A Guanidine-Degrading Enzyme Controls Genomic Stability of Ethylene-Producing Cyanobacteria

Wang, Bo; Xu, Yao; Wang, Xin; Yuan, Joshua S.; Johnson, Carl H.; Young, Jamey D.; Yu, Jianping

doi:10.1038/s41467-021-25369-x

Title: A Guanidine-Degrading Enzyme Controls Genomic Stability of Ethylene-Producing Cyanobacteria

Journal Article · Thu Aug 26 00:00:00 EDT 2021 · Nature Communications

DOI:https://doi.org/10.1038/s41467-021-25369-x· OSTI ID:1819078

Wang, Bo ^[1]; Xu, Yao ^[2]; Wang, Xin ^[3]; Yuan, Joshua S. ^[3]; Johnson, Carl H. ^[2]; Young, Jamey D. ^[2]; Yu, Jianping ^[4]

National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center; Vanderbilt Univ., Nashville, TN (United States)
Vanderbilt Univ., Nashville, TN (United States)
Texas A & M Univ., College Station, TX (United States). Synthetic and Systems Biology Innovation Hub
National Renewable Energy Lab. (NREL), Golden, CO (United States). Biosciences Center

Recent studies have revealed the prevalence and biological significance of guanidine metabolism in nature. However, the metabolic pathways used by microbes to degrade guanidine or mitigate its toxicity have not been widely studied. Here, via comparative proteomics and subsequent experimental validation, we demonstrate that Sll1077, previously annotated as an agmatinase enzyme in the model cyanobacterium Synechocystis sp. PCC 6803, is more likely a guanidinase as it can break down guanidine rather than agmatine into urea and ammonium. The model cyanobacterium Synechococcus elongatus PCC 7942 strain engineered to express the bacterial ethylene-forming enzyme (EFE) exhibits unstable ethylene production due to toxicity and genomic instability induced by accumulation of the EFE-byproduct guanidine. Co-expression of EFE and Sll1077 significantly enhances genomic stability and enables the resulting strain to achieve sustained high-level ethylene production. These findings expand our knowledge of natural guanidine degradation pathways and demonstrate their biotechnological application to support ethylene bioproduction.

View Accepted Manuscript (DOE)

Cite

Export

Save

Research Organization:: National Renewable Energy Lab. (NREL), Golden, CO (United States)

Sponsoring Organization:: USDOE Office of Energy Efficiency and Renewable Energy (EERE), Sustainable Transportation Office. Bioenergy Technologies Office; National Institutes of Health (NIH)

Grant/Contract Number:: AC36-08GO28308; SC0019388; SC0019404; SC0018344; AR0000203; R37 GM067152; R01 GM107434

OSTI ID:: 1819078

Report Number(s):: NREL/JA-2700-79641; MainId:35862; UUID:b8d5d784-4bc7-4c56-b0e1-dc16e3299570; MainAdminID:61709

Journal Information:: Nature Communications, Vol. 12; ISSN 2041-1723

Publisher:: Nature Publishing GroupCopyright Statement

Country of Publication:: United States

Language:: English

References (47)

