Article
Chemistry, Physical
Scott J. Novick, Nikki Dellas, Ravi Garcia, Charlene Ching, Abigail Bautista, David Homan, Oscar Alvizo, David Entwistle, Florian Kleinbeck, Thierry Schlama, Thomas Ruch
Summary: The engineered amine transaminase CDX-043 showed high productivity and selectivity for the desired diastereoisomer, allowing for efficient and cost-effective large-scale production of sacubitril. The structural analysis of enzyme variants provided insight into how the mutations affected activity and selectivity. The variant exhibited high process robustness and could tolerate various reaction conditions, making it a promising candidate for industrial applications.
Article
Biochemistry & Molecular Biology
Rakesh Saroay, Gheorghe-Doru Roiban, Lona M. Alkhalaf, Gregory L. Challis
Summary: The variant R59C of TxtE has a broader substrate scope, allowing it to nitrate a range of tryptamine analogues. This presents a new direction for developing TxtE into a useful aromatic nitration biocatalyst.
Article
Chemistry, Applied
Jesse T. C. Brown, Ryan M. Phelan
Summary: Chiral amines are important in the synthesis of pharmaceutical and agrochemical compounds, and amine transaminases have emerged as efficient and stereoselective catalysts. However, the compatibility between the transaminase and amine donors is a concern. In this study, we compare different amine donors and identify those that are compatible with various amine transaminases, providing insights into the preferences of engineered and wild-type enzymes.
ORGANIC PROCESS RESEARCH & DEVELOPMENT
(2023)
Article
Biochemistry & Molecular Biology
Iris S. Teixeira, Andre B. Farias, Bruno A. C. Horta, Humberto M. S. Milagre, Rodrigo O. M. A. de Souza, Uwe T. Bornscheuer, Cintia D. F. Milagre
Summary: This study investigated the influence of an alpha,beta-unsaturated system on the enzymatic reductive amination of ketones catalyzed by amine transaminases. It was found that the wild-type enzymes' reactivity was dramatically reduced by the presence of the double bond, while an engineered variant still maintained high conversion rates and optical purity.
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES
(2022)
Article
Chemistry, Physical
Fulong Li, Yan Du, Youxiang Liang, Yuwen Wei, Yukun Zheng, Huimin Yu
Summary: This study redesigned a trans-aminase enzyme to enhance its catalytic efficiency towards bulky N-heterocyclic compounds. A variant with significant activity enhancement was obtained by fine-tuning the substrate-binding pocket with a simplified amino acid alphabet. The most active variant exhibited 1064-fold greater catalytic efficiency and improved activity towards a series of cyclic and bulky heterocyclic ketones. It also showed successful gram-scale synthesis of a chiral N-heterocyclic amine.
Article
Chemistry, Multidisciplinary
Chenghai Sun, Bao-Di Ma, Guangjun Li, Wenya Tian, Lu Yang, Haidong Peng, Zhi Lin, Zixin Deng, Xu-Dong Kong, Xudong Qu
Summary: Through genome mining and study of the sequence-product relationship, we identified three key residues (F387, F388, and E73) that play pivotal roles in selecting different diketopiperazine (DKP) substrates. Engineering these residues in Nas(F5053) greatly expanded its substrate specificity, enabling the biosynthesis of 12 self-dimerized and at least 81 cross-dimerized HTDKPs. Structural and molecular dynamics analysis of F387G and E73S revealed that they control substrate specificity by reducing steric hindrance and regulating substrate tunnels.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Chemistry, Applied
Jeffrey T. Kohrt, Peter H. Dorff, Michael Burns, Chewah Lee, Steven O'Neil, Robert J. Maguire, Rajesh Kumar, Melissa Wagenaar, Loren Price, Manjinder S. Lall
Summary: In this study, a continuous three-step flow process was used to scale up the formation and reduction of an azide intermediate, and a transaminase was utilized to prepare the desired enantiomer efficiently.
ORGANIC PROCESS RESEARCH & DEVELOPMENT
(2022)
Review
Chemistry, Physical
Sam Mathew, Dominik Renn, Magnus Rueping
Summary: Amine transaminases play a crucial role in the synthesis of chiral amines, which are essential building blocks in pharmaceuticals and fine chemicals. Advances in enzyme discovery, process engineering, and protein engineering have significantly enhanced the use of transaminases in organic synthesis. Enzymatic cascade engineering has emerged as an efficient and environmentally friendly approach for chiral amine synthesis, allowing the generation of valuable products from inexpensive starting materials. This review focuses on the utilization of amine transaminases in enzymatic and chemoenzymatic cascades for chiral amine synthesis.
