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Hruzova, K., Patel, A., Masák, J., Maťátková, O., Rova, U., Christakopoulos, P. & Matsakas, L. (2020). A novel approach for the production of green biosurfactant from Pseudomonas aeruginosa using renewable forest biomass. Science of the Total Environment, 711, Article ID 135099.
Open this publication in new window or tab >>A novel approach for the production of green biosurfactant from Pseudomonas aeruginosa using renewable forest biomass
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2020 (English)In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026, Vol. 711, article id 135099Article in journal (Refereed) Published
Abstract [en]

The rising demand for surfactants by the pharmaceuticals and cosmetic industries has generated vast amounts of petroleum-based synthetic surfactants, which are often toxic and non-degradable. Owing to their low toxicity, stability in extreme conditions, and biodegradability, biosurfactants could represent a sustainable alternative. The present study aimed to maximize the production of rhamnolipids (RL) from Pseudomonas aeruginosa by optimizing glucose concentration, temperature, and C/N and C/P ratios. After 96 h of cultivation at 37 °C, the final RL concentration was 4.18 ± 0.19 g/L with a final yield of 0.214 ± 0.010 g/gglucose when pure glucose was used as a carbon source. At present, the main obstacle towards commercialization of RL production is economic sustainability, due to the high cost of downstream processes and media components. For this reason, a renewable source such as wood hydrolysates (from birch and spruce woodchips) was examined here as a possible source of glucose for RL production. Both hydrolysates proved to be adequate, resulting in 2.34 ± 0.17 and 2.31 ± 0.10 g/L of RL, respectively, and corresponding yields of 0.081 ± 0.006 and 0.089 ± 0.004 g/gsugar after 96 h. These results demonstrate the potential of using renewable biomass for the production of biosurfactants and, to the best of our knowledge, they constitute the first report on the use of wood hydrolysates for RL production.

Place, publisher, year, edition, pages
Elsevier, 2020
Keywords
Rhamnolipid, Biosurfactants, Pseudomonas, Wood hydrolysate, Organosolv fractionation
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-76888 (URN)10.1016/j.scitotenv.2019.135099 (DOI)000509344700013 ()2-s2.0-85076241604 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-02-25 (johcin)

Available from: 2019-11-27 Created: 2019-11-27 Last updated: 2020-03-13Bibliographically approved
Patel, A., Mu, L., Shi, Y., Rova, U., Christakopoulos, P. & Matsakas, L. (2020). A novel biorefinery approach aimed at vegetarians reduces the dependency on marine fish stocks for obtaining squalene and DHA. ACS Sustainable Chemistry & Engineering
Open this publication in new window or tab >>A novel biorefinery approach aimed at vegetarians reduces the dependency on marine fish stocks for obtaining squalene and DHA
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2020 (English)In: ACS Sustainable Chemistry & Engineering, E-ISSN 2168-0485Article in journal (Refereed) Accepted
Abstract [en]

Squalene and docosahexaenoic acid (DHA) have gained substantial market share as dietary supplements and vital nutraceuticals due to their beneficial effects on human health. Marine fish are the main commercial source of these nutraceuticals, but a growing global demand, issues of sustainability, and an expanding vegan and vegetarian population has prompted the search for alternatives. Oils obtained from oleaginous microorganisms such as microalgae, diatoms, certain fungi, and thraustochytrids are alternatives to fish oils for omega-3 fatty acids. Among these, DHA is now being mined from thraustochytrids due to its highest proportion in their lipids, however, this strategy is not cost-effective. One way to offset such elevated production costs is to simultaneously extract other high value-added biological products from these oleaginous microorganisms. Here, we propose a novel biorefinery process based on single-step purification of squalene from total lipids extracted from an oleaginous thraustochytrid cultivated on non-edible forest biomass. To render the process economically feasible and sustainable, additional squalene-free lipids were exploited for enrichment of DHA; whereas leftover lipids generated as by-product during the process were tested as a novel biolubricant.

