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Research Project

Biodiversity and Bioprospection of Biosurfactants In Saline environments

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Valorisation of microalga Tetradesmus obliquus grown in brewery wastewater using subcritical water extraction towards zero waste
Publication . Ferreira, Alice; Molnar Jazić, Jelena; Gouveia, Luisa; Maletić, Snežana; Tomić, Milan; Agbaba, Jasmina; Vladić, Jelena
n this study, green technology was applied for extraction of compounds from wastewater-grown microalga biomass with the final goal of obtaining microbiologically safe products within a sustainable biorefinery process with zero waste. Tetradesmus obliquus biomass resulting from brewery wastewater treatment, with (To-CO2) and without CO2 supplementation (To), was submitted to subcritical water extraction (SWE) at temperatures 120-220 degrees C for 10 min. The impact of the different SWE conditions in the obtained liquid extracts and solid residues were investigated for metal content and chemical and microbiological profiles. Gas chromatography-mass spectrometry analysis indicated that of T. obliquus extracts and residues are valuable sources of aliphatic saturated, unsaturated, and alkylated (mostly methylated) hydrocarbons, phenols, esters, and ketones. Polyphenolic content and antioxidant activity were enhanced approximately 4 times by increasing the temperature from 120 to 220 degrees C. Also, the content of polyphenols doubled when T. obliquus was supplemented with CO2 for all the tested temperatures (To: 0.249-1.016 mg GAE mL(-1); To-CO2: 0.437 - 1.767 mg GAE mL(-1)). The microbiological analysis determined that liquid extracts and residues represent safe sources of bioactive components that can be used in different industries. In addition, the lower content of heavy metals in residues suggests the possibility of using the solid waste as animal feed or soil conditioner in agricultural applications.
Application of green technology to extract clean and safe bioactive compounds from tetradesmus obliquus biomass grown in poultry wastewater
Publication . Vladić, Jelena; Jazić, Jelena Molnar; Ferreira, Alice; Maletić, Snežana; Cvetković, Dragoljub; Agbaba, Jasmina; Vidović, Senka; Gouveia, Luisa
Microalgae are capable of assimilating nutrients from wastewater (WW), producing clean water and biomass rich in bioactive compounds that need to be recovered from inside the microalgal cell. This work investigated subcritical water (SW) extraction to collect high-value compounds from the microalga Tetradesmus obliquus after treating poultry WW. The treatment efficiency was evaluated in terms of total Kjeldahl nitrogen (TKN), phosphate, chemical oxygen demand (COD) and metals. T. obliquus was able to remove 77% TKN, 50% phosphate, 84% COD, and metals (48-89%) within legislation values. SW extraction was performed at 170 degrees C and 30 bar for 10 min. SW allowed the extraction of total phenols (1.073 mg GAE/mL extract) and total flavonoids (0.111 mg CAT/mL extract) with high antioxidant activity (IC50 value, 7.18 mu g/mL). The microalga was shown to be a source of organic compounds of commercial value (e.g., squalene). Finally, the SW conditions allowed the removal of pathogens and metals in the extracts and residues to values in accordance with legislation, assuring their safety for feed or agriculture applications.
Supercritical CO2 extract from Microalga Tetradesmus obliquus: the effect of high-pressure pre-treatment
Publication . Vladić, Jelena; Jerković, Igor; Radman, Sanja; Molnar Jazić, Jelena; Ferreira, Alice; Maletić, Snežana; Gouveia, Luisa
High-pressure pre-treatment followed by supercritical carbon dioxide (ScCO2 ) extraction (300 bar, 40 ◦C) was applied for the attainment of the lipophilic fraction of microalga Tetradesmus obliquus. The chemical profile of supercritical extracts of T. obliquus was analyzed by ultra-highperformance liquid chromatography–high-resolution mass spectrometry with electrospray ionization (UHPLC-ESI-HRMS). Moreover, the impact of ScCO2 on the microbiological and metal profile of the biomass was monitored. The application of the pre-treatment increased the extraction yield approximately three-fold compared to the control. In the obtained extracts (control and pre-treated extracts), the identified components belonged to triacylglyceroles, fatty acid derivatives, diacylglycerophosphocholines and diacylglycerophosphoserines, pigments, terpenes, and steroids. Triacylglycerols (65%) were the most dominant group of compounds in the control extract. The pre-treatment decreased the percentage of triacylglycerols to 2%, while the abundance of fatty acid derivatives was significantly increased (82%). In addition, the pre-treatment led to an increase in the percentages of carotenoids, terpenoids, and steroids. Furthermore, it was determined that ScCO2 extraction reduced the number of microorganisms in the biomass. Considering its microbiological and metal profiles, the biomass after ScCO2 can potentially be used as a safe and important source of organic compounds.
Green approach for the valorization of microalgae Tetradesmus obliquus
Publication . Gouveia, Luisa; Molnar Jazić, Jelena; Ferreira, Alice; Maletić, Snežana; Cvetković, Dragoljub; Vidović, Senka; Vladić, Jelena
The main goal of this study was to develop an efficient, green approach for the valorization of Tetradesmus obliquus biomass, with zero waste. This microalga was selected because it is widespread, resistant, easy for cultivation, and fast-growing. In the first step, supercritical carbon dioxide (ScCO2) extraction followed by rapid gas decompression was used for the extraction of biomass. The following step was to apply ultrasound-assisted (UA), microwave-assisted (MA), and subcritical water (SW) extraction on the ScCO2-treated biomass to determine the most efficient processing technology. SW demonstrated to be a superior technique over MW and UA with regard to extraction yield and antioxidant content. Moreover, the chemical and microbiological profiles of SW extracts were determined to evaluate their potential and safety. In addition, to create a procedure with zero waste, the solid waste after SW extraction (residue) was analyzed. The organic profile of extracts and residues contained compounds that belong to groups of aliphatic saturated hydrocarbons, aliphatic unsaturated hydrocarbons, alkylated hydrocarbons, ketones, phenols, and esters. Furthermore, these compounds can be applied in different industries including the pharmaceutical and cosmetic industries. Additionally, the content of metals in residues indicated that this material can be used as animal feed and in agriculture. Finally, a complete reduction of microorganisms present in the initial biomass was obtained for the extracts and residues, indicating their safety.

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Funding agency

Fundação para a Ciência e a Tecnologia

Funding programme

3599-PPCDT

Funding Award Number

PTDC/BII-BIO/5554/2020

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