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Patel, A. & Matsakas, L. (2019). A comparative study on de novo and ex novo lipid fermentation by oleaginous yeast using glucose and sonicated waste cooking oil. Ultrasonics sonochemistry
Open this publication in new window or tab >>A comparative study on de novo and ex novo lipid fermentation by oleaginous yeast using glucose and sonicated waste cooking oil
2019 (English)In: Ultrasonics sonochemistry, ISSN 1350-4177, E-ISSN 1873-2828Article in journal (Refereed) Epub ahead of print
Abstract [en]

There are only a few reports available about the assimilation of hydrophobic substrates by microorganisms, however, it is well known that oleaginous microorganisms are capable of utilizing both hydrophilic and hydrophobic substrates and accumulate lipids via two different pathways namely de novo and ex novo lipid synthesis, respectively. In the present study, an oleaginous yeast, Cryptococcus curvatus, was investigated for its potentials to utilize a waste substrate of hydrophobic nature (waste cooking oil – WCO) and compared with its ability to utilize a hydrophilic carbon source (glucose). To facilitate the utilization of WCO by C. curvatus, the broth was sonicated to form a stable oil-in-water emulsion without adding any emulsifier, which was then compared with WCO samples without any ultrasound treatment (unsonicated) for the yeast cultivation. Ultrasonication reduces the size of hydrophobic substrates and improves their miscibility in an aqueous broth making them easily assimilated by oleaginous yeast. Under de novo lipid fermentation, the yeast synthesized 9.93 ± 0.84 g/L of cell dry weight and 5.23 ± 0.49 g/L lipids (lipid content of 52.66 ± 0.93% w/w) when cultivated on 40 g/L of glucose (C/N ratio of 40). The amount of cell dry weight, lipid concentration, and lipid content were considerably higher during the ex novo lipid synthesis. More specifically, the highest lipid content achieved was 70.13 ± 1.65% w/w with a corresponding dry cell weight and lipid concentration of 18.62 ± 0.76 g/L and 13.06 ± 0.92 g/L respectively, when grown on 20 g/L sonicated WCO. The highest lipid concentration, however, was observed when the yeast was cultivated on 40 g/L sonicated WCO. Under these conditions, 20.34 g/L lipids were produced with a lipid content of 57.05% w/w. On the other hand, lipid production with unsonicated WCO was significant lower, reaching 11.16 ± 1.02 g/L (69.14 ± 1.34% w/w of lipid content) and 12.21 ± 1.34 g/L (47.39 ± 1.67% w/w of lipid content) for 20 g/L and 40 g/L of WCO, respectively. This underpins the significance of the sonication treatment, especially at elevated WCO concentrations, to improve the accessibility of the yeast to the WCO. Sonication treatment that was used in this study assisted the utilization of WCO without the need to add emulsifiers, thus reducing the need for chemicals and in turn has a positive impact on the production costs. The microbial lipids produced presented a different fatty acid composition compared to the WCO, making them more suitable for biodiesel production as suggested by the theoretical estimation of the biodiesel properties.

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
Oleaginous yeast, Sonicated waste cooking oil, De novo lipid accumulation, Ex novo lipid accumulation, LipidsFatty acid methyl esters
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-72671 (URN)10.1016/j.ultsonch.2018.12.010 (DOI)
Available from: 2019-01-24 Created: 2019-01-24 Last updated: 2019-01-24
Patel, A., Matsakas, L., Rova, U. & Christakopoulos, P. (2019). A perspective on biotechnological applications of thermophilic microalgae and cyanobacteria. Bioresource Technology, 278, 424-434
Open this publication in new window or tab >>A perspective on biotechnological applications of thermophilic microalgae and cyanobacteria
2019 (English)In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 278, p. 424-434Article in journal (Refereed) Published
Abstract [en]

The importance of expanding our knowledge on microorganisms derived from extreme environments stems from the development of novel and sustainable technologies for our health, food, and environment. Microalgae and cyanobacteria represent a group of diverse microorganisms that inhabit a wide range of environments, are capable of oxygenic photosynthesis, and form a thick microbial mat even at extreme environments. Studies of thermophilic microorganisms have shown a considerable biotechnological potential due to their optimum growth and metabolisms at high temperatures (≥50 °C), which is supported by their thermostable enzymes. Microalgal and cyanobacterial communities present in high-temperature ecosystems account for a large part of the total ecosystem biomass and productivity, and can be exploited to generate several value-added products of agricultural, pharmaceutical, nutraceutical, and industrial relevance. This review provides an overview on the current status of biotechnological applications of thermophilic microalgae and cyanobacteria, with an outlook on the challenges and future prospects.

