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Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone
Triacetic acid lactone (TAL) is a promising renewable platform polyketide with broad biotechnological applications. In this study, we constructed an engineered Pichia pastoris strain for the production of TAL. We first introduced a heterologous TAL biosynthetic pathway by integrating the 2-pyrone sy...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10145311/ https://www.ncbi.nlm.nih.gov/pubmed/37108948 http://dx.doi.org/10.3390/jof9040494 |
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author | Feng, Linjuan Xu, Junhao Ye, Cuifang Gao, Jucan Huang, Lei Xu, Zhinan Lian, Jiazhang |
author_facet | Feng, Linjuan Xu, Junhao Ye, Cuifang Gao, Jucan Huang, Lei Xu, Zhinan Lian, Jiazhang |
author_sort | Feng, Linjuan |
collection | PubMed |
description | Triacetic acid lactone (TAL) is a promising renewable platform polyketide with broad biotechnological applications. In this study, we constructed an engineered Pichia pastoris strain for the production of TAL. We first introduced a heterologous TAL biosynthetic pathway by integrating the 2-pyrone synthase encoding gene from Gerbera hybrida (Gh2PS). We then removed the rate-limiting step of TAL synthesis by introducing the posttranslational regulation-free acetyl-CoA carboxylase mutant encoding gene from S. cerevisiae (ScACC1*) and increasing the copy number of Gh2PS. Finally, to enhance intracellular acetyl-CoA supply, we focused on the introduction of the phosphoketolase/phosphotransacetylase pathway (PK pathway). To direct more carbon flux towards the PK pathway for acetyl-CoA generation, we combined it with a heterologous xylose utilization pathway or endogenous methanol utilization pathway. The combination of the PK pathway with the xylose utilization pathway resulted in the production of 825.6 mg/L TAL in minimal medium with xylose as the sole carbon source, with a TAL yield of 0.041 g/g xylose. This is the first report on TAL biosynthesis in P. pastoris and its direct synthesis from methanol. The present study suggests potential applications in improving the intracellular pool of acetyl-CoA and provides a basis for the construction of efficient cell factories for the production of acetyl-CoA derived compounds. |
format | Online Article Text |
id | pubmed-10145311 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-101453112023-04-29 Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone Feng, Linjuan Xu, Junhao Ye, Cuifang Gao, Jucan Huang, Lei Xu, Zhinan Lian, Jiazhang J Fungi (Basel) Article Triacetic acid lactone (TAL) is a promising renewable platform polyketide with broad biotechnological applications. In this study, we constructed an engineered Pichia pastoris strain for the production of TAL. We first introduced a heterologous TAL biosynthetic pathway by integrating the 2-pyrone synthase encoding gene from Gerbera hybrida (Gh2PS). We then removed the rate-limiting step of TAL synthesis by introducing the posttranslational regulation-free acetyl-CoA carboxylase mutant encoding gene from S. cerevisiae (ScACC1*) and increasing the copy number of Gh2PS. Finally, to enhance intracellular acetyl-CoA supply, we focused on the introduction of the phosphoketolase/phosphotransacetylase pathway (PK pathway). To direct more carbon flux towards the PK pathway for acetyl-CoA generation, we combined it with a heterologous xylose utilization pathway or endogenous methanol utilization pathway. The combination of the PK pathway with the xylose utilization pathway resulted in the production of 825.6 mg/L TAL in minimal medium with xylose as the sole carbon source, with a TAL yield of 0.041 g/g xylose. This is the first report on TAL biosynthesis in P. pastoris and its direct synthesis from methanol. The present study suggests potential applications in improving the intracellular pool of acetyl-CoA and provides a basis for the construction of efficient cell factories for the production of acetyl-CoA derived compounds. MDPI 2023-04-20 /pmc/articles/PMC10145311/ /pubmed/37108948 http://dx.doi.org/10.3390/jof9040494 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Feng, Linjuan Xu, Junhao Ye, Cuifang Gao, Jucan Huang, Lei Xu, Zhinan Lian, Jiazhang Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone |
title | Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone |
title_full | Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone |
title_fullStr | Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone |
title_full_unstemmed | Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone |
title_short | Metabolic Engineering of Pichia pastoris for the Production of Triacetic Acid Lactone |
title_sort | metabolic engineering of pichia pastoris for the production of triacetic acid lactone |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10145311/ https://www.ncbi.nlm.nih.gov/pubmed/37108948 http://dx.doi.org/10.3390/jof9040494 |
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