Guanidine sulphate: Slow release of mineral nitrogen during incubation in soil Zhang, Z.; Nyborg, M.; Worsley, M. Communications in Soil Science and Plant Analysis, Vol. 23, Issue 5-6 https://doi.org/10.1080/00103629209368601	journal	March 1992
Proposed mechanism for urea nitrogen re-utilization: relationship between urea and proposed guanidine cycles. Natelson, S.; Sherwin, J. E. Clinical Chemistry, Vol. 25, Issue 7 https://doi.org/10.1093/clinchem/25.7.1343	journal	July 1979
Metabolism of Free Guanidine in Bacteria Is Regulated by a Widespread Riboswitch Class Nelson, James W.; Atilho, Ruben M.; Sherlock, Madeline E. Molecular Cell, Vol. 65, Issue 2 https://doi.org/10.1016/j.molcel.2016.11.019	journal	January 2017
The Nonenzymatic Decomposition of Guanidines and Amidines Lewis, Charles A.; Wolfenden, Richard Journal of the American Chemical Society, Vol. 136, Issue 1 https://doi.org/10.1021/ja411927k	journal	December 2013
Biodegradation of guanidinium ion in aerobic soil samples Mitchell, Wayne R. Bulletin of Environmental Contamination and Toxicology, Vol. 39, Issue 6 https://doi.org/10.1007/BF01689587	journal	December 1987
Biochemical Validation of a Second Guanidine Riboswitch Class in Bacteria Sherlock, Madeline E.; Malkowski, Sarah N.; Breaker, Ronald R. Biochemistry, Vol. 56, Issue 2 https://doi.org/10.1021/acs.biochem.6b01270	journal	January 2017
Biochemical Validation of a Third Guanidine Riboswitch Class in Bacteria Sherlock, Madeline E.; Breaker, Ronald R. Biochemistry, Vol. 56, Issue 2 https://doi.org/10.1021/acs.biochem.6b01271	journal	January 2017
Discovery and characterization of a fourth class of guanidine riboswitches Lenkeit, Felina; Eckert, Iris; Hartig, Jörg S. Nucleic Acids Research, Vol. 48, Issue 22 https://doi.org/10.1093/nar/gkaa1102	journal	November 2020
Biochemical Validation of a Fourth Guanidine Riboswitch Class in Bacteria Salvail, Hubert; Balaji, Aparaajita; Yu, Diane Biochemistry, Vol. 59, Issue 49 https://doi.org/10.1021/acs.biochem.0c00793	journal	November 2020
Guanidinium export is the primal function of SMR family transporters Kermani, Ali A.; Macdonald, Christian B.; Gundepudi, Roja Proceedings of the National Academy of Sciences, Vol. 115, Issue 12 https://doi.org/10.1073/pnas.1719187115	journal	March 2018
Solving the Conundrum: Widespread Proteins Annotated for Urea Metabolism in Bacteria Are Carboxyguanidine Deiminases Mediating Nitrogen Assimilation from Guanidine Schneider, Nicholas O.; Tassoulas, Lambros J.; Zeng, Danyun Biochemistry, Vol. 59, Issue 35 https://doi.org/10.1021/acs.biochem.0c00537	journal	August 2020
Sequence Analysis of the Genome of the Unicellular Cyanobacterium Synechocystis sp. Strain PCC6803. II. Sequence Determination of the Entire Genome and Assignment of Potential Protein-coding Regions Kaneko, T.; Sato, S.; Kotani, H. DNA Research, Vol. 3, Issue 3 https://doi.org/10.1093/dnares/3.3.109	journal	January 1996
Phylogeny of related functions: the case of polyamine biosynthetic enzymes Sekowska, Agnieszka; Danchin, Antoine; Risler, Jean-Loup Microbiology, Vol. 146, Issue 8 https://doi.org/10.1099/00221287-146-8-1815	journal	August 2000
Ethylene-forming enzyme and bioethylene production Eckert, Carrie; Xu, Wu; Xiong, Wei Biotechnology for Biofuels, Vol. 7, Issue 1 https://doi.org/10.1186/1754-6834-7-33	journal	January 2014
Ethylene Synthesis and Regulated Expression of Recombinant Protein in Synechocystis sp. PCC 6803 Guerrero, Fernando; Carbonell, Verónica; Cossu, Matteo PLoS ONE, Vol. 7, Issue 11 https://doi.org/10.1371/journal.pone.0050470	journal	November 2012
Sustained photosynthetic conversion of CO₂ to ethylene in recombinant cyanobacterium Synechocystis 6803 Ungerer, Justin; Tao, Ling; Davis, Mark Energy & Environmental Science, Vol. 5, Issue 10, p. 8998-9006 https://doi.org/10.1039/c2ee22555g	journal	January 2012
Enhancing photosynthetic production of ethylene in genetically engineered Synechocystis sp. PCC 6803 Zhu, Tao; Xie, Xiaoman; Li, Zhimin Green Chemistry, Vol. 17, Issue 1 https://doi.org/10.1039/C4GC01730G	journal	January 2015
The plasticity of cyanobacterial metabolism supports direct CO2 conversion to ethylene Xiong, Wei; Morgan, John A.; Ungerer, Justin Nature Plants, Vol. 1, Issue 5 https://doi.org/10.1038/nplants.2015.53	journal	April 2015
Metabolic engineering for ethylene production by inserting the ethylene-forming enzyme gene (efe) at the 16S rDNA sites of Pseudomonas putida KT2440 Wang, Jian-Ping; Wu, Li-Xian; Xu, Feng Bioresource Technology, Vol. 101, Issue 16 https://doi.org/10.1016/j.biortech.2010.03.030	journal	August 2010
Photosynthetic conversion of carbon dioxide to ethylene by the recombinant cyanobacterium, Synechococcus sp. PCC 7942, which harbors a gene for the ethylene-forming enzyme of Pseudomonas syringae Sakai, Miho; Ogawa, Takahira; Matsuoka, Masayoshi Journal of Fermentation and Bioengineering, Vol. 84, Issue 5, p. 434-443 https://doi.org/10.1016/S0922-338X(97)82004-1	journal	January 1997
Construction and analysis of a recombinant cyanobacterium expressing a chromosomally inserted gene for an ethylene-forming enzyme at the psbAI locus Takahama, Kazutaka; Matsuoka, Masayoshi; Nagahama, Kazuhiro Journal of Bioscience and Bioengineering, Vol. 95, Issue 3, p. 302-305 https://doi.org/10.1016/S1389-1723(03)80034-8	journal	January 2003
Photosynthetic Co-production of Succinate and Ethylene in a Fast-Growing Cyanobacterium, Synechococcus elongatus PCC 11801 Sengupta, Annesha; Pritam, Prem; Jaiswal, Damini Metabolites, Vol. 10, Issue 6 https://doi.org/10.3390/metabo10060250	journal	June 2020
Nitrogen chlorosis in unicellular cyanobacteria – a developmental program for surviving nitrogen deprivation Forchhammer, Karl; Schwarz, Rakefet Environmental Microbiology, Vol. 21, Issue 4 https://doi.org/10.1111/1462-2920.14447	journal	November 2018
