Optimizing Eicosapentaenoic Acid Production by Grafting a Heterologous Polyketide Synthase Pathway in the Thraustochytrid <i>Aurantiochytrium</i>

Wang, Sen; Lan, Chuanzeng; Wang, Zhuojun; Wan, Weijian; Zhang, Huidan; Cui, Qiu; Song, Xiaojin

doi:10.1021/acs.jafc.0c04299

Cited by 37 publications

(34 citation statements)

References 39 publications

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“…SK4 showed that the fatty acid content of the engineered strain was 44% higher than that of the wild type [ 11 ]. Furthermore, a recent study showed that the whole polyketide synthase (PKS) pathway from Shewanella japonica was expressed in Aurantiochytrium , resulting in a 26.9% increase in the lipid yield [ 12 ]. Up to date, nine genera of thraustochytrids have been recognized [ 13 ], however, efficient gene editing tools suitable for editing thraustochytrids have not been established.…”

Section: Introductionmentioning

confidence: 99%

An emerging simple and effective approach to increase the productivity of thraustochytrids microbial lipids by regulating glycolysis process and triacylglycerols’ decomposition

Wang

Nong

et al. 2021

Biotechnol Biofuels

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Background The oleaginous microorganism Schizochytrium sp. is widely used in scientific research and commercial lipid production processes. However, low glucose-to-lipid conversion rate (GLCR) and low lipid productivity of Schizochytrium sp. restrict the feasibility of its use. Results Orlistat is a lipase inhibitor, which avoids triacylglycerols (TAGs) from hydrolysis by lipase. TAGs are the main storage forms of fatty acids in Schizochytrium sp. In this study, the usage of orlistat increased the GLCR by 21.88% in the middle stage of fermentation. Whereas the productivity of lipid increased 1.34 times reaching 0.73 g/L/h, the saturated fatty acid and polyunsaturated fatty acid yield increased from 21.2 and 39.1 to 34.9 and 48.5 g/L, respectively, indicating the advantages of using a lipase inhibitor in microbial lipids fermentation. Similarly, the system was also successful in Thraustochytrid Aurantiochytrium. The metabolic regulatory mechanisms stimulated by orlistat in Schizochytrium sp. were further investigated using transcriptomics and metabolomics. The results showed that orlistat redistributed carbon allocation and enhanced the energy supply when inhibiting the TAGs’ degradation pathway. Therefore, lipase in Schizochytrium sp. prefers to hydrolyze saturated fatty acid TAGs into the β-oxidation pathway. Conclusions This study provides a simple and effective approach to improve lipid production, and makes us understand the mechanism of lipid accumulation and decomposition in Schizochytrium sp., offering new guidance for the exploitation of oleaginous microorganisms.

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Section: Introductionmentioning

confidence: 99%

An emerging simple and effective approach to increase the productivity of thraustochytrids microbial lipids by regulating glycolysis process and triacylglycerols’ decomposition

Wang

Nong

et al. 2021

Biotechnol Biofuels

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“…Recently, Wang et al reported that Aurantiochytrium sp. transformed with a bacterial EPA-producing PUFA-S gene cluster produced 2.7 g/L EPA 42 . To date, the highest yield of EPA was achieved by engineered Yarrowia lipolytica , which was transformed with several heterologous genes that constitute the ELO/DES pathway required for EPA synthesis 43 .…”

