Fatty acid production in genetically modified cyanobacteria

Liu, Xinyao; Sheng, Jie; Curtiss, Roy

doi:10.1073/pnas.1103014108

Cited by 472 publications

(413 citation statements)

References 42 publications

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“…1 and Table S1). By using the Km R ∕sacB intermediate double cross-over recombination method (13), we inserted synthesized genes fol, shl, gpl, and an artificial operon fol ribosome binding site (RBS) gpl into SD100 after P cmp and before the start codon of the cmpA gene, resulting in strains SD256, SD257, SD258, and SD237, respectively. The gpl gene was inserted into wild type and SD237 after P sbt and before the ATG of the sbtA gene to result in SD252 and SD253, respectively.…”

Section: Resultsmentioning

confidence: 99%

“…Recently, we reported our results in constructing SD strains that secrete FFAs into the culture media by introducing acyl-acyl carrier protein (acyl-ACP) thioesterase genes into SD100 and other genetic modifications (13). The FFA-secretion strains that harbor the Green Recovery system (SD239, SD254, and SD262 in Table 1) are still able to release membrane lipids as FFAs at rates faster than non-FFAsecretion strains following CO 2 limitation.…”

Section: Resultsmentioning

confidence: 99%

“…Green Recovery exhibits other advantages when combined with the previously described cyanobacterial FFA-secretion system (13). The FFA-secretion system avoids the energy-intensive biomass processes such as concentration and extraction by directly recovering the secreted FFA from the culture medium.…”

Section: Discussionmentioning

confidence: 99%

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CO ₂ -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass

Liu

Fallon

Sheng

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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Using genetically modified cyanobacterial strains, we engineered a Green Recovery strategy to convert membrane lipids into fatty acids for economical and environmentally sustainable biofuel production. The Green Recovery strategy utilizes lipolytic enzymes under the control of promoters induced by CO 2 limitation. Data indicate that strains of the cyanobacterium Synechocystis sp. PCC6803 engineered for Green Recovery underwent degradation of membrane diacylglycerols upon CO 2 limitation, leading to release of fatty acids into the culture medium. Recovered fatty acid yields of 36.1 × 10 -12 mg/cell were measured in one of the engineered strains (SD239). Green Recovery can be incorporated into previously constructed fatty-acid-secretion strains, enabling fatty acid recovery from the remaining cyanobacterial biomass that will be generated during fatty acid biofuel production in photobioreactors.

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

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CO ₂ -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass

Liu

Fallon

Sheng

et al. 2011

Proc. Natl. Acad. Sci. U.S.A.

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“…Cyanobacteria are the evolutionary ancestors of plant plastids, therefore FAS machineries of both are similar (Vothknecht and Westhoff, 2001). In a recent study, genetically modified cyanobacteria were used to produce and secrete fatty acids mimicking a process that occurs in plants, where FFAs are synthesized in plastids and subsequently exported to the cytosol (Liu et al, 2011).…”