Place, publisher, year, edition, pages
American Chemical Society (ACS), 2020
Keywords
DHA, squalene, biolubricant, biorefinery, thraustochytrids
National Category
Bioprocess Technology Tribology (Interacting Surfaces including Friction, Lubrication and Wear)
Research subject
Biochemical Process Engineering; Machine Elements
Identifiers
urn:nbn:se:ltu:diva-79161 (URN)10.1021/acssuschemeng.0c02752 (DOI)
Available from: 2020-06-03 Created: 2020-06-03 Last updated: 2020-06-03
Patel, A., Karageorgou, D., Rova, E., Katapodis, P., Rova, U., Christakopoulos, P. & Matsakas, L. (2020). An Overview of Potential Oleaginous Microorganisms and Their Role in Biodiesel and Omega-3 Fatty Acid-Based Industries. Microorganisms, 8(3), Article ID 434.
Open this publication in new window or tab >>An Overview of Potential Oleaginous Microorganisms and Their Role in Biodiesel and Omega-3 Fatty Acid-Based Industries
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2020 (English)In: Microorganisms, ISSN 2076-2607, Vol. 8, no 3, article id 434Article, review/survey (Refereed) Published
Abstract [en]

Microorganisms are known to be natural oil producers in their cellular compartments. Microorganisms that accumulate more than 20% w/w of lipids on a cell dry weight basis are considered as oleaginous microorganisms. These are capable of synthesizing vast majority of fatty acids from short hydrocarbonated chain (C6) to long hydrocarbonated chain (C36), which may be saturated (SFA), monounsaturated (MUFA), or polyunsaturated fatty acids (PUFA), depending on the presence and number of double bonds in hydrocarbonated chains. Depending on the fatty acid profile, the oils obtained from oleaginous microorganisms are utilized as feedstock for either biodiesel production or as nutraceuticals. Mainly microalgae, bacteria, and yeasts are involved in the production of biodiesel, whereas thraustochytrids, fungi, and some of the microalgae are well known to be producers of very long-chain PUFA (omega-3 fatty acids). In this review article, the type of oleaginous microorganisms and their expertise in the field of biodiesel or omega-3 fatty acids, advances in metabolic engineering tools for enhanced lipid accumulation, upstream and downstream processing of lipids, including purification of biodiesel and concentration of omega-3 fatty acids are reviewed.

Place, publisher, year, edition, pages
MDPI, 2020
Keywords
oleaginous microorganisms, lipid accumulation, fatty acid profile, biodiesel production, microalgae, nutraceuticals, omega-3 fatty acid
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-78497 (URN)10.3390/microorganisms8030434 (DOI)32204542 (PubMedID)
Note

Validerad;2020;Nivå 2;2020-04-20 (alebob)

Available from: 2020-04-15 Created: 2020-04-15 Last updated: 2020-04-20Bibliographically approved
Patel, A., Rova, U., Christakopoulos, P. & Matsakas, L. (2020). Assessment of Fatty Acids Profile and Omega-3 Polyunsaturated Fatty Acid Production by the Oleaginous Marine Thraustochytrid Aurantiochytrium sp. T66 Cultivated on Volatile Fatty Acids. Biomolecules, 10(5), Article ID 694.
Open this publication in new window or tab >>Assessment of Fatty Acids Profile and Omega-3 Polyunsaturated Fatty Acid Production by the Oleaginous Marine Thraustochytrid Aurantiochytrium sp. T66 Cultivated on Volatile Fatty Acids
2020 (English)In: Biomolecules, E-ISSN 2218-273X, Vol. 10, no 5, article id 694Article in journal (Refereed) Published
Abstract [en]

Thraustochytrids are considered natural producers of omega-3 fatty acids as they can synthesize up to 70% docosahexaenoic acids (DHA) of total lipids. However, commercial and sustainable production of microbial DHA is limited by elevated cost of carbon substrates for thraustochytrids cultivation. This problem can be addressed by utilizing low-cost renewable substrates. In the present study, growth, lipid accumulation and fatty acid profiles of the marine thraustochytrid Aurantiochytrium sp. T66 (ATCC-PRA-276) cultivated on volatile fatty acids (C1, formic acid; C2, acetic acid; C3, propionic acid; C4, butyric acid; C5, valeric acid and C6, caproic acid) and glucose as control were evaluated for the first time. This strain showed an inability to utilize C3, C5 and C6 as a substrate when provided at >2 g/L, while efficiently utilizing C2 and C4 up to 40 g/L. The highest cell dry weight (12.35 g/L) and total lipid concentration (6.59 g/L) were attained when this strain was cultivated on 40 g/L of butyric acid, followed by cultivation on glucose (11.87 g/L and 5.34 g/L, respectively) and acetic acid (8.70 g/L and 3.43 g/L, respectively). With 40 g/L butyric acid, the maximum docosahexaenoic acid content was 2.81 g/L, corresponding to 42.63% w/w of total lipids and a yield of 0.23 g/gcell dry weight (CDW). This marine oleaginous microorganism showed an elevated potential for polyunsaturated fatty acids production at higher acetic and butyric acid concentrations than previously reported. Moreover, fluorescence microscopy revealed that growth on butyric acid caused cell size to increase to 45 µm, one of the largest values reported for oleaginous microorganisms, as well as the presence of numerous tiny lipid droplets.