Article
Chemistry, Physical
Laurine Ducrot, Megan Bennett, Gwenaelle Andre-Leroux, Eddy Elisee, Sacha Marynberg, Aurelie Fossey-Jouenne, Anne Zaparucha, Gideon Grogan, Carine Vergne-Vaxelaire
Summary: Inspired by the diversity of residues, mutations F140A and W145A were introduced in CfusAmDH, leading to improved conversion and enantioselectivity for certain substrates. The mutant CfusAmDH-W145A showed synthetic potential and crystal structure analysis provided insights into cofactor and substrate specificity, as well as structural dynamics of this enzyme family.
Article
Biotechnology & Applied Microbiology
Feng Cheng, Ming-You Li, Dian-Ju Wei, Xiao-Jian Zhang, Dong-Xu Jia, Zhi-Qiang Liu, Yu-Guo Zheng
Summary: Biocatalysis in high-concentration organic solvents presents advantages but challenges, with modifications in enzyme gate region successfully improving enzyme activity and DMSO resistance, thus enhancing efficiency in handling hydrophobic substrates.
BIOTECHNOLOGY AND BIOENGINEERING
(2022)
Article
Chemistry, Physical
Moritz Voss, Sean Hueppi, Daniela Schaub, Takahiro Hayashi, Mathieu Ligibel, Emine Sager, Kirsten Schroer, Radka Snajdrova, Rebecca Buller
Summary: Enzymatic late-stage diversification of small molecules is a rapid method for generating diversity in compound libraries for drug discovery. The Fe(II)/alpha-ketoglutarate-dependent halogenase enzyme family has become of interest for its ability to selectively halogenate unactivated C(sp(3))-H bonds. In this study, two engineered variants of the halogenase WelO5* were developed for the racemic resolution of a mixture of stereoisomers generated during the synthesis of a bioactive martinelline-derived fragment.
Article
Biochemistry & Molecular Biology
Chao Xiang, Shuke Wu, Uwe T. Bornscheuer
Summary: By performing directed evolution, a new enzyme variant VF-8M-E with significantly increased specific activity was obtained, which was successfully utilized to produce the key chiral amine intermediate in enantiopure form for Apremilast, representing a new enzymatic approach towards the synthesis of Apremilast.
BIOORGANIC & MEDICINAL CHEMISTRY
(2021)
Article
Chemistry, Physical
Nils Janson, Tobias Heinks, Tobias Beuel, Sarfaraz Alam, Matthias Hoehne, Uwe T. Bornscheuer, Gabriele Fischer von Mollard, Norbert Sewald
Summary: The aldehyde tag is used for selective labeling of proteins and preparing antibody-drug conjugates, or immobilizing enzymes or antibodies. A new solid support, modified agarose with an aryl substituted pyrazolone, allows site-selective and efficient immobilization of aldehyde-containing molecules under physiological pH without the need for additives or catalysts. Compared to reductive amination, the modified agarose achieves higher loadings and activities in various buffers at different concentrations and temperatures.
Article
Chemistry, Applied
Mathias Pickl, Markus Ebner, Samantha Gittings, Pere Clapes, Wolfgang Kroutil
Summary: Although optical pure amino alcohols are highly demanded, their synthesis remains challenging due to the requirement of elaborate protection strategies. This study presents a multi-enzymatic methodology that allows the synthesis of enantioenriched 1,3-amino alcohols from commodity chemicals. By utilizing a Type I aldolase and enzymatic transamination, both diastereomers of the 1,3-amino alcohol motif were accessible, resulting in the desired amino alcohols with high enantioselectivity.
