A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study
Abstract Background The climate crisis and depleting fossil fuel reserves have led to a drive for ‘green’ alternatives to the way we manufacture chemicals, and the formation of a bioeconomy that reduces our reliance on petrochemical-based feedstocks. Advances in Synthetic biology have provided the o...
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BMC
2025-01-01
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| Series: | Biotechnology for Biofuels and Bioproducts |
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| Online Access: | https://doi.org/10.1186/s13068-024-02599-x |
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| author | Alec Banner Joseph Webb Nigel Scrutton |
| author_facet | Alec Banner Joseph Webb Nigel Scrutton |
| author_sort | Alec Banner |
| collection | DOAJ |
| description | Abstract Background The climate crisis and depleting fossil fuel reserves have led to a drive for ‘green’ alternatives to the way we manufacture chemicals, and the formation of a bioeconomy that reduces our reliance on petrochemical-based feedstocks. Advances in Synthetic biology have provided the opportunity to engineer micro-organisms to produce compounds from renewable feedstocks, which could play a role in replacing traditional, petrochemical based, manufacturing routes. However, there are few examples of bio-manufactured products achieving commercialisation. This may be partially due to a disparity between academic and industrial focus, and a greater emphasis needs to be placed on economic feasibility at an earlier stage. Terpenoids are a class of compounds with diverse use across fuel, materials and pharmaceutical industries and can be manufactured biologically from the key intermediate mevalonate. Results Here, we report on a method of utilising parallel bioreactors to rapidly map the growth-coupling relationship between the specific product formation rate, specific substrate utilisation rate and specific growth rate. Using mevalonate as an example product, a maximum product yield coefficient of 0.18 gp/gs was achieved at a growth rate ( $$\mu$$ μ ) of 0.34 h−1. However, this process also led to the formation of the toxic byproduct acetate, which can slow growth and cause problems during downstream processing. By using gene editing to knock out the ackA-pta operon and poxB from E. coli BW25113, we were able to achieve the same optimum production rate, without the formation of acetate. Conclusions We demonstrated the power of using parallel bioreactors to assess productivity and the growth-coupling relationship between growth rate and product yield coefficient of mevalonate production. Using genetic engineering, our resultant strain demonstrated rapid mevalonate formation without the unwanted byproduct acetate. Mevalonate production is quantified and reported in industrially relevant units, including key parameters like conversion efficiency that are often omitted in early-stage publications reporting only titre in g/L. |
| format | Article |
| id | doaj-art-7129a10add36472a85cf5be12ff1cdc8 |
| institution | Kabale University |
| issn | 2731-3654 |
| language | English |
| publishDate | 2025-01-01 |
| publisher | BMC |
| record_format | Article |
| series | Biotechnology for Biofuels and Bioproducts |
| spelling | doaj-art-7129a10add36472a85cf5be12ff1cdc82025-01-19T12:13:47ZengBMCBiotechnology for Biofuels and Bioproducts2731-36542025-01-0118111010.1186/s13068-024-02599-xA parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case studyAlec Banner0Joseph Webb1Nigel Scrutton2Manchester Institute of Biotechnology, The University of ManchesterManchester Institute of Biotechnology, The University of ManchesterManchester Institute of Biotechnology, The University of ManchesterAbstract Background The climate crisis and depleting fossil fuel reserves have led to a drive for ‘green’ alternatives to the way we manufacture chemicals, and the formation of a bioeconomy that reduces our reliance on petrochemical-based feedstocks. Advances in Synthetic biology have provided the opportunity to engineer micro-organisms to produce compounds from renewable feedstocks, which could play a role in replacing traditional, petrochemical based, manufacturing routes. However, there are few examples of bio-manufactured products achieving commercialisation. This may be partially due to a disparity between academic and industrial focus, and a greater emphasis needs to be placed on economic feasibility at an earlier stage. Terpenoids are a class of compounds with diverse use across fuel, materials and pharmaceutical industries and can be manufactured biologically from the key intermediate mevalonate. Results Here, we report on a method of utilising parallel bioreactors to rapidly map the growth-coupling relationship between the specific product formation rate, specific substrate utilisation rate and specific growth rate. Using mevalonate as an example product, a maximum product yield coefficient of 0.18 gp/gs was achieved at a growth rate ( $$\mu$$ μ ) of 0.34 h−1. However, this process also led to the formation of the toxic byproduct acetate, which can slow growth and cause problems during downstream processing. By using gene editing to knock out the ackA-pta operon and poxB from E. coli BW25113, we were able to achieve the same optimum production rate, without the formation of acetate. Conclusions We demonstrated the power of using parallel bioreactors to assess productivity and the growth-coupling relationship between growth rate and product yield coefficient of mevalonate production. Using genetic engineering, our resultant strain demonstrated rapid mevalonate formation without the unwanted byproduct acetate. Mevalonate production is quantified and reported in industrially relevant units, including key parameters like conversion efficiency that are often omitted in early-stage publications reporting only titre in g/L.https://doi.org/10.1186/s13068-024-02599-xMevalonateParallel bioreactorGrowth-couplingFermentationScale-up |
| spellingShingle | Alec Banner Joseph Webb Nigel Scrutton A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study Biotechnology for Biofuels and Bioproducts Mevalonate Parallel bioreactor Growth-coupling Fermentation Scale-up |
| title | A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study |
| title_full | A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study |
| title_fullStr | A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study |
| title_full_unstemmed | A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study |
| title_short | A parallel bioreactor strategy to rapidly determine growth-coupling relationships for bioproduction: a mevalonate case study |
| title_sort | parallel bioreactor strategy to rapidly determine growth coupling relationships for bioproduction a mevalonate case study |
| topic | Mevalonate Parallel bioreactor Growth-coupling Fermentation Scale-up |
| url | https://doi.org/10.1186/s13068-024-02599-x |
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