Machine-Directed Evolution of an Imine Reductase for Activity and Stereoselectivity

Eric, J Gilbert; Siirola, Elina; Moore, Charles M.; Kummer, Arkadij; Stoeckli, Markus; Faller, Michael; Bouquet, Caroline; Eggimann, Fabian; Ligibel, Mathieu; Huynh, Dan; Cutler, Geoffrey J.; Siegrist, Luca; Lewis, Richard A.; Acker, Anne-Christine; Freund, Ernst; Koch, Elke; Vogel, Markus; Schlingensiepen, Holger; Oakeley, Edward J.; Šnajdrová, Radka

doi:10.1021/acscatal.1c02786

Cited by 69 publications

(72 citation statements)

References 51 publications

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“…The development of IRED-catalyzed asymmetric reductive aminations from laboratory discovery to industrial scale Scheme 21 Synthesis of ZPL389 54b using enzyme IRED88 Q194L/ S220T/H230Y evolved by Novartis. 81 Scheme 22 Reductive amination reactions investigated by Pfizer. 40,82,83 application illustrates the rapid progress that can now be made with biocatalytic reactions once useful transformations have been identied.…”

Section: Discussionmentioning

confidence: 99%

“…Researchers at Novartis have combined genome mining with recent advances in machine learning for the evolution of an IRED for the stereoselective reductive amination of ketone 54 with methylamine b to molecule ZPL389 54b ( Scheme 21 ), an H4 receptor antagonist used in the treatment of atopic dermatitis. 81 Library screening plus genome mining was used to identify the enzyme ‘IRED88’ as being able to catalyze the amination of 54 to the product 54b with 70% conversion, and to the required ( R )-enantiomer, but with only 30% e.e. A deep mutation scanning (DMS) approach was taken first, in which a library of mutants composed of all amino acid positions mutated to the 19 alternative residues was screened for activity by mass spectrometry.…”

Section: Scalable Reactions and Industrial Examplesmentioning

confidence: 99%

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Reductive aminations by imine reductases: from milligrams to tons

et al. 2022

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Section: Discussionmentioning

confidence: 99%

Section: Scalable Reactions and Industrial Examplesmentioning

confidence: 99%

Reductive aminations by imine reductases: from milligrams to tons

et al. 2022

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“…One mutant was suggested to enable a selectivity factor of E =250, which was experimentally validated ( E =253) [35b] . Further examples of applying ML to enzyme stereoselectivity was reported thereafter, which involved an imine reductase for producing chiral amines of high value in the pharmaceutical sector, [36a] and a Fe/ß‐ketoglutarate dependent halogenase for the functionalization of pharmaceutically important soraphens [36b] . For the future, many more examples of ML for predicting stereoselective mutants as catalysts in organic chemistry can be anticipated [35c] …”

Section: The Crucial Role Of Methodology Development In Directed Evol...mentioning

confidence: 99%

Witnessing the Birth of Directed Evolution of Stereoselective Enzymes as Catalysts in Organic Chemistry

Reetz

2022

Adv Synth Catal

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This invited essay outlines how the idea of directed evolution of stereoselective enzymes was born and implemented experimentally at the Max-Planck-Institut für Kohlenforschung in Mülheim/ Germany during the period 1994-1998, a time when Andreas Pfaltz was present in the Institute. As the new and sole director of the MPI, I initiated new research projects, and also started to restructure the Institute with the establishment of five departments, all dedicated to some form of catalysis and to be led by five independent directors. Andreas Pfaltz was the first director whom I hired, heading the Department of Homogeneous Catalysis. During his stay in Mülheim until 1998, the Pfaltz group invented effective chiral ligands for a number of particularly challenging enantioselective transformations. During this period, Andreas Pfaltz witnessed the birth and development of directed evolution of stereoselective enzymes as a new research direction in the Reetz group. Indeed, he was one of the very few organic chemists who realized at the time that a door to a novel research area had been opened. The widespread application of enzymes was hampered for decades due to limited enantio-, diastereo-, and regioselectivity, which was the reason why organic chemists were not interested in biocatalysis. This attitude slowly changed with the advent of directed evolution of stereoselective enzymes in 1997, in a publication from the Reetz group. Methodology development with emphasis on stereo-and regioselectivity as well as activity followed, the techniques and strategies becoming more and more rational. Today, semirational approaches and so-called rational enzyme design have merged, as evidenced, inter alia, by the development of focused rational iterative site-specific mutagenesis (FRISM). The toolbox of organic chemists now includes enzymes, primarily because the possibility of controlling stereoselectivity by protein engineering has ensured reliability when facing synthetic challenges. 1. Introductory Remarks 2. The Birth of Directed Evolution of Stereoselective Enzymes 3. The Crucial Role of Methodology Development in Directed Evolution of Selective Enzymes as Catalysts in Organic Chemistry 4. Final Message

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“…Finally, researchers at Novartis have recently used the IRED‐mediated reductive amination reaction to access the H4 receptor antagonist ZPL389 to compare machine‐learning guided directed evolution with classical approaches such as deep mutational scanning (DMS) and error‐prone PCR (epPCR) [210] . The best performing variant obtained via DMS and epPCR was used at a gram scale obtaining the desired product in 72 % isolated yield and >99 % ee .…”

Section: Biocatalytic C−n Bond Formation Via Reductive Aminationmentioning

confidence: 99%

New Trends and Future Opportunities in the Enzymatic Formation of C−C, C−N, and C−O bonds

et al. 2021

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Organic chemistry provides society with fundamental products we use daily. Concerns about the impact that the chemical industry has over the environment is propelling major changes in the way we manufacture chemicals. Biocatalysis offers an alternative to other synthetic approaches as it employs enzymes, Nature's catalysts, to carry out chemical transformations. Enzymes are biodegradable, come from renewable sources, operate under mild reaction conditions, and display high selectivities in the processes they catalyse. As a highly multidisciplinary field, biocatalysis benefits from advances in different areas, and developments in the fields of molecular biology, bioinformatics, and chemical engineering have accelerated the extension of the range of available transformations (E. L. Bell et al., Nat. Rev. Meth. Prim. 2021, 1, 1-21). Recently, we surveyed advances in the expansion of the scope of biocatalysis via enzyme discovery and protein engineering (J. R. Marshall et al., Tetrahedron 2021, 82, 131926). Herein, we focus on novel enzymes currently available to the broad synthetic community for the construction of new CÀ C, CÀ N and CÀ O bonds, with the purpose of providing the non-specialist with new and alternative tools for chiral and sustainable chemical synthesis.

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Machine-Directed Evolution of an Imine Reductase for Activity and Stereoselectivity

Cited by 69 publications

References 51 publications

Reductive aminations by imine reductases: from milligrams to tons

Reductive aminations by imine reductases: from milligrams to tons

Witnessing the Birth of Directed Evolution of Stereoselective Enzymes as Catalysts in Organic Chemistry

New Trends and Future Opportunities in the Enzymatic Formation of C−C, C−N, and C−O bonds

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