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
Thermophilic microorganisms, Microalgae, Cyanobacteria, Biotechnological applications, Biofuels, Biologically active compounds, Pigments
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-73043 (URN)10.1016/j.biortech.2019.01.063 (DOI)000457852400049 ()30685131 (PubMedID)2-s2.0-85060269950 (Scopus ID)
Note

Validerad;2019;Nivå 2;2019-02-27 (johcin)

Available from: 2019-02-27 Created: 2019-02-27 Last updated: 2019-02-27Bibliographically approved
Patel, A., Matsakas, L., Hruzova, K., Rova, U. & Christakopoulos, P. (2019). Biosynthesis of Nutraceutical Fatty Acids by the Oleaginous Marine Microalgae Phaeodactylum tricornutum Utilizing Hydrolysates from Organosolv-Pretreated Birch and Spruce Biomass. Marine Drugs, 17(12), Article ID 119.
Open this publication in new window or tab >>Biosynthesis of Nutraceutical Fatty Acids by the Oleaginous Marine Microalgae Phaeodactylum tricornutum Utilizing Hydrolysates from Organosolv-Pretreated Birch and Spruce Biomass
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2019 (English)In: Marine Drugs, ISSN 1660-3397, E-ISSN 1660-3397, Vol. 17, no 12, article id 119Article in journal (Refereed) Published
Abstract [en]

Polyunsaturated fatty acids (PUFAs) are essential for human function, however they have to be provided through the diet. As their production from fish oil is environmentally unsustainable, there is demand for new sources of PUFAs. The aim of the present work was to establish the microalgal platform to produce nutraceutical-value PUFAs from forest biomass. To this end, the growth of Phaeodactylum tricornutum on birch and spruce hydrolysates was compared to autotrophic cultivation and glucose synthetic media. Total lipid generated by P. tricornutum grown mixotrophically on glucose, birch, and spruce hydrolysates was 1.21, 1.26, and 1.29 g/L, respectively. The highest eicosapentaenoic acid (EPA) production (256 mg/L) and productivity (19.69 mg/L/d) were observed on spruce hydrolysates. These values were considerably higher than those obtained from the cultivation without glucose (79.80 mg/L and 6.14 mg/L/d, respectively) and also from the photoautotrophic cultivation (26.86 mg/L and 2.44 mg/L/d, respectively). To the best of our knowledge, this is the first report describing the use of forest biomass as raw material for EPA and docosapentaenoic acid (DHA) production.

Place, publisher, year, edition, pages
MDPI, 2019
Keywords
polyunsaturated fatty acids, EPA, DHA, marine algae, Phaeodactylum tricornutum, forest biomass
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-73136 (URN)10.3390/md17020119 (DOI)30781416 (PubMedID)2-s2.0-85061857091 (Scopus ID)
Note

Validerad;2019;Nivå 2;2019-03-07 (johcin)

Available from: 2019-03-07 Created: 2019-03-07 Last updated: 2019-03-07Bibliographically approved
Matsakas, L., Raghavendran, V., Yakimenko, O., Persson, G., Olsson, E., Rova, U., . . . Christakopoulos, P. (2019). Lignin-first biomass fractionation using a hybrid organosolv: Steam explosion pretreatment technology improves the saccharification and fermentability of spruce biomass. Bioresource Technology, 273, 521-528
Open this publication in new window or tab >>Lignin-first biomass fractionation using a hybrid organosolv: Steam explosion pretreatment technology improves the saccharification and fermentability of spruce biomass
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2019 (English)In: Bioresource Technology, ISSN 0960-8524, E-ISSN 1873-2976, Vol. 273, p. 521-528Article in journal (Refereed) Published
Abstract [en]