Photosynthetic production of the nitrogen-rich compound guanidine Wang, Bo; Dong, Tao; Myrlie, Aldon Green Chemistry, Vol. 21, Issue 11 https://doi.org/10.1039/C9GC01003C	journal	January 2019
Purification and properties of agmatine ureohydrolyase, a putrescine biosynthetic enzyme in Escherichia coli. Satishchandran, C.; Boyle, S. M. Journal of Bacteriology, Vol. 165, Issue 3 https://doi.org/10.1128/jb.165.3.843-848.1986	journal	March 1986
Genomics of Urea Transport and Catabolism in Cyanobacteria: Biotechnological Implications Veaudor, Théo; Cassier-Chauvat, Corinne; Chauvat, Franck Frontiers in Microbiology, Vol. 10 https://doi.org/10.3389/fmicb.2019.02052	journal	September 2019
Purification and properties of an ethylene-forming enzyme from Pseudomonas syringae pv. phaseolicola PK2 Nagahama, K.; Ogawa, T.; Fujii, T. Journal of General Microbiology, Vol. 137, Issue 10 https://doi.org/10.1099/00221287-137-10-2281	journal	October 1991
Overexpression and in vitro reconstitution of the ethylene-forming enzyme from Pseudomonas syringae Ishihara, Katsuya; Matsuoka, Masayoshi; Inoue, Yorinao Journal of Fermentation and Bioengineering, Vol. 79, Issue 3 https://doi.org/10.1016/0922-338X(95)90604-X	journal	January 1995
A Genetic Toolbox for Modulating the Expression of Heterologous Genes in the Cyanobacterium Synechocystis sp. PCC 6803 Wang, Bo; Eckert, Carrie; Maness, Pin-Ching ACS Synthetic Biology, Vol. 7, Issue 1 https://doi.org/10.1021/acssynbio.7b00297	journal	August 2017
Homospermidine biosynthesis in the cyanobacterium Anabaena requires a deoxyhypusine synthase homologue and is essential for normal diazotrophic growth Burnat, Mireia; Li, Bin; Kim, Sok Ho Molecular Microbiology, Vol. 109, Issue 6 https://doi.org/10.1111/mmi.14006	journal	September 2018
Catabolic pathway of arginine in Anabaena involves a novel bifunctional enzyme that produces proline from arginine Burnat, Mireia; Picossi, Silvia; Valladares, Ana Molecular Microbiology, Vol. 111, Issue 4 https://doi.org/10.1111/mmi.14203	journal	February 2019
The cyanobacterial ornithine–ammonia cycle involves an arginine dihydrolase Zhang, Hao; Liu, Yujie; Nie, Xiaoqun Nature Chemical Biology, Vol. 14, Issue 6 https://doi.org/10.1038/s41589-018-0038-z	journal	April 2018
Arginine Catabolism in the CyanobacteriumSynechocystis sp. Strain PCC 6803 Involves the Urea Cycle and Arginase Pathway Quintero, María José; Muro-Pastor, Alicia María; Herrero, Antonia Journal of Bacteriology, Vol. 182, Issue 4 https://doi.org/10.1128/JB.182.4.1008-1015.2000	journal	February 2000
Ethylene production in Synechocystis sp. PCC 6803 promotes phototactic movement Kuchmina, Ekaterina; Klähn, Stephan; Jakob, Annik Microbiology, Vol. 163, Issue 12 https://doi.org/10.1099/mic.0.000564	journal	December 2017
Guanidinium can both Cause and Prevent the Hydrophobic Collapse of Biomacromolecules Heyda, Jan; Okur, Halil I.; Hladílková, Jana Journal of the American Chemical Society, Vol. 139, Issue 2 https://doi.org/10.1021/jacs.6b11082	journal	January 2017
Probing the Action of Chemical Denaturant on an Intrinsically Disordered Protein by Simulation and Experiment Zheng, Wenwei; Borgia, Alessandro; Buholzer, Karin Journal of the American Chemical Society, Vol. 138, Issue 36 https://doi.org/10.1021/jacs.6b05443	journal	September 2016
Inactivation during denaturation of ribonuclease A by guanidinium chloride is accompanied by unfolding at the active site Yang, H. J.; Tsou, C. L. Biochemical Journal, Vol. 305, Issue 2 https://doi.org/10.1042/bj3050379	journal	January 1995
Biochemistry of nitrification in soil Lees, H.; Quastel, J. H. Biochemical Journal, Vol. 40, Issue 5-6 https://doi.org/10.1042/bj0400824	journal	January 1946
Enhanced stable production of ethylene in photosynthetic cyanobacterium Synechococcus elongatus PCC 7942 Carbonell, Veronica; Vuorio, Eerika; Aro, Eva-Mari World Journal of Microbiology and Biotechnology, Vol. 35, Issue 5 https://doi.org/10.1007/s11274-019-2652-7	journal	May 2019
Systematic identification and elimination of flux bottlenecks in the aldehyde production pathway of Synechococcus elongatus PCC 7942 Cheah, Yi Ern; Xu, Yao; Sacco, Sarah A. Metabolic Engineering, Vol. 60 https://doi.org/10.1016/j.ymben.2020.03.007	journal	July 2020
Premethylation of Foreign DNA Improves Integrative Transformation Efficiency in Synechocystis sp. Strain PCC 6803 Wang, Bo; Yu, Jianping; Zhang, Weiwen Applied and Environmental Microbiology, Vol. 81, Issue 24 https://doi.org/10.1128/AEM.02575-15	journal	October 2015
Isotopically nonstationary 13C flux analysis of cyanobacterial isobutyraldehyde production Jazmin, Lara J.; Xu, Yao; Cheah, Yi Ern Metabolic Engineering, Vol. 42 https://doi.org/10.1016/j.ymben.2017.05.001	journal	July 2017
Enhanced limonene production in cyanobacteria reveals photosynthesis limitations Wang, Xin; Liu, Wei; Xin, Changpeng Proceedings of the National Academy of Sciences, Vol. 113, Issue 50 https://doi.org/10.1073/pnas.1613340113	journal	November 2016
ProLuCID: An improved SEQUEST-like algorithm with enhanced sensitivity and specificity Xu, T.; Park, S. K.; Venable, J. D. Journal of Proteomics, Vol. 129 https://doi.org/10.1016/j.jprot.2015.07.001	journal	November 2015
DTASelect and Contrast: Tools for Assembling and Comparing Protein Identifications from Shotgun Proteomics Tabb, David L.; McDonald, W. Hayes; Yates, John R. Journal of Proteome Research, Vol. 1, Issue 1, p. 21-26 https://doi.org/10.1021/pr015504q	journal	February 2002
PatternLab for proteomics: a tool for differential shotgun proteomics Carvalho, Paulo C.; Fischer, Juliana SG; Chen, Emily I. BMC Bioinformatics, Vol. 9, Issue 1 https://doi.org/10.1186/1471-2105-9-316	journal	July 2008
2016 update of the PRIDE database and its related tools Vizcaíno, Juan Antonio; Csordas, Attila; del-Toro, Noemi Nucleic Acids Research, Vol. 44, Issue 22 https://doi.org/10.1093/nar/gkw880	journal	September 2016

