Divergent metabolic and transcriptomic responses of Synechocystis sp. PCC 6803 to salt stress after adaptive laboratory evolution

Hu, Lang; He, Jianzhong; Dong, Mingjie; Tang, Xing; Jiang, Panpan; Lei, Anping; Wang, Jiangxin

doi:10.1016/j.algal.2020.101856

Cited by 12 publications

(6 citation statements)

References 50 publications

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“…In sharp contrast, no mutation in the mannitol cassette was found in the ΔCS_M strain during the entire prolonged cultivation under salt stress. This observation corroborates the result that cells also maintained a continuous mannitol producing phenotype under such conditions (Fig.6a) and leads us to conclude that mannitol production in Synechocystis can be stabilized by salt stress, in mutants lacking the pathways for synthesis of the two endogenous osmoprotectants In addition, by purposely weakening the native abilities of cells to tolerate salt stress (24,25), this strategy could allow more mannitol to be synthesized to compensate the loss of salt tolerance ability. Such could allow that mannitol production could be even further sustainably enhanced.…”

Section: Characterization Of Mutations That Are Selected By the Phenosupporting

confidence: 89%

Using Osmotic Stress to Stabilize Mannitol Production in Synechocystis sp. PCC6803

Du²,

Gallego³

et al. 2020

Preprint

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Abstract Background Mannitol is a C(6) polyol that is used in the food and medical sector as a sweetener and antioxidant, respectively. The sustainable production of mannitol, especially via the direct conversion of CO 2 by photosynthetic cyanobacteria, has become increasingly appealing. However, previous work aiming to achieve mannitol production in the marine Synechococcus sp. PCC 7002 via heterologous expression of mannitol-1-phosphate-5-dehydrogenase ( mtlD ) and mannitol-1-phosphatase ( m1p , in short: a ‘mannitol cassette’), proved to be genetically unstable. Results Here, we explore the stabilizing effect that mannitol production may have on cells faced with osmotic stress, in the freshwater cyanobacterium Synechocystis sp. PCC 6803. We first validated that mannitol can function as a compatible solute in Synechocystis sp. PCC 6803, and in derivative strains in which the ability to produce one or both of the native compatible solutes was impaired. Wild type Synechocystis , complemented with a mannitol cassette, indeed showed increased salt tolerance, which was even more evident in Synechocystis strains in which the ability to synthesize the endogenous compatible solutes was impaired. Next we tested the genetic stability of all these strains with respect to their mannitol productivity, with and without salt stress, during prolonged turbidostat cultivations. The obtained results show that mannitol production under salt stress conditions in the Synechocystis strain that cannot synthesize its endogenous compatible solutes is remarkably stable, while the control strain completely loses this ability in only 6 days. DNA sequencing results of the control groups that lost the ability to synthesize mannitol revealed that multiple types of mutation occurred in the mtlD gene that can explain the disruption of mannitol production. Conclusions Mannitol production in freshwater Synechocsytis sp. PCC6803 confers it with increased salt tolerance. Under this strategy, genetically instability which was the major challenge for mannitol production in cyanobacteria is tackled. This paper marks the first report of utilization of the response to salt stress as a factor that can increase the stability of mannitol production in a cyanobacterial cell factory.

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Section: Characterization Of Mutations That Are Selected By the Phenosupporting

confidence: 89%

Using Osmotic Stress to Stabilize Mannitol Production in Synechocystis sp. PCC6803

Du²,

Gallego³

et al. 2020

Preprint

View full text Add to dashboard Cite

show abstract

“…6a) and leads us to conclude that mannitol production in Synechocystis can be stabilized by salt stress, in mutants lacking the pathways for synthesis of the two endogenous osmoprotectants. In addition, by purposely weakening the native abilities of cells to tolerate salt stress [25,26], this strategy could allow more mannitol to be synthesized to compensate the loss of salt tolerance ability. Such could allow that mannitol production could be even further sustainably enhanced.…”

Section: Characterization Of Mutations That Are Selected By the Phenomentioning

confidence: 99%

Using osmotic stress to stabilize mannitol production in Synechocystis sp. PCC6803

