5-Thiohistidine N-Acetyltransferase from Proteiniphilum Saccharofermentans Cangsong Liao, David Lim, Gladwin Suryatin Alim, Florian P. Seebeck Chembiochem, 2025 Ovothiol A is a 5‐thiohistidine derivative biosynthesized by a broad range of prokaryotic and eukaryotic organisms. Its redox‐active mercaptoimidazole side chain is believed to protect cells from oxidative stress. The three enzymes that produce ovothiol A from histidine, cysteine, and S‐adenosyl methionine have been identified and characterized. In contrast, no enzymes are known that produce other 5‐thiohistidine derivatives. Here, a small family of acetyl‐coenzyme A‐dependent transferases is described that produce N‐acetyl‐5‐thiohistidine. The discovery of these enzymes from Proteiniphilum saccharofermentans and related Bacteroidota provides evidence that the 5‐thiohistidine class may be structurally and functionally more diverse than previously thought.
Biocatalytic Reduction of Heterocyclic Imines in Continuous Flow with Immobilized Enzymes Ana I. Benítez-Mateos, David Lim, David Roura Padrosa, Valentina Marchini, Hao Wu, Frederic Buono, Francesca Paradisi ACS Sustainable Chemistry and Engineering, 2025 Heterocyclic amines are a key structural motif for the synthesis of pharmaceuticals (e.g., antibiotics) as well as pesticides and flavors. In this regard, imine reductases (IREDs) have recently emerged as a highly selective and sustainable alternative for asymmetric reductive amination reactions. Herein, we have applied six IREDs, two of which were newly identified, in the reduction of heterocyclic imines with either a N, S, or O substitution at C-4. Since IREDs are NADPH-dependent enzymes, a commercially available, supported glucose dehydrogenase was added as a cofactor-regenerating system. IREDs were then immobilized on porous microparticles to further improve the efficiency and sustainability of the system. The strategic combination of bioinformatic analysis and immobilization screening resulted in immobilized biocatalysts with 95% retained activity. This enabled the integration of the bienzymatic system into a continuous-flow reactor leading to >90% conversion of 50 mM of the S-heterocyclic amine, 5-methyl-3,6-dihydro-2H-1,4-thiazine, with a residence time of 30 min, and reaching space-time yields up to 14.3 g L–1 h–1. In addition, (S)- or (R)-stereoselectivity of the biocatalytic reduction of the 1,4-disubstituted heterocyclic imines was achieved by using the newly identified IREDs fromGoodfellowiella coeruleoviolaceaandLabilithrix luteola, respectively.
Hybrid Catalytic Systems: Integrating Biocatalysis in the Chemical Space David Lim, Francesca Paradisi ACS Catalysis, 2025 Achieving stereocontrol in chemical transformations remains one of the cornerstones of synthesis. Traditional methods to achieve stereoselectivity are based on organo- or metal catalysts, which may raise environmental concerns. As the global demand to create more renewable processes escalates, there is an increased need to develop procedures that minimize waste buildup and the use of toxic byproducts. Over the past few decades, the use of enzymes for achieving stereocontrol has gained traction. Enzymatic reactions are typically performed under aqueous conditions using benign and often recyclable substrates. These conditions, however, often limit their applicability and have relegated biocatalysis to a niche field for quite a long time. More recently, the integration of biocatalysis with other catalytic approaches has allowed multistep transformations, which would not have been possible with the use of either technique alone. In this perspective, we highlight recent advances in the field that have successfully blended biocatalysis with electro-, photo-, metallo-, and organocatalysis.
Investigations into the Aqueous Synthesis of Selenoglycoconjugates David Lim, Francesca Paradisi European Journal of Organic Chemistry, 2023 Glycosylation of bioactive molecules has been found to improve the pharmacokinetic properties of the parent molecule. However, their syntheses often require tedious protecting group manipulations. The development of methodologies which allow direct aqueous conversion of unprotected sugars into glycosides is therefore an ambitious goal. Herein, we present a broadly applicable method for the synthesis of selenoglycosides in water. We show the ease of direct conjugation of unprotected glycosyl diselenides with various biomolecules, including resorcinol, resveratrol, and the antitumor agent, gimeracil, furnishing the corresponding selenoglycoconjugates in up to 96 % yield. We also demonstrate the oxidatively‐triggered release of the bioactive drug from the sugar, priming these molecules for medicinal applications. The generality and broad substrate scope of this novel transformation will provide access to various selenium‐containing glycomimetics and glycoconjugates.
