1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO
Alcohol oxidase (AOX) oxidizes alcohols to produce carbonyl compounds and peroxides. Its promoter (PAOX1) is widely used in methylotrophic yeasts. A promising yeast expression system (Pichia sp. strain SO) was developed for bacterial lipase expression regulated by PAOX1 of Komagataella phaffii (prev...
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| Format: | Article |
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Springer Nature
2024
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| Online Access: | http://psasir.upm.edu.my/id/eprint/115144/ |
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| author | Mahyon, Nur Iznida Sabri, Suriana Jijew, George Crisol Salleh, Abu Bakar Leow, Thean Chor Lim, Si Jie Oslan, Siti Nur Hazwani Masomian, Malihe Oslan, Siti Nurbaya |
| author_facet | Mahyon, Nur Iznida Sabri, Suriana Jijew, George Crisol Salleh, Abu Bakar Leow, Thean Chor Lim, Si Jie Oslan, Siti Nur Hazwani Masomian, Malihe Oslan, Siti Nurbaya |
| author_sort | Mahyon, Nur Iznida |
| building | UPM Institutional Repository |
| collection | Online Access |
| description | Alcohol oxidase (AOX) oxidizes alcohols to produce carbonyl compounds and peroxides. Its promoter (PAOX1) is widely used in methylotrophic yeasts. A promising yeast expression system (Pichia sp. strain SO) was developed for bacterial lipase expression regulated by PAOX1 of Komagataella phaffii (previously known as Pichia pastoris). However, its unidentified AOX gene and the protein structure have deterred the search for the best inducer. This study was aimed to identify the yeast species and determine the best inducer for PAOX1 upregulation using in silico AOX protein analysis. Morphological (scanning and transmission electron microscopies) and carbon assimilation analyses confirmed isolate SO as Meyerozyma guilliermondii (previously known as Pichia guilliiermondii). Using Hidden-Markov model and degenerate PCR, the LCAO gene (2091 bp) was discovered in M. guilliermondii strain SO. The enzyme, MgFAO1 shared 14% similarity to K. phaffii AOX1 protein (KpAOX1). Molecular docking of MgFAO1 three-dimensional structure predicted using AlphaFold2 showed its preference toward long-chain 1-dodecanol as the substrate unlike KpAOX1 (short-chain methanol). While the alcohol-binding pocket in MgFAO1 was more hydrophobic compared to KpAOX1, 1-dodecanol could be a better inducer for protein expression in M. guilliermondii strain SO. Thus, in silico pipeline employed in this study can help identify homologous proteins in other expression hosts and their preferred substrates for promoter upregulation. However, the computational analyses were merely predictions and further wet-lab validation is required. Yet, this strategy allows cost-efficient screening of potential inducers for microbe-based protein production in the industries, reducing the production cost and offering cheaper options for consumers. |
| first_indexed | 2025-11-15T14:24:43Z |
| format | Article |
| id | upm-115144 |
| institution | Universiti Putra Malaysia |
| institution_category | Local University |
| last_indexed | 2025-11-15T14:24:43Z |
| publishDate | 2024 |
| publisher | Springer Nature |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | upm-1151442025-02-24T06:51:04Z http://psasir.upm.edu.my/id/eprint/115144/ 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO Mahyon, Nur Iznida Sabri, Suriana Jijew, George Crisol Salleh, Abu Bakar Leow, Thean Chor Lim, Si Jie Oslan, Siti Nur Hazwani Masomian, Malihe Oslan, Siti Nurbaya Alcohol oxidase (AOX) oxidizes alcohols to produce carbonyl compounds and peroxides. Its promoter (PAOX1) is widely used in methylotrophic yeasts. A promising yeast expression system (Pichia sp. strain SO) was developed for bacterial lipase expression regulated by PAOX1 of Komagataella phaffii (previously known as Pichia pastoris). However, its unidentified AOX gene and the protein structure have deterred the search for the best inducer. This study was aimed to identify the yeast species and determine the best inducer for PAOX1 upregulation using in silico AOX protein analysis. Morphological (scanning and transmission electron microscopies) and carbon assimilation analyses confirmed isolate SO as Meyerozyma guilliermondii (previously known as Pichia guilliiermondii). Using Hidden-Markov model and degenerate PCR, the LCAO gene (2091 bp) was discovered in M. guilliermondii strain SO. The enzyme, MgFAO1 shared 14% similarity to K. phaffii AOX1 protein (KpAOX1). Molecular docking of MgFAO1 three-dimensional structure predicted using AlphaFold2 showed its preference toward long-chain 1-dodecanol as the substrate unlike KpAOX1 (short-chain methanol). While the alcohol-binding pocket in MgFAO1 was more hydrophobic compared to KpAOX1, 1-dodecanol could be a better inducer for protein expression in M. guilliermondii strain SO. Thus, in silico pipeline employed in this study can help identify homologous proteins in other expression hosts and their preferred substrates for promoter upregulation. However, the computational analyses were merely predictions and further wet-lab validation is required. Yet, this strategy allows cost-efficient screening of potential inducers for microbe-based protein production in the industries, reducing the production cost and offering cheaper options for consumers. Springer Nature 2024-04-18 Article PeerReviewed Mahyon, Nur Iznida and Sabri, Suriana and Jijew, George Crisol and Salleh, Abu Bakar and Leow, Thean Chor and Lim, Si Jie and Oslan, Siti Nur Hazwani and Masomian, Malihe and Oslan, Siti Nurbaya (2024) 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO. Arabian Journal for Science and Engineering, 49. pp. 9133-9147. ISSN 2193-567X; eISSN: 2191-4281 https://link.springer.com/article/10.1007/s13369-024-09018-1?error=cookies_not_supported&code=2a5c7032-689a-4809-9c90-0e2573e2c376 10.1007/s13369-024-09018-1 |
| spellingShingle | Mahyon, Nur Iznida Sabri, Suriana Jijew, George Crisol Salleh, Abu Bakar Leow, Thean Chor Lim, Si Jie Oslan, Siti Nur Hazwani Masomian, Malihe Oslan, Siti Nurbaya 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO |
| title | 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO |
| title_full | 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO |
| title_fullStr | 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO |
| title_full_unstemmed | 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO |
| title_short | 1-dodecanol as potential inducer for the FAO1 promoter (PFAO1) in morphologically identified Meyerozyma guilliermondii strain SO |
| title_sort | 1-dodecanol as potential inducer for the fao1 promoter (pfao1) in morphologically identified meyerozyma guilliermondii strain so |
| url | http://psasir.upm.edu.my/id/eprint/115144/ http://psasir.upm.edu.my/id/eprint/115144/ http://psasir.upm.edu.my/id/eprint/115144/ |