scholarly journals Untargeted Lipidomics Analysis of the Cyanobacterium Synechocystis sp. PCC 6803: Lipid Composition Variation in Response to Alternative Cultivation Setups and to Gene Deletion

2020 ◽  
Vol 21 (23) ◽  
pp. 8883
Author(s):  
Weronika Hewelt-Belka ◽  
Ágata Kot-Wasik ◽  
Paula Tamagnini ◽  
Paulo Oliveira
Keyword(s):  
Lipid Composition ◽  
Gene Deletion ◽  
Total Lipid Content ◽  
Outer Membrane Vesicles ◽  
Cell Factories ◽  
Composition Variation ◽  
Lipid Species ◽  
Alternative Cultivation ◽  
Synechocystis Sp ◽  
Pcc 6803

Cyanobacteria play an important role in several ecological environments, and they are widely accepted to be the ancestors of chloroplasts in modern plants and green algae. Cyanobacteria have become attractive models for metabolic engineering, with the goal of exploring them as microbial cell factories. However, the study of cyanobacterial lipids’ composition and variation, and the assessment of the lipids’ functional and structural roles have been largely overlooked. Here, we aimed at expanding the cyanobacterial lipidomic analytical pipeline by using an untargeted lipidomics approach. Thus, the lipid composition variation of the model cyanobacterium Synechocystis sp. PCC 6803 was investigated in response to both alternative cultivation setups and gene deletion. This approach allowed for detecting differences in total lipid content, alterations in fatty-acid unsaturation level, and adjustments of specific lipid species among the identified lipid classes. The employed method also revealed that the cultivation setup tested in this work induced a deeper alteration of the cyanobacterial cell lipidome than the deletion of a gene that results in a dramatic increase in the release of lipid-rich outer membrane vesicles. This study further highlights how growth conditions must be carefully selected when cyanobacteria are to be engineered and/or scaled-up for lipid or fatty acids production.

scholarly journals Photosystem I oligomerization affects lipid composition in Synechocystis sp. PCC 6803

2019 ◽  
Vol 1864 (10) ◽  
pp. 1384-1395 ◽  
Author(s):  
Terezia Kovacs ◽  
Balazs Szalontai ◽  
Kinga Kłodawska ◽  
Radka Vladkova ◽  
Przemysław Malec ◽  
...  
Keyword(s):  
Photosystem I ◽  
Lipid Composition ◽  
Synechocystis Sp ◽  
Pcc 6803

Quantitative and Qualitative Analyses of Triacylglycerol Production in the Wild-Type Cyanobacterium Synechocystis sp. PCC 6803 and the Strain Expressing AtfA from Acinetobacter baylyi ADP1

2020 ◽  
Vol 61 (9) ◽  
pp. 1537-1547 ◽  
Author(s):  
Motoki Tanaka ◽  
Toshiki Ishikawa ◽  
So Tamura ◽  
Yujiro Saito ◽  
Maki Kawai-Yamada ◽  
...  
Keyword(s):  
Fatty Acids ◽  
Neutral Lipid ◽  
Saturated Fatty Acids ◽  
Wax Ester ◽  
Wild Type ◽  
Acinetobacter Baylyi ◽  
Lipid Species ◽  
Acinetobacter Baylyi Adp1 ◽  
Synechocystis Sp ◽  
Pcc 6803

Abstract Although cyanobacteria do not possess wax ester synthase/acyl-CoA:diacylglycerol acyltransferase (WS/DGAT), the bacterial enzyme for triacylglycerol (TAG) production, there have been several studies reporting the accumulation of TAG-like compounds in cyanobacteria. In this study, we aimed to evaluate TAG productivity of the ΔrecJ::atfA strain of Synechocystis sp. PCC 6803 generated by inserting atfA encoding WS/DGAT from Acinetobacter baylyi ADP1 into recJ (sll1354), together with the wild type (WT) and the gene-disrupted strain of slr2103 having homology with eukaryotic DGAT2 gene family (Δ2103). Thin-layer chromatography (TLC) of neutral lipids or isolation of the neutral lipid-enriched fraction followed by gas chromatography or liquid chromatography–tandem mass spectrometry was employed for analyses. The ΔrecJ::atfA strain accumulated 0.508 nmol ml−1OD730−1 of TAG after a week of incubation at 100 μmol photons m−2 s−1. The saturated fatty acids C16:0 and C18:0 accounted for about 50% and 20% of the TAG fatty acids, respectively, suggesting that de novo-synthesized fatty acids were preferentially incorporated into TAG molecules. When the neutral lipid profile of the lipid extracts was examined by TLC, a spot located in a slightly lower position compared with the TAG standard was detected in WT but not in the Δ2103 strain. TAG accumulation levels of both strains was only 0.01–0.03 nmol ml−1OD730−1, but the fatty acid composition was substantially different from that of the background. These results suggest that trace amounts of TAG can be produced in Synechocystis cells by enzymes other than Slr2103, and major constituents of the TAG-like spot are unknown lipid species produced by Slr2103.

