Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN

Solomon, Joseph B.; Lee, Chi Chung; Jasniewski, Andrew J.; Rasekh, Mahtab F.; Ribbe, Markus W.; Hu, Yilin

doi:10.1002/ange.201916598

Title: Heterologous Expression and Engineering of the Nitrogenase Cofactor Biosynthesis Scaffold NifEN

Journal Article · 03 March 2020 · Angewandte Chemie

DOI: https://doi.org/10.1002/ange.201916598 · OSTI ID:1602600

Solomon, Joseph B. ^[1]; Lee, Chi Chung ^[2]; Jasniewski, Andrew J. ^[2]; Rasekh, Mahtab F. ^[1];

^[1]; Hu, Yilin ^[2]

Department of Molecular Biology &, Biochemistry University of California, Irvine Irvine CA 92697-3900 USA, Department Chemistry University of California, Irvine Irvine CA 92697-2025 USA
Department of Molecular Biology &, Biochemistry University of California, Irvine Irvine CA 92697-3900 USA

Abstract NifEN plays a crucial role in the biosynthesis of nitrogenase, catalyzing the final step of cofactor maturation prior to delivering the cofactor to NifDK, the catalytic component of nitrogenase. The difficulty in expressing NifEN, a complex, heteromultimeric metalloprotein sharing structural/functional homology with NifDK, is a major challenge in the heterologous expression of nitrogenase. Herein, we report the expression and engineering of Azotobacter vinelandii NifEN in Escherichia coli . Biochemical and spectroscopic analyses demonstrate the integrity of the heterologously expressed NifEN in composition and functionality and, additionally, the ability of an engineered NifEN variant to mimic NifDK in retaining the matured cofactor at an analogous cofactor‐binding site. This is an important step toward piecing together a viable pathway for the heterologous expression of nitrogenase and identifying variants for the mechanistic investigation of this enzyme.

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Sponsoring Organization:: USDOE

OSTI ID:: 1602600

Journal Information:: Angewandte Chemie, Journal Name: Angewandte Chemie Vol. 132 Journal Issue: 17; ISSN 0044-8249

Publisher:: Wiley Blackwell (John Wiley & Sons)Copyright Statement

Country of Publication:: Germany

Language:: English

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