Combinatorial Assembly Platform Enabling Engineering of Genetically Stable Metabolic Pathways in Cyanobacteria

Overview

Journal Nucleic Acids Res

Publisher Oxford University Press

Specialty Biochemistry

Date 2021 Sep 23

PMID 34554258

Citations 8

Authors

George M Taylor

Andrew Hitchcock

John T Heap

Affiliations

Soon will be listed here.

Abstract

Cyanobacteria are simple, efficient, genetically-tractable photosynthetic microorganisms which in principle represent ideal biocatalysts for CO2 capture and conversion. However, in practice, genetic instability and low productivity are key, linked problems in engineered cyanobacteria. We took a massively parallel approach, generating and characterising libraries of synthetic promoters and RBSs for the cyanobacterium Synechocystis sp. PCC 6803, and assembling a sparse combinatorial library of millions of metabolic pathway-encoding construct variants. Genetic instability was observed for some variants, which is expected when variants cause metabolic burden. Surprisingly however, in a single combinatorial round without iterative optimisation, 80% of variants chosen at random and cultured photoautotrophically over many generations accumulated the target terpenoid lycopene from atmospheric CO2, apparently overcoming genetic instability. This large-scale parallel metabolic engineering of cyanobacteria provides a new platform for development of genetically stable cyanobacterial biocatalysts for sustainable light-driven production of valuable products directly from CO2, avoiding fossil carbon or competition with food production.

Citing Articles

C-metabolic flux analysis of respiratory chain disrupted strain ΔndhF1 of Synechocystis sp. PCC 6803.

Wada K, Toya Y, Matsuda F, Shimizu H Appl Biochem Biotechnol. 2025; .

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Construction of a Calibration Curve for Lycopene on a Liquid-Handling Platform─Wider Lessons for the Development of Automated Dilution Protocols.

Bultelle M, Casas A, Kitney R ACS Synth Biol. 2024; 13(8):2357-2375.

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Nutraceutical prospects of genetically engineered cyanobacteria- technological updates and significance.

Tiwari D, Kumar N, Bongirwar R, Shukla P World J Microbiol Biotechnol. 2024; 40(9):263.

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