Identification of genes involved in cadmium-ion tolerance in evolutionary Synechocystis sp. PCC 6803 tolerant to both cadmium and high light

doi:10.1016/j.cjche.2024.05.023

中国化学工程学报 ›› 2024, Vol. 74 ›› Issue (10): 74-82.DOI: 10.1016/j.cjche.2024.05.023

• Full Length Article • 上一篇下一篇

Identification of genes involved in cadmium-ion tolerance in evolutionary Synechocystis sp. PCC 6803 tolerant to both cadmium and high light

Jie Xiong^1,2, Zhengxin Dong^1,2, Yaru Xie^1,2, Weiwen Zhang^1,2,3, Tao Sun^1,2,3, Lei Chen^1,2

1 Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, China;
2 Frontier Science Center for Synthetic Biology & Key Laboratory of Systems Bioengineering, Ministry of Education of China, Tianjin 300072, China;
3 Center for Biosafety Research and Strategy, Tianjin University, Tianjin 300072, China

收稿日期:2024-04-06 修回日期:2024-05-26 接受日期:2024-05-27 出版日期:2024-10-28 发布日期:2024-06-24
通讯作者: Tao Sun, Tel:+86 22 2740 6394;Fax:+86 22 2740 6364.E-mail:tsun@tju.edu.cn;Lei Chen,E-mail:lchen@tju.edu.cn
基金资助:
This research was supported by grants from the National Key Research and Development Program of China (2018YFA0903600) as well as the National Natural Science Foundation of China (32371486 and 32270091).

Identification of genes involved in cadmium-ion tolerance in evolutionary Synechocystis sp. PCC 6803 tolerant to both cadmium and high light

Jie Xiong^1,2, Zhengxin Dong^1,2, Yaru Xie^1,2, Weiwen Zhang^1,2,3, Tao Sun^1,2,3, Lei Chen^1,2

1 Laboratory of Synthetic Microbiology, School of Chemical Engineering & Technology, Tianjin University, Tianjin 300072, China;
2 Frontier Science Center for Synthetic Biology & Key Laboratory of Systems Bioengineering, Ministry of Education of China, Tianjin 300072, China;
3 Center for Biosafety Research and Strategy, Tianjin University, Tianjin 300072, China

Received:2024-04-06 Revised:2024-05-26 Accepted:2024-05-27 Online:2024-10-28 Published:2024-06-24
Contact: Tao Sun, Tel:+86 22 2740 6394;Fax:+86 22 2740 6364.E-mail:tsun@tju.edu.cn;Lei Chen,E-mail:lchen@tju.edu.cn
Supported by:
This research was supported by grants from the National Key Research and Development Program of China (2018YFA0903600) as well as the National Natural Science Foundation of China (32371486 and 32270091).

摘要/Abstract

摘要： Photosynthetic cyanobacteria have shown great potential as “autotrophic cell factories” for the synthesis of fuels and chemicals. However, poor tolerance to various environmental stressors such as high light and heavy metals is an important factor limiting their economic viability. While numerous studies have focused on the tolerance mechanism of cyanobacteria to individual stressors, their response to simultaneous stresses remains to be recovered. To investigate the mechanism of cross tolerance to heavy-metal Cd²⁺ and high light, the model cyanobacterium Synechocystis sp. PCC 6803 tolerant to both Cd²⁺ and high light was obtained via about 800 days’ cross-adaptive laboratory evolution. Three evolutionary strains capable of tolerating both 5.5 μmol·L^-1 Cd²⁺ and 600 μmol·m^-2·s^-1 high light were successfully obtained, achieving about 83% enhancement of Cd²⁺ tolerance compared with the parent strain. The different response of parent and evolutionary strains to Cd²⁺ was elucidated via metabolomics. Furthermore, a total of 15 genes that were mutated during evolution were identified by whole-genome re-sequencing. Finally, by single-gene knockout and complementation analysis, four genes including ssl2615, sll1732, ssr1480, and sll1659 involved in the improvement of Cd²⁺ tolerance under high-light condition were successfully identified. This work explored the tolerance mechanism of Synechocystis sp. PCC 6803 to cadmium under high-light condition and provided valuable reference for deciphering multi-tolerance mechanism of cyanobacteria in the future.