scholarly journals Expanding the toolbox for Synechocystis sp. PCC 6803: validation of replicative vectors and characterization of a novel set of promoters

2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Eunice A Ferreira ◽  
Catarina C Pacheco ◽  
Filipe Pinto ◽  
José Pereira ◽  
Pedro Lamosa ◽  
...  
Keyword(s):  
Low Cost ◽  
Carbon Sources ◽  
Proof Of Concept ◽  
Gene Encoding ◽  
Metabolic Plasticity ◽  
Cell Factories ◽  
Wide Range ◽  
Synechocystis Sp ◽  
Pcc 6803

Abstract Cyanobacteria are promising ‘low-cost’ cell factories since they have minimal nutritional requirements, high metabolic plasticity and can use sunlight and CO2 as energy and carbon sources. The unicellular Synechocystis sp. PCC 6803, already considered the ‘green’ Escherichia coli, is the best studied cyanobacterium but to be used as an efficient and robust photoautotrophic chassis it requires a customized and well-characterized toolbox. In this context, we evaluated the possibility of using three self-replicative vectors from the Standard European Vector Architecture (SEVA) repository to transform Synechocystis. Our results demonstrated that the presence of the plasmid does not lead to an evident phenotype or hindered Synechocystis growth, being the vast majority of the cells able to retain the replicative plasmid even in the absence of selective pressure. In addition, a set of heterologous and redesigned promoters were characterized exhibiting a wide range of activities compared to the reference PrnpB, three of which could be efficiently repressed. As a proof-of-concept, from the expanded toolbox, one promoter was selected and assembled with the ggpS gene [encoding one of the proteins involved in the synthesis of the native compatible solute glucosylglycerol (GG)] and the synthetic device was introduced into Synechocystis using one of the SEVA plasmids. The presence of this device restored the production of the GG in a ggpS deficient mutant validating the functionality of the tools/device developed in this study.

scholarly journals Enhanced Production of Photosynthetic Pigments and Various Metabolites and Lipids in the Cyanobacteria Synechocystis sp. PCC 7338 Culture in the Presence of Exogenous Glucose

Biomolecules ◽  
2021 ◽  
Vol 11 (2) ◽  
pp. 214
Author(s):  
YuJin Noh ◽  
Hwanhui Lee ◽  
Myeongsun Kim ◽  
Seong-Joo Hong ◽  
Hookeun Lee ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Cell Growth ◽  
Photosynthetic Pigments ◽  
Industrial Applications ◽  
Gas Chromatography Mass Spectrometry ◽  
Enhanced Production ◽  
Exogenous Glucose ◽  
Nanoelectrospray Ionization ◽  
Lipid Species ◽  
Synechocystis Sp

Synechocystis strains are cyanobacteria that can produce useful biomaterials for biofuel and pharmaceutical resources. In this study, the effects of exogenous glucose (5-mM) on cell growth, photosynthetic pigments, metabolites, and lipids in Synechocystis sp. PCC 7338 (referred to as Synechocystis 7338) were investigated. Exogenous glucose increased cell growth on days 9 and 18. The highest production (mg/L) of chlorophyll a (34.66), phycocyanin (84.94), allophycocyanin (34.28), and phycoerythrin (6.90) was observed on day 18 in Synechocystis 7338 culture under 5-mM glucose. Alterations in metabolic and lipidomic profiles under 5-mM glucose were investigated using gas chromatography-mass spectrometry (MS) and nanoelectrospray ionization-MS. The highest production (relative intensity/L) of aspartic acid, glutamic acid, glycerol-3-phosphate, linolenic acid, monogalactosyldiacylglycerol (MGDG) 16:0/18:1, MGDG 16:0/20:2, MGDG 18:1/18:2, neophytadiene, oleic acid, phosphatidylglycerol (PG) 16:0/16:0, and PG 16:0/17:2 was achieved on day 9. The highest production of pyroglutamic acid and sucrose was observed on day 18. We suggest that the addition of exogenous glucose to Synechocystis 7338 culture could be an efficient strategy for improving growth of cells and production of photosynthetic pigments, metabolites, and intact lipid species for industrial applications.

scholarly journals Cloning of the psbK gene from Synechocystis sp. PCC 6803 and characterization of photosystem II in mutants lacking PSII-K

1991 ◽  
Vol 266 (17) ◽  
pp. 11111-11115
Author(s):  
M. Ikeuchi ◽  
B. Eggers ◽  
G.Z. Shen ◽  
A. Webber ◽  
J.J. Yu ◽  
...  
Keyword(s):  
Photosystem Ii ◽  
Synechocystis Sp ◽  
Pcc 6803
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