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

Ishibashi, Yoshihiro; Goda, Hatsumi M.; Hamaguchi, Rie; Sakaguchi, Koichiro; Sekiguchi, Takayoshi; Ishiwata, Y; Okita, Yuji; Mochinaga, Seiya; Ikeuchi, Satoru; Mizobuchi, Takahiro; Takao, Yoshimi; Mori, K.; Tashiro, Kosuke; Okino, Nozomu; Honda, Daiske; Hayashi, Masahiro; Ito, Makoto

doi:10.1038/s42003-021-02857-w

Cited by 16 publications

(13 citation statements)

References 52 publications

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“…Results suggested that electroporation might not be suitable for transformation of Schizochytrium sp. M326, although electroporation was the most widely used method for thraustochytrid transformation. ,,, The same situations have been described for a few strains of Parietichytrium sp . and Schizochytrium sp …”

Section: Resultsmentioning

confidence: 66%

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Modulating DHA-Producing Schizochytrium sp. toward Astaxanthin Biosynthesis via a Seamless Genome Editing System

Liu

Lin

et al. 2022

ACS Synth. Biol.

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Schizochytrium sp. is commercially used for the production of docosahexaenoic acid (DHA). Some strains of Schizochytrium sp. are also known to produce low amounts of carotenoids, including astaxanthin and β-carotene. In order to enhance the production of astaxanthin in Schizochytrium sp., we established a seamless genome editing system with a dual selection marker for rapid screening of positive transformants. By using this system, we strengthened the endogenous mevalonate pathway, enhanced the supply of geranylgeranyl diphosphate and β-carotene, upregulated endogenous β-carotene hydroxylase, and introduced the algal astaxanthin pathway. The highest astaxanthin production in the engineered Schizochytrium sp. was achieved at 8.1 mg/L (307.1 μg/g dry cell weight) under shake-flask conditions, which was 2.6-fold higher than that in the start strain. Meanwhile, the percentage of DHA to total fatty acids was not obviously affected. We then eliminated the dual selection marker by using the Cre-loxP recombination system, and the engineered strain was ready for iterative editing. The developed system could be applied to seamlessly engineer DHA-producing Schizochytrium sp. toward astaxanthin and other value-added terpenoids, which broadens the application of this strain.

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Section: Resultsmentioning

confidence: 66%

“…16,20,25,26 The same situations have been described for a few strains of Parietichytrium sp. 36 and Schizochytrium sp. 19 Next, more constructed vectors were evaluated by using particle bombardment-mediated transformation.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

Modulating DHA-Producing Schizochytrium sp. toward Astaxanthin Biosynthesis via a Seamless Genome Editing System

Liu

Lin

et al. 2022

ACS Synth. Biol.

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“… Strain EPA of TFA (%) EPA (g/L) DHA of TFA (%) DHA (g/L) Reference Hondae a gen. nov. 0.41 ± 0.01 0.01 ± 0.01 50.12 ± 1.23 0.06 ± 0.01 9 Parietichytrium sp. 2.11 ± 0.17 0.08 ± 0.01 5.1 ± 0.3 0.19 ± 0.02 10 Schizochytrium sp. A-2 2.01 ± 0.20 0.13 ± 0.01 53.2 ± 0.30 3.44 ± 0.02 42 Nannochloropsis oceanica 4.12 ± 0.35 0.02 ± 0.01 2.37 ± 0.40 0.01 ± 0.01 47 Phaeodactylum tricornutum and Aurantiochytrium limacinum 17.96 ± 1.23 0.25 ± 0.01 18.42 ± 0.45 0.26 ± 0.01 48 Rhodomonas baltica NIVA-5/91 12.1 ± 1.07 0.88 ± 0.07 12.8 ± 0.28 0.95 ± 0.03 49 Phaeodactylum tricornutum CCMP 2561 26.2 ± 0.36 1.64 ± 0.05 4.93 ± 0.09 0.25 ± 0.02 49 Schizochytrium sp.…”

Section: Resultsmentioning

confidence: 99%

“…The reported proportion of DHA in total fatty acids (TFAs) is 5.1% in Parietichytrium sp. and 54.6% in Thraustochytrium roseum ATCC 28210 10 , 11 . Presently, thraustochytrids are preferred being used in scientific research and DHA commercial production due to their high lipid content (>50% of dry weight) and ease of cultivation.…”

Section: Introductionmentioning

confidence: 97%

Efficient co-production of EPA and DHA by Schizochytrium sp. via regulation of the polyketide synthase pathway