关键词: Synechocystis, Adaptive laboratory evolution, Cross tolerance, Cadmium-ion tolerance

Abstract: Photosynthetic cyanobacteria have shown great potential as “autotrophic cell factories” for the synthesis of fuels and chemicals. However, poor tolerance to various environmental stressors such as high light and heavy metals is an important factor limiting their economic viability. While numerous studies have focused on the tolerance mechanism of cyanobacteria to individual stressors, their response to simultaneous stresses remains to be recovered. To investigate the mechanism of cross tolerance to heavy-metal Cd²⁺ and high light, the model cyanobacterium Synechocystis sp. PCC 6803 tolerant to both Cd²⁺ and high light was obtained via about 800 days’ cross-adaptive laboratory evolution. Three evolutionary strains capable of tolerating both 5.5 μmol·L^-1 Cd²⁺ and 600 μmol·m^-2·s^-1 high light were successfully obtained, achieving about 83% enhancement of Cd²⁺ tolerance compared with the parent strain. The different response of parent and evolutionary strains to Cd²⁺ was elucidated via metabolomics. Furthermore, a total of 15 genes that were mutated during evolution were identified by whole-genome re-sequencing. Finally, by single-gene knockout and complementation analysis, four genes including ssl2615, sll1732, ssr1480, and sll1659 involved in the improvement of Cd²⁺ tolerance under high-light condition were successfully identified. This work explored the tolerance mechanism of Synechocystis sp. PCC 6803 to cadmium under high-light condition and provided valuable reference for deciphering multi-tolerance mechanism of cyanobacteria in the future.

Key words: Synechocystis, Adaptive laboratory evolution, Cross tolerance, Cadmium-ion tolerance

Jie Xiong, Zhengxin Dong, Yaru Xie, Weiwen Zhang, Tao Sun, Lei Chen. Identification of genes involved in cadmium-ion tolerance in evolutionary Synechocystis sp. PCC 6803 tolerant to both cadmium and high light[J]. 中国化学工程学报, 2024, 74(10): 74-82.

