Fed-batch is the most commonly used cultivation mode for industrial production of recombinant proteins with Pichia pastoris. On a laboratory scale, fed-batch culture provides a way to control the specific biomass growth rates at any pre-set value, allowing the conditions of biomass growth and recombinant product formation to be systematically studied.In this chapter, we present an accessible and versatile approach for designing, performing, and evaluating a fed-batch cultivation in laboratory-scale stirred tank bioreactors.
- Klíčová slova
- Bioprocess design and development, Bioreactor, Data evaluation, Fed-batch culture, Feed rate, Fermentation, Komagataella phaffii, Pichia pastoris, Specific growth rate, Specific productivity,
- MeSH
- biomasa MeSH
- bioreaktory mikrobiologie MeSH
- fermentace MeSH
- kultivační média MeSH
- Pichia * růst a vývoj MeSH
- rekombinantní proteiny genetika biosyntéza MeSH
- Saccharomycetales * růst a vývoj metabolismus MeSH
- techniky vsádkové kultivace * metody MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- kultivační média MeSH
- rekombinantní proteiny MeSH
Laboratory automation is transforming biotechnology, yet current microalgal phenotyping platforms lack the integration and scalability needed for comprehensive trait analysis under diverse conditions. To address this, PhenoSelect was developed, an automated system combining robotics, spectroscopy, fluorometry, flow cytometry, and data analytics for high-throughput, multi-trait phenotyping. Five algal species were profiled across 96 environmental and chemical conditions, quantifying photosynthetic efficiency, growth rate, and cell size. Phenotypic plasticity was quantified using convex hull volume, with trait patterns visualized through Ranked Spider Plots and heatmaps. Haematococcus pluvialis exhibited the largest phenome size, indicating broad plasticity, while Nannochloropsis australis showed the smallest. Optimal growth and photosynthetic performance varied by species, with low light and nutrient-rich media as key drivers. PhenoSelect enables precise, reproducible phenotyping with minimal manual input, supporting applications in biofuels, bioremediation, and nutraceuticals. By accelerating strain screening and optimisation, PhenoSelect bridges phenotyping gaps and drives scalable microalgal biotechnology.
- Klíčová slova
- Automation, Biotechnology, Phenome size, Phenotypic plasticity, Robotics in research,
- MeSH
- biotechnologie metody MeSH
- fenotyp MeSH
- fotosyntéza MeSH
- mikrořasy * růst a vývoj cytologie MeSH
- rychlé screeningové testy * metody MeSH
- Publikační typ
- časopisecké články MeSH
Over the past two decades, advances in molecular and microbiological methods have broadened the range of microorganisms used in biotechnology. Among them, phototrophic bacteria - especially cyanobacteria - are gaining attention for their potential in tackling climate change and producing biopharmaceuticals. While traditional strains such as Escherichia coli and Bacillus subtilis dominate the field, cyanobacteria offer unique features that present both challenges and opportunities, such as complex gene regulation linked to photosynthesis and carbon fixation, protein sorting, and secretion, as well as the ability to establish novel symbiotic partnerships. This review highlights key developments in engineering cyanobacteria and outlines a vision for a future 'supercyanobacterium' that combines the best traits of current strains, unlocking new possibilities in heterotrophy-dominated biotechnology.
- Klíčová slova
- CRISPR systems, RM systems, biotechnology, carbon fixation, carbon fixing, cyanobacteria, photosynthesis, polyploidy, synthetic biology,
- MeSH
- biotechnologie * metody MeSH
- fotosyntéza MeSH
- metabolické inženýrství * metody MeSH
- sinice * genetika metabolismus fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
A new Escherichia coli laboratory evolution screen for detecting plant ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) mutations with enhanced CO2-fixation capacity has identified substitutions that can enhance plant productivity. Selected were a large subunit catalytic (Met-116-Leu) mutation that increases the kcatc of varying plant Rubiscos by 25% to 40% and a solubility (Ala-242-Val) mutation that improves plant Rubisco biogenesis in E. coli 2- to 10-fold. Plastome transformation of either mutation into the tobacco plastome rbcL gene had no impact on leaf Rubisco production, photosynthesis or plant growth. However, tobacco transformed with low-abundance hybrid Arabidopsis Rubisco coding M116L improved plant exponential growth rate by ~75% relative to unmutated hybrid enzyme, with the A242V substitution increasing both hybrid Rubisco production and plant growth by ~50%. Our identification of mutations with the potential to enhance plant growth bodes well for broadening the survey of Rubisco sequence space for catalytic switches that can impart more substantive plant productivity improvements.
