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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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WRAP-Metabolic-engineering-Rhodococcus-jostii-RHA1-lignin-2021.pdf - Published Version - Requires a PDF viewer. Available under License Creative Commons Attribution 4.0. Download (1415Kb) | Preview |
Official URL: https://doi.org/10.1186/s12934-020-01504-z
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 | |||||||||
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| Subjects: | Q Science > QD Chemistry Q Science > QK Botany Q Science > QP Physiology T Technology > TA Engineering (General). Civil engineering (General) |
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| Divisions: | Faculty of 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: |
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| Volume: | 20 | |||||||||
| Number: | 1 | |||||||||
| Article Number: | 15 | |||||||||
| DOI: | 10.1186/s12934-020-01504-z | |||||||||
| Status: | Peer Reviewed | |||||||||
| Publication Status: | Published | |||||||||
| Publisher Statement: | ||||||||||
| Access rights to Published version: | Restricted or Subscription Access | |||||||||
| RIOXX Funder/Project Grant: |
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