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Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin

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Spence, Edward M., Calvo-Bado, Leonides, Mines, Paul and Bugg, Timothy D. H. (2021) Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin. Microbial Cell Factories, 20 (1). 15. doi:10.1186/s12934-020-01504-z

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Official URL: https://doi.org/10.1186/s12934-020-01504-z

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Abstract

Genetic modification of Rhodococcus jostii RHA1 was carried out in order to optimise the production of pyridine-2,4-dicarboxylic acid and pyridine-2,5-dicarboxylic acid bioproducts from lignin or lignocellulose breakdown, via insertion of either the Sphingobium SYK-6 ligAB genes or Paenibacillus praA gene respectively. Insertion of inducible plasmid pTipQC2 expression vector containing either ligAB or praA genes into a ΔpcaHG R. jostii RHA1 gene deletion strain gave 2–threefold higher titres of PDCA production from lignocellulose (200–287 mg/L), compared to plasmid expression in wild-type R. jostii RHA1. The ligAB genes were inserted in place of the chromosomal pcaHG genes encoding protocatechuate 3,4-dioxygenase, under the control of inducible Picl or PnitA promoters, or a constitutive Ptpc5 promoter, producing 2,4-PDCA products using either wheat straw lignocellulose or commercial soda lignin as carbon source. Insertion of Amycolatopsis sp. 75iv2 dyp2 gene on a pTipQC2 expression plasmid led to enhanced titres of 2,4-PDCA products, due to enhanced rate of lignin degradation. Growth in minimal media containing wheat straw lignocellulose led to the production of 2,4-PDCA in 330 mg/L titre in 40 h, with > tenfold enhanced productivity, compared with plasmid-based expression of ligAB genes in wild-type R. jostii RHA1. Production of 2,4-PDCA was also observed using several different polymeric lignins as carbon sources, and a titre of 240 mg/L was observed using a commercially available soda lignin as feedstock.

Item Type: Journal Article
Subjects: Q Science > QD Chemistry
Q Science > QK Botany
Q Science > QP Physiology
T Technology > TA Engineering (General). Civil engineering (General)
Divisions: Faculty of Science, Engineering and Medicine > Science > Chemistry
SWORD Depositor: Library Publications Router
Library of Congress Subject Headings (LCSH): Lignin , Lignin -- Biodegradation , Pyridine, Metabolism, Bioengineering, Rhodococcus , Genetic engineering
Journal or Publication Title: Microbial Cell Factories
Publisher: BioMed Central
ISSN: 1475-2859
Official Date: 19 January 2021
Dates:
DateEvent
19 January 2021Published
29 December 2020Accepted
Volume: 20
Number: 1
Article Number: 15
DOI: 10.1186/s12934-020-01504-z
Status: Peer Reviewed
Publication Status: Published
Reuse Statement (publisher, data, author rights):
Access rights to Published version: Restricted or Subscription Access
RIOXX Funder/Project Grant:
Project/Grant IDRIOXX Funder NameFunder ID
TS/M008282/1[EPSRC] Engineering and Physical Sciences Research Councilhttp://dx.doi.org/10.13039/501100000266
BB/T010622/1[BBSRC] Biotechnology and Biological Sciences Research Councilhttp://dx.doi.org/10.13039/501100000268
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