Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i>
Being essential intermediates for the biosynthesis of heme, chlorophyll, and several other biologically critical compounds, porphyrins have wide practical applications. However, up till now, their bio-based production remains challenging. In this study, we identified potential metabolic factors limi...
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2025-01-01
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| author | Bahareh Arab Murray Moo-Young Yilan Liu C. Perry Chou |
| author_facet | Bahareh Arab Murray Moo-Young Yilan Liu C. Perry Chou |
| author_sort | Bahareh Arab |
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| description | Being essential intermediates for the biosynthesis of heme, chlorophyll, and several other biologically critical compounds, porphyrins have wide practical applications. However, up till now, their bio-based production remains challenging. In this study, we identified potential metabolic factors limiting the biosynthesis of type-III stereoisomeric porphyrins in <i>Escherichia coli</i>. To alleviate this limitation, we developed bioprocessing strategies by redirecting more dissimilated carbon flux toward the HemD-enzymatic pathway to enhance the production of type-III uroporphyrin (UP-III), which is a key precursor for heme biosynthesis. Our approaches included the use of antioxidant reagents and strain engineering. Supplementation with ascorbic acid (up to 1 g/L) increased the UP-III/UP-I ratio from 0.62 to 2.57. On the other hand, overexpression of ROS-scavenging genes such as <i>sod-</i> and <i>kat</i>-genes significantly enhanced UP production in <i>E. coli</i>. Notably, overexpression of <i>sodA</i> alone led to a 72.9% increase in total porphyrin production (1.56 g/L) while improving the UP-III/UP-I ratio to 1.94. Our findings highlight the potential of both antioxidant supplementation and strain engineering to mitigate ROS-induced oxidative stress and redirect more dissimilated carbon flux toward the biosynthesis of type-III porphyrins in <i>E. coli</i>. This work offers an effective platform to enhance the bio-based production of porphyrins. |
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| institution | Kabale University |
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| spelling | doaj-art-bd79fb3de3cc475ca488ab22f7b5309d2025-01-24T13:23:12ZengMDPI AGBioengineering2306-53542025-01-011218310.3390/bioengineering12010083Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i>Bahareh Arab0Murray Moo-Young1Yilan Liu2C. Perry Chou3Department of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, CanadaDepartment of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, CanadaDepartment of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, CanadaDepartment of Chemical Engineering, University of Waterloo, 200 University Avenue West, Waterloo, ON N2L 3G1, CanadaBeing essential intermediates for the biosynthesis of heme, chlorophyll, and several other biologically critical compounds, porphyrins have wide practical applications. However, up till now, their bio-based production remains challenging. In this study, we identified potential metabolic factors limiting the biosynthesis of type-III stereoisomeric porphyrins in <i>Escherichia coli</i>. To alleviate this limitation, we developed bioprocessing strategies by redirecting more dissimilated carbon flux toward the HemD-enzymatic pathway to enhance the production of type-III uroporphyrin (UP-III), which is a key precursor for heme biosynthesis. Our approaches included the use of antioxidant reagents and strain engineering. Supplementation with ascorbic acid (up to 1 g/L) increased the UP-III/UP-I ratio from 0.62 to 2.57. On the other hand, overexpression of ROS-scavenging genes such as <i>sod-</i> and <i>kat</i>-genes significantly enhanced UP production in <i>E. coli</i>. Notably, overexpression of <i>sodA</i> alone led to a 72.9% increase in total porphyrin production (1.56 g/L) while improving the UP-III/UP-I ratio to 1.94. Our findings highlight the potential of both antioxidant supplementation and strain engineering to mitigate ROS-induced oxidative stress and redirect more dissimilated carbon flux toward the biosynthesis of type-III porphyrins in <i>E. coli</i>. This work offers an effective platform to enhance the bio-based production of porphyrins.https://www.mdpi.com/2306-5354/12/1/83antioxidant<i>Escherichia coli</i>porphyrinstrain engineeringreactive oxygen species |
| spellingShingle | Bahareh Arab Murray Moo-Young Yilan Liu C. Perry Chou Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i> Bioengineering antioxidant <i>Escherichia coli</i> porphyrin strain engineering reactive oxygen species |
| title | Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i> |
| title_full | Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i> |
| title_fullStr | Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i> |
| title_full_unstemmed | Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i> |
| title_short | Manipulating Intracellular Oxidative Conditions to Enhance Porphyrin Production in <i>Escherichia coli</i> |
| title_sort | manipulating intracellular oxidative conditions to enhance porphyrin production in i escherichia coli i |
| topic | antioxidant <i>Escherichia coli</i> porphyrin strain engineering reactive oxygen species |
| url | https://www.mdpi.com/2306-5354/12/1/83 |
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