Use of Escherichia coli add/ade mutant and Saccharomyces cerevisiae WSH2 to construct a highly efficient coupled system for glutathione production

How to supply ATP efficiently and economically is one of the key issues to achieve the commercialization of the enzymatic production of glutathione (GSH). In this work, a highly efficient coupled system for GSH enzymatic production was constructed with Escherichia coli Δadd/ade and Saccharomyces cerevisiae WSH2. In this system, ATP-consuming reactions for GSH synthesis are coupled with ATP-generating reactions. The results indicated that the irreversible transformation from ATP into hypoxanthine (Hx) was completely blocked in E. coli Δadd/ade, and ATP was mainly transformed into adenosine (Ado) and adenine (Ade). Due to the facts that Ado and Ade could be both used to generate ATP by S. cerevisiae WSH2, ATP-regenerating reaction was established in this coupled system. As a result, GSH production in this system reached 10.88 mM within 6 h, which was 4.03-fold of the coupled system of E. coli BW25113 and S. cerevisiae WSH2. The results are helpful for investigating the enzymatic production of GSH and other valuable ATP-dependent products.