Protein Engineering: Methods and Protocols, Methods. Mol. Biol., Vol. 1685

U.T. Bornscheuer / Höhne M. (Eds.) (Humana Press, 2017)

About this book

This volume details basic and advanced protocols for both stages of protein engineering: the library design phase and the identification of improved variants by screening and selection. Chapters focus on enzyme engineering using rational and semi-rational approaches. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, Protein Engineering: Methods and Protocols aims to aid scientists in the planning and performance of their experiments. 

Available here


52. Kollipara M, Matzel P, Bornscheuer U, Höhne M (2022), Activity levels of amine transaminases correlate with active site hydrophobicity, Chem. Ing. Tech. DOI:10.1002/cite.202200062

51. Kollipara M, Matzel P, Sowa M, Brott S, Bornscheuer U, Höhne M (2022), Characterization of proteins from the 3N5M family reveals an operationally stable amine transaminase, Appl. Microbiol. Biotechnol. 106, 5563–5574 here

50. Janson N, Heinks T, Beuel T, Alam S, Höhne M, Bornscheuer UT, Fischer von Mollard G, Sewald N (2022), Efficient site-selective immobilization of aldehyde-tagged peptides and proteins by Knoevenagel ligation, ChemCatChem, 14, e202101485 here

49. Heinks T, Paulus J, Koopmeiners S, Beuel T, Sewald N, Höhne M, Bornscheuer UT, Fischer von Mollard G (2022), Recombinant L-Amino acid oxidase with broad substrate spectrum for co-substrate recycling in (S)-selective transaminase-catalyzed kinetic resolutions, ChemBioChem 23, e2022003 here

48. Yi D, Bayer T, Badenhorst CPS, Wu SK, Doerr M, Höhne M, Bornscheuer UT (2021), Recent trends in biocatalysis. Chem. Soc. Rev. 50, 8003-8049 here

47. Marx L, Rios-Lombardia N, Süss P, Höhne M, Moris F, Gonzales-Sabin J, Berglund P (2020), Chemoenzymatic Synthesis of Sertraline. Eur. J. Org. Chem, 2020, 510-513 here

46. Voss M, Xiang C, Esque J, Nobili A, Menke MJ, André I, Höhne M, Bornscheuer UT (2020), Creation of (R)‐amine transaminase activity within an α‐amino acid transaminase scaffold, ACS Chem. Biol.15, 416-424 here

45. Matassa C, Ormerod D, Bornscheuer UT, Höhne M, Satyawali Y (2020), Three‐liquid‐phase spinning reactor for the transaminase‐catalyzed synthesis and recovery of a chiral amine. ChemCatChem, 12, 1288-1291 here

44. Matassa C,  Romani A, Ormerod D, Bornscheuer UT, Höhne M, Satyawali Y (2020), Jeffamine (R) ED-600: a polyether amine donor for enzymatic transamination in organic solvent/solvent-free medium with membrane-assisted product extraction. J. Chem. Technol. Biotechnol. 95, 604-613 here

43. Höhne M (2019), Engineering imine reductases for industrial applications. Nat. Catal. 2, 841-842 here

42. Sviatenko O, Ríos‐Lombardía N, Morís F, González‐Sabín J, Manideep KV, Merdivan S, Günther S, Süss P, Höhne M (2019), One‐pot synthesis of 4‐aminocyclohexanol isomers by combining a keto reductase and an amine transaminase. ChemCatChem11, 5794-5799 here

41. Schwendenwein D, Ressmann AK, Dörr M, Höhne M, Bornscheuer UT, Mihovilovic MD, Rudroff F, Winkler M (2019), Random mutagenesis‐driven improvement of carboxylate reductase activity using an amino benzamidoxime‐mediated high‐throughput assay. Adv. Synth. Catal. 361, 2544-2549 here

40. Matzel P, Wenske S, Merdivan S, Günther S, Höhne M (2019), Synthesis of β‐chiral amines by dynamic kinetic resolution of α‐branched aldehydes applying imine reductases. ChemCatChem, 11, 4281–4285 here