For a transition to a sustainable society, fuels, chemicals, and materials should be produced from renewable resources. Lignocellulosic biomass constitutes an abundant and renewable feedstock; however, its successful application in a biorefinery requires efficient fractionation into its components; cellulose, hemicellulose and lignin. Here, we demonstrate that a newly established hybrid organosolv – steam explosion pretreatment can effectively fractionate spruce biomass to yield pretreated solids with high cellulose (72% w/w) and low lignin (delignification up to 79.4% w/w) content. The cellulose-rich pretreated solids present high saccharification yields (up to 61% w/w) making them ideal for subsequent bioconversion processes. Moreover, under high-gravity conditions (22% w/w) we obtained an ethanol titer of 61.7 g/L, the highest so far reported for spruce biomass. Finally, the obtained high-purity lignin is suitable for various advanced applications. In conclusion, hybrid organosolv pretreatment could offer a closed-loop biorefinery while simultaneously adding value to all biomass components.

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
Softwood, Organosolv explosion, High gravity fermentation, Fractionation, Biorefinery
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-71766 (URN)10.1016/j.biortech.2018.11.055 (DOI)000453742100064 ()30471644 (PubMedID)2-s2.0-85056772752 (Scopus ID)
Funder
Swedish Energy Agency, 2015-006989
Note

Validerad;2018;Nivå 2;2018-11-27 (johcin)

Available from: 2018-11-27 Created: 2018-11-27 Last updated: 2019-01-30Bibliographically approved
Lage, S., Kudahettige, N. P., Ferro, L., Matsakas, L., Funk, C., Rova, U. & Gentili, F. G. (2019). Microalgae Cultivation for the Biotransformation of Birch Wood Hydrolysate and Dairy Effluent. Catalysts, 9(2), Article ID 150.
Open this publication in new window or tab >>Microalgae Cultivation for the Biotransformation of Birch Wood Hydrolysate and Dairy Effluent
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2019 (English)In: Catalysts, ISSN 2073-4344, Vol. 9, no 2, article id 150Article in journal (Refereed) Published
Abstract [en]

In order to investigate environmentally sustainable sources of organic carbon and nutrients, four Nordic green microalgal strains, Chlorella sorokiniana, Chlorella saccharophila, Chlorella vulgaris, and Coelastrella sp., were grown on a wood (Silver birch, Betula pendula) hydrolysate and dairy effluent mixture. The biomass and lipid production were analysed under mixotrophic, as well as two-stage mixotrophic/heterotrophic regimes. Of all of the species, Coelastrella sp. produced the most total lipids per dry weight (~40%) in the mixture of birch hydrolysate and dairy effluent without requiring nutrient (nitrogen, phosphorus, and potassium-NPK) supplementation. Overall, in the absence of NPK, the two-stage mixotrophic/heterotrophic cultivation enhanced the lipid concentration, but reduced the amount of biomass. Culturing microalgae in integrated waste streams under mixotrophic growth regimes is a promising approach for sustainable biofuel production, especially in regions with large seasonal variation in daylight, like northern Sweden. To the best of our knowledge, this is the first report of using a mixture of wood hydrolysate and dairy effluent for the growth and lipid production of microalgae in the literature.

Keywords
Birch hydrolysate, Chlorella, Coelastrella, Dairy wastewater, Fatty acid methyl esters, Green algae, Heterotrophic, Lipids, Mixotrophic
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-73233 (URN)10.3390/catal9020150 (DOI)2-s2.0-85062564536 (Scopus ID)
Note

Validerad;2019;Nivå 2;2019-03-18 (svasva)

