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Paul, C.E. (author), Hanefeld, U. (author), Hollmann, F. (author), Qu, Ge (author), Yuan, Bo (author), Sun, Zhoutong (author)Contemporary Biocatalysis heavily relies on enzyme engineering as natural enzymes frequently lack the requisite attributes for effective organic synthesis. The inherent limitations in stability, catalytic activity, and selectivity of wild-type enzymes often hinder their suitability for chemical synthesis. Over the past 25 years, there has...journal article 2024
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Abdelraheem, E.M.M. (author), Kuijpers, Robin (author), Hagedoorn, P.L. (author), Hollmann, F. (author), Hanefeld, U. (author)This study presents a three-step one pot enzymatic cascade for the synthesis of a δ-lactone. Utilising acetaldehyde, combining 2-deoxyribose-5-phosphate aldolase (DERA) with an alcohol dehydrogenase (ADH) and a cofactor regeneration system this δ-lactone is synthesised with the same stereochemistry as the statin side chain precursor. The...journal article 2024
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Coloma Hurel, J.L. (author), Hagedoorn, P.L. (author), Bento, Isabel (author), Hanefeld, U. (author)Granulicella tundricola hydroxynitrile lyase (GtHNL) is a manganese dependent cupin that catalyzes the enantioselective synthesis of cyanohydrins. The analysis of its active site shows high similarity with the active site of cupin Tm1459 from Thermotoga maritima, an enzyme that catalyzes the oxidative cleavage of styrene derivatives. GtHNL ...journal article 2023
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Stradomska, Dominika (author), Świętochowska, Daria (author), Kubica, Robert (author), Hanefeld, U. (author), Szymańska, Katarzyna (author)Effectiveness of catalytic processes using heterogeneous biocatalysts depends not only on the activity of the enzyme, but also on the efficiency of the used reactor. In this paper, we present a novel design of a basket reactor with a stationary catalyst bed (StatBioChem). The developed design was compared to a commercially available rotating...journal article 2023
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Abdelraheem, E.M.M. (author), Jockmann, Emely (author), Li, Jianyu (author), Günther, Stefan (author), Andexer, Jennifer N. (author), Hagedoorn, P.L. (author), Hanefeld, U. (author)S-Adenosyl-l-methionine (SAM)-dependent methyltransferases (MTs) are highly chemoselective enzymes grouped in C-, N-, O-, S- and halide MTs, depending on the (hetero) atom that acts as the methyl group acceptor. So far, OMTs present the largest group, including many well investigated candidates. The catechol OMT from mammals such as from...journal article 2023
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Kowalczykiewicz, Daria (author), Przypis, Marta (author), Mestrom, L. (author), Kumpf, Antje (author), Tischler, Dirk (author), Hagedoorn, P.L. (author), Hanefeld, U. (author), Jarzębski, Andrzej (author), Szymańska, Katarzyna (author)Here, we present a two-step continuous flow enzymatic synthesis process in monolithic microreactors using basic sugars as substrates. In the first step UDP-glucose pyrophosphorylase (TaGalU) catalyses the synthesis of uridine-diphosphate-glucose (UDP-Glc) using uridine triphosphate (UTP) and glucose-1-phosphate (Glc-1-P). This is followed by...journal article 2022
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Stradomska, Dominika (author), Coloma Hurel, J.L. (author), Hanefeld, U. (author), Szymańska, Katarzyna (author)Enantiomerically pure cyanohydrins are of great importance in the chemical and pharmaceutical industries. Their synthesis is possible through the use of highly selective hydroxynitrile lyases. In this work, an R-selective hydroxynitrile lyase (AtHNL) from Arabidopsis thaliana was immobilized inside a silica microreactor with a tortuous and...journal article 2022
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Schober, Lukas (author), Tonin, F. (author), Hanefeld, U. (author), Gröger, Harald (author)A sequential-type as well as a tandem-type chemoenzymatic flow cascade combining an organocatalytic aldol reaction and a biocatalytic reduction to form stereoselectively a 1,3-diol with two stereogenic centers were developed. Initially, a comprehensive screening of 24 alcohol dehydrogenases was carried out and the identified candidates were...journal article 2022
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Marsden, S.R. (author), Wijma, Hein J. (author), Mohr, M.K.F. (author), Justo, Inês (author), Hagedoorn, P.L. (author), Laustsen, Jesper (author), Mestrom, L. (author), McMillan, D.G.G. (author), Hanefeld, U. (author)Regulation of enzyme activity is vital for living organisms. In metalloenzymes, far-reaching rearrangements of the protein scaffold are generally required to tune the metal cofactor's properties by allosteric regulation. Here structural analysis of hydroxyketoacid aldolase from Sphingomonas wittichii RW1 (SwHKA) revealed a dynamic movement of...journal article 2022
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Varela, Romina Fernández (author), Valino, Ana Laura (author), Abdelraheem, E.M.M. (author), Medici, R. (author), Sayé, Melisa (author), Pereira, Claudio A. (author), Hagedoorn, P.L. (author), Hanefeld, U. (author), Iribarren, Adolfo (author), Lewkowicz, Elizabeth (author)In nature 2-deoxy-D-ribose-5-phosphate aldolase (DERA) catalyses the reversible formation of 2-deoxyribose 5-phosphate from D-glyceraldehyde 3-phosphate and acetaldehyde. In addition, this enzyme can use acetaldehyde as the sole substrate, resulting in a tandem aldol reaction, yielding 2,4,6-trideoxy-D-erythro-hexapyranose, which...journal article 2022