ADVANCED SYNTHESIS & CATALYSIS
(2023)
Article
Chemistry, Multidisciplinary
Allwin D. McDonald, Samantha K. Bruffy, Aadhishre T. Kasat, Andrew R. Buller
Summary: In this study, we used a substrate multiplexed screening approach to improve the substrate scope overlap in biocatalytic cascades for the synthesis of chiral 1,2-amino alcohols. By engineering the activity of enzymes, we achieved selective and efficient synthesis of enantiopure products in a one-pot cascade reaction.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2022)
Article
Chemistry, Multidisciplinary
Ramapada Dolai, Rahul Kumar, Benedict J. Elvers, Pradeep Kumar Pal, Benson Joseph, Rina Sikari, Mithilesh Kumar Nayak, Avijit Maiti, Tejender Singh, Nicolas Chrysochos, Arumugam Jayaraman, Ivo Krummenacher, Jagannath Mondal, U. Deva Priyakumar, Holger Braunschweig, Cem B. Yildiz, Carola Schulzke, Anukul Jana
Summary: A new type of carbodicarbene (CDC) with two different classes of carbenes has been reported, which shows promising potential as organic electron donors and n-dopants. These compounds exhibit notable redox properties and can form carbon-carbon sigma bonds through reactions with [NO][SbF6].
CHEMISTRY-A EUROPEAN JOURNAL
(2023)
Article
Chemistry, Physical
Tobias Heinks, Nicolai Montua, Michelle Teune, Jan Liedtke, Matthias Hoehne, Uwe T. Bornscheuer, Gabriele Fischer von Mollard
Summary: This study aims to compare the stability of soluble and immobilized transaminases from different species. Immobilization of transaminases enabled their reusability for at least 10 cycles and maintained full or high activity. Upscaled kinetic resolutions demonstrated high conversion, maintained enantioselectivity, and high product yields, showing their applicability.
Article
Chemistry, Multidisciplinary
Yu-Fei Ao, Shuxin Pei, Chao Xiang, Marian J. Menke, Lin Shen, Chenghai Sun, Mark Dorr, Stefan Born, Matthias Hohne, Uwe T. Bornscheuer
Summary: We created variants of ATA from Ruegeria sp. (3FCR) with improved catalytic activity and reversed stereoselectivity, and collected a high-quality dataset. We designed a modified one-hot code to describe the effects of substrates and residues within ATAs. Finally, we built a predictor for catalytic activity and stereoselectivity, and applied it for the data-driven design of optimized variants which showed improved activity. We also demonstrated that the model can predict the catalytic activity for ATA variants of another origin by retraining with additional data.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Biotechnology & Applied Microbiology
Stefan Brott, Ki Hyun Nam, Francois Thomas, Theresa Dutschei, Lukas Reisky, Maike Behrens, Hanna C. C. Grimm, Gurvan Michel, Thomas Schweder, Uwe T. T. Bornscheuer
Summary: Marine algae produce complex polysaccharides that can be degraded by marine heterotrophic bacteria using carbohydrate-active enzymes. The degradation of the red algal polysaccharide porphyran involves oxidative demethylation, catalyzed by cytochrome P450 monooxygenase and its redox partners. Genes encoding zinc-dependent alcohol dehydrogenases (ADHs) were found near the genes for key enzymes in this process. The physiological role of these marine ADHs was investigated and it was found that they are required for the utilization of a specific substrate, G6Me.
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
(2023)
Article
Biochemistry & Molecular Biology
Henrik Terholsen, Kamela Myrtollari, Mirna Larva, Christina Moeller, Andreas Taden, Robert Kourist, Uwe T. Bornscheuer, Daniel Kracher
Summary: Biocatalytic decarboxylation of hydroxycinnamic acids yields phenolic styrenes, which are important precursors for antioxidants, epoxy coatings, adhesives, and other polymeric materials. This study presents two robust and sensitive assays based on photometry and fluorimetry to track decarboxylation reactions with high sensitivity while avoiding product extraction and long analysis times. The optimized assay procedures were used to measure BsPAD activity and determine the kinetic constants of the purified enzyme for different hydroxycinnamic acids, showing substrate inhibition for caffeic acid.
Article
Chemistry, Organic
Siva Sankar Murthy Bandaru, Carola Schulzke
Summary: The novel organic salt was synthesized through a Lewis acid and Lewis-base reaction and analyzed by various spectroscopic and crystallographic techniques.
