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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...

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Autores principales: Feng, Linjuan, Xu, Junhao, Ye, Cuifang, Gao, Jucan, Huang, Lei, Xu, Zhinan, Lian, Jiazhang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
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.
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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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