Gallego

et al. 2020

Biotechnol Biofuels

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Background: Mannitol is a C(6) polyol that is used in the food and medical sector as a sweetener and antioxidant, respectively. The sustainable production of mannitol, especially via the direct conversion of CO 2 by photosynthetic cyanobacteria, has become increasingly appealing. However, previous work aiming to achieve mannitol production in the marine Synechococcus sp. PCC7002 via heterologous expression of mannitol-1-phosphate-5-dehydrogenase (mtlD) and mannitol-1-phosphatase (m1p, in short: a 'mannitol cassette'), proved to be genetically unstable. In this study, we aim to overcome this genetic instability by conceiving a strategy to stabilize mannitol production using Synechocystis sp. PCC6803 as a model cyanobacterium. Results: Here, we explore the stabilizing effect that mannitol production may have on cells faced with osmotic stress, in the freshwater cyanobacterium Synechocystis sp. PCC6803. We first validated that mannitol can function as a compatible solute in Synechocystis sp. PCC6803, and in derivative strains in which the ability to produce one or both of the native compatible solutes was impaired. Wild-type Synechocystis, complemented with a mannitol cassette, indeed showed increased salt tolerance, which was even more evident in Synechocystis strains in which the ability to synthesize the endogenous compatible solutes was impaired. Next we tested the genetic stability of all these strains with respect to their mannitol productivity, with and without salt stress, during prolonged turbidostat cultivations. The obtained results show that mannitol production under salt stress conditions in the Synechocystis strain that cannot synthesize its endogenous compatible solutes is remarkably stable, while the control strain completely loses this ability in only 6 days. DNA sequencing results of the control groups that lost the ability to synthesize mannitol revealed that multiple types of mutation occurred in the mtlD gene that can explain the disruption of mannitol production. Conclusions: Mannitol production in freshwater Synechocsytis sp. PCC6803 confers it with increased salt tolerance. Under this strategy, genetically instability which was the major challenge for mannitol production in cyanobacteria is tackled. This paper marks the first report of utilization of the response to salt stress as a factor that can increase the stability of mannitol production in a cyanobacterial cell factory.

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“…ALE researchers have revealed the adaptive effects under various conditions: flue gas (mainly includes CO 2 , NO x , and SO x ) (Cheng et al, 2019), ultraviolet radiation (UVR) (Korkaric et al, 2015), light intensity (Deblois et al, 2013), organic contaminants (Cho et al, 2016;Wang et al, 2016;Li et al, 2021), wastewater (the main stressors were high concentrations of heavy metals, oxygen, and ammonium nitrogen) (Osundeko et al, 2014), landfill leachate (the main stressors were high concentrations of heavy metals, and ammonium nitrogen) (Okurowska et al, 2021), heavy metals (Muyssen and Janssen, 2001;Mikulic and Beardall, 2014;Yu et al, 2020), nutrient stress (Helliwell et al, 2015;Li T. et al, 2016), sludge extract (Zhao and Huang, 2020). (Wang et al, 2018), CO 2 (Collins and Bell, 2004;Lohbeck et al, 2012;Schlüter et al, 2014;Li et al, 2015;Cheng et al, 2016;Schaum et al, 2017;Listmann et al, 2020), and high salinity (Perrineau et al, 2014b;Lachapelle et al, 2015;Kato et al, 2017;Meijer et al, 2017;Li X. et al, 2018;Sun et al, 2018a;Hu et al, 2020). The cultivation of microalgae under abiotic stress conditions may increase the synthesis of high-value metabolites.…”

Section: Ale Application In Improving Stress Resistance Of Microalgaementioning

confidence: 99%

“…Many research processes have "adaptively evolved" their laboratory microorganisms merely through the inevitable growth or plate inoculation cycle in cell culture. Profiting from the development of proteomics (Wang et al, 2018), transcriptomics (Perrineau et al, 2014b;Li K. et al, 2017;Qi et al, 2017;Zhou et al, 2017;Li X. et al, 2018;Sun et al, 2018a;Cheng et al, 2019;Zhu et al, 2020;Zhang et al, 2021), genomics (Perrineau et al, 2014a;Helliwell et al, 2015;Shin et al, 2017;Rossoni and Weber, 2019;Yu et al, 2020), and metabolomics (Lohbeck et al, 2014;Li X. et al, 2017;Hu et al, 2020), the application of appropriate bioinformatics techniques will contribute to identification of the key mutation mechanisms in ALE (Sandberg et al, 2019).…”

Section: Introductionmentioning

confidence: 99%

Adaptive Laboratory Evolution of Microalgae: A Review of the Regulation of Growth, Stress Resistance, Metabolic Processes, and Biodegradation of Pollutants

Zhang

Meng

2021

Front. Microbiol.

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Adaptive laboratory evolution (ALE) experiments are a serviceable method for the industrial utilization of the microalgae, which can improve the phenotype, performance, and stability of microalgae to obtain strains containing beneficial mutations. In this article, we reviewed the research into the microalgae ALE test and assessed the improvement of microalgae growth, tolerance, metabolism, and substrate utilization by ALE. In addition, the principles of ALE and the key factors of experimental design, as well as the issues and drawbacks of the microalgae ALE method were discussed. In general, improving the efficiency of ALE and verifying the stability of ALE resulting strains are the primary problems that need to be solved in future research, making it a promising method for the application of microalgae biotechnology.

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Divergent metabolic and transcriptomic responses of Synechocystis sp. PCC 6803 to salt stress after adaptive laboratory evolution

Cited by 12 publications

References 50 publications

Using Osmotic Stress to Stabilize Mannitol Production in Synechocystis sp. PCC6803

Using Osmotic Stress to Stabilize Mannitol Production in Synechocystis sp. PCC6803

Using osmotic stress to stabilize mannitol production in Synechocystis sp. PCC6803

Adaptive Laboratory Evolution of Microalgae: A Review of the Regulation of Growth, Stress Resistance, Metabolic Processes, and Biodegradation of Pollutants

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