journal article·2021-06Open access

An engineered, non-diazotrophic cyanobacterium and its application in bioelectrochemical nitrogen fixation

Fangyuan Dong, Yoo Seok Lee, Erin M. Gaffney, Matteo Grattieri, Helena Haddadin, Shelley D. Minteer, Hui Chen

Cell Reports Physical Science· Volume 2· Issue 6

Abstract

The reduction of chemically inert nitrogen to ammonia is a critical step in the global nitrogen cycle. Microbial nitrogen fixation is a promising way to realize nitrogen reduction and ammonia production at mild conditions. Here, we report an engineered, non-diazotrophic Synechococcus elongatus PCC 7942 strain with nitrogen fixation activity that is constructed by integrating a modified nitrogenase gene cluster into the genome. The engineered S. elongatus PCC 7942 strain is employed in a bioelectrochemical nitrogen-fixation (e-BNF) system for ammonia production. Because the e-BNF system supplies adequate external electrons for the turnover of nitrogenase, the nitrogen fixation activity of the engineered S. elongatus PCC 7942 strain is significantly improved. After 48 h of reaction, the e-BNF system accumulates 173 μM of NH3, which is 21 times higher than that generated from solely photosynthesis-driven nitrogen fixation, with faradaic efficiency of 6.85%. This work may provide new insight into biological nitrogen-fixation systems and ammonium production.
Source: openalex_inverted_index · Quality: baseline

Research context

OpenAlex topics: Ammonia Synthesis and Nitrogen Reduction · Microbial Fuel Cells and Bioremediation · Advanced Photocatalysis Techniques

SciVX discovery tags are terminology-based. OpenAlex topics are source-provided classifications.

References (67)

