Biology subjects

Cerveny, J.

Publications and source records attributed to Cerveny, J..

9 recordsLinked to original sources

bioRxiv2026

Glycogen removal enhances ethylene production in Synechocystis sp. PCC 6803 but exposes a 2-oxoglutarate-GS-GOGAT-phosphate constraint

Cyanobacteria are promising platforms for converting CO2 directly into value-added chemicals. However, redirecting fixed carbon toward a product requires understanding of how cells balance the energetic and redox supplies and demands - and how efficiently metabolic flux can be channelled into product formation rather than being dissipated or excreted. Glycogen, synthesized via ADP-glucose pyrophosphorylase (GlgC), is the major native carbon and energy sink, and its removal forces cells to redirect excess carbon and reducing power into alternative pathways. As a target product we used ethylene, a major industrial building block currently derived almost entirely from fossil feedstocks, which can be produced photoautotrophically by the ethylene-forming enzyme (EFE) at the expense of 2-oxoglutarate (2-OG), a central node linking carbon and nitrogen metabolism. Here, we compared wild-type Synechocystis sp. PCC 6803 with strains carrying either a complete ({Delta}glgC) or attenuated, light-responsive (LGS; psbA2-driven glgC) glycogen synthesis block, with an EFE-expressing strain (EFE), and with the combined {Delta}glgC+EFE strain. All strains were cultivated across a light gradient (35-300 mol photons m-2 s-1) under nitrogen-replete conditions. Integrating measurements of growth and ethylene production rates, chlorophyll-fluorescence analysis, quantitative proteomics, FTIR profiling, and overflow-metabolite analysis revealed that partial suppression of glycogen synthesis (LGS) enhanced growth under low irradiance but became detrimental above a critical light threshold. EFE expression in the {Delta}glgC background increased ethylene production in a light-dependent manner but further compromised growth. Both strains with EFE increased photosynthetic electron transport consistent with the carbon-sink effect, yet this electron surplus was not channelled proportionally into product or biomass. Instead, continuous 2-OG drain by EFE constrained flux through the GS-GOGAT node and induced a secondary inorganic phosphate limitation in {Delta}glgC+EFE that activated photoprotection and limited productivity. These results point to the 2-OG/GS-GOGAT/Pi axis as a key design constraint determining whether the surplus photosynthetic flux gets converted into product, dissipated through photoprotection, or released as overflow metabolites.

plant biology↗

bioRxiv2026

Metabolic cost as a determinant of light quality acclimation: a full-PAR characterization of the CA3 cyanobacterium Nostoc sp. CCAP 1453/38

Light quality acclimation is one of the key drivers of cyanobacterial physiology, ecology, and productivity. Although chromatic acclimation type 3 (CA3) is the canonical example of light quality-driven phycobilisome remodeling, full-PAR physiological characterization of CA3 strains has so far been limited. Here, we characterize the spectral acclimation strategies of the cyanobacterium Nostoc sp. CCAP 1453/38 across the full photosynthetically active radiation (PAR) range. Genomic analysis confirmed Nostoc as a CA3 strain capable of dynamically adjusting phycoerythrin (PE) and phycocyanin (PC) content in its phycobilisome (PBS) rods. During cultivation under narrow-band LEDs, PE was upregulated under violet, blue and green light (435-555 nm), optimizing light harvesting primarily in the blue-green part of the PAR spectrum, while PC was upregulated under red light (633-687 nm). Beyond canonical CA3 pigment switching, Nostoc responded to wavelengths poorly absorbed by PBS in two qualitatively different ways. Under growth-constraining blue light (465 nm), the strain upregulated total PBS and photosystem II (PSII) levels and biased phycobilisome coupling toward PSII (an increased PBS-PSII/PBS-PSI ratio). However, this metabolically costly response could not overcome the underlying excitonic imbalance caused by preferential PSI excitation, resulting in a low cell division rate. Under near far-red light (687 nm), PBS-PSII coupling itself was enhanced, yet total PBS content was reduced rather than increased. Specific growth rates remained as high as under red light, suggesting that this PBS-PSII reorganization avoided the metabolic burden of antenna upregulation. These results indicate that the same underlying challenge of PSII under-excitation can trigger qualitatively different acclimation responses, only some of which are energetically affordable. Spectral acclimation thus depends not only on matching pigment composition to incident wavelengths, but also on the metabolic cost of the response. Compared with parallel datasets on CA1 and non-CA strains obtained under identical conditions, our findings extend CA3 characterization beyond the canonical green/red framework, highlight bottlenecks and advantages of light quality acclimation in Nostoc, and provide a physiological basis for optimizing light regimes in controlled cyanobacterial cultivations.

microbiology↗

bioRxiv2025

Assessing the redox state of the plastoquinone pool in algae and cyanobacteria via OJIP fluorescence: perspectives and limitations