Glycosyl benzoates as novel substrates for glycosynthases Sabrina de Lorenzo, Lauriane Pillet, David Lim, Francesca Paradisi Organic and Biomolecular Chemistry, 2023 An engineered glycosidase from H. orenii showcases for the first time that glycosyl benzoates are better donors than p-nitrophenyl glucoside in enzyme-catalysed thioglucosylations. The thioglycoside products were formed with conversions up to 94%.
In Vitro Production of Ergothioneine Isotopologues Mariia A. Beliaeva, Reto Burn, David Lim, Florian P. Seebeck Angewandte Chemie International Edition, 2021 Ergothioneine is an emerging component of the redox homeostasis system in human cells and in microbial pathogens, such as Mycobacterium tuberculosis or Burkholderia pseudomallei. Synthesis of stable isotope labelled ergothioneine derivatives may provide important tools for deciphering the distribution, function and metabolism of this compound in vivo. We describe a general protocol for the production of ergothioneine isotopologues with programmable 2H, 15N 13C, 34S and 33S isotope labelling patterns. This enzyme-based approach makes efficient use of commercial isotope reagents and is also directly applicable for the synthesis of radio-isotopologues.
Selenoimidazolium Salts as Supramolecular Reagents for Protein Alkylation David Lim, Xiaojin Wen, Florian P. Seebeck Chembiochem, 2020 Se‐benzyl selenoimidazolium salts are characterized by remarkable alkyl‐transfer potential under physiological conditions. Structure‐activity relationship studies show that selective monoalkylation of primary amines depends on supramolecular interactions between the selenoimidazole leaving group and the target nucleophile. We demonstrate that these reagents can be used for site‐selective and nearly quantitative modification of the model protein lysozyme on Lys13, bypassing the higher intrinsic reactivities of Lys1 and Lys33. These observations introduce selenoimidazolium salts as novel class of electrophiles for selective N‐alkylation of native proteins.
Selenocysteine as a Substrate, an Inhibitor and a Mechanistic Probe for Bacterial and Fungal Iron-Dependent Sulfoxide Synthases Kristina V. Goncharenko, Sebastian Flückiger, Cangsong Liao, David Lim, Anja R. Stampfli, Florian P. Seebeck Chemistry A European Journal, 2020 Sulfoxide synthases are non-heme iron enzymes that participate in the biosynthesis of thiohistidines such as ergothioneine and ovothiol A. The sulfoxide synthase EgtB from Chloracidobacterium thermophilum ( Cth EgtB) catalyzes oxidative coupling between the side chains of N-α-trimethyl histidine (TMH) and cysteine (Cys) in a reaction that entails complete reduction of molecular oxygen, carbon-sulfur (C-S) and sulfur-oxygen (S-O) bond formation and carbon-hydrogen (C-H) bond cleavage. In this report we show that bacterial sulfoxide synthases cannot efficiently turnover selenocysteine (SeCys) as an alternative substrate because the sulfur-to-selenium substitution. In contrast, the sulfoxide synthase from the filamentous fungus Chaetomium thermophilum ( Cth Egt1) catalyzes C-S and C-Se bond formation at almost equal efficiency. We discuss evidence suggesting that this difference emerges from different modes of oxygen-activation.
Total Synthesis and Functional Characterization of Selenoneine David Lim, Dirk Gründemann, Florian P. Seebeck Angewandte Chemie International Edition, 2019 The N- α-trimethyl 2-selenohistidine selenoneine is the selenium isolog of the natural antioxidant ergothioneine. Sulfur-to-selenium substitutions are known to endow proteins and nucleic acids with special activities. In contrast, secondary metabolites that exploit selenium-specific chemistry are rare. Selenoneine therefore provides a unique opportunity to study how natural organoselenides interact with cellular processes. In this report we describe the chemical synthesis of selenoneine and other 2-selenoimidazoles. With synthetic selenoneine at hand we discovered a set of reactivities that distinguish selenoneine from ergothioneine, showing that the two compounds can fill distinct functional niches. Synthetic access to 2-selenoimidazoles should pave the way to explore the pharmaceutical potential and physiological function of this heretofore inaccessible class of compounds.