Section: Accase In Bacteria and Algae-based Biotech Projectsmentioning

confidence: 99%

OPINIONS Acetyl-coenzyme A carboxylase – an attractive enzyme for biotechnology

Podkowiński¹,

Tworak²

2011

bta

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Acetyl-CoA carboxylase catalyzes the carboxylation of acetyl-CoA to malonyl-CoA. This reaction constitutes the first committed step of fatty acid synthesis in most organisms, while its product also serves as a universal precursor for various other high-value compounds. The important regulatory and rate-limiting role of acetyl-CoA carboxylase makes this enzyme a powerful tool in a variety of biotechnological and medical projects. This review presents the current knowledge on structural and functional features of the enzyme and focuses on its divergent applications. Different metabolic engineering attempts that lead to the production of various compounds, such as fatty acids, polyketides or flavonoids, are presented and their advantages and limitations discussed. The importance of studies on acetyl-CoA carboxylase activity for design and development of new herbicides and antibiotics as well as for medical intervention is also highlighted. Acetyl-coenzyme A carboxylase -enzyme architecture, biochemistry and its role in cell physiologyA variety of biotechnological projects targeted to modulate acetyl-coenzyme A carboxylase activity in bacteria, plants and humans have been proposed and experimentally tested. Here, we would like to present a comprehensive review of these strategies together with a survey of the potential of acetyl-CoA carboxylase for biotechnology.Acetyl-coenzyme A carboxylase (ACC, E.C.6.4.1.2), recognized as an essential enzyme for all Eukaryotes and the majority of Bacteria, is responsible for carboxylation of acetyl-CoA that results in malonyl-coenzyme A formation (Fig. 1). Malonyl-CoA is a major building block for compounds such as fatty acids, polyketides and flavonoids -important components for cell growth, as well as for food, pharmaceuticals and energy production.The malonyl-CoA synthesis consists of two half-reactions (Nikolau et al., 2003). The first one, adenosine triphosphate-dependent carboxylation of biotin prosthetic group covalently bound to a module named biotin carboxyl carrier protein (BCCP), is catalyzed by ACCase segment of biotin carboxylase activity (BC) (Fig. 2). This reaction is immediately followed by the second half-re action: the transfer of a carboxyl group from carboxylated biotin to acetyl-CoA, resulting in malonyl-CoA formation. Carboxyl transferase (CT) is a module that catalyzes the second reaction and provides the required specificity in recognition of the carboxyl acceptor (acetyl-CoA). The ACCase model usually presents BCCP, a module with covalently attached biotin, as a beam responsible for biotin dislocation between two active centers -one with BT and the other with CT activity. The three (Fig. 3). The prokaryotic and eukaryotic types of ACCases are evolutionary related and the phylogeny of the modules provides some hints about their cenancestor, a split between Archaea and Bacteria, and arising of Eukaryota (Lombard and Moreira, 2011).Phylogeny is out of scope of this manuscript, but acetyl-coenzyme A carboxylases from various organisms representing v...

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“…nutrients and light; Kim et al, 2011;Lea-Smith et al, 2014;Burnap, 2015;Touloupakis et al, 2015;van Alphen and Hellingwerf, 2015). Understanding of the factors controlling the limitation of Synechocystis growth would facilitate the use of this strain as a cell factory (Yu et al, 2013) for the production of biomass (Joseph et al, 2014), pigments (Sekar and Chandramohan, 2008), secondary metabolite natural products (Frommeyer et al, 2016), biofuel (Dexter and Fu, 2009;Baebprasert et al, 2011;Liu et al, 2011), and other high-value compounds.…”

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confidence: 99%

A Novel Mechanism, Linked to Cell Density, Largely Controls Cell Division in Synechocystis

et al. 2017

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ORCID IDs: 0000-0001-7618-4748 (M.I.); 0000-0001-8931-7722 (T.O.); 0000-0002-6113-9570 (R.S.).Many studies have investigated the various genetic and environmental factors regulating cyanobacterial growth. Here, we investigated the growth and metabolism of Synechocystis sp. PCC 6803 under different nitrogen sources, light intensities, and CO 2 concentrations. Cells grown on urea showed the highest growth rates. However, for all conditions tested, the daily growth rates in batch cultures decreased steadily over time, and stationary phase was obtained with similar cell densities. Unexpectedly, metabolic and physiological analyses showed that growth rates during log phase were not controlled primarily by the availability of photoassimilates. Further physiological investigations indicated that nutrient limitation, quorum sensing, light quality, and light intensity (self-shading) were not the main factors responsible for the decrease in the growth rate and the onset of the stationary phase. Moreover, cell division rates in fed-batch cultures were positively correlated with the dilution rates. Hence, not only light, CO 2 , and nutrients can affect growth but also a cell-cell interaction. Accordingly, we propose that cell-cell interaction may be a factor responsible for the gradual decrease of growth rates in batch cultures during log phase, culminating with the onset of stationary phase.

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Fatty acid production in genetically modified cyanobacteria

Cited by 472 publications

References 42 publications

CO ₂ -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass

CO ₂ -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass

OPINIONS Acetyl-coenzyme A carboxylase – an attractive enzyme for biotechnology

A Novel Mechanism, Linked to Cell Density, Largely Controls Cell Division in Synechocystis

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Fatty acid production in genetically modified cyanobacteria

Cited by 472 publications

References 42 publications

CO 2 -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass

CO 2 -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass

OPINIONS Acetyl-coenzyme A carboxylase – an attractive enzyme for biotechnology

A Novel Mechanism, Linked to Cell Density, Largely Controls Cell Division in Synechocystis

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Product

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CO ₂ -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass

CO ₂ -limitation-inducible Green Recovery of fatty acids from cyanobacterial biomass