Kinetic compartmentalization by unnatural reaction for itaconate production
Ye, Dae-Yeol (Pohang University of Science and Technology. Department of Chemical Engineering)
Noh, Myung Hyun (Pohang University of Science and Technology. Department of Chemical Engineering)
Moon, Jo Hyun (Pohang University of Science and Technology. Department of Chemical Engineering)
Milito, Alfonsina (Centre de Recerca en Agrigenòmica)
Kim, Minsun (Pohang University of Science and Technology. School of Interdisciplinary Bioscience and Bioengineering)
Lee, Jeong Wook (Pohang University of Science and Technology. Department of Chemical Engineering)
Yang, Jae-Seong (Centre de Recerca en Agrigenòmica)
Jung, Gyoo Yeol (Pohang University of Science and Technology. School of Interdisciplinary Bioscience and Bioengineering)

Date:	2022
Abstract:	Physical compartmentalization of metabolism using membranous organelles in eukaryotes is helpful for chemical biosynthesis to ensure the availability of substrates from competitive metabolic reactions. Bacterial hosts lack such a membranous system, which is one of the major limitations for efficient metabolic engineering. Here, we employ kinetic compartmentalization with the introduction of an unnatural enzymatic reaction by an engineered enzyme as an alternative strategy to enable substrate availability from competitive reactions through kinetic isolation of metabolic pathways. As a proof of concept, we kinetically isolate the itaconate synthetic pathway from the tricarboxylic acid cycle in Escherichia coli, which is natively separated by mitochondrial membranes in Aspergillus terreus. Specifically, 2-methylcitrate dehydratase is engineered to alternatively catalyze citrate and kinetically secure cis -aconitate for efficient production using a high-throughput screening system. Itaconate production can be significantly improved with kinetic compartmentalization and its strategy has the potential to be widely applicable. Unlike eukaryotic system, bacterial hosts lack membranous system, which is one of the limitations for efficient metabolic engineering. Here, the authors report a kinetic compartmentalization strategy to increase substrate availability from competitive reactions for the efficient production of itaconate in E. coli.
Grants:	Ministerio de Economía y Competitividad SEV-2015-0533 Ministerio de Economía y Competitividad CEX2019-000902-S European Commission 945043
Note:	Altres ajuts: CERCA Programme/Generalitat de Catalunya
Rights:	Aquest document està subjecte a una llicència d'ús Creative Commons. Es permet la reproducció total o parcial, la distribució, la comunicació pública de l'obra i la creació d'obres derivades, fins i tot amb finalitats comercials, sempre i quan es reconegui l'autoria de l'obra original.
Language:	Anglès
Document:	Article ; recerca ; Versió publicada
Subject:	Metabolic engineering ; Synthetic biology ; Applied microbiology
Published in:	Nature communications, Vol. 13 (September 2022) , art. 5353, ISSN 2041-1723

DOI: 10.1038/s41467-022-33033-1
PMID: 36097012

10 p, 944.4 KB

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Articles > Research articles
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