Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoids
Abstract Background Phenylpropanoids are a large group of plant secondary metabolites with various biological functions, derived from aromatic amino acids. Cyanobacteria are promising host organisms for sustainable production of plant phenylpropanoids. We have previously engineered Synechocystis sp....
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| Format: | Article |
| Language: | English |
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BMC
2024-02-01
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| Series: | Microbial Cell Factories |
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| Online Access: | https://doi.org/10.1186/s12934-024-02330-3 |
| _version_ | 1797272316704456704 |
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| author | Kateryna Kukil Pia Lindberg |
| author_facet | Kateryna Kukil Pia Lindberg |
| author_sort | Kateryna Kukil |
| collection | DOAJ |
| description | Abstract Background Phenylpropanoids are a large group of plant secondary metabolites with various biological functions, derived from aromatic amino acids. Cyanobacteria are promising host organisms for sustainable production of plant phenylpropanoids. We have previously engineered Synechocystis sp. PCC 6803 to produce trans-cinnamic acid (tCA) and p-coumaric acid (pCou), the first intermediates of phenylpropanoid pathway, by overexpression of phenylalanine- and tyrosine ammonia lyases. In this study, we aimed to enhance the production of the target compounds tCA and pCou in Synechocystis. Results We eliminated the 4-hydroxyphenylpyruvate dioxygenase (HPPD) activity, which is a competing pathway consuming tyrosine and, possibly, phenylalanine for tocopherol synthesis. Moreover, several genes of the terminal steps of the shikimate pathway were overexpressed alone or in operons, such as aromatic transaminases, feedback insensitive cyclohexadienyl dehydrogenase (TyrC) from Zymomonas mobilis and the chorismate mutase (CM) domain of the fused chorismate mutase/prephenate dehydratase enzyme from Escherichia coli. The obtained engineered strains demonstrated nearly 1.5 times enhanced tCA and pCou production when HPPD was knocked out compared to the parental production strains, accumulating 138 ± 3.5 mg L−1 of tCA and 72.3 ± 10.3 mg L−1 of pCou after seven days of photoautotrophic growth. However, there was no further improvement when any of the pathway genes were overexpressed. Finally, we used previously obtained AtPRM8 and TsPRM8 Synechocystis strains with deregulated shikimate pathway as a background for the overexpression of synthetic constructs with ppd knockout. Conclusions HPPD elimination enhances the tCA and pCou productivity to a similar extent. The use of PRM8 based strains as a background for overexpression of synthetic constructs, however, did not promote tCA and pCou titers, which indicates a tight regulation of the terminal steps of phenylalanine and tyrosine synthesis. This work contributes to establishing cyanobacteria as hosts for phenylpropanoid production. |
| first_indexed | 2024-03-07T14:25:37Z |
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| id | doaj.art-d4abe4fddbfe4eacadb21655aa3ecc5f |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-03-07T14:25:37Z |
| publishDate | 2024-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-d4abe4fddbfe4eacadb21655aa3ecc5f2024-03-06T08:06:14ZengBMCMicrobial Cell Factories1475-28592024-02-0123111410.1186/s12934-024-02330-3Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoidsKateryna Kukil0Pia Lindberg1Microbial Chemistry, Department of Chemistry - Ångström, Uppsala UniversityMicrobial Chemistry, Department of Chemistry - Ångström, Uppsala UniversityAbstract Background Phenylpropanoids are a large group of plant secondary metabolites with various biological functions, derived from aromatic amino acids. Cyanobacteria are promising host organisms for sustainable production of plant phenylpropanoids. We have previously engineered Synechocystis sp. PCC 6803 to produce trans-cinnamic acid (tCA) and p-coumaric acid (pCou), the first intermediates of phenylpropanoid pathway, by overexpression of phenylalanine- and tyrosine ammonia lyases. In this study, we aimed to enhance the production of the target compounds tCA and pCou in Synechocystis. Results We eliminated the 4-hydroxyphenylpyruvate dioxygenase (HPPD) activity, which is a competing pathway consuming tyrosine and, possibly, phenylalanine for tocopherol synthesis. Moreover, several genes of the terminal steps of the shikimate pathway were overexpressed alone or in operons, such as aromatic transaminases, feedback insensitive cyclohexadienyl dehydrogenase (TyrC) from Zymomonas mobilis and the chorismate mutase (CM) domain of the fused chorismate mutase/prephenate dehydratase enzyme from Escherichia coli. The obtained engineered strains demonstrated nearly 1.5 times enhanced tCA and pCou production when HPPD was knocked out compared to the parental production strains, accumulating 138 ± 3.5 mg L−1 of tCA and 72.3 ± 10.3 mg L−1 of pCou after seven days of photoautotrophic growth. However, there was no further improvement when any of the pathway genes were overexpressed. Finally, we used previously obtained AtPRM8 and TsPRM8 Synechocystis strains with deregulated shikimate pathway as a background for the overexpression of synthetic constructs with ppd knockout. Conclusions HPPD elimination enhances the tCA and pCou productivity to a similar extent. The use of PRM8 based strains as a background for overexpression of synthetic constructs, however, did not promote tCA and pCou titers, which indicates a tight regulation of the terminal steps of phenylalanine and tyrosine synthesis. This work contributes to establishing cyanobacteria as hosts for phenylpropanoid production.https://doi.org/10.1186/s12934-024-02330-3Synechocystis sp. PCC 6803Phenylalanine ammonia lyaseTrans-cinnamic acidp-coumaric acidPhenylpropanoids4-hydroxyphenylpyruvate dioxygenase |
| spellingShingle | Kateryna Kukil Pia Lindberg Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoids Microbial Cell Factories Synechocystis sp. PCC 6803 Phenylalanine ammonia lyase Trans-cinnamic acid p-coumaric acid Phenylpropanoids 4-hydroxyphenylpyruvate dioxygenase |
| title | Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoids |
| title_full | Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoids |
| title_fullStr | Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoids |
| title_full_unstemmed | Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoids |
| title_short | Metabolic engineering of Synechocystis sp. PCC 6803 for the improved production of phenylpropanoids |
| title_sort | metabolic engineering of synechocystis sp pcc 6803 for the improved production of phenylpropanoids |
| topic | Synechocystis sp. PCC 6803 Phenylalanine ammonia lyase Trans-cinnamic acid p-coumaric acid Phenylpropanoids 4-hydroxyphenylpyruvate dioxygenase |
| url | https://doi.org/10.1186/s12934-024-02330-3 |
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