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Coloma Hurel, J.L. (author), Teeuwisse, Lidwien (author), Afendi, Muhammad (author), Hagedoorn, P.L. (author), Hanefeld, U. (author)Granulicella tundricola hydroxynitrile lyase (GtHNL) catalyses the synthesis of chiral (R)‐ cyanohydrins and (R)‐β‐nitro alcohols. The triple variant GtHNL‐A40H/V42T/Q110H (GtHNL‐3V) was immobilised on Celite R‐633 and used in monophasic MTBE saturated with 100 mM KPi buffer pH 7 for the synthesis of (R)‐2‐nitro‐1‐phenylethanol (NPE) in batch...journal article 2022
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Coloma Hurel, J.L. (author), Guiavarc'h, Y.P. (author), Hagedoorn, P.L. (author), Hanefeld, U. (author)The merger of enzyme immobilisation and flow chemistry has attracted the attention of the scientific community during recent years. Immobilisation enhances enzyme stability and enables recycling, flow chemistry allows process intensification. Their combination is desirable for the development of more efficient and environmentally friendly...journal article 2021
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Bouchaut, B.F.H.J. (author), Asveld, L. (author), Hanefeld, U. (author), Vlierboom, A.C.E. (author)Although both the Inherent Safety Principles (ISPs) and the Safe-by-Design (SbD) approach revolve around the central value of safety, they have a slightly different focus in terms of developing add-on features or considering initial design choices. This paper examines the differences between these approaches and analyses which approach is more...journal article 2021
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Marsden, S.R. (author), Mestrom, L. (author), Wijma, Hein J. (author), Noordam, Sander J. (author), McMillan, D.G.G. (author), Hanefeld, U. (author)Diastereomers are characterised by an intrinsic energy difference, and thermodynamics dictate their distribution within a dynamic equilibrium. The characteristic mechanistic reversibility and non-ideal stereoselectivity of catalysts therefore simultaneously promote both synthesis and epimerization of products during the formation of...journal article 2021
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de Martino, M. Teresa (author), Tonin, F. (author), Bloemendal, Victor R.L.J. (author), Hanefeld, U. (author), Rutjes, Floris P.J.T. (author), van Hest, Jan C.M. (author)A new immobilization strategy using compartmentalized nanoreactors is herein reported for two biocatalytic processes: (1)N-acetylneuraminate lyase (NAL) is internalized in NAL-c-CLEnAs and used in a continuous flow aldol condensation ofN-acetyl-d-mannosamine with sodium pyruvate toN-acetylneuraminic acid; (2) two hydroxysteroid dehydrogenases...journal article 2021
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Coloma Hurel, J.L. (author), Guiavarc'h, Y.P. (author), Hagedoorn, P.L. (author), Hanefeld, U. (author)Granulicella tundricolahydroxynitrile lyase (GtHNL) is a manganese dependent cupin which catalyses the enantioselective synthesis of (R)-cyanohydrins. TheGtHNL triple variant A40H/V42T/Q110H, previously reported to exhibit a high activity and stability, was immobilised on Celite R-633 by adsorption. The synthesis of (R)-mandelonitrile...journal article 2020
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van Amelrooy, E.F. (author), Schut, H. (author), Egger, Werner (author), Dickmann, Marcel (author), Hugenschmidt, Christoph (author), Mallée, L. (author), Hanefeld, U. (author), McMillan, D.G.G. (author), Eijt, S.W.H. (author)A new method is developed to produce mesoporous titania thin films at room temperature using the enzyme papain in a dip-coating procedure, providing low-cost titania films in a sustainable manner. Quartz crystal microbalance, positron annihilation Doppler broadening and lifetime spectroscopy, scanning electron microscopy, and X-ray...journal article 2020
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Tonin, F. (author), Marti, E. (author), Arends, I.W.C.E. (author), Hanefeld, U. (author)The specific oxidation of 12α-OH group of hydroxysteroids is required for the preparation of cheno-and ursodeoxycholic acid (CDCA and UDCA, respectively). The C<sub>12</sub> oxidation of hydroxysteroids into their 12-oxo derivatives can selectively be performed by employing 12α-hydroxysteroid dehydrogenases. These enzymes use NAD(P)<sup>+<...journal article 2020
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Bracco Garcia, M.P. (author), van Midden, Nelleke (author), Arango, Epifanía (author), Torrelo, Guzman (author), Ferrario, Valerio (author), Gardossi, Lucia (author), Hanefeld, U. (author)The question of how to distinguish between lipases and esterases is about as old as the definition of the subclassification is. Many different criteria have been proposed to this end, all indicative but not decisive. Here, the activity of lipases in dry organic solvents as a criterion is probed on a minimal α/β hydrolase fold enzyme, the...journal article 2020
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De Martino, M. Teresa (author), Tonin, F. (author), Yewdall, N. Amy (author), Abdelghani, Mona (author), Williams, David S. (author), Hanefeld, U. (author), Rutjes, Floris P.J.T. (author), Abdelmohsen, Loai K.E.A. (author), Van Hest, Jan C.M. (author)Nano-sized enzyme aggregates, which preserve their catalytic activity are of great interest for flow processes, as these catalytic species show minimal diffusional issues, and are still sizeable enough to be effectively separated from the formed product. The realization of such catalysts is however far from trivial. The stable formation of a...journal article 2020