Growth of Cyanobacteria Is Constrained by the Abundance of Light and Carbon Assimilation Proteins

Jahn, Michael; Vialás, Vital; Karlsen, Jan; Maddalo, Gianluca; Edfors, Fredrik; Forsström, Björn; Uhlén, Mathias; Käll, Lukas; Hudson, Elton P.

doi:10.1016/j.celrep.2018.09.040

Cited by 109 publications

(134 citation statements)

References 49 publications

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“…Hence, within the limits of the Hi3 technique, it appears that the L-, P-, T- and H-protein are present in a molar ratio of approximately 1L 2 -0.5P 2 -0.6T-2.4H in Synechocystis cells and collectively represent at least ∼0.051 mole %, corresponding to ∼0.07% mass % of the total protein. With a protein content of roughly 300 mg ml -1 for Synechocystis (Jahn et al , 2018), this value corresponds to about 0.2 mg ml -1 total GCS protein, which is distinctly above the pea leaf GDC dissociation threshold of about 0.08 mg ml -1 determined by (Oliver et al , 1990).…”

Section: Resultsmentioning

confidence: 76%

Stoichiometry of two plant glycine decarboxylase complexes and comparison with a cyanobacterial glycine cleavage system

Wittmiß

Mikkat

Hagemann

et al. 2020

Preprint

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ABSTRACTThe multienzyme glycine cleavage system (GCS) converts glycine and tetrahydrofolate to the one-carbon compound 5,10-methylenetetrahydrofolate, which is of vital importance for most if not all organisms. Photorespiring plant mitochondria contain very high levels of GCS proteins organised as a fragile glycine decarboxylase complex (GDC). The aim of this study is to provide mass spectrometry-based stoichiometric data for the plant leaf GDC and examine whether complex formation could be a general property of the GCS in photosynthesizing organisms. The molar ratios of the leaf GDC component proteins are 1L2-4P2-8T-26H and 1L2-4P2-8T-20H for pea and Arabidopsis, respectively, as determined by mass spectrometry. The minimum mass of the plant leaf GDC ranges from 1,550-1,650 kDa, which is larger than previously assumed. The Arabidopsis GDC contains four times more of the isoforms GCS-P1 and GCS-L1 in comparison with GCS-P2 and GCS-L2, respectively, whereas the H-isoproteins GCS-H1 and GCS-H3 are fully redundant as indicated by their about equal amounts. Isoform GCS-H2 is not present in leaf mitochondria. In the cyanobacterium Synechocystis sp. PCC 6803, GCS proteins are present at low concentration but above the complex formation threshold reported for pea leaf GDC. Indeed, formation of a cyanobacterial GDC from the individual recombinant GCS proteins in vitro could be demonstrated. Presence and metabolic significance of a Synechocystis GDC in vivo remain to be examined but could involve multimers of the GCS H-protein that dynamically crosslink the three GCS enzyme proteins, facilitating glycine metabolism by the formation of multienzyme metabolic complexes.

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Section: Resultsmentioning

confidence: 76%

Stoichiometry of two plant glycine decarboxylase complexes and comparison with a cyanobacterial glycine cleavage system

Wittmiß

Mikkat

Hagemann

et al. 2020

Preprint

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“…2c). Cyanobacteria invest most of their resources into synthesizing proteins from these groups 17,18 . Interestingly, nearly all genes with a low fitness score are also among proteins found by Jahn et al to be regulated with growth rate (Supplemental Fig.…”

Section: Gene Fitness During Photoautotrophic Growthmentioning

confidence: 99%

“…The Synechocystis sgRNA library was cultivated in an 8-tube photobioreactor (Multi-Cultivator MC-1000-OD, Photon System Instruments, Drasov, CZ). The system was customized to perform turbidostat cultivation as described previously 17 . Reactors (65 mL) were bubbled with 1% v/v CO 2 in air at 2.5 mL/min, and light intensity was controlled by a computer program.…”

