Kazuki Tanifuji1, Chi Chung Lee1, Yasuhiro Ohki2, Kazuyuki Tatsumi2, Yilin Hu3, Markus W Ribbe4,5. 1. Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697-3900 (USA). 2. Department of Chemistry, Graduate School of Science and Research Center for Materials Science, Nagoya University, Furo-cho, Chikusa-ku, Nagoya 464-8602 (Japan). 3. Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697-3900 (USA). yilinh@uci.edu. 4. Department of Molecular Biology and Biochemistry, University of California, Irvine, Irvine, CA 92697-3900 (USA). mribbe@uci.edu. 5. Department of Chemistry, University of California, Irvine, Irvine, CA 92697-2025 (USA). mribbe@uci.edu.
Abstract
Nitrogenase catalyzes substrate reduction at its cofactor center ([(Cit)MoFe7S9C](n-); designated M-cluster). Here, we report the formation of an artificial, nitrogenase-mimicking enzyme upon insertion of a synthetic model complex ([Fe6S9(SEt)2](4-); designated Fe6(RHH)) into the catalytic component of nitrogenase (designated NifDK(apo)). Two Fe6(RHH) clusters were inserted into NifDK(apo), rendering the conformation of the resultant protein (designated NifDK(Fe)) similar to the one upon insertion of native M-clusters. NifDK(Fe) can work together with the reductase component of nitrogenase to reduce C2H2 in an ATP-dependent reaction. It can also act as an enzyme on its own in the presence of Eu(II)DTPA, displaying a strong activity in C2H2 reduction while demonstrating an ability to reduce CN(-) to C1-C3 hydrocarbons in an ATP-independent manner. The successful outcome of this work provides the proof of concept and underlying principles for continued search of novel enzymatic activities based on this approach.
Nitrogenase catalyzes substrate reduction at its n class="Chemical">cofactor center ([(Cit)MoFe7S9C](n-); designated M-cluster). Here, we report the formation of an artificial, nitrogenase-mimicking enzyme upon insertion of a synthetic model complex ([Fe6S9(SEt)2](4-); designated Fe6(RHH)) into the catalytic component of nitrogenase (designated NifDK(apo)). Two Fe6(RHH) clusters were inserted into NifDK(apo), rendering the conformation of the resultant protein (designated NifDK(Fe)) similar to the one upon insertion of native M-clusters. NifDK(Fe) can work together with the reductase component of nitrogenase to reduce C2H2 in an ATP-dependent reaction. It can also act as an enzyme on its own in the presence of Eu(II)DTPA, displaying a strong activity in C2H2 reduction while demonstrating an ability to reduce CN(-) to C1-C3hydrocarbons in an ATP-independent manner. The successful outcome of this work provides the proof of concept and underlying principles for continued search of novel enzymatic activities based on this approach.
Authors: Benedikt Schmid; Markus W Ribbe; Oliver Einsle; Mika Yoshida; Leonard M Thomas; Dennis R Dean; Douglas C Rees; Barbara K Burgess Journal: Science Date: 2002-04-12 Impact factor: 47.728
Authors: Thomas Spatzal; Müge Aksoyoglu; Limei Zhang; Susana L A Andrade; Erik Schleicher; Stefan Weber; Douglas C Rees; Oliver Einsle Journal: Science Date: 2011-11-18 Impact factor: 47.728
Authors: Kazuki Tanifuji; Andrew J Jasniewski; Chi Chung Lee; Joseph B Solomon; Takayuki Nagasawa; Yasuhiro Ohki; Kazuyuki Tatsumi; Britt Hedman; Keith O Hodgson; Yilin Hu; Markus W Ribbe Journal: Chembiochem Date: 2022-08-25 Impact factor: 3.461