39. Matassa C, Ormeroda D, Bornscheuer UT, Höhne M, Satyawali Y (2019), Application of novel high molecular weight amine donors in chiral amine synthesis facilitates integrated downstream processing and provides in situ product recovery opportunities.  Process Biochem. 80, 17-25 here

38. Zhang W, Fueyo EF, Hollmann F, Martin LL, Pesic M, Wardenga R, Höhne M, Schmidt S (2019), Combining photo-organo redox- and enzyme catalysis facilitates asymmetric C-H bond functionalization. Eur. J. Org. Chem. 2019, 80-84  here

37. Mystkowska AA, Robb C, Vidal-Melgosa S, Vanni C, Fernandez-Guerra A, Höhne M, Hehemann J-H (2018), Molecular recognition of branched laminarin by a SusD-like glycan binding protein of a marine Bacteroidetes, FEBS J. 285, 4465-4481 here

36. Voss M, Das D, Genz M, Kumar A, Kulkarni N, Kustosz J, Kumar P, Bornscheuer UT, Höhne M (2018), A quantum mechanics based engineering approach to improve transaminases for the conversion of bulky substrates, ACS Catal. 8, 11524–11533.

35. Calvelage S, Dörr M, Höhne M, Bornscheuer UT (2017), A systematic analysis of the substrate scope of (S)- and (R)-selective amine transaminases, Adv. Synth. Catal. 359, 4235-4243 here

34. Matzel P, Krautschick L, Höhne M (2017), Photometric characterization of the reductive amination scope of the imine reductases from Streptomyces tsukubaensis and Streptomyces ipomoeae, ChemBioChem 18, 2022-2027 here

33. Matzel P, Gand M, Höhne M (2017), One-step asymmetric synthesis of (R)- and (S)-rasagiline by reductive amination applying imine reductases, Green Chem. 19, 385-389 here

32. Gand M, Thöle C, Müller H, Brundiek H, Bashiri G, Höhne M (2016), A NADH- accepting imine reductase variant: Immobilization and cofactor regeneration by oxidative deamination, J. Biotechnol. 230, 11-18 here

31. Cuetos A, Steffen-Munsberg F, Mangas Sanchez J, Frese A, Bornscheuer UT, Höhne M, Grogan G (2016), Structural basis for phospholyase activity of a Class III transaminase homolog, ChemBioChem 17, 2308–2311 here

30. Wetzl D, Gand M, Ross A, Müller H, Matzel P, Hanlon SP, Müller M, Wirz B, Höhne M, Iding H (2016), Asymmetric reductive amination of ketones catalyzed by imine reductases, ChemCatChem 8, 2023-2026 here

29. Höhne M, Kabisch J (2016), Brewing painkillers: a yeast cell factory for the production of opioids from sugar, Angew. Chem. Int. Ed. 55, 1248-1250 here; Schmerzmittel brauen: Eine Hefe-Zellfabrik produziert Opiate aus Zucker, Angew. Chem. 128, 1266-1268 here

28. Steffen-Munsberg F, Matzel P, Sowa M, Berglund P, Bornscheuer UT, Höhne M (2016), Bacillus anthracis ω-amino acid:pyruvate transaminase employs a different mechanism for dual substrate recognition than other amine transaminases, Appl. Microb. Biotechnol. 100, 4511–4521 here

27. Kohls H, Anderson M, Dickerhoff J, Weisz K, Córdova A, Berglund P, Brundiek H, Bornscheuer UT, Höhne M (2015), Selective access to all four diastereomers of an 1,3-amino alcohol by combination of a keto reductase- and an amine transaminase-catalysed reaction, Adv. Synth. Catal. 357, 1808-1814 here

26. Skalden L, Peters C, Dickerhoff J, Nobili A, Joosten HJ, Weisz K, Höhne M, Bornscheuer UT (2015) Two subtle amino acid changes in a transaminase substantially enhance or invert enantiopreference in cascade syntheses, ChemBioChem 16, 1041-1045 here