参考文献

[1] N. Quintana, F. Van der Kooy, M.D. Van de Rhee, G.P. Voshol, R. Verpoorte, Renewable energy from Cyanobacteria: energy production optimization by metabolic pathway engineering, Appl. Microbiol. Biotechnol. 91 (3) (2011) 471-490.
[2] M. Mehdizadeh Allaf, H. Peerhossaini, Cyanobacteria: model microorganisms and beyond, Microorganisms 10 (4) (2022) 696.
[3] C. Halfmann, L.P. Gu, W. Gibbons, R.B. Zhou, Genetically engineering cyanobacteria to convert CO₂, water, and light into the long-chain hydrocarbon farnesene, Appl. Microbiol. Biotechnol. 98 (23) (2014) 9869-9877.
[4] S. Roussou, A. Albergati, F.Y. Liang, P. Lindblad, Engineered cyanobacteria with additional overexpression of selected Calvin-Benson-Bassham enzymes show further increased ethanol production, Metab. Eng. Commun. 12 (2021) e00161.
[5] R. Miao, H. Xie, F.M. Ho, P. Lindblad, Protein engineering of α-ketoisovalerate decarboxylase for improved isobutanol production in Synechocystis PCC 6803, Metab. Eng. 47 (2018) 42-48.
[6] S. Gudmundsson, J. Nogales, Cyanobacteria as photosynthetic biocatalysts: a systems biology perspective, Mol. Biosyst. 11 (1) (2015) 60-70.
[7] P.K. Babele, J. Kumar, V. Chaturvedi, Proteomic de-regulation in cyanobacteria in response to abiotic stresses, Front. Microbiol. 10 (2019) 1315.
[8] M. Muramatsu, Y. Hihara, Acclimation to high-light conditions in cyanobacteria: from gene expression to physiological responses, J. Plant Res. 125 (1) (2012) 11-39.
[9] K. Ogawa, K. Yoshikawa, F. Matsuda, Y. Toya, H. Shimizu, Transcriptome analysis of the cyanobacterium Synechocystis sp. PCC 6803 and mechanisms of photoinhibition tolerance under extreme high light conditions, J. Biosci. Bioeng. 126 (5) (2018) 596-602.
[10] M. Dann, E.M. Ortiz, M. Thomas, A. Guljamow, M. Lehmann, H. Schaefer, D. Leister, Enhancing photosynthesis at high light levels by adaptive laboratory evolution, Nat. Plants 7 (5) (2021) 681-695.
[11] B. Nowicka, Heavy metal-induced stress in eukaryotic algae-mechanisms of heavy metal toxicity and tolerance with particular emphasis on oxidative stress in exposed cells and the role of antioxidant response, Environ. Sci. Pollut. Res. Int. 29 (12) (2022) 16860-16911.
[12] D. Kalaivanan, A.N. Ganeshamurthy, Mechanisms of Heavy Metal Toxicity in Plants. Rao N, Shivashankara K, Laxman R, Abiotic Stress Physiology of Horticultural Crops. New Delhi: Springer, 2016: 85-102.
[13] H. Kupper, E. Andresen, Mechanisms of metal toxicity in plants, Metallomics 8 (3) (2016) 269-285.
[14] S.Z. Wang, R. Wufuer, J. Duo, W.F. Li, X.L. Pan, Cadmium caused different toxicity to photosystem I and photosystem II of freshwater unicellular algae chlorella pyrenoidosa (chlorophyta), Toxics 10 (7) (2022) 352.
[15] C.X. Xu, T. Sun, S.B. Li, L. Chen, W.W. Zhang, Adaptive laboratory evolution of cadmium tolerance in Synechocystis sp. PCC 6803, Biotechnol. Biofuels 11 (1) (2018) 205.
[16] T. Sun, L. Xu, L.N. Wu, Z.D. Song, L. Chen, W.W. Zhang, Identification of a new target slr0946 of the response regulator Sll0649 involving cadmium tolerance in Synechocystis sp. PCC 6803, Front. Microbiol. 8 (2017) 1582.
[17] R. Mittler, Abiotic stress, the field environment and stress combination, Trends Plant Sci. 11 (1) (2006) 15-19.
[18] N. Murata, S. Takahashi, Y. Nishiyama, S.I. Allakhverdiev, Photoinhibition of photosystem II under environmental stress, Biochim. Biophys. Acta 1767 (6) (2007) 414-421.
[19] Y. Yamamoto, R. Aminaka, M. Yoshioka, M. Khatoon, K. Komayama, D. Takenaka, A. Yamashita, N. Nijo, K. Inagawa, N. Morita, T. Sasaki, Y. Yamamoto, Quality control of photosystem II: impact of light and heat stresses, Photosynth. Res. 98 (1-3) (2008) 589-608.
[20] M.P.V.V.B. Singh, S.M. Prasad, V.P. Singh, M. Singh, Antioxidant system against active oxygen species in cyanobacterium aphanothece stagnina: response to excess light under cadmium stress, Proc. Natl. Acad. Sci. India B Biol. Sci. 85 (2) (2015) 535-543.
[21] J. Du, B. Qiu, M. Pedrosa Gomes, P. Juneau, G. Dai, Influence of light intensity on cadmium uptake and toxicity in the cyanobacteria Synechocystis sp. PCC6803, Aquat. Toxicol. 211 (2019) 163-172.
[22] D. Kaczmarzyk, J. Anfelt, A. Sarnegrim, E.P. Hudson, Overexpression of sigma factor SigB improves temperature and butanol tolerance of Synechocystis sp. PCC6803, J. Biotechnol. 182-183 (2014) 54-60.
[23] T. Sun, S.B. Li, X.Y. Song, G.S. Pei, J.J. Diao, J.Y. Cui, M.L. Shi, L. Chen, W.W. Zhang, re-direction of carbon flux to key precursor malonyl-CoA via artificial small RNAs in photosynthetic Synechocystis sp. PCC 6803, Biotechnol. Biofuels 11 (2018) 26.
[24] Y.P. Wu, D.W. Krogmann, The orange carotenoid protein of Synechocystis PCC 6803, Biochim. Biophys. Acta 1322 (1) (1997) 1-7.