Liu

Zhang

et al. 2022

Commun Biol

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Presently, the supply of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) traditionally produced by marine fisheries will be insufficient to meet their market demand in food industry. Thus a sustainable alternative source is urgently required. Schizochytrium sp. is an ideal producer of DHA; however, its ability to co-produce DHA and EPA has not yet been proved. Herein, we first described a cobalamin-independent methionine synthase-like (MetE-like) complex, which contains independent acyltransferase and 3-ketoacyl synthase domains, independent of the traditional polyketide synthase (PKS) system. When the MetE-like complex was activated, the EPA content was increased from 1.26% to 7.63%, which is 6.06-folds higher than that in the inactivated condition. Through lipidomics, we find that EPA is more inclined to be stored as triglyceride. Finally, the EPA production was enhanced from 4.19 to 29.83 (mg/g cell dry weight) using mixed carbon sources, and the final yield reached 2.25 g/L EPA and 9.59 g/L DHA, which means that Schizochytrium sp. has great market potential for co-production of EPA and DHA.

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“…Moreover, PUFAs functionally attenuated inflammatory bowel disease (IBD) through regulatory effects on gut bacterial diversity, which included the effective proliferation of probiotics, such as Bifidobacterium ( 10 – 12 ). However, PUFAs in mammals are less efficiently synthesized due to restrictions of certain essential factors including precursors preparation, fatty acids construction, α-linolenic acid (ALA, C18:3), and several elongations and desaturation chemical reactions ( 13 ). Fortunately, increasing potency of PUFA content could be achieved by dietary management such as supplementation with marine organisms.…”

Section: Introductionmentioning

confidence: 99%

Metagenomic insights into the modulatory effects of kelp powder (Thallus laminariae)-Treated dairy milk on growth performances and physiological lipometabolic processes of kunming mice

Xue

Mo²,

Ma³

et al. 2022

Front. Nutr.

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Kelp powder, supplemented with a dairy cow diet, effectively improved the milk polyunsaturated fatty acids (PUFAs) content. However, little information exists on the downstream effects of the kelp-treated milk on body health, gut microbiota, and nutrient metabolism. For this purpose, 48 3-week old Kunming (KM) male mice with an average body weight of 16.1 g ± 0.2 g were randomly divided into the control treatment (CON, fed with standard chow), the common milk supplement treatment (Milk), and the kelp powder-treated milk supplement treatment (KPM). The experiment lasted for 35 days, with a 7-day long adaptive period and a 28-day long main trial. Phenotypic parameters including growth performances and serum lipids-related parameters were first measured, and results indicated that Milk and KPM supplement significantly promoted the total body weight gain (P < 0.05), while significantly decreasing the feed conversion ratio compared with CON (P < 0.05). No significant differences were observed in the blood lipids content among all three treatments, however, the triglyceride content showed a decreasing trend after KPM supplement treatment. Further, activities of liver lipometabolic-related enzymes were investigated to determine the underlying factors that impacted physiological lipid metabolism. KPM treatment showed a significant reductive effect on the activity of lipogenesis-related enzymes, such as FAS and ACC, while a significant stimulative effect on the activity of lipolysis-related enzymes included the ATGL and CPT1 compared with CON (P < 0.05). Finally, gastrointestinal tract development and cecal microbiota community that correlated with body lipid degradation and absorption were measured to determine the underlying mechanism of KPM supplementation on physiological lipid metabolism. Results indicated that supplementation with KPM significantly enhanced cecal bacteria diversity which was reflected in the significant increase of Chao1 and ACE indexes. Besides, starch-degraded bacteria such as Faecalibacterium, Ruminococcaceae, and Streptococcus are significant decreased (P < 0.05), while cellulose-degraded bacteria including Parabacteroides, Prevotella, Lactobacillus, Clostridium, and Bifidobacterium are significantly increased (P < 0.05) after KPM supplement, which may further restrict the energy generation and therefore reduce the lipid deposition. In summary, kelp supplement helped increase the milk PUFAs content, enhance the bacterial diversity and relative abundances of probiotics, which finally modulated physiological lipid metabolism, and promote growth performances.

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

Cited by 16 publications

References 52 publications

Modulating DHA-Producing Schizochytrium sp. toward Astaxanthin Biosynthesis via a Seamless Genome Editing System

Modulating DHA-Producing Schizochytrium sp. toward Astaxanthin Biosynthesis via a Seamless Genome Editing System

Efficient co-production of EPA and DHA by Schizochytrium sp. via regulation of the polyketide synthase pathway

Metagenomic insights into the modulatory effects of kelp powder (Thallus laminariae)-Treated dairy milk on growth performances and physiological lipometabolic processes of kunming mice

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