Place, publisher, year, edition, pages
MDPI, 2020
Keywords
oleaginous thraustochytrids, polyunsaturated fatty acids, volatile fatty acids
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-78850 (URN)10.3390/biom10050694 (DOI)32365742 (PubMedID)2-s2.0-85084276592 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-05-12 (alebob)

Available from: 2020-05-11 Created: 2020-05-11 Last updated: 2020-05-18Bibliographically approved
Patel, A., Liefeldt, S., Rova, U., Christakopoulos, P. & Matsakas, L. (2020). Co-production of DHA and squalene by thraustochytrid from forest biomass. Scientific Reports, 10, Article ID 1992.
Open this publication in new window or tab >>Co-production of DHA and squalene by thraustochytrid from forest biomass
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2020 (English)In: Scientific Reports, ISSN 2045-2322, E-ISSN 2045-2322, Vol. 10, article id 1992Article in journal (Refereed) Published
Abstract [en]

Omega-3 fatty acids, and specifically docosahexaenoic acid (DHA), are important and essential nutrients for human health. Thraustochytrids are recognised as commercial strains for nutraceuticals production, they are group of marine oleaginous microorganisms capable of co-synthesis of DHA and other valuable carotenoids in their cellular compartment. The present study sought to optimize DHA and squalene production by the thraustochytrid Schizochytrium limacinum SR21. The highest biomass yield (0.46 g/gsubstrate) and lipid productivity (0.239 g/gsubstrate) were observed with 60 g/L of glucose, following cultivation in a bioreactor, with the DHA content to be 67.76% w/wtotal lipids. To reduce costs, cheaper feedstocks and simultaneous production of various value-added products for pharmaceutical or energy use should be attempted. To this end, we replaced pure glucose with organosolv-pretreated spruce hydrolysate and assessed the simultaneous production of DHA and squalene from S. limacinum SR21. After the 72 h of cultivation period in bioreactor, the maximum DHA content was observed to 66.72% w/wtotal lipids that was corresponded to 10.15 g/L of DHA concentration. While the highest DHA productivity was 3.38 ± 0.27 g/L/d and squalene reached a total of 933.72 ± 6.53 mg/L (16.34 ± 1.81 mg/gCDW). In summary, we show that the co-production of DHA and squalene makes S. limacinum SR21 appropriate strain for commercial-scale production of nutraceuticals.

Place, publisher, year, edition, pages
Springer Nature, 2020
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-78518 (URN)10.1038/s41598-020-58728-7 (DOI)32029800 (PubMedID)2-s2.0-85079041429 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-04-16 (alebob)

Available from: 2020-04-16 Created: 2020-04-16 Last updated: 2020-04-16Bibliographically approved
Patel, A., Rova, U., Christakopoulos, P. & Matsakas, L. (2020). Mining of squalene as a value-added byproduct from DHA producing marine thraustochytrid cultivated on food waste hydrolysate. Science of the Total Environment
Open this publication in new window or tab >>Mining of squalene as a value-added byproduct from DHA producing marine thraustochytrid cultivated on food waste hydrolysate
2020 (English)In: Science of the Total Environment, ISSN 0048-9697, E-ISSN 1879-1026Article in journal (Refereed) Epub ahead of print
Abstract [en]

The commercial production of docosahexaenoic acid (DHA) from oleaginous microorganisms is getting more attention due to several advantages over fish oils. The processing cost became a major bottleneck for commercialization of DHA from microorganisms. The most of cost shares in the feedstock to cultivate the microorganisms and downstream processing. The cost of feedstock can be compensated with the utilization of substrate from waste stream whereas production of value-added chemicals boosts the economic viability of nutraceutical production. In the present study, the docosahexaenoic acid (DHA)-producing marine protist Aurantiochytrium sp. T66 was cultivated on post-consumption food waste hydrolysate for the mining of squalene. After 120 h of cultivation, cell dry weight was 14.7 g/L, of which 6.34 g/L (43.13%; w/w) were lipids. DHA accounted for 2.15 g/L (34.05%) of total extracted lipids or 0.15 g/gCDW. Maximum squalene concentration and yield were 1.05 g/L and 69.31 mg/gCDW, respectively. Hence, utilization of food waste represents an excellent low-cost strategy for cultivating marine oleaginous thraustochytrids and produce squalene as a byproduct of DHA.