The redox state of the plastoquinone pool (PQ-redox) acts as a central element in a variety of intracellular signal pathways. Several methods for determining PQ-redox have been established. Although some of these methods may be quantitative, such as those based on liquid chromatography, they are typically sensitive to sample preparation. Here, we critically evaluate the use of fast chlorophyll a fluorescence induction kinetics (the so-called OJIP transient) for semi-quantitative PQ-redox estimation in green algae (Chlorella vulgaris) and cyanobacteria (Synechocystis sp. PCC 6803). The method, based on the evaluation of relative fluorescence yield at the J-step of the OJIP transient (VJ, VJ), has already been reported; however, thus far, it has been used mostly for studying dark-acclimated leaves, which limits its range of application. Here, we show that the OJIP transient can be used for semi-quantitative estimation of PQ-redox in algal and cyanobacterial cell cultures, in addition to plants. We further show that it can reflect PQ-redox in both dark-acclimated and light-acclimated samples. Our systematic comparison of Multi-Color PAM, AquaPen, and FL 6000 fluorometers demonstrates that accurate measurement of VJ and VJ parameters in suspension cultures requires low culture density and a high-intensity saturation pulse. We further show that with increasing light intensity to which the cells are exposed, the state of photosystem II (PSII) changes due to light-induced reduction of quinone A (QA-) and conformational changes, which in turn influence both the sensitivity and dynamic range of the VJ parameter towards PQ-redox estimation. A comparison of fluorescence transients in Chlorella and Synechocystis revealed high homeostatic control over PQ-redox in Synechocystis, maintained by terminal oxidases present at the thylakoid membrane. While we discuss certain limitations, our systematic assessment suggests that the OJIP method has great potential to become a routine tool for semi-quantitative PQ-redox estimation under a wide range of experimental conditions in green algae and cyanobacteria.

microbiology↗

bioRxiv2024

The new phycobilisome linker protein ApcI regulates high light adaptation in Synechocystis sp. PCC 6803

Phycobilisomes are versatile cyanobacterial antenna complexes that harvest light energy to drive photosynthesis. These complexes can also adapt to various light conditions, dismantling under high light to prevent photo-oxidation and arranging in rows under low light to increase light harvesting efficiency. Light quality also influences phycobilisome structure and function, as observed under far-red light exposure. Here we describe a new, phycobilisome linker protein, ApcI (previously hypothetical protein sll1911), expressed specifically under red light. We characterized ApcI in Synechocystis sp. PCC 6803 using mutant strain analyses, phycobilisome binding experiments, and protein interaction studies. Mutation of apcI conferred high light tolerance to Synechocystis sp. PCC 6803 compared to wild type with reduced energy transfer from phycobilisomes to the photosystems. Binding experiments revealed that ApcI replaces the linker protein ApcG at the membrane-facing side of the phycobilisome core using a paralogous C-terminal domain. Additionally, the N-terminal extension of ApcI was found to interact with photosystem II. Our findings highlight the importance of phycobilisome remodeling for adaptation under different light conditions. The characterization of ApcI provides new insights into the mechanisms by which cyanobacteria optimize light-harvesting in response to varying light environments.

plant biology↗

bioRxiv2023

Shedding Light On Blue-Green Photosynthesis: A Wavelength-Dependent Mathematical Model Of Photosynthesis In Synechocystis sp. PCC 6803

Cyanobacteria hold great potential to revolutionize conventional industries and farming practices with their light-driven chemical production. To fully exploit their photosynthetic capacity and enhance product yield, it is crucial to investigate their intricate interplay with the environment including the light intensity and spectrum. Mathematical models provide valuable insights for optimizing strategies in this pursuit. In this study, we present an ordinary differential equation-based model for the cyanobacterium Synechocystis sp. PCC 6803 to assess its performance under various light sources, including monochromatic light. Our model can reproduce a variety of physiologically measured quantities, e.g. experimentally reported partitioning of electrons through four main pathways, O2 evolution, and the rate of carbon fixation for ambient and saturated CO2. By capturing the interactions between different components of a photosynthetic system, our model helps in understanding the underlying mechanisms driving system behavior. Our model qualitatively reproduces fluorescence emitted under various light regimes, replicating Pulse-amplitude modulation (PAM) fluorometry experiments with saturating pulses. Using our model, we test four hypothesized mechanisms of cyanobacterial state transitions. Moreover, we evaluate metabolic control for biotechnological production under diverse light colors and irradiances. By offering a comprehensive computational model of cyanobacterial photosynthesis, our work enhances the basic understanding of light-dependent cyanobacterial behavior and sets the first wavelength-dependent framework to systematically test their producing capacity for biocatalysis.

plant biology↗

bioRxiv2023

Photo-physiological Acclimation in Synechocystis sp. PCC 6803 Provides Insight into Growth Limitation in Underwater Spectra

