Engineering Strategies in Microorganisms for the Enhanced Production of Squalene: Advances, Challenges and Opportunities

Gohil, Nisarg; Bhattacharjee, Gargi; Khambhati, Khushal; Braddick, Darren; Singh, Vijai

doi:10.3389/fbioe.2019.00050

Cited by 78 publications

(74 citation statements)

References 189 publications

(318 reference statements)

Supporting

Mentioning

Contrasting

Order By: Relevance

“…erefore, we recommend the consumption of foods containing SQ, such as shark liver oil and olive oil, because oral administration of SQ is well absorbed (60-85%) in humans [32] and minimize the SQ solubility and bioavailability problems of several pharmacological activities.…”

Section: Resultsmentioning

confidence: 99%

“…Transforming growth factor β-activated kinase 1 activates MAPK kinases, and they subsequently phosphorylate extracellular signal-regulated kinase, p38, and c-Jun N-terminal kinase. Moreover, phosphorylation of activator protein-1, ETS Like-1 protein Elk-1, and transcription factors initiate the transcription of inflammatory genes [32]. In the future, the effect of HEHSQ on NF-κB and MAPK signaling, which are the key mediators of inflammatory signaling, should be evaluated.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Synthesis of 2-(2-Hydroxyethoxy)-3-hydroxysqualene and Characterization of Its Anti-Inflammatory Effects

Sasaki

Inami

Tominaga

et al. 2020

BioMed Research International

View full text Add to dashboard Cite

Squalene (SQ), a natural precursor of many steroids, can inhibit tumor progression and decrease serum cholesterol levels. However, it is difficult to discern the effect of highly active molecules in the treatment of diseases because not enough active compounds reach the site of pathology in crowded biosystems. Therefore, it is necessary to design artificial probes that work effectively within crowded systems. In this study, to facilitate cell penetration, the ethylene glycol moiety (used as a probe) was chemically added to SQ to form 2-(2-hydroxyethoxy)-3-hydroxysqualene (HEHSQ). HEHSQ was prepared from 2,3-epoxysqualene and characterized by Rf, FT-IR, 1H NMR, 13C NMR, and high-resolution mass spectrometry. We then evaluated the anti-inflammatory effects of SQ and HEHSQ on lipopolysaccharide- (LPS-) stimulated RAW264.7 murine macrophages. To determine the effect of SQ and HEHSQ on the viability of RAW264.7 cells, an MTT assay was performed. To quantify the anti-inflammatory effect of SQ and HEHSQ, we measured nitric oxide (NO) production, gene expression, and secretion of the proinflammatory cytokine tumor necrosis factor α (TNF-α) and chemokine C-C motif chemokine 2 (CCL2) in LPS-stimulated RAW264.7 cells using an in vitro inflammatory model. 2,3-Epoxysqualene was prepared according to a reported methodology. The reaction of 2,3-epoxysqualene and ethylene glycol in 2-propanol produced 49% HEHSQ. MTT results showed that 10 and 100 µg/mL HEHSQ treatment decreased cell viability, whereas SQ treatment (1–100 µg/mL) did not have any effect on viability. SQ (100 µg/mL) and HEHSQ (1 µg/mL) treatment significantly reduced the production of LPS-stimulated NO and decreased the expression and secretion of proinflammatory TNF-α and CCL2. Therefore, our results suggested that the anti-inflammatory effects of HEHSQ are 100 times higher than that of unmodified SQ. To the best of our knowledge, this study has demonstrated for the first time that HEHSQ can be potentially used as a safe alternative treatment to anti-inflammatory drugs.

show abstract

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Synthesis of 2-(2-Hydroxyethoxy)-3-hydroxysqualene and Characterization of Its Anti-Inflammatory Effects

Sasaki

Inami

Tominaga

et al. 2020

BioMed Research International

View full text Add to dashboard Cite

show abstract

“…Most thraustochytrids produce several antioxidants, such as β-carotene, squalene, and astaxanthin 15 . Aurantiochytrium/Schizochytrium species produce the polyunsaturated hydrocarbon triterpenoid squalene (2,6,10,15,19,23-hexamethyl-6,6,10,14,18,20-tetracosahexane) 16 . The most common source of squalene is liver oil of deep-sea sharks and whales; however, contamination with heavy metals and polychlorinated biphenyls poses a safety risk, while the oil's often putrid odor and unpleasant taste diminish its appeal to consumers 17 .…”

