Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin
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 Paenibac...
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| Format: | Article |
| Language: | English |
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BMC
2021-01-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-020-01504-z |
| _version_ | 1818722231246651392 |
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| author | Edward M. Spence Leonides Calvo-Bado Paul Mines Timothy D. H. Bugg |
| author_facet | Edward M. Spence Leonides Calvo-Bado Paul Mines Timothy D. H. Bugg |
| author_sort | Edward M. Spence |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-12-17T20:51:20Z |
| format | Article |
| id | doaj.art-674f161b95d443b495e63e36aa267cb0 |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-12-17T20:51:20Z |
| publishDate | 2021-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-674f161b95d443b495e63e36aa267cb02022-12-21T21:33:01ZengBMCMicrobial Cell Factories1475-28592021-01-0120111210.1186/s12934-020-01504-zMetabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from ligninEdward M. Spence0Leonides Calvo-Bado1Paul Mines2Timothy D. H. Bugg3Department of Chemistry, University of WarwickDepartment of Chemistry, University of WarwickBiome Bioplastics LtdDepartment of Chemistry, University of WarwickAbstract 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.https://doi.org/10.1186/s12934-020-01504-zLignin degradationPyridine dicarboxylic acidMetabolic engineeringRhodococcus jostii RHA1Gene promoter |
| spellingShingle | Edward M. Spence Leonides Calvo-Bado Paul Mines Timothy D. H. Bugg Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin Microbial Cell Factories Lignin degradation Pyridine dicarboxylic acid Metabolic engineering Rhodococcus jostii RHA1 Gene promoter |
| title | Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin |
| title_full | Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin |
| title_fullStr | Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin |
| title_full_unstemmed | Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin |
| title_short | Metabolic engineering of Rhodococcus jostii RHA1 for production of pyridine-dicarboxylic acids from lignin |
| title_sort | metabolic engineering of rhodococcus jostii rha1 for production of pyridine dicarboxylic acids from lignin |
| topic | Lignin degradation Pyridine dicarboxylic acid Metabolic engineering Rhodococcus jostii RHA1 Gene promoter |
| url | https://doi.org/10.1186/s12934-020-01504-z |
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