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Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast
3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS) is a key enzyme in the shikimate pathway for the biosynthesis of aromatic compounds. (L)-Phe and (L)-Tyr bind to the two main DAHPS isoforms and inhibit their enzyme activities, respectively. Synthetic biologists aim to relieve such inhibi...
Autores principales: | , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Nature Publishing Group UK
2023
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10349828/ https://www.ncbi.nlm.nih.gov/pubmed/37454208 http://dx.doi.org/10.1038/s42004-023-00946-x |
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author | Liu, Huayi Xiao, Qingjie Wu, Xinxin Ma, He Li, Jian Guo, Xufan Liu, Zhenyu Zhang, Yan Luo, Yunzi |
author_facet | Liu, Huayi Xiao, Qingjie Wu, Xinxin Ma, He Li, Jian Guo, Xufan Liu, Zhenyu Zhang, Yan Luo, Yunzi |
author_sort | Liu, Huayi |
collection | PubMed |
description | 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS) is a key enzyme in the shikimate pathway for the biosynthesis of aromatic compounds. (L)-Phe and (L)-Tyr bind to the two main DAHPS isoforms and inhibit their enzyme activities, respectively. Synthetic biologists aim to relieve such inhibitions in order to improve the productivity of aromatic compounds. In this work, we reported a point mutant of yeast DHAPS, Aro3(D154N), which retains the wild type enzyme activity but converts it highly inert to the inhibition by (L)-Phe. The Aro3 crystal structure along with the molecular dynamics simulations analysis suggests that the D154N mutation distant from the inhibitor binding cavity may reduce the binding affinity of (L)-Phe. Growth assays demonstrated that substitution of the conserved D154 with asparagine suffices to relieve the inhibition of (L)-Phe on Aro3, (L)-Tyr on Aro4, and the inhibitions on their corresponding homologues from diverse yeasts. The importance of our discovery is highlighted by the observation of 29.1% and 43.6% increase of yield for the production of tyrosol and salidroside respectively upon substituting ARO3 with ARO3(D154N). We anticipate that this allele would be used broadly to increase the yield of various aromatic products in metabolically diverse microorganisms. |
format | Online Article Text |
id | pubmed-10349828 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2023 |
publisher | Nature Publishing Group UK |
record_format | MEDLINE/PubMed |
spelling | pubmed-103498282023-07-17 Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast Liu, Huayi Xiao, Qingjie Wu, Xinxin Ma, He Li, Jian Guo, Xufan Liu, Zhenyu Zhang, Yan Luo, Yunzi Commun Chem Article 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase (DAHPS) is a key enzyme in the shikimate pathway for the biosynthesis of aromatic compounds. (L)-Phe and (L)-Tyr bind to the two main DAHPS isoforms and inhibit their enzyme activities, respectively. Synthetic biologists aim to relieve such inhibitions in order to improve the productivity of aromatic compounds. In this work, we reported a point mutant of yeast DHAPS, Aro3(D154N), which retains the wild type enzyme activity but converts it highly inert to the inhibition by (L)-Phe. The Aro3 crystal structure along with the molecular dynamics simulations analysis suggests that the D154N mutation distant from the inhibitor binding cavity may reduce the binding affinity of (L)-Phe. Growth assays demonstrated that substitution of the conserved D154 with asparagine suffices to relieve the inhibition of (L)-Phe on Aro3, (L)-Tyr on Aro4, and the inhibitions on their corresponding homologues from diverse yeasts. The importance of our discovery is highlighted by the observation of 29.1% and 43.6% increase of yield for the production of tyrosol and salidroside respectively upon substituting ARO3 with ARO3(D154N). We anticipate that this allele would be used broadly to increase the yield of various aromatic products in metabolically diverse microorganisms. Nature Publishing Group UK 2023-07-15 /pmc/articles/PMC10349828/ /pubmed/37454208 http://dx.doi.org/10.1038/s42004-023-00946-x Text en © The Author(s) 2023 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . |
spellingShingle | Article Liu, Huayi Xiao, Qingjie Wu, Xinxin Ma, He Li, Jian Guo, Xufan Liu, Zhenyu Zhang, Yan Luo, Yunzi Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast |
title | Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast |
title_full | Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast |
title_fullStr | Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast |
title_full_unstemmed | Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast |
title_short | Mechanistic investigation of a D to N mutation in DAHP synthase that dictates carbon flux into the shikimate pathway in yeast |
title_sort | mechanistic investigation of a d to n mutation in dahp synthase that dictates carbon flux into the shikimate pathway in yeast |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10349828/ https://www.ncbi.nlm.nih.gov/pubmed/37454208 http://dx.doi.org/10.1038/s42004-023-00946-x |
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