Place, publisher, year, edition, pages
Elsevier, 2020
Keywords
Marine oleaginous thraustochytrid, Food waste, Docosahexaenoic acid, Squalene, Nutraceuticals
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-78997 (URN)10.1016/j.scitotenv.2020.139691 (DOI)
Available from: 2020-05-26 Created: 2020-05-26 Last updated: 2020-05-26
Najjarzadeh, N., Krige, A., Pamidi, T. R., Johansson, Ö., Enman, J., Matsakas, L., . . . Christakopoulos, P. (2020). Numerical modeling and verification of a sonobioreactor and its application on two model microorganisms. PLoS ONE, 15(3), Article ID e0229738.
Open this publication in new window or tab >>Numerical modeling and verification of a sonobioreactor and its application on two model microorganisms
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2020 (English)In: PLoS ONE, E-ISSN 1932-6203, Vol. 15, no 3, article id e0229738Article in journal (Refereed) Published
Abstract [en]

Ultrasound has many uses, such as in medical imaging, monitoring of crystallization, characterization of emulsions and suspensions, and disruption of cell membranes in the food industry. It can also affect microbial cells by promoting or slowing their growth and increasing the production of some metabolites. However, the exact mechanism explaining the effect of ultrasound has not been identified yet. Most equipment employed to study the effect of ultrasound on microorganisms has been designed for other applications and then only slightly modified. This results in limited control over ultrasound frequency and input power, or pressure distribution in the reactor. The present study aimed to obtain a well-defined reactor by simulating the pressure distribution of a sonobioreactor. Specifically, we optimized a sonotrode to match the bottle frequency and compared it to measured results to verify the accuracy of the simulation. The measured pressure distribution spectrum presented the same overall trend as the simulated spectrum. However, the peaks were much less intense, likely due to non-linear events such as the collapse of cavitation bubbles. To test the application of the sonobioreactor in biological systems, two biotechnologically interesting microorganisms were assessed: an electroactive bacterium, Geobacter sulfurreducens, and a lignocellulose-degrading fungus, Fusarium oxysporum. Sonication resulted in increased malate production by Gsulfurreducens, but no major effect on growth. In comparison, morphology and growth of Foxysporum were more sensitive to ultrasound intensity. Despite considerable morphological changes at 4 W input power, the growth rate was not adversely affected; however, at 12 W, growth was nearly halted. The above findings indicate that the novel sonobioreactor provides an effective tool for studying the impact of ultrasound on microorganisms.

Place, publisher, year, edition, pages
PLOS, 2020
National Category
Bioprocess Technology Fluid Mechanics and Acoustics
Research subject
Biochemical Process Engineering; Engineering Acoustics
Identifiers
urn:nbn:se:ltu:diva-78111 (URN)10.1371/journal.pone.0229738 (DOI)32160222 (PubMedID)2-s2.0-85081204531 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-03-26 (alebob)

Available from: 2020-03-19 Created: 2020-03-19 Last updated: 2020-03-26Bibliographically approved
Hruzova, K., Matsakas, L., Sand, A., Rova, U. & Christakopoulos, P. (2020). Organosolv lignin hydrophobic micro- and nanoparticles as a low-carbon footprint biodegradable flotation collector in mineral flotation. Bioresource Technology, 306, Article ID 123235.
Open this publication in new window or tab >>Organosolv lignin hydrophobic micro- and nanoparticles as a low-carbon footprint biodegradable flotation collector in mineral flotation
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2020 (English)In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 306, article id 123235Article in journal (Refereed) Published
Abstract [en]

Flotation is a key step during mineral separation. Xanthates are the most commonly used collectors for recovering Cu, Ni, and Zn from sulphide ores. However, xanthates are fossil-based and toxic for the environment. The aim of this study was to evaluate the use of lignin nanoparticles and microparticles as sustainable and environmentally friendly collectors. Lignin particles demonstrated good selectivity toward Cu (chalcopyrite), with total recoveries exceeding 80% and grades of up to 8.6% w/w from a Cu-Ni ore in rougher flotation tests. When floating Zn-Pb-Cu ore, lignin nanoparticles could reduce the use of xanthates by 50%. Moreover, they outperformed xanthates alone, achieving total recoveries of up to 91%, 85%, and 98% for Cu, Pb, and Zn, respectively. These results prove the potential of lignin as a flotation collector.

