Identification, heterologous expression and characterization of a new unspecific peroxygenase from Marasmius fiardii PR-910
Unspecific peroxygenases (UPOs) are glycosylated enzymes that provide an efficient method for oxyfunctionalizing a variety of substrates using only hydrogen peroxide ($ H_{2} $$ O_{2} $) as the oxygen donor. However, their poor heterologous expression has hindered their practical application. Here, a novel UPO from Marasmius fiardii PR910 (MfiUPO) was identified and heterologously expressed in Pichia pastoris. By employing a two-copy expression cassette, the protein titer reached 1.18 g $ L^{−1} $ in a 5 L bioreactor, marking the highest record. The glycoprotein rMfiUPO exhibited a smeared band in the 40 to 55 kDa range and demonstrated hydroxylation, epoxidation and alcohol oxidation. Moreover, the peroxidative activity was enhanced by 150% after exposure to 50% (v/v) acetone for 40 h. A semi-preparative production of 4-OH-β-ionone on a 100 mL scale resulted in a 54.2% isolated yield with 95% purity. With its high expression level, rMfiUPO is a promising candidate as an excellent parental template for enhancing desirable traits such as increased stability and selectivity through directed evolution, thereby meeting the necessary criteria for practical application. Graphical Abstract.
| Medienart: |
E-Artikel |
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| Erscheinungsjahr: |
2024 |
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| Erschienen: |
2024 |
| Enthalten in: |
Zur Gesamtaufnahme - volume:11 |
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| Enthalten in: |
Bioresources and Bioprocessing - 11(2024), 1 vom: 27. März |
| Sprache: |
Englisch |
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| Beteiligte Personen: |
Fu, Xin [VerfasserIn] |
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| Links: |
Volltext [kostenfrei] |
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| Themen: |
Characterization |
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| Anmerkungen: |
© The Author(s) 2024 |
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| doi: |
10.1186/s40643-024-00751-x |
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| funding: |
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| Förderinstitution / Projekttitel: |
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| PPN (Katalog-ID): |
SPR055329098 |
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| 245 | 1 | 0 | |a Identification, heterologous expression and characterization of a new unspecific peroxygenase from Marasmius fiardii PR-910 |
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| 520 | |a Unspecific peroxygenases (UPOs) are glycosylated enzymes that provide an efficient method for oxyfunctionalizing a variety of substrates using only hydrogen peroxide ($ H_{2} $$ O_{2} $) as the oxygen donor. However, their poor heterologous expression has hindered their practical application. Here, a novel UPO from Marasmius fiardii PR910 (MfiUPO) was identified and heterologously expressed in Pichia pastoris. By employing a two-copy expression cassette, the protein titer reached 1.18 g $ L^{−1} $ in a 5 L bioreactor, marking the highest record. The glycoprotein rMfiUPO exhibited a smeared band in the 40 to 55 kDa range and demonstrated hydroxylation, epoxidation and alcohol oxidation. Moreover, the peroxidative activity was enhanced by 150% after exposure to 50% (v/v) acetone for 40 h. A semi-preparative production of 4-OH-β-ionone on a 100 mL scale resulted in a 54.2% isolated yield with 95% purity. With its high expression level, rMfiUPO is a promising candidate as an excellent parental template for enhancing desirable traits such as increased stability and selectivity through directed evolution, thereby meeting the necessary criteria for practical application. Graphical Abstract | ||
| 650 | 4 | |a Unspecific peroxygenase |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Heterologous expression |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Characterization |7 (dpeaa)DE-He213 | |
| 650 | 4 | |a Semi-preparative |7 (dpeaa)DE-He213 | |
| 700 | 1 | |a Lin, Kexin |4 aut | |
| 700 | 1 | |a Zhang, Xiaodong |4 aut | |
| 700 | 1 | |a Guo, Zhiyong |4 aut | |
| 700 | 1 | |a Kang, Lixin |4 aut | |
| 700 | 1 | |a Li, Aitao |0 (orcid)0000-0001-6036-4058 |4 aut | |
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