eprintid: 10125446 rev_number: 14 eprint_status: archive userid: 608 dir: disk0/10/12/54/46 datestamp: 2021-04-01 13:27:39 lastmod: 2021-09-25 23:35:00 status_changed: 2021-04-01 13:27:39 type: article metadata_visibility: show creators_name: Popadić, D creators_name: Mhaindarkar, D creators_name: Dang Thai, MHN creators_name: Hailes, HC creators_name: Mordhorst, S creators_name: Andexer, JN title: A bicyclic S-adenosylmethionine regeneration system applicable with different nucleosides or nucleotides as cofactor building blocks ispublished: inpress divisions: UCL divisions: B04 divisions: C06 divisions: F56 note: This article is licensed under a Creative Commons Attribution-NonCommercial 3.0 Unported Licence (http://creativecommons.org/licenses/by-nc/3.0/). abstract: The ubiquitous cofactor S-adenosyl-L-methionine (SAM) is part of numerous biochemical reactions in metabolism, epigenetics, and cancer development. As methylation usually improves physiochemical properties of compounds relevant for pharmaceutical use, the sustainable use of SAM as a methyl donor in biotechnological applications is an important goal. SAM-dependent methyltransferases are consequently an emerging biocatalytic tool for environmentally friendly and selective alkylations. However, SAM shows undesirable characteristics such as degradation under mild conditions and its stoichiometric use is economically not reasonable. Here, we report an optimised biomimetic system for the regeneration of SAM and SAM analogues consisting of effective nucleoside triphosphate formation and an additional L-methionine regeneration cycle without by-product accumulation. The bicyclic system uses seven enzymes, S-methylmethionine as methyl donor and a surplus of inorganic polyphosphate, along with catalytic amounts of L-methionine and cofactor building block reaching conversions of up to 99% (up to 200 turnovers). We also show that the cycle can be run with cofactor building blocks containing different purine and pyrimidine nucleobases, which can be fed in at the nucleoside or nucleotide stage. These alternative cofactors are in turn converted to the corresponding SAM analogues, which are considered to be a key for the development of bioorthogonal systems. In addition to purified enzymes, the bicyclic system can also be used with crude lysates highlighting its broad biocatalytic applicability. date: 2021-03-22 date_type: published publisher: Royal Society of Chemistry (RSC) official_url: https://doi.org/10.1039/d1cb00033k oa_status: green full_text_type: pub language: eng primo: open primo_central: open_green verified: verified_manual elements_id: 1855787 doi: 10.1039/d1cb00033k lyricists_name: Hailes, Helen lyricists_id: HCHAI24 actors_name: Kalinowski, Damian actors_id: DKALI47 actors_role: owner full_text_status: public publication: RSC Chemical Biology citation: Popadić, D; Mhaindarkar, D; Dang Thai, MHN; Hailes, HC; Mordhorst, S; Andexer, JN; (2021) A bicyclic S-adenosylmethionine regeneration system applicable with different nucleosides or nucleotides as cofactor building blocks. RSC Chemical Biology 10.1039/d1cb00033k <https://doi.org/10.1039/d1cb00033k>. (In press). Green open access document_url: https://discovery.ucl.ac.uk/id/eprint/10125446/1/Hailes_A%20bicyclic%20S-adenosylmethionine%20regeneration%20system%20applicable%20with%20different%20nucleosides%20or%20nucleotides%20as%20cofactor%20building%20block_AOP.pdf