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Background  1,3-propanediol (PDO) is a substantially industrial metabolite used in the polymer industry. Although several natural PDO production hosts exist, e.g.  Klebsiella  sp.,  Citrobacter  sp. and  Clostridium  sp., the PDO yield on glycerol is insufficient for an economically viable bio-process. Enhancing this yield via strain improvement can be achieved by disconnecting the production and growth pathways. In the case of PDO formation, this approach results in a microorganism metabolizing glycerol strictly for PDO production, while catabolizing a co-substrate for growth and maintenance. We applied this strategy to improve the PDO production with  Citrobacter werkmanii  DSM17579 .Results  Genetic tools were developed and used to create  Citrobacter werkmanii  DSM17579 ∆ dhaD  in which  dhaD,  encoding for glycerol dehydrogenase, was deleted. Since this strain was unable to grow on glycerol anaerobically, both pathways were disconnected. The knock-out strain was perturbed with 13 different co-substrates for growth and maintenance. Glucose was the most promising, although a competition between NADH-consuming enzymes and 1,3-propanediol dehydrogenase emerged.    Conclusion  Due to the deletion of  dhaD  in  Citrobacter werkmanii  DSM17579, the PDO production and growth pathway were split. As a consequence, the PDO yield on glycerol was improved 1,5 times, strengthening the idea that  Citrobacter werkmanii  DSM17579 could become an industrially interesting host for PDO production.

1,3-propanediol production with Citrobacter werkmanii DSM17579: effect of a dhaD knock-out