Place, publisher, year, edition, pages
Elsevier, 2020
Keywords
Lignin, Nanoparticles, Flotation, Collector, Organosolv
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-78359 (URN)10.1016/j.biortech.2020.123235 (DOI)32229063 (PubMedID)2-s2.0-85082530221 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-05-04 (alebob)

Available from: 2020-04-06 Created: 2020-04-06 Last updated: 2020-05-04Bibliographically approved
Matsakas, L., Gerber, M., Yu, L., Rova, U. & Christakopoulos, P. (2020). Preparation of low carbon impact lignin nanoparticles with controllable size by using different strategies for particles recovery. Industrial crops and products (Print), 147, Article ID 112243.
Open this publication in new window or tab >>Preparation of low carbon impact lignin nanoparticles with controllable size by using different strategies for particles recovery
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2020 (English)In: Industrial crops and products (Print), ISSN 0926-6690, E-ISSN 1872-633X, Vol. 147, article id 112243Article in journal (Refereed) Published
Abstract [en]

Lignin still remains an underutilized plentiful resource whose conversion to high-added value products is a cornerstone towards establishing a viable biomass biorefinery. Bio-materials in the form of nanoparticles represent promising high-value products with numerous downstream applications. The aim of the current work was to develop a method that would allow controlling the size of (birch and spruce) lignin nano- and micro-particles for their subsequent recovery into a solid product. We tested different two-step and one-step isolation processes and demonstrated that particle size could be easily controlled to meet different ranges (<100 nm, <500 nm, and>1 μm). In general, two-step isolation methods, i.e. a step of decrease of solvent concentration followed by isolation of lignin particles, were better for the isolation of well-defined spherical particles. In particular, the rate at which ethanol concentration was decreased played a significant role in determining the size of lignin particles. Moreover, when lignin concentration was increased from 1 % to 5 % and 10 % (w/v), particle size and homogeneity decreased slightly, but productivity augmented. The present study demonstrates that different isolation methods can be applied to obtain renewable, customarily sized, lignin spherical micro- and nano-particles.

Place, publisher, year, edition, pages
Elsevier, 2020
Keywords
Lignin, Nanoparticles, Organosolv, Birch, Spruce
National Category
Bioprocess Technology Chemical Process Engineering
Research subject
Biochemical Process Engineering; Chemical Technology
Identifiers
urn:nbn:se:ltu:diva-77832 (URN)10.1016/j.indcrop.2020.112243 (DOI)2-s2.0-85079670850 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-02-24 (alebob)

Available from: 2020-02-24 Created: 2020-02-24 Last updated: 2020-04-22Bibliographically approved
Paulsen Thoresen, P., Matsakas, L., Rova, U. & Christakopoulos, P. (2020). Recent advances in organosolv fractionation: Towards biomass fractionation technology of the future. Bioresource Technology, 306, Article ID 123189.
Open this publication in new window or tab >>Recent advances in organosolv fractionation: Towards biomass fractionation technology of the future
2020 (English)In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 306, article id 123189Article, review/survey (Refereed) Published
Abstract [en]

Organosolv treatment is among the most promising strategies for valorising lignocellulosic biomass and could facilitate the transition towards enhanced utilization of renewable feedstocks. However, issues such as inefficient solvent recycle and fractionation has to be overcome. The present review aims to address these issues and discuss the role of the components present during organosolv treatment and their influence on the overall process. Thus, the review focuses not only on how the choice of solvent and catalyst affects lignocellulosic fractionation, but also on how the choice of treatment liquor influences the possibility for solvent recycling and product isolation. Several organic solvents have been investigated in combination with water and acid/base catalysts; however, the lack of a holistic approach often compromises the performance of the different operational units. Thus, an economically viable organosolv process should optimize biomass fractionation, product isolation, and solvent recycling.

Place, publisher, year, edition, pages
Elsevier, 2020
Keywords
Biomass, Lignocellulose, Organosolv, Ionic liquids, Biorefinery
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-78374 (URN)10.1016/j.biortech.2020.123189 (DOI)32220471 (PubMedID)2-s2.0-85082462286 (Scopus ID)
Note

Validerad;2020;Nivå 2;2020-05-05 (alebob)

Available from: 2020-04-07 Created: 2020-04-07 Last updated: 2020-05-05Bibliographically approved
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ORCID iD: ORCID iD iconorcid.org/0000-0001-7500-2367

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