Available from: 2019-03-18 Created: 2019-03-18 Last updated: 2019-03-18Bibliographically approved
Mu, L., Wu, J., Matsakas, L., Chen, M., Rova, U., Christakopoulos, P., . . . Shi, Y. (2019). Two important factors of selecting lignin as efficient lubricating additives in poly (ethylene glycol): Hydrogen bond and molecular weight. International Journal of Biological Macromolecules, 129, 564-570
Open this publication in new window or tab >>Two important factors of selecting lignin as efficient lubricating additives in poly (ethylene glycol): Hydrogen bond and molecular weight
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2019 (English)In: International Journal of Biological Macromolecules, ISSN 0141-8130, E-ISSN 1879-0003, Vol. 129, p. 564-570Article in journal (Refereed) Published
Abstract [en]

Lignin, one of the most abundant natural polymers, has been successfully used as an effective lubricant additive with high value. The chemical structure of lignin is very diverse and strongly affected by both the source of lignin (i.e. plant species) and the lignin extraction process. In this work, a series of lignin from different biomass sources (hard or soft wood) and extraction process (organosolv with or without acid catalyst) has been successfully incorporated into poly(ethylene glycol) (PEG) and fortified lubricating properties were achieved. The effects of different lignin on the rheological, thermal and tribological properties of the lignin/EG lubricants were systematically investigated by different characterization techniques. Lignin in PEG significantly improves the lubricating property, where a wear reduction of 93.8% was observed. The thermal and lubrication properties of the PEG lubricants filled with different kinds of lignin are tightly related to the synergistic state of hydrogen bonding and molecular weight distribution. Lignin with broader molecular weight distribution and higher hydroxyl content shows better adhesion on metal surfaces and strengthened lubricating film, which could be used as the efficient lubricating additives. This work provides a criterion for selecting appropriate lignin as the efficient lubricant additive and accelerates the application of lignin.

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
Lignin, Lubrication, Poly (ethylene glycol), Hydrogen bonding, Molecular weight
National Category
Bioprocess Technology Tribology (Interacting Surfaces including Friction, Lubrication and Wear)
Research subject
Machine Elements; Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-72899 (URN)10.1016/j.ijbiomac.2019.01.175 (DOI)2-s2.0-85061540788 (Scopus ID)
Note

Validerad;2019;Nivå 2;2019-02-20 (svasva)

Available from: 2019-02-14 Created: 2019-02-14 Last updated: 2019-02-26Bibliographically approved
Raghavendran, V., Nitsos, C., Matsakas, L., Rova, U., Christakopoulos, P. & Olsson, L. (2018). A comparative study of the enzymatic hydrolysis of batch organosolv-pretreated birch and spruce biomass. AMB Express, 8(1), Article ID 114.
Open this publication in new window or tab >>A comparative study of the enzymatic hydrolysis of batch organosolv-pretreated birch and spruce biomass
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2018 (English)In: AMB Express, ISSN 2191-0855, E-ISSN 2191-0855, Vol. 8, no 1, article id 114Article in journal (Refereed) Published
Abstract [en]

A shift towards a sustainable and green society is vital to reduce the negative effects of climate change associated with increased CO2 emissions. Lignocellulosic biomass is both renewable and abundant, but is recalcitrant to deconstruction. Among the methods of pretreatment available, organosolv (OS) delignifies cellulose efficiently, significantly improving its digestibility by enzymes. We have assessed the hydrolysability of the cellulose-rich solid fractions from OS-pretreated spruce and birch at 2% w/v loading (dry matter). Almost complete saccharification of birch was possible with 80 mg enzyme preparation/gsolids (12 FPU/gsolids), while the saccharification yield for spruce was only 70%, even when applying 60 FPU/gsolids. As the cellulose content is enriched by OS, the yield of glucose was higher than in their steam-exploded counterparts. The hydrolysate was a transparent liquid due to the absence of phenolics and was also free from inhibitors. OS pretreatment holds potential for use in a large-scale, closed-loop biorefinery producing fuels from the cellulose fraction and platform chemicals from the hemicellulose and lignin fractions respectively.

