Crystal Contact Engineering to Enhance Protein Crystallization Processes

Phillip Grob, Dissertation Technische Universität München, 2020

The crystal contacts of an alcohol dehydrogenase from Lactobacillus brevis, recombinantly available with Escherichia coli, were analyzed to subsequently exchange individual amino acids in a targeted approach. The aim was to enhance crystal contact interactions. Many of the purified enzyme mutants crystallized faster than the non-modified enzyme. The generation of electrostatic interactions was particularly successful, enabling protein crystallization from cell lysate as an efficient purification method.

Publikationen

• Hermann J, Bischoff D, Grob P, Janowski R, Hekmat D, Niessing D, Zacharias M, Weuster-Botz D (2021): Controlling protein crystallization by free energy guided design of interactions at crystal contacts. Crystals 11: 588.
• Grob P, Huber M, Walla B, Hermann J, Janowski R, Niessing D, Hekmat D, Weuster-Botz D (2020) Crystal contact engineering enables efficient capture and purification of an oxidoreductase by technical crystallization. Biotechnol J 15: 2000010.
• Nowotny P, Hermann J, Li J, Krautenbacher AE, Klöpfer K, Hekmat D, Weuster-Botz D (2019): Rational crystal contact engineering of Lactobacillus brevis alcohol dehydrogenase to promote technical protein crystallization. Cryst Growth Des 19: 2380–2387.
• Hermann J, Nowotny P, Schrader TE, Biggel P, Hekmat D, Weuster-Botz D (2018): Neutron and X-ray crystal structures of Lactobacillus brevis alcohol dehydrogenase reveal new insights on hydrogen bonding pathways. Acta Cryst F74: 754-764.