Article
Chemistry, Inorganic & Nuclear
Siva Sankar Murthy Bandaru, Claudia Schindler, Felix Wenzek, Carola Schulzke
Summary: Several synthetic cross-coupling procedures were evaluated for tethering pyrazine to dithiocarbonates. The resulting compounds can serve as pro-ligands and can be converted into ene-dithiolate ligand systems resembling molybdopterin by removing the C=O moiety. The coupling of 2-iodopyrazine with stannylated dithiocarbonate units mediated by copper(I)-thiophene-2-carboxylate in stoichiometric equivalents proved to be the most efficient and reliable method for synthesizing the targeted compounds.
Article
Chemistry, Multidisciplinary
Priyanka Saha, Nicolas Chrysochos, Benedict J. Elvers, Sebastian Paetsch, Sk Imraj Uddin, Ivo Krummenacher, Muneshwar Nandeshwar, Anshika Mishra, Karthik V. Raman, Gopalan Rajaraman, Ganesan Prabusankar, Holger Braunschweig, Prince Ravat, Carola Schulzke, Anukul Jana
Summary: A modular approach for the synthesis of isolable crystalline Schlenk hydrocarbon diradicals from m-phenylene bridged electron-rich bis-triazaalkenes as synthons is demonstrated. The diradical nature and triplet electronic structure of the synthesized compounds are confirmed through EPR spectroscopy and computational analysis. The potential of the modular methodology is further illustrated through the utilization of 4,6-dimethyl-m-phenylene as a coupling unit.
ANGEWANDTE CHEMIE-INTERNATIONAL EDITION
(2023)
Article
Biochemistry & Molecular Biology
Tobias Heinks, Simon Koopmeiners, Nicolai Montua, Norbert Sewald, Matthias Hoehne, Uwe T. Bornscheuer, Gabriele Fischer von Mollard
Summary: An enzyme cascade consisting of l-amino acid oxidase (hcLAAO4) and catalase (hCAT) was immobilized together with the (S)-selective amine transaminase (ATA-Vfl) to achieve higher reaction rates and more efficient H2O2-removal. The co-immobilized enzyme cascade showed good stability and high enantiomeric purity in preparative kinetic resolutions. However, further recycling was limited due to the instability of ATA-Vfl. The use of an engineered ATA-Vfl-8M in the immobilized enzyme cascade allowed a significant reduction in the amount of co-substrate required.
Article
Biochemistry & Molecular Biology
Yannik Brack, Chenghai Sun, Dong Yi, Uwe T. Bornscheuer
Summary: Researchers have discovered new substrate switch motifs in aromatic ammonia lyases and successfully altered the enzyme's substrate specificity. In particular, introducing previously unknown residue combinations enabled a switch from tyrosine to phenylalanine in tyrosine ammonia lyases.
Article
Chemistry, Physical
Andreas Kunzendorf, Bastian Zirpel, Lars Milke, Jakob P. Ley, Uwe T. Bornscheuer
Summary: Directed evolution of ZgOMT resulted in highly regioselective biocatalysts for the preparation of hesperetin dihydrochalcone and related compounds, providing an attractive synthesis route for para-methylation of catechol scaffolds. These enzyme variants are challenging to perform with high regioselectivity using wild-type O-methyltransferases.
Article
Chemistry, Multidisciplinary
Mithilesh Kumar Nayak, Benedict J. Elvers, Debdeep Mandal, Ayan Das, Raghunathan Ramakrishnan, Kaustubh R. Mote, Carola Schulzke, Cem Burak Yildiz, Anukul Jana
Summary: The reduction of 2-H-substituted pyrrolinium cations results in dimerisation or H-abstracted products, depending on the N-substituents. The dimerised 2,2'-bipyrrolidine derivatives with a central carbon-carbon single bond are stable and can be oxidised chemically or electrochemically. These dimers, which are air and moisture-stable, can serve as a source of two electrons in various chemical transformations.
CHEMICAL COMMUNICATIONS
(2023)
Article
Chemistry, Multidisciplinary
Nicolas Chrysochos, Sebastian Paetsch, Benedict J. Elvers, Ivo Krummenacher, Muneshwar Nandeshwar, Ganesan Prabusankar, Holger Braunschweig, Carola Schulzke, Prince Ravat, Anukul Jana
Summary: The synthesis of a twisted, push-pull, and electron-rich alkene is reported, providing a unique combination for alkene compounds. This newly synthesized alkene motif was used as a donor for the synthesis of a boron-containing pi-conjugated compound with zwitterionic properties through intramolecular charge transfer.
CHEMICAL COMMUNICATIONS
(2023)