Show source-provided references
1. Recent developments in nitrogen reduction catalysts: A virtual issue10.1021/acsenergylett.8b02197 ↗
2. Bioelectrochemical Haber-Bosch process: an ammonia-producing H2/N2 fuel cell10.1002/anie.201612500 ↗
3. Ammonia synthesis on wool-like Au, Pt, Pd, Ag, or Cu electrode catalysts in nonthermal atmospheric-pressure plasma of N2 and H210.1021/acscatal.7b01624 ↗
4. Fe-Mediated Nitrogen Fixation with a metallocene mediator: exploring pKa effects and demonstrating electrocatalysis10.1021/jacs.8b02335 ↗
5. Light-driven dinitrogen reduction catalyzed by a CdS:nitrogenase MoFe protein biohybrid10.1126/science.aaf2091 ↗
6. Defining intermediates of nitrogenase MoFe protein during N2 reduction under photochemical electron delivery from CdS quantum dots10.1021/jacs.0c06343 ↗
7. Catalysts for nitrogen reduction to ammonia10.1038/s41929-018-0092-7 ↗
8. A review of the existing and alternative methods for greener nitrogen fixation10.1016/j.cep.2015.02.004 ↗
9. Evaluating the thermodynamics of electrocatalytic N2 reduction in acetonitrile10.1021/acsenergylett.6b00319 ↗
10. Energy transduction in nitrogenase10.1021/acs.accounts.8b00112 ↗
11. Enzymatic bioelectrosynthetic ammonia production: recent electrochemistry of nitrogenase, nitrate reductase, and nitrite reductase10.1002/cplu.201600442 ↗
12. Electroenzymatic nitrogen fixation using an organic redox polymer-immobilized MoFe protein system10.1002/anie.202007198 ↗
13. Nitrogenase bioelectrocatalysis: heterogeneous ammonia and hydrogen production by MoFe protein10.1039/C6EE01432A ↗
14. The in vivo potential-regulated protective protein of nitrogenase in Azotobacter vinelandii supports aerobic bioelectrochemical dinitrogen reduction in vitro10.1021/jacs.7b04893 ↗
15. Pyrene-based noncovalent immobilization of nitrogenase on carbon surfaces10.1002/cbic.201900697 ↗
16. Distribution of nitrogen fixation and nitrogenase-like sequences amongst microbial genomes10.1186/1471-2164-13-162 ↗
17. Role of nitrogenase and ferredoxin in the mechanism of bioelectrocatalytic nitrogen fixation by the cyanobacteria Anabaena variabilis SA-1 mutant immobilized on indium tin oxide (ITO) electrodes10.1016/j.electacta.2017.02.148 ↗
18. The role of HetN in maintenance of the heterocyst pattern in Anabaena sp. PCC 712010.1046/j.1365-2958.2001.02437.x ↗
19. Effect of light intensity and inhibitors of nitrogen assimilation on NH4+ inhibition of nitrogenase activity in Rhodospirillum rubrum and Anabaena sp10.1128/jb.151.2.800-806.1982 ↗
20. Heterocyst formation in Anabaena
21. Ambient nitrogen reduction cycle using a hybrid inorganic-biological system10.1073/pnas.1706371114 ↗
22. Heterologous expression and engineering of the nitrogenase cofactor biosynthesis scaffold NifEN10.1002/anie.201916598 ↗
23. Using synthetic biology to overcome barriers to stable expression of nitrogenase in eukaryotic organelles10.1073/pnas.2002307117 ↗
24. State of the art in eukaryotic nitrogenase engineering10.1093/femsle/fnx274 ↗
25. Refactoring the nitrogen fixation gene cluster from Klebsiella oxytoca10.1073/pnas.1120788109 ↗
26. Genetic transfer of nitrogen fixation from Klebsiella pneumoniae to Escherichia coli10.1038/237102a0 ↗
27. A minimal nitrogen fixation gene cluster from Paenibacillus sp. WLY78 enables expression of active nitrogenase in Escherichia coli10.1371/journal.pgen.1003865 ↗
28. Reconstruction and minimal gene requirements for the alternative iron-only nitrogenase in Escherichia coli10.1073/pnas.1411185111 ↗
29. Functional optimization of gene clusters by combinatorial design and assembly10.1038/nbt.3063 ↗
30. Engineering nitrogen fixation activity in an oxygenic phototroph10.1128/mBio.01029-18 ↗
31. The Klebsiella pneumoniae nitrogenase Fe protein gene (nifH) functionally substitutes for the chlL gene in Chlamydomonas reinhardtii10.1016/j.bbrc.2005.02.064 ↗
32. Formation of nitrogenase NifDK tetramers in the mitochondria of Saccharomyces cerevisiae10.1021/acssynbio.6b00371 ↗
33. Expression of a functional oxygen-labile nitrogenase component in the mitochondrial matrix of aerobically grown yeast10.1038/ncomms11426 ↗
34. Adaptation of the GoldenBraid modular cloning system and creation of a toolkit for the expression of heterologous proteins in yeast mitochondria10.1186/s12896-017-0393-y ↗