[25] O. Sozer, J. Komenda, B. Ughy, I. Domonkos, H. Laczko-Dobos, P. Malec, Z. Gombos, M. Kis, Involvement of carotenoids in the synthesis and assembly of protein subunits of photosynthetic reaction centers of Synechocystis sp. PCC 6803, Plant Cell Physiol. 51 (5) (2010) 823-835.
[26] J. Nielsen, It is all about MetabolicFluxes, J. Bacteriol. 185 (24) (2003) 7031-7035.
[27] T. Obata, A.R. Fernie, The use of metabolomics to dissect plant responses to abiotic stresses, Cell. Mol. Life Sci. 69 (19) (2012) 3225-3243.
[28] V. Shulaev, D. Cortes, G. Miller, R. Mittler, Metabolomics for plant stress response, Physiol. Plantarum 132 (2) (2008) 199-208.
[29] A. Srivastava, S. Biswas, S. Yadav, S. Kumar, V. Srivastava, Y. Mishra, Acute cadmium toxicity and post-stress recovery: insights into coordinated and integrated response/recovery strategies of Anabaena sp. PCC 7120, J. Hazard Mater. 411 (2021) 124822.
[30] P.P. Zhang, N. Battchikova, T. Jansen, J. Appel, T. Ogawa, E.M. Aro, Expression and functional roles of the two distinct NDH-1 complexes and the carbon acquisition complex NdhD3/NdhF3/CupA/Sll1735 in Synechocystis sp PCC 6803, Plant Cell 16 (12) (2004) 3326-3340.
[31] S.M.E. Daley, A.D. Kappell, M.J. Carrick, R.L. Burnap, Regulation of the cyanobacterial CO₂-concentrating mechanism involves internal sensing of NADP+ and α-ketogutarate levels by transcription factor CcmR, PLoS One 7 (7) (2012) e41286.
[32] A.P. Eker, P. Kooiman, J.K. Hessels, A. Yasui, DNA photoreactivating enzyme from the cyanobacterium Anacystis nidulans, J. Biol. Chem. 265 (14) (1990) 8009-8015.
[33] U. Fels, K. Gevaert, P. van Damme, Bacterial genetic engineering by means of recombineering for reverse genetics, Front. Microbiol. 11 (2020) 548410.
[34] N.J. Atkinson, P.E. Urwin, The interaction of plant biotic and abiotic stresses: from genes to the field, J. Exp. Bot. 63 (10) (2012) 3523-3543.
[35] S. Rasmussen, P. Barah, M.C. Suarez-Rodriguez, S. Bressendorff, P. Friis, P. Costantino, A.M. Bones, H.B. Nielsen, J. Mundy, Transcriptome responses to combinations of stresses in Arabidopsis, Plant Physiol. 161 (4) (2013) 1783-1794.
[36] W.J. Lou, X.M. Tan, K. Song, S.S. Zhang, G.D. Luan, C. Li, X.F. Lu, A specific single nucleotide polymorphism in the ATP synthase gene significantly improves environmental stress tolerance of synechococcus elongatus PCC 7942, Appl. Environ. Microbiol. 84 (18) (2018) e01222-e01218.
[37] G. Bernat, J. Appel, T. Ogawa, M. Rogner, Distinct roles of multiple NDH-1 complexes in the cyanobacterial electron transport network as revealed by kinetic analysis of P700+ reduction in various Ndh-deficient mutants of Synechocystis sp. strain PCC6803, J. Bacteriol. 193 (1) (2011) 292-295.
[38] T. Zeller, G. Klug, Thioredoxins in bacteria: functions in oxidative stress response and regulation of thioredoxin genes, Naturwissenschaften 93 (6) (2006) 259-266.
[39] F. Rollin-Genetet, C. Berthomieu, A.H. Davin, E. Quemeneur, Escherichia coli thioredoxin inhibition by cadmium: two mutually exclusive binding sites involving Cys32 and Asp26, Eur. J. Biochem. 271 (7) (2004) 1299-1309.
[40] M. Muthusamy, J.H. Kim, J.A. Kim, S.I. Lee, Plant RNA binding proteins as critical modulators in drought, high salinity, heat, and cold stress responses: an updated overview, Int. J. Mol. Sci. 22 (13) (2021) 6731.
[41] C. Greening, F.H. Ahmed, A.E. Mohamed, B.M. Lee, G. Pandey, A.C. Warden, C. Scott, J.G. Oakeshott, M.C. Taylor, C.J. Jackson, Physiology, biochemistry, and applications of F420- and fo-dependent redox reactions, Microbiol. Mol. Biol. Rev. 80 (2) (2016) 451-493.
[42] B.N.P. Kumar, S. Mahaboobi, S. Satyam, Cyanobacteria: a potential natural source for drug discovery and bioremediation, Ind. Pollut. Control 32 (2) (2016) 508-517.
[43] R. Mota, F. Rossi, L. Andrenelli, S.B. Pereira, R. De Philippis, P. Tamagnini, Released polysaccharides (RPS) from Cyanothece sp. CCY 0110 as biosorbent for heavy metals bioremediation: interactions between metals and RPS binding sites, Appl. Microbiol. Biotechnol. 100 (17) (2016) 7765-7775.
[44] S. Ozturk, B. Aslim, Z. Suludere, Cadmium(II) sequestration characteristics by two isolates of Synechocystis sp. in terms of exopolysaccharide (EPS) production and monomer composition, Bioresour. Technol. 101 (24) (2010) 9742-9748.
[45] L. Shen, Z.F. Li, J.J. Wang, A.J. Liu, Z.H. Li, R.L. Yu, X.L. Wu, Y.D. Liu, J.K. Li, W.M. Zeng, Characterization of extracellular polysaccharide/protein contents during the adsorption of Cd(II) by Synechocystis sp. PCC6803, Environ. Sci. Pollut. Res. Int. 25 (21) (2018) 20713-20722.

Identification of genes involved in cadmium-ion tolerance in evolutionary Synechocystis sp. PCC 6803 tolerant to both cadmium and high light

Identification of genes involved in cadmium-ion tolerance in evolutionary Synechocystis sp. PCC 6803 tolerant to both cadmium and high light

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