Place, publisher, year, edition, pages
Springer, 2018
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-70136 (URN)10.1186/s13568-018-0643-y (DOI)000438255300003 ()29992363 (PubMedID)2-s2.0-85049690419 (Scopus ID)
Note

Validerad;2018;Nivå 2;2018-07-19 (inah)

Available from: 2018-07-19 Created: 2018-07-19 Last updated: 2018-09-14Bibliographically approved
Matsakas, L., Nitsos, C., Raghavendran, V., Yakimenko, O., Persson, G., Olsson, E., . . . Christakopoulos, P. (2018). A novel hybrid organosolv: steam explosion method for the efficient fractionation and pretreatment of birch biomass. Biotechnology for Biofuels, 11(1), Article ID 160.
Open this publication in new window or tab >>A novel hybrid organosolv: steam explosion method for the efficient fractionation and pretreatment of birch biomass
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2018 (English)In: Biotechnology for Biofuels, ISSN 1754-6834, E-ISSN 1754-6834, Vol. 11, no 1, article id 160Article in journal (Refereed) Published
Abstract [en]

The main role of pretreatment is to reduce the natural biomass recalcitrance and thus enhance saccharification yield. A further prerequisite for efficient utilization of all biomass components is their efficient fractionation into well-defined process streams. Currently available pretreatment methods only partially fulfill these criteria. Steam explosion, for example, excels as a pretreatment method but has limited potential for fractionation, whereas organosolv is excellent for delignification but offers poor biomass deconstruction.

Results

In this article, a hybrid method combining the cooking and fractionation of conventional organosolv pretreatment with the implementation of an explosive discharge of the cooking mixture at the end of pretreatment was developed. The effects of various pretreatment parameters (ethanol content, duration, and addition of sulfuric acid) were evaluated. Pretreatment of birch at 200 °C with 60% v/v ethanol and 1% w/wbiomass H2SO4 was proven to be the most efficient pretreatment condition yielding pretreated solids with 77.9% w/w cellulose, 8.9% w/w hemicellulose, and 7.0 w/w lignin content. Under these conditions, high delignification of 86.2% was demonstrated. The recovered lignin was of high purity, with cellulose and hemicellulose contents not exceeding 0.31 and 3.25% w/w, respectively, and ash to be < 0.17% w/w in all cases, making it suitable for various applications. The pretreated solids presented high saccharification yields, reaching 68% at low enzyme load (6 FPU/g) and complete saccharification at high enzyme load (22.5 FPU/g). Finally, simultaneous saccharification and fermentation (SSF) at 20% w/w solids yielded an ethanol titer of 80 g/L after 192 h, corresponding to 90% of the theoretical maximum.

Conclusions

The novel hybrid method developed in this study allowed for the efficient fractionation of birch biomass and production of pretreated solids with high cellulose and low lignin contents. Moreover, the explosive discharge at the end of pretreatment had a positive effect on enzymatic saccharification, resulting in high hydrolyzability of the pretreated solids and elevated ethanol titers in the following high-gravity SSF. To the best of our knowledge, the ethanol concentration obtained with this method is the highest so far for birch biomass.

Place, publisher, year, edition, pages
BioMed Central, 2018
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-69455 (URN)10.1186/s13068-018-1163-3 (DOI)000434952000002 ()29930706 (PubMedID)2-s2.0-85048400180 (Scopus ID)
Note

Validerad;2018;Nivå 2;2018-06-13 (andbra)

Available from: 2018-06-13 Created: 2018-06-13 Last updated: 2018-08-08Bibliographically approved
Kalogiannis, K. G., Matsakas, L., Aspden, J., Lappas, A. A., Rova, U. & Christakopoulos, P. (2018). Acid Assisted Organosolv Delignification of Beechwood and Pulp Conversion towards High Concentrated Cellulosic Ethanol via High Gravity Enzymatic Hydrolysis and Fermentation. Molecules, 23(7), Article ID 1647.
Open this publication in new window or tab >>Acid Assisted Organosolv Delignification of Beechwood and Pulp Conversion towards High Concentrated Cellulosic Ethanol via High Gravity Enzymatic Hydrolysis and Fermentation
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2018 (English)In: Molecules, ISSN 1420-3049, E-ISSN 1420-3049, Vol. 23, no 7, article id 1647Article in journal (Refereed) Published
Abstract [en]

