Overexpression of an endogenous type 2 diacylglycerol acyltransferase in the marine diatom Phaeodactylum tricornutum enhances lipid production and omega-3 Long Chain Polyunsaturated Fatty Acid content Article Swipe
Richard P. Haslam
,
Mary Lynn Hamilton
,
Chloe K. Economou
,
Richard T. Smith
,
Kirsty L. Hassall
,
Johnathan A. Napier
,
Olga Sayanova
·
YOU?
·
· 2020
· Open Access
·
· DOI: https://doi.org/10.21203/rs.2.21513/v2
YOU?
·
· 2020
· Open Access
·
· DOI: https://doi.org/10.21203/rs.2.21513/v2
Background: Oleaginous microalgae represent a valuable resource for the production of high value molecules. Considering the importance of omega-3 long chain polyunsaturated fatty acids (LC-PUFAs) for human health and nutrition the yields of high value eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) require significant improvement to meet demand, however, the current cost of production remains high. A promising approach is to metabolically engineer strains with enhanced levels of triacylglycerols (TAGs) enriched in EPA and DHA. Results: Recently we have engineered the marine diatom Phaeodactylum tricornutum to accumulate enhanced levels of DHA in TAG. To further improve the incorporation of omega-3 LC-PUFAs in TAG we focused our effort on the identification of a type 2 acyl-CoA:diacylglycerol acyltransferase (DGAT) capable of improving lipid production and the incorporation of DHA in TAG. DGAT is a key enzyme in lipid synthesis. Following a diatom based in vivo screen of candidate DGATs, a native P. tricornutum DGAT2B was taken forward for detailed characterisation. Overexpression of the endogenous P. tricornutum DGAT2B was confirmed by qRT-PCR and the transgenic strain grew successfully in comparison to wildtype. PtDGAT2B has broad substrate specificity with preferences for C16 and LC-PUFAs acyl groups. Moreover, the overexpression of an endogenous DGAT2B resulted in higher lipid yields and enhanced levels of DHA in TAG. Furthermore, a combined overexpression of the endogenous DGAT2B and ectopic expression of a Δ5-elongase showed how iterative metabolic engineering can be used to increase DHA and TAG content, irrespective of nitrogen treatment. Conclusion: This study provides further insight into lipid metabolism in P. tricornutum and suggests a metabolic engineering approach for the efficient production of EPA and DHA in microalgae.
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Metadata
- Type
- preprint
- Language
- en
- Landing Page
- https://doi.org/10.21203/rs.2.21513/v2
- https://www.researchsquare.com/article/rs-12104/v2.pdf?c=1631848956000
- OA Status
- green
- Cited By
- 3
- References
- 31
- Related Works
- 10
- OpenAlex ID
- https://openalex.org/W4247255298
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Raw OpenAlex JSON
- OpenAlex ID
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https://openalex.org/W4247255298Canonical identifier for this work in OpenAlex
- DOI
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https://doi.org/10.21203/rs.2.21513/v2Digital Object Identifier
- Title
-
Overexpression of an endogenous type 2 diacylglycerol acyltransferase in the marine diatom Phaeodactylum tricornutum enhances lipid production and omega-3 Long Chain Polyunsaturated Fatty Acid contentWork title
- Type
-
preprintOpenAlex work type
- Language
-
enPrimary language
- Publication year
-
2020Year of publication
- Publication date
-
2020-05-19Full publication date if available
- Authors
-
Richard P. Haslam, Mary Lynn Hamilton, Chloe K. Economou, Richard T. Smith, Kirsty L. Hassall, Johnathan A. Napier, Olga SayanovaList of authors in order
- Landing page
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https://doi.org/10.21203/rs.2.21513/v2Publisher landing page
- PDF URL
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https://www.researchsquare.com/article/rs-12104/v2.pdf?c=1631848956000Direct link to full text PDF
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YesWhether a free full text is available
- OA status
-
greenOpen access status per OpenAlex
- OA URL
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https://www.researchsquare.com/article/rs-12104/v2.pdf?c=1631848956000Direct OA link when available
- Concepts
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Phaeodactylum tricornutum, Polyunsaturated fatty acid, Chemistry, Diatom, Diacylglycerol kinase, Biochemistry, Endogeny, Acyltransferase, Cell biology, Fatty acid, Biology, Enzyme, Botany, Protein kinase CTop concepts (fields/topics) attached by OpenAlex
- Cited by
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3Total citation count in OpenAlex
- Citations by year (recent)
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2025: 1, 2024: 1, 2021: 1Per-year citation counts (last 5 years)
- References (count)
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31Number of works referenced by this work
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10Other works algorithmically related by OpenAlex
