Study of up- and downstream processes in Microcystis aeruginosa cultivation - One approach, two distinct objectives

BioTech 2017 and 7th Czech-Swiss Symposium with Exhibition[Excerpt] The cyanobacterium Microcystis aeruginosa and the accumulation of its cyanotoxin microcystin (MC) have been responsible for several human/animal deaths and intoxication incidents. Therefore, the World Health Organization established recommendation values for MC in water, givingrisetoanincreasingdemandforMC’s analytical standards to be used as laboratory standards both in human and environmental risk assessment studies. These Cyanotoxinsarealsoconsideredpromising anticancer/antitumor drugs as well as antifungal, antialgal and insecticide agents. Despite the interest, commercial MC availability is still limited due to constraints found in production, which inflate the final price to values as high as 28000 e/mg. [...]info:eu-repo/semantics/publishedVersio

Optimization of Microcystis aeruginosa cell disruption to enhance microcystin extraction and purification processes

[Excerpt] Worldwide occurrence of cyanobacterium Microcystis aeruginosa and accumulation of its hepatotoxin microcystin (MC) have been responsible for several incidents, leading the World Health Organization to implement guideline values for this toxin in water thus boosting the demand for MC’s analytical standards. Furthermore, cyanotoxins are also considered promising anticancer/antitumor drugs as well as antifungal, antialgal and insecticide agents. Consequently, the U.S. Environmental Protection Agency has introduced cyanotoxins in its list of substances to be studied as a precursor to regulatory action between 2018 and 2020. [...]info:eu-repo/semantics/publishedVersio

Assessment of synergistic interactions between environmental factors on Microcystis aeruginosa growth and microcystin production

The combined effect of four abiotic factors on Microcystis aeruginosa growth and toxin production was assessed by culturing the cyanobacterium under different light intensities (10190 mol photons·m 2·s 1), CO2 concentrations (010% (v/v)), temperatures (1540 °C), and pH values (6.59.5). Results indicate a significant influence caused by the synergistic effect of environmental factors over growth-related parameters and cyanobacteria toxicity. The combined use of low to medium light intensities (50120 mol photons·m 2·s 1) and CO2 concentration (16% v/v) led to higher cell concentrations, while specific growth rate and biomass productivity were favoured by medium to high light intensities (110190 mol photons·m 2·s 1), CO2 concentrations (49.5% v/v) and temperatures (2939 °C). Regarding microcystin (MC) production, higher concentrations were obtained at low light intensities and low CO2 concentrations while approximately 2000-fold lower MC concentrations were achieved by simultaneous use of high values of light intensity, CO2 concentration and temperature.info:eu-repo/semantics/publishedVersio

Pathway for cyanotoxin valorization: microscystin as case study

Book of Abstracts of CEB Annual Meeting 2017info:eu-repo/semantics/publishedVersio

Valorization of toxic cyanobacteria biomass - disruption efficiency assessment and consequent bioproduct availability using different disruption techniques

The worldwide occurrence of hepatotoxic cyanobacterium Microcystis aeruginosa and accumulation of its toxin microcystin-LR, have been responsible for several human deaths and animal intoxication incidents. In recognition to its toxicity, the World Health Organization and several national governments established guideline values for this toxin in water, which gave rise to an increasing demand for microcystin′s analytical standards. These standards might be useful either as laboratory standards to apply in human and environmental risk assessment or as tools for molecular and cell biology studies. However, their availability is still limited due to constraints found in production and purification processes, which inflate the final price to values as high as 28000 €/mg. As an example of the increasing interest observed over the last years, U.S. Environmental Protection Agency has recently announced that cyanotoxins became part of its list of substances to be studied as a precursor to regulatory action between 2018 and 2020. Consequently, the optimization of this cyanobacterium cultivation and toxin purification techniques is needed to decrease the production cost of such high added-value product. In biotechnological industrial scale processes, the costs associated with downstream processing often represent more than 60 % of the overall expenses. The aim of this work is therefore to provide an insight regarding the development of a costeffective process for obtaining high-quality and affordable microcystin-LR by evaluating the efficiency of three different methodologies (microwave, freeze-thaw cycles and bead mill) on the disruption of M. aeruginosa and consequent availability of bioproducts. For that purpose, several parameters including time, power, and temperature were tested. The best conditions determined for each extraction method were the following: i) 1.5 minutes at 800 W (microwave), ii) three 12- hour cycles at -20 ºC (freeze-thaw cycles), and iii) 7 minutes using 20 % (v/v) of glass beads (bead mill). According to cell counting and intracellular organic matter release determining techniques, freeze-thaw cycles have shown to be the best disruption method presenting an overall efficiency around 97 %

