Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate)
Abstract Background Coenzyme A (CoA) is a carrier of acyl groups. This cofactor is synthesized from pantothenic acid in five steps. The phosphorylation of pantothenate is catalyzed by pantothenate kinase (CoaA), which is a key step in the CoA biosynthetic pathway. To determine whether the enhancemen...
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| Format: | Article |
| Language: | English |
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BMC
2023-04-01
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| Series: | Microbial Cell Factories |
| Subjects: | |
| Online Access: | https://doi.org/10.1186/s12934-023-02083-5 |
| _version_ | 1797840781509132288 |
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| author | Hirotaka Kudo Sho Ono Kenta Abe Mami Matsuda Tomohisa Hasunuma Tomoyasu Nishizawa Munehiko Asayama Hirofumi Nishihara Shigeru Chohnan |
| author_facet | Hirotaka Kudo Sho Ono Kenta Abe Mami Matsuda Tomohisa Hasunuma Tomoyasu Nishizawa Munehiko Asayama Hirofumi Nishihara Shigeru Chohnan |
| author_sort | Hirotaka Kudo |
| collection | DOAJ |
| description | Abstract Background Coenzyme A (CoA) is a carrier of acyl groups. This cofactor is synthesized from pantothenic acid in five steps. The phosphorylation of pantothenate is catalyzed by pantothenate kinase (CoaA), which is a key step in the CoA biosynthetic pathway. To determine whether the enhancement of the CoA biosynthetic pathway is effective for producing useful substances, the effect of elevated acetyl-CoA levels resulting from the introduction of the exogenous coaA gene on poly(3-hydroxybutyrate) [P(3HB)] synthesis was determined in Escherichia coli, which express the genes necessary for cyanobacterial polyhydroxyalkanoate synthesis (phaABEC). Results E. coli containing the coaA gene in addition to the pha genes accumulated more P(3HB) compared with the transformant containing the pha genes alone. P(3HB) production was enhanced by precursor addition, with P(3HB) content increasing from 18.4% (w/w) to 29.0% in the presence of 0.5 mM pantothenate and 16.3%–28.2% by adding 0.5 mM β-alanine. Strains expressing the exogenous coaA in the presence of precursors contained acetyl-CoA in excess of 1 nmol/mg of dry cell wt, which promoted the reaction toward P(3HB) formation. The amount of acetate exported into the medium was three times lower in the cells carrying exogenous coaA and pha genes than in the cells carrying pha genes alone. This was attributed to significantly enlarging the intracellular pool size of CoA, which is the recipient of acetic acid and is advantageous for microbial production of value-added materials. Conclusions Enhancing the CoA biosynthetic pathway with exogenous CoaA was effective at increasing P(3HB) production. Supplementing the medium with pantothenate facilitated the accumulation of P(3HB). β-Alanine was able to replace the efficacy of adding pantothenate. |
| first_indexed | 2024-04-09T16:20:16Z |
| format | Article |
| id | doaj.art-5375cfa8eb5c4f3d86d31981d33055bc |
| institution | Directory Open Access Journal |
| issn | 1475-2859 |
| language | English |
| last_indexed | 2024-04-09T16:20:16Z |
| publishDate | 2023-04-01 |
| publisher | BMC |
| record_format | Article |
| series | Microbial Cell Factories |
| spelling | doaj.art-5375cfa8eb5c4f3d86d31981d33055bc2023-04-23T11:32:11ZengBMCMicrobial Cell Factories1475-28592023-04-0122111310.1186/s12934-023-02083-5Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate)Hirotaka Kudo0Sho Ono1Kenta Abe2Mami Matsuda3Tomohisa Hasunuma4Tomoyasu Nishizawa5Munehiko Asayama6Hirofumi Nishihara7Shigeru Chohnan8Department of Food and Life Sciences, Ibaraki University College of AgricultureDepartment of Food and Life Sciences, Ibaraki University College of AgricultureDepartment of Food and Life Sciences, Ibaraki University College of AgricultureGraduate School of Science, Technology and Innovation, Kobe UniversityGraduate School of Science, Technology and Innovation, Kobe UniversityDepartment of Food and Life Sciences, Ibaraki University College of AgricultureDepartment of Food and Life Sciences, Ibaraki University College of AgricultureDepartment of Food and Life Sciences, Ibaraki University College of AgricultureDepartment of Food and Life Sciences, Ibaraki University College of AgricultureAbstract Background Coenzyme A (CoA) is a carrier of acyl groups. This cofactor is synthesized from pantothenic acid in five steps. The phosphorylation of pantothenate is catalyzed by pantothenate kinase (CoaA), which is a key step in the CoA biosynthetic pathway. To determine whether the enhancement of the CoA biosynthetic pathway is effective for producing useful substances, the effect of elevated acetyl-CoA levels resulting from the introduction of the exogenous coaA gene on poly(3-hydroxybutyrate) [P(3HB)] synthesis was determined in Escherichia coli, which express the genes necessary for cyanobacterial polyhydroxyalkanoate synthesis (phaABEC). Results E. coli containing the coaA gene in addition to the pha genes accumulated more P(3HB) compared with the transformant containing the pha genes alone. P(3HB) production was enhanced by precursor addition, with P(3HB) content increasing from 18.4% (w/w) to 29.0% in the presence of 0.5 mM pantothenate and 16.3%–28.2% by adding 0.5 mM β-alanine. Strains expressing the exogenous coaA in the presence of precursors contained acetyl-CoA in excess of 1 nmol/mg of dry cell wt, which promoted the reaction toward P(3HB) formation. The amount of acetate exported into the medium was three times lower in the cells carrying exogenous coaA and pha genes than in the cells carrying pha genes alone. This was attributed to significantly enlarging the intracellular pool size of CoA, which is the recipient of acetic acid and is advantageous for microbial production of value-added materials. Conclusions Enhancing the CoA biosynthetic pathway with exogenous CoaA was effective at increasing P(3HB) production. Supplementing the medium with pantothenate facilitated the accumulation of P(3HB). β-Alanine was able to replace the efficacy of adding pantothenate.https://doi.org/10.1186/s12934-023-02083-5Coenzyme AAcetyl-CoAPantothenate kinasePoly(3-hydroxybutyrate)Escherichia coliPantothenic acid |
| spellingShingle | Hirotaka Kudo Sho Ono Kenta Abe Mami Matsuda Tomohisa Hasunuma Tomoyasu Nishizawa Munehiko Asayama Hirofumi Nishihara Shigeru Chohnan Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate) Microbial Cell Factories Coenzyme A Acetyl-CoA Pantothenate kinase Poly(3-hydroxybutyrate) Escherichia coli Pantothenic acid |
| title | Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate) |
| title_full | Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate) |
| title_fullStr | Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate) |
| title_full_unstemmed | Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate) |
| title_short | Enhanced supply of acetyl-CoA by exogenous pantothenate kinase promotes synthesis of poly(3-hydroxybutyrate) |
| title_sort | enhanced supply of acetyl coa by exogenous pantothenate kinase promotes synthesis of poly 3 hydroxybutyrate |
| topic | Coenzyme A Acetyl-CoA Pantothenate kinase Poly(3-hydroxybutyrate) Escherichia coli Pantothenic acid |
| url | https://doi.org/10.1186/s12934-023-02083-5 |
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