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| best_oa_location.raw_type | posted-content |
| best_oa_location.license_id | https://openalex.org/licenses/cc-by |
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| best_oa_location.is_published | False |
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| best_oa_location.landing_page_url | https://doi.org/10.21203/rs.2.21513/v2 |
| primary_location.id | doi:10.21203/rs.2.21513/v2 |
| primary_location.is_oa | True |
| primary_location.source.id | https://openalex.org/S4306402450 |
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| primary_location.source.type | repository |
| primary_location.source.is_oa | False |
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| primary_location.source.is_core | False |
| primary_location.source.is_in_doaj | False |
| primary_location.source.display_name | Research Square (Research Square) |
| primary_location.source.host_organization | https://openalex.org/I4210096694 |
| primary_location.source.host_organization_name | Research Square (United States) |
| primary_location.source.host_organization_lineage | https://openalex.org/I4210096694 |
| primary_location.license | cc-by |
| primary_location.pdf_url | https://www.researchsquare.com/article/rs-12104/v2.pdf?c=1631848956000 |
| primary_location.version | acceptedVersion |
| primary_location.raw_type | posted-content |
| primary_location.license_id | https://openalex.org/licenses/cc-by |
| primary_location.is_accepted | True |
| primary_location.is_published | False |
| primary_location.raw_source_name | |
| primary_location.landing_page_url | https://doi.org/10.21203/rs.2.21513/v2 |
| publication_date | 2020-05-19 |
| publication_year | 2020 |
| referenced_works | https://openalex.org/W2004718580, https://openalex.org/W6758923043, https://openalex.org/W2948728919, https://openalex.org/W6677678175, https://openalex.org/W2050116705, https://openalex.org/W6922268357, https://openalex.org/W2102450255, https://openalex.org/W4213396071, https://openalex.org/W2025366392, https://openalex.org/W634700418, https://openalex.org/W2531068756, https://openalex.org/W2073256396, https://openalex.org/W2038039984, https://openalex.org/W2028819747, https://openalex.org/W2152250226, https://openalex.org/W2022690406, https://openalex.org/W1593117641, https://openalex.org/W2418628744, https://openalex.org/W2050386030, https://openalex.org/W2012393980, https://openalex.org/W2561533418, https://openalex.org/W4237876224, https://openalex.org/W2914590810, https://openalex.org/W2156778513, https://openalex.org/W2258753467, https://openalex.org/W2027921745, https://openalex.org/W2791495298, https://openalex.org/W2965815217, https://openalex.org/W2294327620, https://openalex.org/W2116382358, https://openalex.org/W2127540098 |
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| abstract_inverted_index.2 | 114 |
| abstract_inverted_index.A | 57 |
| abstract_inverted_index.a | 5, 112, 132, 139, 148, 213, 224, 258 |
| abstract_inverted_index.P. | 150, 163, 254 |
| abstract_inverted_index.To | 94 |
| abstract_inverted_index.an | 197 |
| abstract_inverted_index.be | 232 |
| abstract_inverted_index.by | 168 |
| abstract_inverted_index.in | 72, 92, 102, 128, 135, 142, 176, 201, 210, 253, 270 |
| abstract_inverted_index.is | 60, 131 |
| abstract_inverted_index.of | 11, 18, 33, 53, 68, 90, 99, 111, 119, 126, 145, 160, 196, 208, 216, 223, 241, 266 |
| abstract_inverted_index.on | 108 |
| abstract_inverted_index.to | 46, 61, 86, 178, 234 |
| abstract_inverted_index.we | 78, 104 |
| abstract_inverted_index.C16 | 188 |
| abstract_inverted_index.DHA | 91, 127, 209, 236, 269 |
| abstract_inverted_index.EPA | 73, 267 |
| abstract_inverted_index.TAG | 103, 238 |
| abstract_inverted_index.and | 29, 39, 74, 123, 170, 189, 205, 220, 237, 256, 268 |
| abstract_inverted_index.can | 231 |
| abstract_inverted_index.for | 8, 26, 156, 187, 262 |
| abstract_inverted_index.has | 181 |
| abstract_inverted_index.how | 227 |
| abstract_inverted_index.key | 133 |
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| abstract_inverted_index.the | 9, 16, 31, 50, 81, 97, 109, 124, 161, 171, 194, 217, 263 |
| abstract_inverted_index.was | 153, 166 |
| abstract_inverted_index.DGAT | 130 |
| abstract_inverted_index.DHA. | 75 |
| abstract_inverted_index.TAG. | 93, 129, 211 |
| abstract_inverted_index.This | 245 |
| abstract_inverted_index.acid | 37, 41 |
| abstract_inverted_index.acyl | 191 |
| abstract_inverted_index.cost | 52 |
| abstract_inverted_index.grew | 174 |
| abstract_inverted_index.have | 79 |
| abstract_inverted_index.high | 12, 34 |
| abstract_inverted_index.into | 250 |
| abstract_inverted_index.long | 20 |
| abstract_inverted_index.meet | 47 |
| abstract_inverted_index.type | 113 |
| abstract_inverted_index.used | 233 |
| abstract_inverted_index.vivo | 143 |
| abstract_inverted_index.with | 65, 185 |
| abstract_inverted_index.(DHA) | 42 |
| abstract_inverted_index.(EPA) | 38 |
| abstract_inverted_index.acids | 24 |
| abstract_inverted_index.based | 141 |