35. Stable chromosomal integration of the entire nitrogen fixation gene cluster from Klebsiella pneumoniae in yeast10.1073/pnas.78.6.3496 ↗
36. Biosynthesis of the nitrogenase active-site cofactor precursor NifB-co in Saccharomyces cerevisiae10.1073/pnas.1904903116 ↗
37. Electron transfer precedes ATP hydrolysis during nitrogenase catalysis10.1073/pnas.1311218110 ↗
38. Pyrene hydrogel for promoting direct bioelectrochemistry: ATP-independent electroenzymatic reduction of N210.1039/C8SC01638K ↗
39. Nitrogenase bioelectrocatalysis: atp-independent ammonia production using a redox polymer/mofe protein system10.1021/acscatal.0c01397 ↗
40. Using synthetic biology to increase nitrogenase activity10.1186/s12934-016-0442-6 ↗
41. Control of electron transfer in nitrogenase10.1016/j.cbpa.2018.08.011 ↗
42. Biohybrid photoheterotrophic metabolism for significant enhancement of biological nitrogen fixation in pure microbial cultures10.1039/C9EE00705A ↗
43. Expression of 16 nitrogenase proteins within the plant mitochondrial matrix10.3389/fpls.2017.00287 ↗
44. Diversity of nitrogenase systems in diazotrophs10.1111/j.1744-7909.2006.00271.x ↗
45. In vitro synthesis of the iron-molybdenum cofactor of nitrogenase from iron, sulfur, molybdenum, and homocitrate using purified proteins10.1073/pnas.0703050104 ↗
46. Reactivity, mechanism, and assembly of the alternative nitrogenases10.1021/acs.chemrev.9b00704 ↗
47. Biosynthesis of nitrogenase cofactors10.1021/acs.chemrev.9b00489 ↗
48. Homocitrate cures the NifV− phenotype in Klebsiella pneumoniae10.1128/jb.170.4.1978-1979.1988 ↗
49. NifX and NifEN exchange NifB cofactor and the VK-cluster, a newly isolated intermediate of the iron-molybdenum cofactor biosynthetic pathway10.1111/j.1365-2958.2006.05514.x ↗
50. P-cluster maturation on nitrogenase MoFe protein10.1073/pnas.0704297104 ↗
51. Suppression of hesA mutation on nitrogenase activity in Paenibacillus polymyxa WLY78 with the addition of high levels of molybdate or cystine10.7717/peerj.6294 ↗
52. Systematic comparison of co-expression of multiple recombinant thermophilic enzymes in Escherichia coli BL21(DE3)10.1007/s00253-017-8206-8 ↗
53. Quantitative analysis of the factors limiting solar power transduction by Synechocystis sp. PCC 6803 in biological photovoltaic devices10.1039/c1ee02531g ↗
54. Time-series resolution of gradual nitrogen starvation and its impact on photosynthesis in the cyanobacterium Synechocystis PCC 680310.1111/j.1399-3054.2012.01585.x ↗
55. Time-of-day-dependent responses of cyanobacterial cellular viability against oxidative stress10.1038/s41598-020-77141-8 ↗
56. Effects of oxidative stress on production of heterologous and native protein, and culture morphology in batch and chemostat cultures of Aspergillus niger (B1-D)10.1016/j.enzmictec.2003.07.008 ↗
57. Inhibitory effects of hypo-osmotic stress on extracellular carbonic anhydrase and photosynthetic efficiency of green alga Dunaliella salina possibly through reactive oxygen species formation10.1016/j.plaphy.2012.01.018 ↗
58. Overexpression of orange carotenoid protein protects the repair of PSII under strong light in Synechocystis sp. PCC 680310.1093/pcp/pcy218 ↗
59. Conductometric nitrate biosensor based on methyl viologen/Nafion/nitrate reductase interdigitated electrodes10.1016/j.talanta.2005.10.014 ↗
60. Construction of an electro-enzymatic bioreactor for the production of (R)-mandelate from benzoylformate10.1023/B:BILE.0000009454.19082.03 ↗
61. Metabolic flexibility of Clostridium acetobutylicum in response to methyl viologen addition10.1007/BF00173928 ↗
62. How neutral red modified carbon and electron flow in Clostridium acetobutylicum grown in chemostat culture at neutral pH10.1111/j.1574-6976.1995.tb00163.x ↗
63. Upgraded bioelectrocatalytic N2 fixation: from N2 to chiral amine intermediates10.1021/jacs.9b00147 ↗
64. Bioelectrochemical Nitrogen fixation (e-BNF): Electro-stimulation of enriched biofilm communities drives autotrophic nitrogen and carbon fixation10.1016/j.bioelechem.2018.10.002 ↗
65. Studies on the mechanism of inhibition of glutamine synthetase by methionine sulfoximine10.1021/bi00831a038 ↗
66. Validation of quantitative NMR10.1016/j.jpba.2005.01.043 ↗
67. Modular engineering for efficient photosynthetic biosynthesis of 1-butanol from CO2 in cyanobacteria10.1039/C9EE01214A ↗