Section: Cultivation In Photobioreactor Turbidostatsmentioning

confidence: 99%

“…To correlate gene fitness (obtained from sgRNA library competition experiments) with variability in gene expression, a previously published proteomics data set was used 17 . The dataset contains mass spectrometry based protein measurements of Synechocystis for 2,000 proteins at 5 different growth rates, each for CO 2 and light limitation.…”

Section: Correlation Between Gene Expression Variability and Fitnessmentioning

confidence: 99%

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Pooled CRISPRi screening of the cyanobacteriumSynechocystissp. PCC 6803 for enhanced growth, tolerance, and chemical production

Yao

Shabestary²,

Björk³

et al. 2019

Preprint

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We developed an inducible CRISPRi gene repression library in the cyanobacterium Synechocystis sp. PCC 6803, where all annotated genes are targeted for repression. We used the library to estimate gene fitness in multiple conditions. The library revealed several mutants with increased specific growth rates (up to 17%), and transcriptomics of these mutants revealed common upregulation of genes within photosynthetic electron flow. We challenged the library with L-lactate stress to find more tolerant mutants. Repression of the peroxiredoxin Bcp2 increased growth rate by 49% in the presence of 0.1 M L-lactate. Finally, the library was transformed into a L-lactate-secreting strain, and droplet microfluidics sorting of top producers enriched sgRNAs targeting nutrient assimilation, redox modulation, and cyclic-electron flow.Several clones showed increased productivity in batch cultivations (up to 75%). In some cases, tolerance or productivity was enhanced by partial repression of essential genes, which are difficult to access by transposon insertion.

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“…Synechcocystis PCC 6803 is an ideal host because it grows rapidly and it is genetically tractable, 32,33 tolerant to abiotic stress, 34 and conditions for optimizing its growth are well understood. 35,36 The photobiological conversion of CO 2 into medium chain-length fatty acids 37 and long chain hydrocarbons 38 in Synechcocystis PCC 6803 has been described, the latter utilising an ADO-or FAP-based system. Thioesterase A from E. coli was incorporated, which catalyses the conversion of fatty acyl-ACP to free fatty acids, in addition to knocking out the native fatty acyl ACP synthase gene (∆aas) to minimise the reverse reaction (Figure 7a).…”

Section: Bio-propane Production By Photosynthetic Carbon Capturementioning

confidence: 99%

Distributed Biomanufacturing of Liquefied Petroleum Gas

Hoeven

Jmx

Amer

et al. 2019

Preprint

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Liquefied Petroleum Gas (LPG) is a major domestic and transport fuel. Its combustion lessens NO x , greenhouse gas and particulates emissions compared to other fuels. Propane -the major constituent of LPG -is a clean, high value 'drop-in' fuel that can help governments develop integrated fuels and energy policies with low carbon burden, providing solutions to the multi-faceted challenges of future energy supply. We show that bio-LPG (bio-propane and bio-butane) can be produced by microbial conversion of waste volatile fatty acids that can be derived from anaerobic digestion, industrial waste, or CO 2 via photosynthesis. Bio-LPG production was achieved photo-catalytically, using biomass propagated from bioengineered bacteria including E. coli, Halomonas (in non-sterile seawater), and Synechocystis (photosynthetic). These fuel generation routes could be implemented rapidly in advanced and developing nations of the world to meet energy needs, global carbon reduction targets and clean air directives.[141words]

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Growth of Cyanobacteria Is Constrained by the Abundance of Light and Carbon Assimilation Proteins

Cited by 109 publications

References 49 publications

Stoichiometry of two plant glycine decarboxylase complexes and comparison with a cyanobacterial glycine cleavage system

Stoichiometry of two plant glycine decarboxylase complexes and comparison with a cyanobacterial glycine cleavage system

Pooled CRISPRi screening of the cyanobacteriumSynechocystissp. PCC 6803 for enhanced growth, tolerance, and chemical production

Distributed Biomanufacturing of Liquefied Petroleum Gas

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