Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of Concept

Testosterone holds significant medical and economic importance, with the global market for testosterone replacement therapies valued at approximately USD 1.9 billion in 2023. This hormone is essential for the development and maintenance of male sexual characteristics as well as bone and muscle healt...

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Main Authors: Lidia Ortega-de los Ríos, Luis Getino, Beatriz Galán, José Luis García, José M. Luengo, Alejandro Chamizo-Ampudia, José M. Fernández-Cañón
Format: Article
Language:English
Published: MDPI AG 2024-11-01
Series:Biomolecules
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Online Access:https://www.mdpi.com/2218-273X/14/12/1502
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author Lidia Ortega-de los Ríos
Luis Getino
Beatriz Galán
José Luis García
José M. Luengo
Alejandro Chamizo-Ampudia
José M. Fernández-Cañón
author_facet Lidia Ortega-de los Ríos
Luis Getino
Beatriz Galán
José Luis García
José M. Luengo
Alejandro Chamizo-Ampudia
José M. Fernández-Cañón
author_sort Lidia Ortega-de los Ríos
collection DOAJ
description Testosterone holds significant medical and economic importance, with the global market for testosterone replacement therapies valued at approximately USD 1.9 billion in 2023. This hormone is essential for the development and maintenance of male sexual characteristics as well as bone and muscle health. It plays a key role in conditions such as hypogonadism, muscle disorders, and andropause. However, the industrial production of testosterone often involves complex chemical processes that result in low yields, high costs, and environmental damage. Microbial biotransformation of steroids presents an eco-friendly alternative to traditional chemical synthesis. A knockout strain of <i>Aspergillus nidulans</i> deficient in steroid 11α-hydroxylase activity was developed, rendering it incapable of hydroxylating androstenedione, progesterone, and testosterone. In these strains, two newly identified CYP450 enzymes, CYP68L1 from <i>A. nidulans</i> and CYP68L8 from <i>Aspergillus ochraceus,</i> were expressed to confirm their roles as steroid 11α-hydroxylases of androstenedione, progesterone, and testosterone. The availability of these 11α-hydroxylases represents significant progress toward achieving efficient single-step steroid fermentation. Furthermore, the <i>A. nidulans</i> knockout strain serves as an effective model for studying the conversion of androstenedione to testosterone upon the expression of the enzyme 17β-hydroxysteroid dehydrogenase, due to its inability to hydroxylate testosterone.
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spelling doaj-art-043c91a0ef0d492f899f415277c98ab02025-08-20T02:00:56ZengMDPI AGBiomolecules2218-273X2024-11-011412150210.3390/biom14121502Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of ConceptLidia Ortega-de los Ríos0Luis Getino1Beatriz Galán2José Luis García3José M. Luengo4Alejandro Chamizo-Ampudia5José M. Fernández-Cañón6Área de Bioquímica y Biología Molecular, Departamento de Biología Molecular, Universidad de León, 24007 León, SpainÁrea de Genética, Departamento de Biología Molecular, Universidad de León, 24007 León, SpainCentro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, SpainCentro de Investigaciones Biológicas, Consejo Superior de Investigaciones Científicas, 28040 Madrid, SpainÁrea de Bioquímica y Biología Molecular, Departamento de Biología Molecular, Universidad de León, 24007 León, SpainÁrea de Bioquímica y Biología Molecular, Departamento de Biología Molecular, Universidad de León, 24007 León, SpainÁrea de Bioquímica y Biología Molecular, Departamento de Biología Molecular, Universidad de León, 24007 León, SpainTestosterone holds significant medical and economic importance, with the global market for testosterone replacement therapies valued at approximately USD 1.9 billion in 2023. This hormone is essential for the development and maintenance of male sexual characteristics as well as bone and muscle health. It plays a key role in conditions such as hypogonadism, muscle disorders, and andropause. However, the industrial production of testosterone often involves complex chemical processes that result in low yields, high costs, and environmental damage. Microbial biotransformation of steroids presents an eco-friendly alternative to traditional chemical synthesis. A knockout strain of <i>Aspergillus nidulans</i> deficient in steroid 11α-hydroxylase activity was developed, rendering it incapable of hydroxylating androstenedione, progesterone, and testosterone. In these strains, two newly identified CYP450 enzymes, CYP68L1 from <i>A. nidulans</i> and CYP68L8 from <i>Aspergillus ochraceus,</i> were expressed to confirm their roles as steroid 11α-hydroxylases of androstenedione, progesterone, and testosterone. The availability of these 11α-hydroxylases represents significant progress toward achieving efficient single-step steroid fermentation. Furthermore, the <i>A. nidulans</i> knockout strain serves as an effective model for studying the conversion of androstenedione to testosterone upon the expression of the enzyme 17β-hydroxysteroid dehydrogenase, due to its inability to hydroxylate testosterone.https://www.mdpi.com/2218-273X/14/12/1502<i>Aspergillus nidulans</i>steroid 11α-hydroxylase<i>Aspergillus ochraceus</i>testosterone17β-hydroxysteroid dehydrogenase (17β-HSD) EC 1.1.1.51androstenedione
spellingShingle Lidia Ortega-de los Ríos
Luis Getino
Beatriz Galán
José Luis García
José M. Luengo
Alejandro Chamizo-Ampudia
José M. Fernández-Cañón
Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of Concept
Biomolecules
<i>Aspergillus nidulans</i>
steroid 11α-hydroxylase
<i>Aspergillus ochraceus</i>
testosterone
17β-hydroxysteroid dehydrogenase (17β-HSD) EC 1.1.1.51
androstenedione
title Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of Concept
title_full Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of Concept
title_fullStr Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of Concept
title_full_unstemmed Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of Concept
title_short Unlocking Testosterone Production by Biotransformation: Engineering a Fungal Model of <i>Aspergillus nidulans</i> Strain Deficient in Steroid 11α-Hydroxylase Activity and Expressing 17β-Hydroxysteroid Dehydrogenase Enzyme as Proof of Concept
title_sort unlocking testosterone production by biotransformation engineering a fungal model of i aspergillus nidulans i strain deficient in steroid 11α hydroxylase activity and expressing 17β hydroxysteroid dehydrogenase enzyme as proof of concept
topic <i>Aspergillus nidulans</i>
steroid 11α-hydroxylase
<i>Aspergillus ochraceus</i>
testosterone
17β-hydroxysteroid dehydrogenase (17β-HSD) EC 1.1.1.51
androstenedione
url https://www.mdpi.com/2218-273X/14/12/1502
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