Comparison of harvesting methods for the cyanobacteria Microcystis aeruginosa

BioTech 2017 and 7th Czech-Swiss Symposium with ExhibitionMicrocystis aeruginosa is a wellknown cyanobacterium that has been spreading all over the world due to increased temperatures and eutrophication of water bodies caused by intensive anthropogenic activities. This toxin-producing microorganism is frequently responsible for diminishing water quality and causing intoxication of humans and animals. Due to this, its intracellular cyanotoxin – microcystin (MC) – is commonly used as tool for molecular and cell biology studies or as a standard in human and environmental risk assessment assays. Moreover, MC is a promising anticancer/antitumor drug candidate and a possible antimicrobial, antifungal, antialgal and insecticide agent. Despite MC’s potential application in several biotechnological fields, its high production costs significantly contribute for the prohibitive selling prices (28000 e/mg). Thus, improvements in process’ cost-effectiveness is needed, especially in terms of downstream processing techniques which are probably the major bottlenecks of cyanobacteria production at large scale, commonly representing 20-30 % of the total costs. Bearing this in mind, this study aimed at optimizing harvesting of M. aeruginosa induced by pH change and compares the optimal conditions obtained with the use of three different flocculant agents: chitosan, ferric chloride, and aluminium chloride. Harvesting induced by pH was assessed by testing pH values ranging between 2 and 14. Despite the fact that harvesting efficiencies above 90 % were obtained for most pH values, pH 2 was the one where higher sedimentation rate was observed and consequently the chosen method to compare with the three flocculants. Aluminium chloride addition was found to be the most efficient method, reaching 93 % of sedimentation efficiency within the first 2 h. These results are in agreement with zeta potential measurements where cells presented nearly neutral (approx. 0 mV) charge, while positive or negative charges where achieved using the other three methodologies.This research work was supported by the grant SFRH/BPD/98694/2013 (Bruno Fernandes) and SFRH/BD/52335/2013 (Pedro Geada) from Fundação para a Ciência e a Tecnologia (Portugal). Luís Loureiro is recipient of a fellowship supported by a doctoral advanced training (call NORTE-69-2015-15) funded by the European Social Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), Project UID/Multi/04423/2013, Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462), FCT Strategic Project of UID/BIO/04469/2013 unit, by the project NOVELMAR (reference NORTE-01-0145-FEDER-000035), co-financed by the North Portugal Regional Operational Programme (Norte 2020) under the National Strategic Reference Framework (NSRF), through the ERDF, and by BioTecNorte operation (NORTE-01-0145-FEDER-000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio

Enhancing extraction of food-grade pigments from the microalgae Chlorella Vulgaris through application of ohmic heating

[Excerpt] Introduction: The use of ohmic heating (OH) with the associated non-thermal effects due to the presence of an electrical field and frequency, has been suggested for extraction of compounds from biological matrices. Microalgae are considered as a very valuable source of compounds of interest for food sector (i.e. pigments, lipids, carbohydrates, and proteins) and the selection of extraction technique to recover these compounds is very challenging due to the intrinsic nature of the microalgae cell walls, which limits the mass transfer through it. The main objective of this study is to investigate the effects of OH on the extraction of pigmented solutes from Chlorella vulgaris. [...

Assessment of cell disruption efficiency of Microcystis aeruginosa using different mechanical techniques

This research work was supported by the grant SFRH/BPD/98694/2013 (Bruno Fernandes) and SFRH/BD/52335/2013 (Pedro Geada) from Fundação para a Ciência e a Tecnologia (Portugal). Luís Loureiro is recipient of a fellowship supported by a doctoral advanced training (call NORTE-69-2015-15) funded by the European Social Fund under the scope of Norte2020 - Programa Operacional Regional do Norte. This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), Project UID/Multi/04423/2013, Project RECI/BBB-EBI/0179/2012 (FCOMP-01-0124-FEDER-027462), FCT Strategic Project of UID/BIO/04469/2013 unit, by the project NOVELMAR (reference NORTE-01-0145-FEDER-000035), co-financed by the North Portugal Regional Operational Programme (Norte 2020) under the National Strategic Reference Framework (NSRF), through the ERDF, and by BioTecNorte operation (NORTE-01-0145-FEDER000004) funded by the European Regional Development Fund under the scope of Norte2020 - Programa Operacional Regional do Norte.info:eu-repo/semantics/publishedVersio