25. Nobili A, Steffen-Munsberg F, Kohls H, Trentin I, Schulzke C, Höhne M, Bornscheuer UT (2015) Engineering the active site of the amine-transaminase from Vibrio fluvialis for the asymmetric synthesis of aryl-alkyl amines and amino alcohols, ChemCatChem 7, 757-760 here

24. Dörr M, Böttcher D, Hummel A, Höhne M, Bornscheuer UT (2015) Eine Roboterplattform für das Hochdurchsatz-Screening von Biokatalysatoren, Biospektrum 21, 230-231 here

23. Steffen-Munsberg F, Vickers C, Kohls H, Land H, Mallin H, Nobili A, Skalden L, van den Bergh T, Joosten HJ, Berglund P, Höhne M, Bornscheuer UT (2015) Bioinformatic analysis of a PLP-dependent enzyme superfamily suitable for biocatalytic applications, Biotechnol. Adv. 33, 566-604 here

22. Skalden L, Thomsen M, Höhne M, Bornscheuer UT, Hinrichs H (2015), Structural and biochemical characterization of the dual substrate recognition of the (R)-selective amine transaminase from Aspergillus fumigatus, FEBS J. 282, 407-415 here

21. Weiss M, Pavlidis I, Vickers C, Höhne M, Bornscheuer UT (2014), A glycine oxidase based high-throughput solid-phase-assay for substrate profiling and directed evolution of (R)- and (S)-selective amine transaminases, Anal. Chem. 86, 11847-11853 here

20. Gand M, Müller H, Wardenga R, Höhne M (2014), Characterization of three novel enzymes with imine reductase activity. J. Mol. Catal. B. Enzym. 110, 126-132 here

19. Mallin H, Höhne M, Bornscheuer UT (2014), Immobilization of (R)- and (S)-amine transaminases on chitosan support and their application for amine synthesis using isopropylamine as donor, J. Biotechnol. 191, 32-37 here

18. Kohls H, Steffen-Munsberg F, Höhne M (2014), Recent achievements in developing the biocatalytic toolbox for chiral amine synthesis. Curr. Opin. Chem. Biol. 19, 180-192 here

17. Thomsen M, Skalden L, Palm GJ, Höhne M, Bornscheuer UT, Hinrichs W (2014), Crystallographic characterization of the (R)-selective amine transaminase from Aspergillus fumigatus, Acta Cryst. Sect D. 70, 1086-1093 here

16. Höhne M, Bornscheuer UT (2014), Protein engineering from 'scratch' is maturing, Angew. Chem. Int. Ed. 53, 1200-1202 here; Protein engineering aus dem "Nichts" wird praktikabel, Angew. Chem. 126, 1222-1224 here

15. Thomsen M, Skalden L, Palm G, Höhne M, Bornscheuer UT, Hinrichs W (2013), Crystallization and preliminary X-ray diffraction studies of the (R)-selective amine transaminase from Aspergillus fumigatus, Acta Cryst. Sect. F. 69, 1415-1417 here

14. Wardenga R, Bednarczyk A, Höhne M (2013), Asymmetric synthesis of chiral amines from ketones. How to apply biocatalysis and find a suitable enzyme. PharmaChem 12, 22-25 here

13. Mallin H, Menyes U, Vorhaben T, Höhne M, Bornscheuer UT (2013), Immobilization of two (R)-amine transaminases on an optimized chitosan support for the enzymatic synthesis of optically pure amines, ChemCatChem 5, 588-593 here

12. Steffen-Munsberg F, Vickers C, Thontowi A, Schätzle S, Meinhardt T, Svedendahl Humble M, Land H, Berglund P, Bornscheuer UT, Höhne M (2013), Revealing the structural basis of promiscuous amine transaminase activity, ChemCatChem 5, 154-157 here