Hybrid Catalytic Systems: Integrating Biocatalysis in the Chemical Space D Lim, F Paradisi ACS Catalysis 15 (18), 16278-16291 , 2025 2025 Citations: 2
Biocatalytic reduction of heterocyclic imines in continuous flow with immobilized enzymes AI Benítez-Mateos, D Lim, D Roura Padrosa, V Marchini, H Wu, F Buono, ... ACS Sustainable Chemistry & Engineering 13 (13), 5009-5018 , 2025 2025 Citations: 8
5‐Thiohistidine N‐acetyltransferase from Proteiniphilum saccharofermentans C Liao, D Lim, G Suryatin Alim, FP Seebeck ChemBioChem, e202400439 , 2025 2025 Citations: 1
Biocatalysis 101–A Chemist's Guide to Starting Biocatalysis P Díaz‐Kruik, D Lim, F Paradisi Enabling Tools and Techniques for Organic Synthesis: A Practical Guide to … , 2023 2023 Citations: 1
Investigations into the Aqueous Synthesis of Selenoglycoconjugates D Lim, F Paradisi European Journal of Organic Chemistry 26 (34), e202300496 , 2023 2023 Citations: 3
Glycosyl benzoates as novel substrates for glycosynthases S de Lorenzo, L Pillet, D Lim, F Paradisi Organic & biomolecular chemistry 21 (31), 6356-6359 , 2023 2023 Citations: 2
Novel triple mutant of an extremophilic glycosyl hydrolase enables the rapid synthesis of thioglycosides L Pillet, D Lim, N Almulhim, AI Benítez-Mateos, F Paradisi Chemical Communications 58 (86), 12118-12121 , 2022 2022 Citations: 6
In vitro production of ergothioneine isotopologues MA Beliaeva, R Burn, D Lim, FP Seebeck Angewandte Chemie International Edition 60 (10), 5209-5212 , 2021 2021 Citations: 20
Selenoimidazolium salts as supramolecular reagents for protein alkylation D Lim, X Wen, FP Seebeck ChemBioChem 21 (24), 3515-3520 , 2020 2020 Citations: 10
Selenocysteine as a substrate, an inhibitor and a mechanistic probe for bacterial and fungal iron‐dependent sulfoxide synthases KV Goncharenko, S Flückiger, C Liao, D Lim, AR Stampfli, FP Seebeck Chemistry–A European Journal 26 (6), 1328-1334 , 2020 2020 Citations: 12
Total synthesis and functional characterization of selenoneine D Lim, D Gründemann, FP Seebeck Angewandte Chemie International Edition 58 (42), 15026-15030 , 2019 2019 Citations: 42
Structure and Mechanism of Ergothionase from Treponema denticola A Maurer, F Leisinger, D Lim, FP Seebeck Chemistry–A European Journal 25 (44), 10298-10303 , 2019 2019 Citations: 29
Selective Transformations of the Anomeric Centre in Water Using DMC and Derivatives D Lim, AJ Fairbanks Coupling and Decoupling of Diverse Molecular Units in Glycosciences, 109-131 , 2017 2017
Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates SR Alexander, D Lim, Z Amso, MA Brimble, AJ Fairbanks Organic & Biomolecular Chemistry 15 (10), 2152-2156 , 2017 2017 Citations: 44
Selective anomeric acetylation of unprotected sugars in water D Lim, AJ Fairbanks Chemical science 8 (3), 1896-1900 , 2017 2017 Citations: 44
Is there a link between dietary phytoestrogens and reproductive health in men? A meta-analysis of data from the USA and China D Lim, IC Shaw International Journal of Food Science and Technology 51 (1), 23-29 , 2016 2016 Citations: 9
Synthesis of O-Linked Glycopeptides Using Enzymatic Catalysis D Lim University of Canterbury , 2015 2015
Protecting‐group‐free one‐pot synthesis of glycoconjugates directly from reducing sugars D Lim, MA Brimble, R Kowalczyk, AJA Watson, AJ Fairbanks Angewandte Chemie 126 (44), 12101-12105 , 2014 2014 Citations: 105
MOST CITED SCHOLAR PUBLICATIONS