Cyanobacteria play a key role in primary production in both oceans and fresh waters and hold great potential for sustainable production of a large number of commodities. During their life, cyanobacteria cells need to acclimate to a multitude of challenges, including shifts in intensity and quality of incident light. Despite our increasing understanding of metabolic regulation under various light regimes, detailed insight into fitness advantages and limitations under shifting light quality has been missing. Here, we study photo-physiological acclimation in the cyanobacterium Synechocystis sp. PCC 6803 through the whole range of photosynthetically active radiation (PAR). Using LEDs with qualitatively different narrow spectra, we describe wavelength dependence of light capture, electron transport and energy transduction to main cellular pools. In addition, we describe processes fine-tuning light capture such as state transitions and efficiency of energy transfer from phycobilisomes to photosystems. We show that growth was the most limited under blue light due to inefficient light harvesting, and that many cellular processes are tightly linked to the redox state of the PQ pool, which was the most reduced under red light. The PSI-to-PSII ratio was low under blue photons, however, it was not the main growth-limiting factor, since it was even more reduced under violet and near far-red lights, where Synechocystis grew faster compared to blue light. Our results provide insight into the spectral dependence of phototrophic growth and can provide the foundation for future studies of molecular mechanisms underlying light acclimation in cyanobacteria, leading to light optimization in controlled cultivations.

plant biology↗

bioRxiv2023

The phycobilisome linker protein ApcG interacts with photosystem II and regulates energy transfer to photosystem I in Synechocystis sp. PCC 6803

Photosynthetic organisms harvest light using pigment-protein super-complexes. In cyanobacteria, these are water-soluble antennae known as phycobilisomes (PBSs). The light absorbed by PBS is transferred to the photosystems in the thylakoid membrane to drive photosynthesis. The energy transfer between these super-complexes implies that protein-protein interactions allow the association of PBS with the photosystems. However, the specific proteins involved in the interaction of PBS with the photosystems are not fully characterized. Here, we show that the newly discovered PBS linker protein ApcG interacts specifically with photosystem II through its N-terminal region. Growth of cyanobacteria is impaired in apcG deletion strains under light-limiting conditions. Furthermore, complementation of these strains using a phospho-mimicking version of ApcG exhibit reduced growth under normal growth conditions. Interestingly, the interaction of ApcG with photosystem II is affected when a phospho-mimicking version of ApcG is used, targeting the positively charged residues interacting with thylakoid membrane suggesting a regulatory role mediated by phosphorylation of ApcG. Low temperature fluorescence measurements showed increased photosystem I fluorescence in apcG deletion and complementation strains. The photosystem I fluorescence was the highest in the phospho-mimicking complementation strain while pull-down experiment showed no interaction of ApcG with PSI under any tested condition. Our results highlight the importance of ApcG for selectively directing energy harvested by the PBS and implies that the phosphorylation status of ApcG plays a role in regulating energy transfer from PSII to PSI.

plant biology↗

bioRxiv2018

Quantitative insights into the cyanobacterial cell economy

Phototrophic microorganisms are promising resources for green biotechnology. Compared to heterotrophic microorganisms, however, the cellular economy of phototrophic growth is still insufficiently understood. We provide a quantitative analysis of light-limited, light-saturated, and light-inhibited growth of the cyanobacterium Synechocystis sp. PCC 6803 using a reproducible cultivation setup. We report key physiological parameters, including growth rate, cell size, and photosynthetic activity over a wide range of light intensities. Intracellular proteins were quantified to monitor proteome allocation as a function of growth rate. Among other physiological adaptations, we identify an upregulation of the translational machinery and downregulation of light harvesting components with increasing light intensity and growth rate. The resulting growth laws are discussed in the context of a coarse-grained model of phototrophic growth and available data obtained by a comprehensive literature search. Our insights into quantitative aspects of cyanobacterial adaptations to different growth rates have implications to understand and optimize photosynthetic productivity.

microbiology↗

bioRxiv2017

A minimal model of protein allocation during phototrophic growth

Photoautotrophic growth depends upon an optimal allocation of finite cellular resources to diverse intracellular processes. Commitment of a certain mass fraction of the proteome to a specific cellular function, typically reduces the proteome available for other cellular functions. Here, we develop a minimal semi-quantitative kinetic model of cyanobacterial phototrophic growth to describe such trade-offs of cellular protein allocation. The model is based on coarse-grained descriptions of key cellular processes, in particular carbon uptake, metabolism, photosynthesis, and protein translation. The model is parametrized using literature data and experimentally obtained growth curves. Of particular interest are the resulting cyanobacterial growth laws as fundamental characteristics of cellular growth. We show that the model gives rise to similar growth laws as observed for heterotrophic organisms, with several important differences due to the distinction between light energy and carbon uptake. We discuss recent experimental data supporting the model results and show that minimal growth models have implications for our understanding of the limits of phototrophic growth and bridge a gap between molecular physiology and ecology.

systems biology↗