPOs from dark-spored agaric Basidiomycota, AaeUPO and CraUPO.
- Enzymes yielding the epoxide (about 55%) to some extent as well as hydroxylating subterminally and at the C2 position (about 20%). Two wild-type UPOs, CeuMS III and CabMS II (both from the genus Candolleomyces).
- A UPO hydroxylating at C2 (34%), C3 (18%) and favorably at the C8 (ω-1; 37%) position. One recombinant enzyme, rCab I (from the genus Candolleomyces).
- Two enzymes favorably hydroxylating close to the carboxylic group at C2 (28–35%), C3 (25–28%), and C4 (4–5%) positions, and yielding the epoxide to minor extent; in addition, they catalyze a second hydroxylation yielding dihydroxylated products. This group includes UPOs from bright-spored agaric Basidiomycota (genus Marasmius), MroUPO and MweUPO.
4. Concluding Remarks
Supplementary Materials
Author Contributions
Funding
Data Availability Statement
Conflicts of Interest
References
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Enzyme (UPO Abbreviation) | Fungal Species | Specific Activity (UVA/mg) * | Isoelectric Point (pI) | Reference |
---|---|---|---|---|
CglUPO | Chaetomium globosum | 3.9 | 4.3–4.7 (5.59 *) | [21] |
CraUPO | Coprinellus radians | 72.0 | 3.8–4.2 | [22] |
AaeUPO | Cyclocybe (Agrocybe) aegerita | 82.0 | 4.9–6.1 | [5] |
CeuMQ V | Candolleomyces eurysporus | 12.5 | 4.25 | [23] |
CeuMS III | C. eurysporus | 11.0 | 7.6 | herein |
CabMS II | C. aberdarensis | 72.0 | 5.1 | herein |
CabMQ II | C. aberdarensis | 172.0 | 4.1 | [23] |
rCab II | C. aberdarensis (heterologously expressed in Pichia pastoris) | 21.0 | 5.1 | [24] |
rCab I | C. aberdarensis (heterologously expressed in in Pichia pastoris) | 21.0 | n.d. | [24] |
MweUPO | Marasmius wettsteinii | 50.0 | 5.1 ** | [2,25] |
MroUPO | M. rotula | 25.4 | 5.0–5.3 | [26] |
Number of Carbon Atoms | Substrates (Unsaturated Short- and Medium-Chain Fatty Acids) | Abbreviation and Double Bond Position (according to IUPAC Nomenclature) | Acetonitrile Concentration |
---|---|---|---|
9 | trans-2-nonenoic acid | t-C9:1(2) | 55% |
3-nonenoic acid | m-C9:1(3) | ||
8-nonenoic acid | C9:1(8) | ||
8 | trans-2-octenoic acid | t-C8:1(2) | 50% |
3-octenoic acid | m-C8:1(3) | ||
7-octenoic acid | C8:1(7) | ||
7 | 2-heptenoic acid | m-C7:1(2) | 40% |
3-heptenoic acid | m-C7:1(3) | ||
6-heptenoic acid | C7:1(6) | ||
6 | cis-2-hexenoic acid | c-C6:1(2) | 30% |
trans-2-hexenoic acid | t-C6:1(2) | ||
trans-3-hexenoic acid | t-C6:1(3) | ||
5-hexenoic acid | C6:1(5) | ||
5 | trans-2-pentenoic acid | t-C5:1(2) | 15% |
3-pentenoic acid | m-C5:1(3) | ||
4-pentenoic acid | C5:1(4) | ||
4 | 3-butenoic acid | C4:1(3) | 7% |
(vinylacetic acid) | |||
3 | 2-propenoic acid | C3:1(2) | 5% |
(acrylic acid) |
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Karich, A.; Salzsieder, F.; Kluge, M.; Alcalde, M.; Ullrich, R.; Hofrichter, M. Conversion of Unsaturated Short- to Medium-Chain Fatty Acids by Unspecific Peroxygenases (UPOs). Appl. Microbiol. 2023, 3, 826-840. https://doi.org/10.3390/applmicrobiol3030057
Karich A, Salzsieder F, Kluge M, Alcalde M, Ullrich R, Hofrichter M. Conversion of Unsaturated Short- to Medium-Chain Fatty Acids by Unspecific Peroxygenases (UPOs). Applied Microbiology. 2023; 3(3):826-840. https://doi.org/10.3390/applmicrobiol3030057
Chicago/Turabian StyleKarich, Alexander, Fabian Salzsieder, Martin Kluge, Miguel Alcalde, René Ullrich, and Martin Hofrichter. 2023. "Conversion of Unsaturated Short- to Medium-Chain Fatty Acids by Unspecific Peroxygenases (UPOs)" Applied Microbiology 3, no. 3: 826-840. https://doi.org/10.3390/applmicrobiol3030057