eprintid: 10190622 rev_number: 9 eprint_status: archive userid: 699 dir: disk0/10/19/06/22 datestamp: 2024-04-12 13:41:42 lastmod: 2024-04-12 13:41:42 status_changed: 2024-04-12 13:41:42 type: article metadata_visibility: show sword_depositor: 699 creators_name: Jiang, W creators_name: Cai, Y creators_name: Sun, S creators_name: Wang, W creators_name: Tišma, M creators_name: Baganz, F creators_name: Hao, J title: Inactivation of hydrogenase-3 leads to enhancement of 1,3-propanediol and 2,3-butanediol production by Klebsiella pneumoniae ispublished: pub divisions: UCL divisions: B04 divisions: C05 divisions: F47 note: This version is the author accepted manuscript. For information on re-use, please refer to the publisher’s terms and conditions. abstract: Klebsiella pneumoniae can use glucose or glycerol as carbon sources to produce 1,3-propanediol or 2,3-butanediol, respectively. In the metabolism of Klebsiella pneumoniae, hydrogenase-3 is responsible for H2 production from formic acid, but it is not directly related to the synthesis pathways for 1,3-propanediol and 2,3-butanediol. In the first part of this research, hycEFG, which encodes subunits of the enzyme hydrogenase-3, was knocked out, so K. pneumoniae ΔhycEFG lost the ability to produce H2 during cultivation using glycerol as a carbon source. As a consequence, the concentration of 1,3-propanediol increased and the substrate (glycerol) conversion ratio reached 0.587 mol/mol. Then, K. pneumoniae ΔldhAΔhycEFG was constructed to erase lactic acid synthesis which led to the further increase of 1,3-propanediol concentration. A substrate (glycerol) conversion ratio of 0.628 mol/mol in batch conditions was achieved, which was higher compared to the wild type strain (0.545 mol/mol). Furthermore, since adhE encodes an alcohol dehydrogenase that catalyzes ethanol production from acetaldehyde, K. pneumoniae ΔldhAΔadhEΔhycEFG was constructed to prevent ethanol production. Contrary to expectations, this did not lead to a further increase, but to a decrease in 1,3-propanediol production. In the second part of this research, glucose was used as the carbon source to produce 2,3-butanediol. Knocking out hycEFG had distinct positive effect on 2,3-butanediol production. Especially in K. pneumoniae ΔldhAΔadhEΔhycEFG, a substrate (glucose) conversion ratio of 0.730 mol/mol was reached, which is higher compared to wild type strain (0.504 mol/mol). This work suggests that the inactivation of hydrogenase-3 may have a global effect on the metabolic regulation of K. pneumoniae, leading to the improvement of the production of two industrially important bulk chemicals, 1,3-propanediol and 2,3-butanediol. date: 2024-06 date_type: published publisher: Elsevier BV official_url: https://doi.org/10.1016/j.enzmictec.2024.110438 full_text_type: other language: eng verified: verified_manual elements_id: 2263495 doi: 10.1016/j.enzmictec.2024.110438 medium: Print-Electronic pii: S0141-0229(24)00045-0 lyricists_name: Baganz, Frank lyricists_id: FBAGA65 actors_name: Baganz, Frank actors_id: FBAGA65 actors_role: owner full_text_status: restricted publication: Enzyme and Microbial Technology volume: 177 article_number: 110438 event_location: United States issn: 0141-0229 citation: Jiang, W; Cai, Y; Sun, S; Wang, W; Tišma, M; Baganz, F; Hao, J; (2024) Inactivation of hydrogenase-3 leads to enhancement of 1,3-propanediol and 2,3-butanediol production by Klebsiella pneumoniae. Enzyme and Microbial Technology , 177 , Article 110438. 10.1016/j.enzmictec.2024.110438 <https://doi.org/10.1016/j.enzmictec.2024.110438>. document_url: https://discovery.ucl.ac.uk/id/eprint/10190622/1/Baganz_Inactivation%20of%20hydrogenase%20MS_Final.pdf