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- W2914950023 abstract "Pyruvate carboxylase is an anaplerotic carbon dioxide-fixing enzyme replenishing the tricarboxylic acid cycle with oxaloacetate during growth on sugars. In this study, we applied a lysine biosensor to identify pyruvate carboxylase variants in Corynebacterium glutamicum that enable improved l-lysine production from glucose. A suitable reporter strain was transformed with a pyc gene library created by error-prone PCR and screened by fluorescence-activated cell sorting (FACS) for cells with increased fluorescence triggered by an elevated cytoplasmic lysine concentration. Two pyruvate carboxylase variants, PCxT343A,I1012S and PCxT132A were identified allowing 9% and 19% increased lysine titers upon plasmid-based expression. Chromosomal expression of PCxT132A and PCxT343A variants led to 6% and 14% higher l-lysine levels. The new PCx variants can be useful also for other microbial strains producing TCA cycle-derived metabolites. Our approach indicates that a biosensor such as pSenLys enables directed evolution of many enzymes involved in converting a carbon source into the target metabolite." @default.
- W2914950023 created "2019-02-21" @default.
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- W2914950023 date "2019-02-01" @default.
- W2914950023 modified "2023-10-03" @default.
- W2914950023 title "Pyruvate Carboxylase Variants Enabling Improved Lysine Production from Glucose Identified by Biosensor-Based High-Throughput Fluorescence-Activated Cell Sorting Screening" @default.
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- W2914950023 doi "https://doi.org/10.1021/acssynbio.8b00510" @default.
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