- MeSH
- Arabidopsis * genetika enzymologie růst a vývoj MeSH
- Escherichia coli genetika MeSH
- fotosyntéza MeSH
- geneticky modifikované rostliny genetika MeSH
- mutace MeSH
- ribulosa-1,5-bisfosfát-karboxylasa * genetika metabolismus chemie MeSH
- řízená evoluce molekul MeSH
- rozpustnost MeSH
- tabák * genetika růst a vývoj enzymologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- ribulosa-1,5-bisfosfát-karboxylasa * MeSH
We examined the effect of light-induced heating of leaves in wild-type (Col-0) and tmm-1 Arabidopsis plants. The results showed that the tmm-1 mutant exhibits accelerated cooling of rosettes in response to changes in light intensity, as demonstrated by thermal imaging measurements. This is evident in the time constants determined for temperature kinetics, which were reduced during the transition from light to dark. Our findings indicate that these dynamics can be reliably assessed under various light conditions. Measurements of gas exchange showed that stomatal conductance, transpiration rate, and net CO2 assimilation rate were increased in the tmm-1 mutant. Furthermore, fluorescence analysis revealed that the tmm-1 mutant exhibited an increased ratio of total energy dissipation to the number of active reaction centers, as well as higher average absorption. We also observed that the performance index (PIABS) was reduced compared to Col-0. Nevertheless, the clustering of stomata did not affect the maximum quantum yield of PSII. Our results demonstrate that the maintenance of lower leaf temperatures and the greater cooling capacity in plants with clustered stomata positively affects overall photosynthetic performance.
- Klíčová slova
- IR thermal imaging, Light-induced thermal kinetics, Photosynthesis, Stomatal patterning,
- MeSH
- Arabidopsis * fyziologie genetika účinky záření MeSH
- fotosyntéza fyziologie MeSH
- listy rostlin * fyziologie účinky záření MeSH
- mutace MeSH
- oxid uhličitý metabolismus MeSH
- průduchy rostlin fyziologie MeSH
- světlo MeSH
- teplota MeSH
- transpirace rostlin fyziologie MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- oxid uhličitý MeSH
Plant water use efficiency (WUE) links physiological processes to ecosystem-scale carbon and water cycles, making it a crucial parameter for climate change adaptation modelling. Climate and stratospheric ozone dynamics expose plants to varying intensity of ultraviolet-B radiation (UV-B), which affects stomatal function and transpiration. This meta-analysis evaluates UV-B effects on WUE using gas exchange and isotopic proxies. While UV-B radiation reduces stomatal conductance and transpiration, it also suppresses photosynthesis, particularly under non-saturating light. As a result, WUE remains unchanged or declines in UV-B exposed plants, depending on the measurement method. Instantaneous gas exchange-based WUE proxies indicate a decrease, whereas isotope-based proxies, integrating long-term fluxes, show no significant UV-B effect. The suppression of photosynthesis due to UV-B occurs only when UV-B lamps are used to increase the UV-B dose; when UV-B is excluded under field conditions there is no significant effect on WUE. Only some field studies report improved WUE under ambient UV-B, suggesting potential adaptive benefits. Overall, the findings challenge the assumption that UV-B-induced decreases in transpiration enhance WUE. Instead, they highlight a complex interplay between UV radiation, photosynthesis, and stomatal regulation, emphasizing the need to reconsider UV-B's role in plant water relations under future climate conditions.