mentioning

confidence: 99%

Co-production of DHA and squalene by thraustochytrid from forest biomass

Patel

Liefeldt

Rova

et al. 2020

Sci Rep

View full text Add to dashboard Cite

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/g substrate) and lipid productivity (0.239 g/g substrate) were observed with 60 g/L of glucose, following cultivation in a bioreactor, with the DHA content to be 67.76% w/w total 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/w total 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/ g CDW). In summary, we show that the co-production of DHA and squalene makes S. limacinum SR21 appropriate strain for commercial-scale production of nutraceuticals. Omega-6 (n-6) and omega-3 (n-3) are polyunsaturated fatty acids (PUFAs), whose precursors include linoleic acid and alpha-linolenic acid. They are considered essential because humans cannot synthesize them due to insufficient levels of elongases and delta-6-desaturases 1,2. Although conversion of alpha-linolenic acid into omega-3 fatty acids, such as eicosapentaenoic acid (C 20:5n−3), docosapentaenoic acid (C 22:5n−3), and docosahexaenoic acid (C 22:6n−3 , DHA) occurs in humans, it happens at a very slow rate 3 and so these fatty acids must be provided by the diet. These PUFAs have several health benefits and have been found to be very effective in the preventions and treatment of fetal diseases 4. DHA plays a major role in cell signalling and is found mainly in the brain and retina tissues 5. Additionally, it acts as an anti-inflammatory agent, a precursor of several metabolites and potent lipid mediators, and could be used to treat several cardiovascular or neurologic disorders, such as hypertension and Alzheimer's 6. Fish of the Salmonidae, Scombridae, and Clupeidae families have been the sole commercial source of DHA as their oil contains approximately 20 to 30% DHA 3. However, DHA purification and concentration from fish oil is costly and the resulting oil quality depends on species, location, and pollution 7. Furthermore, fish-derived DHA is unsuitable for vegetarians, and diminishing fish stocks together with marine pollution limit the increasing demand for DHA 7. Oils from genetically engineered plant oilseeds, such as Brassica juncea, Arabidopsis thaliana, an...

show abstract

“…According to our previous results, this ratio is dramatically different between pdr18 Δ and parental strain cells (Cabrito et al ., 2011). Interestingly, squalene is a molecule of great medical and cosmetic interest, and cells accumulating high squalene levels might be of high biotechnological potential (Gohil et al ., 2019).…”

Section: Discussionmentioning

confidence: 99%

The ABC transporter Pdr18 is required for yeast thermotolerance due to its role in ergosterol transport and plasma membrane properties

Godinho

Costa

Sá‐Correia

2020

Environmental Microbiology

View full text Add to dashboard Cite

Among the mechanisms by which yeast overcomes multiple stresses is the expression of genes encoding ATP-binding cassette (ABC) transporters required for resistance to a wide range of toxic compounds. These substrates may include weak acids, alcohols, agricultural pesticides, polyamines, metal cations, as in the case of Pdr18. This pleotropic drug resistance transporter was previously proposed to transport ergosterol at the plasma membrane (PM) level contributing to the maintenance of PM lipid organization and reduced diffusional permeation induced by lipophilic compounds. The present work reports a novel phenotype associated with the putative drug/xenobiotic-efflux-pump transporter Pdr18: the resistance to heat shock and to long-term growth at supra-optimal temperatures. Cultivation at 40 C was demonstrated to lead to higher PM permeabilization of a pdr18Δ cell population with the PDR18 gene deleted compared with the parental strain population, as indicated by flow cytometry analysis of propidium iodide stained cells. Cells of pdr18Δ grown at 40 C also exhibited increased transcription levels from genes of the ergosterol biosynthetic pathway, compared with parental cells. However, this adaptive response at 40 C was not enough to maintain PM physiological ergosterol levels in the population lacking the Pdr18 transporter and free ergosterol precursors accumulate in the deletion mutant cells.

show abstract

Engineering Strategies in Microorganisms for the Enhanced Production of Squalene: Advances, Challenges and Opportunities

Cited by 78 publications

References 189 publications

Synthesis of 2-(2-Hydroxyethoxy)-3-hydroxysqualene and Characterization of Its Anti-Inflammatory Effects

Synthesis of 2-(2-Hydroxyethoxy)-3-hydroxysqualene and Characterization of Its Anti-Inflammatory Effects

Co-production of DHA and squalene by thraustochytrid from forest biomass

The ABC transporter Pdr18 is required for yeast thermotolerance due to its role in ergosterol transport and plasma membrane properties

Contact Info

Product

Resources

About