AI Article Synopsis
- ATP-dependent multi-enzymatic reactions typically require large amounts of ATP and extra enzymes for recycling, making them costly and inefficient.
- A new method has been developed to create inexpensive recombinant E. coli extracts that can carry out these reactions using only small amounts of ATP.
- These extracts utilize both specially overexpressed enzymes and the bacteria's natural kinases to recycle ATP, effectively streamlining the biotransformation process for various enzymatic reactions.
Article Abstract
The utility of ATP-dependent multi-enzymatic reactions is limited by their requirement for stoichiometric amounts of this expensive cofactor or additional purified enzymes for its recycling. Here we describe a simple method for the production of recombinant cell-free extracts (or lysates) of E. coli that support ATP-dependent biotransformations. The inexpensive preparation described is obtained with modest processing from a single recombinant bacterial culture of E. coli. In addition to recombinantly overexpressed enzymes that catalyze the primary ATP-dependent reactions of interest, endogenous kinases that are naturally present in the extract catalyze recycling of the requisite ATP. This means that only catalytic amounts of cofactor are necessary to drive the biotransformation, and without the requirement for additional purified enzymes. This approach has been applied successfully to an array of in vitro enzymatic cascades with multiple ATP-dependent steps.
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| http://dx.doi.org/10.1007/978-1-0716-2269-8_18 | DOI Listing |
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