11. Steffen-Munsberg F, Vickers C, Thontowi A, Schätzle S, Tumlirsch T, Svedendahl Humble M, Land H, Berglund P, Bornscheuer UT, Höhne M (2013), Connecting unexplored protein crystal structures to enzymatic function, ChemCatChem 5, 150-153 here

10. Schätzle S, Steffen-Munsberg F, Thontowi A, Höhne M, Robins K, Bornscheuer UT (2011), Enzymatic asymmetric synthesis of enantiomerically pure aliphatic, aromatic and arylaliphatic amines with (R)-selective amine transaminases, Adv. Synth. Catal. 353, 2439-2445 here

9. Höhne M, Schätzle S, Jochens H, Robins K, Bornscheuer UT (2010) Rational assignment of key motifs for function guides in silico enzyme identification, Nature Chem. Biol. 6, 807-813 here

8. Soltwedel O, Ivanova O, Höhne M, Gopinadhan M, Helm CA (2010) Aggregation and rearrangement within a silver nanoparticle layer during polyelectrolyte multilayer formation, Langmuir 26, 15219-152284 here

7. Schätzle S, Höhne M, Robins K, Bornscheuer UT (2010), A conductometric method for the rapid characterization of the substrate specificity of amine-transaminases, Anal. Chem. 82, 2082-2086 here

6. Schätzle S, Höhne M, Redestad E, Robins K, Bornscheuer UT (2009), A rapid and sensitive kinetic assay for characterization of omega-transaminases, Anal. Chem, 81, 8244-8248 here

5. Höhne M, Bornscheuer UT (2009), Biocatalytic routes to optically active amines, ChemCatChem 1, 42-51 here

4. Kourist R, Höhne M, Bornscheuer UT (2009), Protein Design: Im Spannungsfeld zwischen gerichteter Evolution und rationalem Design, Chemie in unserer Zeit 43, 132-142 here

3. Höhne M, Robins K, Bornscheuer UT (2008), A protection strategy substantially enhances rate and enantioselectivity in transaminase-catalyzed kinetic resolutions, Adv. Synth. Catal. 350, 807-812 here

2. Höhne M, Kühl S, Robins K, Bornscheuer UT (2008), Efficient asymmetric synthesis of chiral amines by combining transaminase and pyruvate decarboxylase, ChemBioChem 9, 363-365 here

1. Schmidt M, Barbayianni E, Fotakopoulou I, Höhne M, Constantinou-Kokotou V, Bornscheuer UT, Kokotos G (2005), Enzymatic removal of carboxyl protecting groups. Part I: Cleavage of the tert-butyl moiety, J. Org. Chem. 70, 3737-3040 here

Book chapters

Höhne M, Matzel P, Gand M (2019). Asymmetric reduction of C=N bonds by imine reductases and reductive aminases. In: Grunwald P (editor). Pharmaceutical Biocatalysis – Chemoenzymatic Synthesis of Active Pharmaceutical Ingredients, 1st Edition, Jenny Stanford Series on Biocatalysis 5, ISBN 978-981-4800-80-8, Jenny Stanford Publishing, Singapore;

Brundiek H, Höhne M (2016). Transaminases – A Biosynthetic Route for Chiral Amines. In: Applied Biocatalysis - From Fundamental Science to Industrial Applications, Liese, A., et al. (Eds.), Wiley-VCH, Weinheim;

Höhne, M., Bornscheuer, U.T., (2011) Application of transaminases in organic synthesis. In: Enzymes in Organic Synthesis, May, O., Gröger, H., Drauz, W. (Eds.), Wiley-VCH, Weinheim


3. Höhne, M., Bornscheuer, U., Robins, K., Schätzle, S. (2011), A process fort he identification and preparation of a (R)-specific omega-transaminase. Intern. Patent PCT WO 2011026556

2. Robins, K., Bornscheuer, U., Höhne, M. (2008), Intern. Patent PCT WO 2008028654.

1. Robins, K., Bornscheuer, U., Höhne, M. (2007), European Patent EP 1818411