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


38. Zhang W, Fueyo EF, Hollmann F, Martin LL, Pesic M, Wardenga R, Höhne M, Schmidt S (2018). Combining photo-organo redox- and enzyme catalysis facilitates asymmetric C-H bond functionalization. Eur J Org Chem,  DOI:10.1002/ejoc.201801692, 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, DOI:10.1111/febs.14674, 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., 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