Background: Future biorefineries will focus on converting low value waste streams to chemical products that are derived from petroleum or refined sugars. Feedstock pretreatment in a simple, cost effective, agnostic manner is a major challenge. Methods: In this work, beechwood sawdust was delignified via an organosolv process, assisted by homogeneous inorganic acid catalysis. Mixtures of water and several organic solvents were evaluated for their performance. Specifically, ethanol (EtOH), acetone (AC), and methyl- isobutyl- ketone (MIBK) were tested with or without the use of homogeneous acid catalysis employing sulfuric, phosphoric, and oxalic acids under relatively mild temperature of 175 °C for one hour. Results: Delignification degrees (DD) higher than 90% were achieved, where both AC and EtOH proved to be suitable solvents for this process. Both oxalic and especially phosphoric acid proved to be good alternative catalysts for replacing sulfuric acid. High gravity simultaneous saccharification and fermentation with an enzyme loading of 8.4 mg/gsolids at 20 wt.% initial solids content reached an ethanol yield of 8.0 w/v%. Conclusions: Efficient delignification combining common volatile solvents and mild acid catalysis allowed for the production of ethanol at high concentration in an efficient manner

Place, publisher, year, edition, pages
MDPI, 2018
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-70135 (URN)10.3390/molecules23071647 (DOI)000445301800150 ()29976912 (PubMedID)2-s2.0-85050509943 (Scopus ID)
Note

Validerad;2018;nivå 2;2018-07-19 (inah)

Available from: 2018-07-19 Created: 2018-07-19 Last updated: 2019-03-06Bibliographically approved
Patel, A., Mikes, F. & Matsakas, L. (2018). An Overview of Current Pretreatment Methods Used to Improve Lipid Extraction from Oleaginous Microorganisms. Molecules, 23(7), Article ID 1562.
Open this publication in new window or tab >>An Overview of Current Pretreatment Methods Used to Improve Lipid Extraction from Oleaginous Microorganisms
2018 (English)In: Molecules, ISSN 1420-3049, E-ISSN 1420-3049, Vol. 23, no 7, article id 1562Article in journal (Refereed) Published
Abstract [en]

Microbial oils, obtained from oleaginous microorganisms are an emerging source of commercially valuable chemicals ranging from pharmaceuticals to the petroleum industry. In petroleum biorefineries, the microbial biomass has become a sustainable source of renewable biofuels. Biodiesel is mainly produced from oils obtained from oleaginous microorganisms involving various upstream and downstream processes, such as cultivation, harvesting, lipid extraction, and transesterification. Among them, lipid extraction is a crucial step for the process and it represents an important bottleneck for the commercial scale production of biodiesel. Lipids are synthesized in the cellular compartment of oleaginous microorganisms in the form of lipid droplets, so it is necessary to disrupt the cells prior to lipid extraction in order to improve the extraction yields. Various mechanical, chemical and physicochemical pretreatment methods are employed to disintegrate the cellular membrane of oleaginous microorganisms. The objective of the present review article is to evaluate the various pretreatment methods for efficient lipid extraction from the oleaginous cellular biomass available to date, as well as to discuss their advantages and disadvantages, including their effect on the lipid yield. The discussed mechanical pretreatment methods are oil expeller, bead milling, ultrasonication, microwave, high-speed and high-pressure homogenizer, laser, autoclaving, pulsed electric field, and non-mechanical methods, such as enzymatic treatment, including various emerging cell disruption techniques.

Place, publisher, year, edition, pages
MDPI, 2018
National Category
Bioprocess Technology
Research subject
Biochemical Process Engineering
Identifiers
urn:nbn:se:ltu:diva-70044 (URN)10.3390/molecules23071562 (DOI)000445301800065 ()29958398 (PubMedID)2-s2.0-85049436891 (Scopus ID)
Note

Validerad;2018;Nivå 2;2018-08-02 (rokbeg)

Available from: 2018-07-03 Created: 2018-07-03 Last updated: 2018-10-22Bibliographically approved
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Identifiers
ORCID iD: ORCID iD iconorcid.org/0000-0002-3687-6173

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