| abstract_inverted_index.broad | 182 |
| abstract_inverted_index.chain | 21 |
| abstract_inverted_index.fatty | 23 |
| abstract_inverted_index.high. | 56 |
| abstract_inverted_index.human | 27 |
| abstract_inverted_index.lipid | 121, 136, 203, 251 |
| abstract_inverted_index.study | 246 |
| abstract_inverted_index.taken | 154 |
| abstract_inverted_index.value | 13, 35 |
| abstract_inverted_index.(DGAT) | 117 |
| abstract_inverted_index.(TAGs) | 70 |
| abstract_inverted_index.DGAT2B | 152, 165, 199, 219 |
| abstract_inverted_index.DGATs, | 147 |
| abstract_inverted_index.diatom | 83, 140 |
| abstract_inverted_index.effort | 107 |
| abstract_inverted_index.enzyme | 134 |
| abstract_inverted_index.health | 28 |
| abstract_inverted_index.higher | 202 |
| abstract_inverted_index.levels | 67, 89, 207 |
| abstract_inverted_index.marine | 82 |
| abstract_inverted_index.native | 149 |
| abstract_inverted_index.screen | 144 |
| abstract_inverted_index.showed | 226 |
| abstract_inverted_index.strain | 173 |
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| abstract_inverted_index.capable | 118 |
| abstract_inverted_index.current | 51 |
| abstract_inverted_index.demand, | 48 |
| abstract_inverted_index.ectopic | 221 |
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| abstract_inverted_index.forward | 155 |
| abstract_inverted_index.further | 95, 248 |
| abstract_inverted_index.groups. | 192 |
| abstract_inverted_index.improve | 96 |
| abstract_inverted_index.insight | 249 |
| abstract_inverted_index.omega-3 | 19, 100 |
| abstract_inverted_index.qRT-PCR | 169 |
| abstract_inverted_index.remains | 55 |
| abstract_inverted_index.require | 43 |
| abstract_inverted_index.strains | 64 |
| abstract_inverted_index.Abstract | 0 |
| abstract_inverted_index.LC-PUFAs | 101, 190 |
| abstract_inverted_index.PtDGAT2B | 180 |
| abstract_inverted_index.Recently | 77 |
| abstract_inverted_index.Results: | 76 |
| abstract_inverted_index.approach | 59, 261 |
| abstract_inverted_index.combined | 214 |
| abstract_inverted_index.content, | 239 |
| abstract_inverted_index.detailed | 157 |
| abstract_inverted_index.engineer | 63 |
| abstract_inverted_index.enhanced | 66, 88, 206 |
| abstract_inverted_index.enriched | 71 |
| abstract_inverted_index.however, | 49 |
| abstract_inverted_index.increase | 235 |
| abstract_inverted_index.nitrogen | 242 |
| abstract_inverted_index.provides | 247 |
| abstract_inverted_index.resource | 7 |
| abstract_inverted_index.resulted | 200 |
| abstract_inverted_index.suggests | 257 |
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| abstract_inverted_index.improving | 120 |
| abstract_inverted_index.iterative | 228 |
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| abstract_inverted_index.promising | 58 |
| abstract_inverted_index.represent | 4 |
| abstract_inverted_index.substrate | 183 |
| abstract_inverted_index.wildtype. | 179 |
| abstract_inverted_index.(LC-PUFAs) | 25 |
| abstract_inverted_index.Oleaginous | 2 |
| abstract_inverted_index.accumulate | 87 |
| abstract_inverted_index.comparison | 177 |
| abstract_inverted_index.endogenous | 162, 198, 218 |
| abstract_inverted_index.engineered | 80 |
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| abstract_inverted_index.importance | 17 |
| abstract_inverted_index.metabolism | 252 |
| abstract_inverted_index.microalgae | 3 |
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| abstract_inverted_index.production | 10, 54, 122, 265 |
| abstract_inverted_index.synthesis. | 137 |
| abstract_inverted_index.transgenic | 172 |
| abstract_inverted_index.treatment. | 243 |
| abstract_inverted_index.Background: | 1 |
| abstract_inverted_index.Conclusion: | 244 |
| abstract_inverted_index.Considering | 15 |
| abstract_inverted_index.engineering | 230, 260 |
| abstract_inverted_index.improvement | 45 |
| abstract_inverted_index.microalgae. | 271 |
| abstract_inverted_index.preferences | 186 |
| abstract_inverted_index.significant | 44 |
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| abstract_inverted_index.Furthermore, | 212 |
| abstract_inverted_index.irrespective | 240 |
| abstract_inverted_index.successfully | 175 |
| abstract_inverted_index.Δ5-elongase | 225 |
| abstract_inverted_index.Phaeodactylum | 84 |
| abstract_inverted_index.incorporation | 98, 125 |
| abstract_inverted_index.metabolically | 62 |
| abstract_inverted_index.Overexpression | 159 |
| abstract_inverted_index.identification | 110 |
| abstract_inverted_index.overexpression | 195, 215 |
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| abstract_inverted_index.eicosapentaenoic | 36 |
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| abstract_inverted_index.acyl-CoA:diacylglycerol | 115 |
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| cited_by_percentile_year.min | 89 |
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| institutions_distinct_count | 7 |
| corresponding_institution_ids | https://openalex.org/I2799553609 |
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