Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production.
Nitrogen-heterocycles are widely found in nature as subunits of compounds from secondary metabolism (vitamins, alkaloids, hormones etc.) and are valuable building blocks to manufacture a wide range of pharmaceuticals, agrochemicals, flavours and fragrances. Currently, N-heterocycles are produced thr...
| Main Author: | |
|---|---|
| Format: | Thesis (University of Nottingham only) |
| Language: | English |
| Published: |
2019
|
| Subjects: | |
| Online Access: | https://eprints.nottingham.ac.uk/56093/ |
| _version_ | 1848799270637928448 |
|---|---|
| author | Anyanwu, Valentine E. |
| author_facet | Anyanwu, Valentine E. |
| author_sort | Anyanwu, Valentine E. |
| building | Nottingham Research Data Repository |
| collection | Online Access |
| description | Nitrogen-heterocycles are widely found in nature as subunits of compounds from secondary metabolism (vitamins, alkaloids, hormones etc.) and are valuable building blocks to manufacture a wide range of pharmaceuticals, agrochemicals, flavours and fragrances. Currently, N-heterocycles are produced through unsustainable chemical processes that rely on petrochemical feedstocks and high-energy consumption. The six-membered simple N-heterocycle Δ¹-piperideine (2,3,4,5-tetrahydropyridine), is a key building block for biosynthesis of some alkaloids, for example, the piperidine, quinolizidine, indolizidine and lycopodium alkaloids. Δ¹-piperideine also has considerable promise as a platform compound for chemical synthesis of functionalized or complex compounds with economic importance. Although the bioproduction of complex, functionalised N-heterocycles have been reported, bioproduction of unsubstituted simple N-heterocycles have not yet been achieved. Advances in metabolic engineering and synthetic biology offers an approach to develop bioprocesses for sustainable production of simple N-heterocycles from renewable sugar feedstocks.
Δ¹-Piperideine can be obtained by oxidation of cadaverine using amine oxidases or transaminases, to yield 5-aminopentanal. This product is known to cyclise spontaneously into Δ¹-piperideine. However, Δ¹-piperideine reacts spontaneously in aqueous solution to form the multimeric products (tetrahydroanabasine, α-tripiperideine and isotripiperideine). Therefore, the suitability of putrescine oxidase from Rhodococcus erythropolis (PuORh) for cell-free and whole-cell bioproduction of Δ¹-piperideine was studied. PuORh was characterised for the first time by monitoring oxygen concentration in a direct substrate consumption assay. Using purified PuORh and PuORh induced whole cells; the conversion of cadaverine to Δ¹-piperideine was demonstrated qualitatively and quantitatively. This study indicates that PuORh is suitable for the bioproduction of Δ¹-piperideine. Finally, a metabolic route has been designed and proposed for whole cell bioproduction of Δ¹-Piperideine from renewable feedstocks. |
| first_indexed | 2025-11-14T20:33:00Z |
| format | Thesis (University of Nottingham only) |
| id | nottingham-56093 |
| institution | University of Nottingham Malaysia Campus |
| institution_category | Local University |
| language | English |
| last_indexed | 2025-11-14T20:33:00Z |
| publishDate | 2019 |
| recordtype | eprints |
| repository_type | Digital Repository |
| spelling | nottingham-560932025-02-28T14:24:07Z https://eprints.nottingham.ac.uk/56093/ Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. Anyanwu, Valentine E. Nitrogen-heterocycles are widely found in nature as subunits of compounds from secondary metabolism (vitamins, alkaloids, hormones etc.) and are valuable building blocks to manufacture a wide range of pharmaceuticals, agrochemicals, flavours and fragrances. Currently, N-heterocycles are produced through unsustainable chemical processes that rely on petrochemical feedstocks and high-energy consumption. The six-membered simple N-heterocycle Δ¹-piperideine (2,3,4,5-tetrahydropyridine), is a key building block for biosynthesis of some alkaloids, for example, the piperidine, quinolizidine, indolizidine and lycopodium alkaloids. Δ¹-piperideine also has considerable promise as a platform compound for chemical synthesis of functionalized or complex compounds with economic importance. Although the bioproduction of complex, functionalised N-heterocycles have been reported, bioproduction of unsubstituted simple N-heterocycles have not yet been achieved. Advances in metabolic engineering and synthetic biology offers an approach to develop bioprocesses for sustainable production of simple N-heterocycles from renewable sugar feedstocks. Δ¹-Piperideine can be obtained by oxidation of cadaverine using amine oxidases or transaminases, to yield 5-aminopentanal. This product is known to cyclise spontaneously into Δ¹-piperideine. However, Δ¹-piperideine reacts spontaneously in aqueous solution to form the multimeric products (tetrahydroanabasine, α-tripiperideine and isotripiperideine). Therefore, the suitability of putrescine oxidase from Rhodococcus erythropolis (PuORh) for cell-free and whole-cell bioproduction of Δ¹-piperideine was studied. PuORh was characterised for the first time by monitoring oxygen concentration in a direct substrate consumption assay. Using purified PuORh and PuORh induced whole cells; the conversion of cadaverine to Δ¹-piperideine was demonstrated qualitatively and quantitatively. This study indicates that PuORh is suitable for the bioproduction of Δ¹-piperideine. Finally, a metabolic route has been designed and proposed for whole cell bioproduction of Δ¹-Piperideine from renewable feedstocks. 2019-07-18 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/56093/1/Anyanwu%20Valentine-Final%20Thesis%20after%20Correction.pdf Anyanwu, Valentine E. (2019) Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. PhD thesis, University of Nottingham. Biotechnology sustainability metabolic engineering cadaverine Δ¹-piperideine N-heterocycles putrescine oxidase biocatalysis bioproduction |
| spellingShingle | Biotechnology sustainability metabolic engineering cadaverine Δ¹-piperideine N-heterocycles putrescine oxidase biocatalysis bioproduction Anyanwu, Valentine E. Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. |
| title | Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. |
| title_full | Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. |
| title_fullStr | Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. |
| title_full_unstemmed | Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. |
| title_short | Design and exploitation of cadaverine metabolic pathway for simple N-heterocyclic chemical production. |
| title_sort | design and exploitation of cadaverine metabolic pathway for simple n-heterocyclic chemical production. |
| topic | Biotechnology sustainability metabolic engineering cadaverine Δ¹-piperideine N-heterocycles putrescine oxidase biocatalysis bioproduction |
| url | https://eprints.nottingham.ac.uk/56093/ |