Similar Records

Photosynthetic production of the nitrogen-rich compound guanidine

Journal Article · Tue Jun 04 00:00:00 EDT 2019 · Green Chemistry · OSTI ID:1819078

Wang, Bo; Dong, Tao; Myrlie, Aldon; +6 more

Phototaxis in a wild isolate of the cyanobacterium Synechococcus elongatus

Journal Article · Fri Dec 14 00:00:00 EST 2018 · Proceedings of the National Academy of Sciences of the United States of America · OSTI ID:1819078

Yang, Yiling; Lam, Vinson; Adomako, Marie; +11 more

Zn²⁺-Inducible Expression Platform for Synechococcus sp. Strain PCC 7002 Based on the smtA Promoter/Operator and smtB Repressor

Journal Article · Tue Jan 17 00:00:00 EST 2017 · Applied and Environmental Microbiology · OSTI ID:1819078

Pérez, Adam A.; Gajewski, John P.; Ferlez, Bryan H.; +4 more

Related Subjects

59 BASIC BIOLOGICAL SCIENCES
ammonia
cyanobacteria
ethylene
guanidine
urea
applied microbiology
biocatalysis
metabolic engineering

Title: A Guanidine-Degrading Enzyme Controls Genomic Stability of Ethylene-Producing Cyanobacteria

Citation Formats

References (47)

Similar Records

Related Subjects