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Lactococcus lactis as a cell factory for high-level diacetyl production
J Hugenholtz1, M Kleerebezem, M Starrenburg
1Wageningen Centre for Food Sciences and NIZO Food Research, 6710 BA Ede, The Netherlands. hugenhol@nizo.nl
Applied and Environmental Microbiology
|August 31, 2000
Summary
We engineered Lactococcus lactis to efficiently convert sugar into diacetyl by overproducing NADH-oxidase and inactivating alpha-acetolactate decarboxylase. This rerouted 80% of carbon flux, enhancing diacetyl production in this microbial cell factory.
Area of Science:
- Biotechnology
- Microbial Engineering
- Metabolic Engineering
Background:
- Lactococcus lactis is a key bacterium in dairy fermentation.
- Diacetyl is a valuable flavor compound in food production.
- Efficient microbial production of diacetyl is industrially relevant.
Purpose of the Study:
- To engineer Lactococcus lactis for enhanced diacetyl production.
- To optimize the metabolic pathway for diacetyl synthesis.
- To investigate the role of NADH-oxidase and alpha-acetolactate decarboxylase.
Main Methods:
- Genetic engineering of Lactococcus lactis.
- Overproduction of NADH-oxidase enzyme.
- Inactivation of alpha-acetolactate decarboxylase gene.
- Metabolic flux analysis to track carbon flow.
Main Results:
- Achieved efficient conversion of sugar to diacetyl.
- Successfully rerouted 80% of carbon flux towards diacetyl production.
- Demonstrated the effectiveness of combining NADH-oxidase overproduction and enzyme inactivation.
Conclusions:
- Engineered Lactococcus lactis serves as an effective cell factory for diacetyl.
- Metabolic engineering strategies can significantly enhance the production of valuable compounds.
- This approach offers a sustainable method for flavor compound generation.