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Type 2 IDI performs better than type 1 for improving lycopene production in metabolically engineered E. coli strains
Sara Abolhassani Rad1, Hossein Shahbani Zahiri, Kambiz Akbari Noghabi
1Department of Molecular Genetics, National Institute of Genetic Engineering and Biotechnology, Shahrak-e Pajoohesh, km 15, Tehran-Karaj Highway, P.O Box: 14965/161, Tehran, I. R. Iran.
World Journal of Microbiology & Biotechnology
|July 19, 2012
Summary
Type 2 isopentenyl diphosphate isomerase (IDI) significantly enhances lycopene production in E. coli. Supplementing with citrate and engineering the mevalonate pathway further boosts isoprenoid accumulation.
Area of Science:
- Metabolic Engineering
- Synthetic Biology
- Biotechnology
Background:
- Lycopene, a valuable carotenoid, is produced via the isoprenoid pathway.
- Isopentenyl diphosphate isomerases (IDI) are crucial enzymes in this pathway, catalyzing the interconversion of isopentenyl diphosphate and dimethylallyl diphosphate.
- Optimizing IDI activity is key to enhancing isoprenoid production in microbial hosts like Escherichia coli.
Purpose of the Study:
- To compare the efficacy of type 1 and type 2 isopentenyl diphosphate isomerases (IDI) in improving lycopene production in Escherichia coli.
- To investigate the impact of heterologous mevalonate pathway introduction and auxiliary substrates on lycopene yield.
- To identify optimal metabolic engineering strategies for increased isoprenoid accumulation in E. coli.
Main Methods:
- Overexpression of Bacillus licheniformis (type 2, i(Bl)) and E. coli (type 1, i(Ec)) IDI genes in lycopene-producing E. coli strains.
- Introduction of a heterologous mevalonate pathway to increase isopentenyl diphosphate (IPP) abundance.
- Evaluation of various auxiliary substrates (fructose, mannose, arabinose, acetate, citrate) with glycerol to assess their effect on lycopene production.
Main Results:
- Overexpression of i(Ec) increased lycopene production from 33 to 68 mg/gDCW, while i(Bl) enhanced it to 80 mg/gDCW.
- Combining i(Bl) with the mevalonate pathway yielded up to 181 mg/gDCW lycopene.
- Citrate as an auxiliary substrate significantly improved lycopene production across all tested strains, reaching a maximum of 198 mg/gDCW.
Conclusions:
- Type 2 IDI from B. licheniformis is more effective than type 1 IDI from E. coli for enhancing isoprenoid production in E. coli.
- Metabolic engineering of the mevalonate pathway and the strategic use of auxiliary substrates like citrate are promising approaches for boosting lycopene and other isoprenoid yields.
- This study provides valuable insights for optimizing microbial cell factories for the sustainable production of valuable isoprenoids.
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