Protecting‐group‐free one‐pot synthesis of glycoconjugates directly from reducing sugars D Lim, MA Brimble, R Kowalczyk, AJA Watson, AJ Fairbanks Angewandte Chemie 126 (44), 12101-12105 , 2014 2014 Citations: 105
Protecting group free synthesis of glycosyl thiols from reducing sugars in water; application to the production of N-glycan glycoconjugates SR Alexander, D Lim, Z Amso, MA Brimble, AJ Fairbanks Organic & Biomolecular Chemistry 15 (10), 2152-2156 , 2017 2017 Citations: 44
Selective anomeric acetylation of unprotected sugars in water D Lim, AJ Fairbanks Chemical science 8 (3), 1896-1900 , 2017 2017 Citations: 44
Total synthesis and functional characterization of selenoneine D Lim, D Gründemann, FP Seebeck Angewandte Chemie International Edition 58 (42), 15026-15030 , 2019 2019 Citations: 42
Structure and Mechanism of Ergothionase from Treponema denticola A Maurer, F Leisinger, D Lim, FP Seebeck Chemistry–A European Journal 25 (44), 10298-10303 , 2019 2019 Citations: 29
In vitro production of ergothioneine isotopologues MA Beliaeva, R Burn, D Lim, FP Seebeck Angewandte Chemie International Edition 60 (10), 5209-5212 , 2021 2021 Citations: 20
Selenocysteine as a substrate, an inhibitor and a mechanistic probe for bacterial and fungal iron‐dependent sulfoxide synthases KV Goncharenko, S Flückiger, C Liao, D Lim, AR Stampfli, FP Seebeck Chemistry–A European Journal 26 (6), 1328-1334 , 2020 2020 Citations: 12
Selenoimidazolium salts as supramolecular reagents for protein alkylation D Lim, X Wen, FP Seebeck ChemBioChem 21 (24), 3515-3520 , 2020 2020 Citations: 10
Is there a link between dietary phytoestrogens and reproductive health in men? A meta-analysis of data from the USA and China D Lim, IC Shaw International Journal of Food Science and Technology 51 (1), 23-29 , 2016 2016 Citations: 9
Biocatalytic reduction of heterocyclic imines in continuous flow with immobilized enzymes AI Benítez-Mateos, D Lim, D Roura Padrosa, V Marchini, H Wu, F Buono, ... ACS Sustainable Chemistry & Engineering 13 (13), 5009-5018 , 2025 2025 Citations: 8
Novel triple mutant of an extremophilic glycosyl hydrolase enables the rapid synthesis of thioglycosides L Pillet, D Lim, N Almulhim, AI Benítez-Mateos, F Paradisi Chemical Communications 58 (86), 12118-12121 , 2022 2022 Citations: 6
Investigations into the Aqueous Synthesis of Selenoglycoconjugates D Lim, F Paradisi European Journal of Organic Chemistry 26 (34), e202300496 , 2023 2023 Citations: 3
Hybrid Catalytic Systems: Integrating Biocatalysis in the Chemical Space D Lim, F Paradisi ACS Catalysis 15 (18), 16278-16291 , 2025 2025 Citations: 2
Glycosyl benzoates as novel substrates for glycosynthases S de Lorenzo, L Pillet, D Lim, F Paradisi Organic & biomolecular chemistry 21 (31), 6356-6359 , 2023 2023 Citations: 2
5‐Thiohistidine N‐acetyltransferase from Proteiniphilum saccharofermentans C Liao, D Lim, G Suryatin Alim, FP Seebeck ChemBioChem, e202400439 , 2025 2025 Citations: 1
Biocatalysis 101–A Chemist's Guide to Starting Biocatalysis P Díaz‐Kruik, D Lim, F Paradisi Enabling Tools and Techniques for Organic Synthesis: A Practical Guide to … , 2023 2023 Citations: 1
Selective Transformations of the Anomeric Centre in Water Using DMC and Derivatives D Lim, AJ Fairbanks Coupling and Decoupling of Diverse Molecular Units in Glycosciences, 109-131 , 2017 2017
Synthesis of O-Linked Glycopeptides Using Enzymatic Catalysis D Lim University of Canterbury , 2015 2015
