Mechanoresponsive Protein Crystals for NADH Recycling in Multicycle Enzyme Reactions
Reza Yekta, Xu Xiong, Jiaxin Li, Bradley S. Heater, Marianne M. Lee, Michael K. Chan
Abstract
High Resolution Image Download MS PowerPoint Slide NAD(H)-dependent enzymes play a crucial role in the biosynthesis of pharmaceuticals and fine chemicals, but the limited recyclability of the NAD(H) cofactor hinders its more general application. Here, we report the generation of mechano-responsive PEI-modified Cry3Aa protein crystals and their use for NADH recycling over multiple reaction cycles. For demonstration of its practical utility, a complementary Cry3Aa protein particle containing genetically encoded and co-immobilized formate dehydrogenase for NADH regeneration and leucine dehydrogenase for catalyzing the NADH-dependent l - tert -leucine ( l - tert -Leu) biosynthesis has been produced. When combined with the PEI-modified Cry3Aa crystal, the resultant reaction system could be used for the efficient biosynthesis of l - tert -Leu for up to 21 days with a 10.5-fold improvement in the NADH turnover number.