Section: Discussionmentioning

confidence: 99%

PUFA synthase-independent DHA synthesis pathway in Parietichytrium sp. and its modification to produce EPA and n-3DPA

et al. 2021

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The demand for n-3 long-chain polyunsaturated fatty acids (n-3LC-PUFAs), such as docosahexaenoic acid (DHA) and eicosapentaenoic acid (EPA), will exceed their supply in the near future, and a sustainable source of n-3LC-PUFAs is needed. Thraustochytrids are marine protists characterized by anaerobic biosynthesis of DHA via polyunsaturated fatty acid synthase (PUFA-S). Analysis of a homemade draft genome database suggested that Parietichytrium sp. lacks PUFA-S but possesses all fatty acid elongase (ELO) and desaturase (DES) genes required for DHA synthesis. The reverse genetic approach and a tracing experiment using stable isotope-labeled fatty acids revealed that the ELO/DES pathway is the only DHA synthesis pathway in Parietichytrium sp. Disruption of the C20 fatty acid ELO (C20ELO) and ∆4 fatty acid DES (∆4DES) genes with expression of ω3 fatty acid DES in this thraustochytrid allowed the production of EPA and n-3docosapentaenoic acid (n-3DPA), respectively, at the highest level among known microbial sources using fed-batch culture.

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“…ONC-T18 [ 55 ], and A. limacinum SR21 [ 52 ], while foot-and-mouth disease virus 2A (F2A) peptides have been used in Aurantiochytrium sp. SD116 [ 41 ]. In Aurantiochytrium sp.…”

Section: The Properties Of the Dna Related To Gene Expression In Thraustochytridsmentioning

confidence: 99%

“…In Aurantiochytrium sp. SD116, the proteins that contain F2A peptides were nearly 100% cleaved [ 41 ]. However, in Thraustochytrium sp.…”

Section: The Properties Of the Dna Related To Gene Expression In Thraustochytridsmentioning

confidence: 99%

“… ( a ) The number of successfully engineered thraustochytrid strains by one or more methods (electroporation [ 20 , 21 , 22 , 23 , 24 , 25 , 26 , 27 , 28 , 29 , 30 , 31 , 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 , 41 , 42 , 43 , 44 , 45 , 46 , 47 , 48 , 49 , 50 , 51 , 52 ], biolistic transformation [ 5 , 27 , 53 , 54 , 55 ], and others [ 56 , 57 , 58 , 59 ]) ( Tables S1 and S2 ). The doughnut pie charts indicate the proportion of different genera.…”

Section: Figurementioning

confidence: 99%

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Method Development Progress in Genetic Engineering of Thraustochytrids

Rau

Ertesvåg

2021

Marine Drugs

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Thraustochytrids are unicellular, heterotrophic marine eukaryotes. Some species are known to store surplus carbon as intracellular lipids, and these also contain the long-chain polyunsaturated fatty acid docosahexaenoic acid (DHA). Most vertebrates are unable to synthesize sufficient amounts of DHA, and this fatty acid is essential for, e.g., marine fish, domesticated animals, and humans. Thraustochytrids may also produce other commercially valuable fatty acids and isoprenoids. Due to the great potential of thraustochytrids as producers of DHA and other lipid-related molecules, a need for more knowledge on this group of organisms is needed. This necessitates the ability to do genetic manipulation of the different strains. Thus far, this has been obtained for a few strains, while it has failed for other strains. Here, we systematically review the genetic transformation methods used for different thraustochytrid strains, with the aim of aiding studies on strains not yet successfully transformed. The designs of transformation cassettes are also described and compared. Moreover, the potential problems when trying to establish transformation protocols in new thraustochytrid species/strains are discussed, along with suggestions utilized in other organisms to overcome similar challenges. The approaches discussed in this review could be a starting point when designing protocols for other non-model organisms.

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Optimizing Eicosapentaenoic Acid Production by Grafting a Heterologous Polyketide Synthase Pathway in the Thraustochytrid Aurantiochytrium

Cited by 37 publications

References 39 publications

An emerging simple and effective approach to increase the productivity of thraustochytrids microbial lipids by regulating glycolysis process and triacylglycerols’ decomposition

An emerging simple and effective approach to increase the productivity of thraustochytrids microbial lipids by regulating glycolysis process and triacylglycerols’ decomposition

PUFA synthase-independent DHA synthesis pathway in Parietichytrium sp. and its modification to produce EPA and n-3DPA

Method Development Progress in Genetic Engineering of Thraustochytrids

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