- Klíčová slova
- photosynthesis, stable carbon isotopes, stomata, ultraviolet radiation,
- MeSH
- fotosyntéza účinky záření MeSH
- klimatické změny MeSH
- průduchy rostlin účinky záření fyziologie MeSH
- rostliny * účinky záření metabolismus MeSH
- transpirace rostlin * účinky záření fyziologie MeSH
- ultrafialové záření * MeSH
- voda * metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- metaanalýza MeSH
- Názvy látek
- voda * MeSH
Desmids are valuable bioindicators in peatland ecosystems due to their sensitivity to environmental changes. In temperate and boreal wetlands, seasonal desiccation of aquatic habitats, which is increasing in frequency and severity due to ongoing climate change, is currently considered a key factor structuring the distribution of individual taxa. In this study, the desiccation tolerance of Micrasterias thomasiana and Staurastrum hirsutum isolated from contrasting hydrological environments in the peatland habitats of the Ore Mountains, Czech Republic, is investigated. Using controlled experimental conditions, we subjected both young, actively growing and old, mature cultures to four different desiccation treatments and evaluated morphology and photosynthetic performance. Our results showed that young and old cultures of both species exhibited a very similar photophysiological response. Severe desiccation led to an irreversible decline in the effective quantum yield of photosystem II in both species, resulting in cell death. Mild drought stress allowed the cultures to recover, indicating that the stress severity determines the recovery potential. Finally, prolonged desiccation resulted in irreversible damage in older cultures of both species, emphasizing the limited desiccation resilience of desmids. We observed similarities in morphology with Zygnema "pre-akinetes," but in contrast to these resilient cells, the old cells of M. thomasiana and S. hirsutum did not survive the harsher desiccation conditions. Long-term mild desiccation revealed a higher resistance of S. hirsutum, probably due to the protective role of its dense mucilage. In nature, these two species usually inhabit localities with low desiccation risk or avoid and mitigate desiccation stress through localized survival strategies.
- Klíčová slova
- Chlorophyll fluorescence, Desiccation tolerance, Desmids, Peat bog, Ultrastructure, Zygnematophyceae,
- MeSH
- fotosyntéza MeSH
- vysoušení * MeSH
- Publikační typ
- časopisecké články MeSH
- srovnávací studie MeSH
Microcontact printing (µCP) is a widely used technique for microscale surface patterning. In this study, we present a polymer-supported µCP method for the patterning of (bioactive) glycosylated surfaces under hydrated conditions. Patterning is achieved by direct contact with a grooved polydimethylsiloxane (PDMS) stamp, whose surface was grafted with a dopamine-containing polymer. The polymer brushes offer an anchor for the boronic acid derivative 6-aminobenzo[c][1,2]oxaborol-1(3H)-ol (ABOB), used as an ink for surface functionalization, to introduce patterns to three different surfaces as substrates: (1) monosaccharide-modified hydrogel surfaces possessing aldose (glucose, fucose, galactose) or ketose (fructose, sorbose) functions; (2) glycosylated surfaces of polymeric microspheres; and (3) the membranes of mammalian cells, such as human primary gastric cells and others. During µCP, ABOB patterns transferred to the target surface through the formation of carbohydrate-ABOB complexes at fully hydrated, neutral pH conditions. Fluorescence microscopy confirmed the successful transfer of ABOB patterns to glycosylated surfaces, with clear "tattoo-like" signatures observed on hydrogels, glycosylated particle surfaces and cellular interfaces.
