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Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization

Enzyme spatial organization is an evolved mechanism for facilitating multi‐step biocatalysis and can play an important role in the regulation of promiscuous enzymes. The latter function suggests that artificial spatial organization can be an untapped avenue for controlling the specificity of bioengi...

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Autores principales: Cheah, Li Chen, Liu, Lian, Plan, Manuel R., Peng, Bingyin, Lu, Zeyu, Schenk, Gerhard, Vickers, Claudia E., Sainsbury, Frank
Formato: Online Artículo Texto
Lenguaje:English
Publicado: John Wiley and Sons Inc. 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10646250/
https://www.ncbi.nlm.nih.gov/pubmed/37750486
http://dx.doi.org/10.1002/advs.202303415
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author Cheah, Li Chen
Liu, Lian
Plan, Manuel R.
Peng, Bingyin
Lu, Zeyu
Schenk, Gerhard
Vickers, Claudia E.
Sainsbury, Frank
author_facet Cheah, Li Chen
Liu, Lian
Plan, Manuel R.
Peng, Bingyin
Lu, Zeyu
Schenk, Gerhard
Vickers, Claudia E.
Sainsbury, Frank
author_sort Cheah, Li Chen
collection PubMed
description Enzyme spatial organization is an evolved mechanism for facilitating multi‐step biocatalysis and can play an important role in the regulation of promiscuous enzymes. The latter function suggests that artificial spatial organization can be an untapped avenue for controlling the specificity of bioengineered metabolic pathways. A promiscuous terpene synthase (nerolidol synthase) is co‐localized and spatially organized with the preceding enzyme (farnesyl diphosphate synthase) in a heterologous production pathway, via translational protein fusion and/or co‐encapsulation in a self‐assembling protein cage. Spatial organization enhances nerolidol production by ≈11‐ to ≈62‐fold relative to unorganized enzymes. More interestingly, striking differences in the ratio of end products (nerolidol and linalool) are observed with each spatial organization approach. This demonstrates that artificial spatial organization approaches can be harnessed to modulate the product profiles of promiscuous enzymes in engineered pathways in vivo. This extends the application of spatial organization beyond situations where multiple enzymes compete for a single substrate to cases where there is competition among multiple substrates for a single enzyme.
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spelling pubmed-106462502023-09-26 Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization Cheah, Li Chen Liu, Lian Plan, Manuel R. Peng, Bingyin Lu, Zeyu Schenk, Gerhard Vickers, Claudia E. Sainsbury, Frank Adv Sci (Weinh) Research Articles Enzyme spatial organization is an evolved mechanism for facilitating multi‐step biocatalysis and can play an important role in the regulation of promiscuous enzymes. The latter function suggests that artificial spatial organization can be an untapped avenue for controlling the specificity of bioengineered metabolic pathways. A promiscuous terpene synthase (nerolidol synthase) is co‐localized and spatially organized with the preceding enzyme (farnesyl diphosphate synthase) in a heterologous production pathway, via translational protein fusion and/or co‐encapsulation in a self‐assembling protein cage. Spatial organization enhances nerolidol production by ≈11‐ to ≈62‐fold relative to unorganized enzymes. More interestingly, striking differences in the ratio of end products (nerolidol and linalool) are observed with each spatial organization approach. This demonstrates that artificial spatial organization approaches can be harnessed to modulate the product profiles of promiscuous enzymes in engineered pathways in vivo. This extends the application of spatial organization beyond situations where multiple enzymes compete for a single substrate to cases where there is competition among multiple substrates for a single enzyme. John Wiley and Sons Inc. 2023-09-26 /pmc/articles/PMC10646250/ /pubmed/37750486 http://dx.doi.org/10.1002/advs.202303415 Text en © 2023 The Authors. Advanced Science published by Wiley‐VCH GmbH https://creativecommons.org/licenses/by/4.0/This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Articles
Cheah, Li Chen
Liu, Lian
Plan, Manuel R.
Peng, Bingyin
Lu, Zeyu
Schenk, Gerhard
Vickers, Claudia E.
Sainsbury, Frank
Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization
title Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization
title_full Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization
title_fullStr Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization
title_full_unstemmed Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization
title_short Product Profiles of Promiscuous Enzymes Can be Altered by Controlling In Vivo Spatial Organization
title_sort product profiles of promiscuous enzymes can be altered by controlling in vivo spatial organization
topic Research Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10646250/
https://www.ncbi.nlm.nih.gov/pubmed/37750486
http://dx.doi.org/10.1002/advs.202303415
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