- Klíčová slova
- Carbohydrate‐boronic acid interactions, Functionalization of cellular interfaces, Glycopolymers, Glycoproteins, Microcontact printing,
- MeSH
- dimethylpolysiloxany chemie MeSH
- glykosylace MeSH
- hydrogely * chemie MeSH
- kyseliny boronové * chemie MeSH
- lidé MeSH
- povrchové vlastnosti MeSH
- tiskařství * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- dimethylpolysiloxany MeSH
- hydrogely * MeSH
- kyseliny boronové * MeSH
BACKGROUND: It is well established that the cancerous transformation of cells is accompanied by profound alterations in glycosylation. In this study, we demonstrate the diagnostic potential of N-glycan profiling in tissue specimens from patients, primarily representing the two major types of lung cancer: non-small cell and small cell lung cancer. METHODS: Lung tissues and biopsies obtained from surgery and bronchoscopy underwent sample processing and enzymatic digestion. After labeling, glycans were analyzed employing matrix-assisted laser desorption/ionization mass spectrometry. Statistical analysis was conducted using methods following principles of compositional data analysis. RESULTS: Comparison of glycan profiles demonstrated an increase in paucimannose and high mannose glycans in most tumor specimens, including those with inflammation and histological negative for malignancy. Cancerous tissues exhibited more profound changes in glycosylation. Despite the high heterogeneity in profiles, two main groups of not detected glycans in peritumoral tissues, considered as controls, were observed to correlate with cancer progression in patients. One with complex polylactosamine multifucosylated glycans frequently harboring terminal N-acetyl-glucosamine residues. These glycans were present in most tumors, with their numbers and intensities increasing as cancer progressed. In contrast, the second group exhibited polylactosamine glycans sporadically. Instead, the biopsies of several patients with rapid progression displayed a significant presence in a set of tri- and tetra-antennary core fucosylated glycans having mostly unoccupied N-acetyl-glucosamine residues unless carrying additional fucose unit(s). CONCLUSIONS: The results imply distinct glycosylation patterns even in patients with the same histological type of lung cancer, supporting trends toward personalized diagnosis and more tailored therapies. Currently, tissue biopsies remain the gold standard for diagnosing premalignant and malignant lesions in the lung. Expanded knowledge on glycosylation in these lesions could contribute to improved diagnostic accuracy and better monitoring of malignant disease progression in clinical practice.
- Klíčová slova
- N-glycans, Branched glycans, Fucosylation, Glycosylation, Lung cancer, MALDI-MS,
- MeSH
- glykosylace MeSH
- lidé středního věku MeSH
- lidé MeSH
- nádory plic * diagnóza metabolismus patologie MeSH
- nemalobuněčný karcinom plic diagnóza metabolismus patologie MeSH
- plíce * patologie metabolismus MeSH
- polysacharidy * metabolismus chemie MeSH
- prognóza MeSH
- prospektivní studie MeSH
- senioři MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- polysacharidy * MeSH
The current interpretation of excitation energy transfer (EET) processes in natural photosynthesis generally relies on Kasha's rule, suggesting that internal conversion (IC) processes usually outpace any EET between higher excited states. It is, however, known from research on artificial systems that Kasha's rule does not apply to many dyes, especially when found in assembled clusters analogous to photosynthetic chlorophyll (Chl)-protein complexes. In this contribution, a semiempirical Förster-type model is applied to otherwise well-investigated pigments of natural photosynthesis (Chls a, b, c1 and various carotenoids). Strong potential for anti-Kasha processes is identified in all investigated pigments, based on their high Coulomb coupling elements, similar to compounds with already known anti-Kasha properties. The pigments are further found to form strongly delocalized excitons, especially between the higher excited states usually responsible for anti-Kasha pathways. Test calculations with different pigment compositions for various natural light harvesting complexes (LHCII, CP24, CP26, CP29, FCP) demonstrate how the higher band EET network and absorbance could be affected by the presence of accessory pigments: Chl a-only networks should perform anti-Kasha EET, but this is suppressed by the presence of accessory pigments via several mechanisms (exciton disruption, spectral competition, energy sinks and fast, non-Chl a IC). The apparent "special" behavior of photosynthetic systems is thus resolved as the result of pigment mixtures.
- MeSH
- biologické pigmenty * chemie MeSH
- chlorofyl chemie MeSH
- fotosyntéza * MeSH
- karotenoidy chemie MeSH
- přenos energie MeSH
- světlosběrné proteinové komplexy chemie metabolismus MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- biologické pigmenty * MeSH
- chlorofyl MeSH
- karotenoidy MeSH
- světlosběrné proteinové komplexy MeSH
