Fermentation performance and metabolomic analysis of an engineered high-yield PUFA-producing strain of Schizochytrium sp.

Geng, Lingjun; Chen, Shenglan; Sun, Xiao‐Man; Hu, Xuechao; Ji, Xiao‐Jun; Huang, He; Ren, Lu‐Jing

doi:10.1007/s00449-018-2015-z

Cited by 25 publications

(10 citation statements)

References 37 publications

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“…Schizochytrium sp. is known to be a microalga predominantly producing polyunsaturated fatty acid (PUFA) such as docosahexaenoic acid (DHA), on an industrial scale . Although Schizochytrium sp.…”

Section: Resultsmentioning

confidence: 99%

Supplementation with Chlorella vulgaris, Chlorella protothecoides, and Schizochytrium sp. increases propionate‐producing bacteria in in vitro human gut fermentation

Jin¹,

Wook²,

Shin

et al. 2020

J Sci Food Agric

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BACKGROUND Gut microbiota are major contributors to host metabolism and are considered as potential targets of novel therapeutics. Microalgae have a strong potential for use as prebiotics because they are a rich source of proteins, fatty acids, fiber, and minerals for nutritional supplementation in humans. Nevertheless, there has been insufficient research into the effect of microalgae on gut microbiota. To investigate the effects of three edible microalgae (Chlorella vulgaris, Chlorella protothecoides, and Schizochytrium sp.) on gut microbiota, simulated digestion and colonic fermentation were examined. RESULTS Following in vitro digestion, the microalgae displayed different levels of bioaccessibility and the nutrient analysis revealed that unabsorbed nutrients during the digestion process could be used for colonic fermentation. Following colonic fermentation, the control, inulin, and microalgae groups displayed different metabolite tendencies when investigated with nuclear magnetic resonance (NMR) spectroscopic analysis. In particular, microalgae supplementation increased the proportion of propionate in the colonic culture (control: 19.14%, Inulin: 18.38%, C. vulgaris: 25.80%, C. protothecoides: 25.46%, and Schizochytrium sp.: 25.56%). Microbial profiling analysis using 16S rRNA gene sequencing also disclosed that the relative abundance of Bacteroides (control: 1.91%, inulin: 2.61%, C. vulgaris: 14.77%, C. protothecoides: 11.17%, and Schizochytrium sp.: 5.51%) and Dialister (control: 0.08%, inulin: 2.06%, C. vulgaris: 6.79%, C. protothecoides: 4.45%, and Schizochytrium sp.: 4.48%), involved in propionate metabolism increased more than in the inulin group. CONCLUSION Our findings suggest the potential use of microalgae as a functional food to increase propionate generation because propionate has been reported to be effective in weight loss and the inhibition of pathogen infection. © 2020 Society of Chemical Industry

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Section: Resultsmentioning

confidence: 99%

Supplementation with Chlorella vulgaris, Chlorella protothecoides, and Schizochytrium sp. increases propionate‐producing bacteria in in vitro human gut fermentation

Jin¹,

Wook²,

Shin

et al. 2020

J Sci Food Agric

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“…The decrease in glycine, L-serine and glycerol in the fluconazole group at 72 h and 96 h was due to the inhibited utilization of glucose, which reduced the synthesis of glyceraldehyde 3-phosphate. Geng et al pointed out that some intermediate metabolites including glyceraldehyde-3-phosphate and phosphoenolpyruvic acid in glycolysis pathway reduced, which promoted the glycolytic pathway to synthesize pyruvate [51]. However, the content of phenylalanine in the fluconazole group was much higher than that in the control group at 72 h, but far less at 96 h and 120 h. This trend was the same as that of squalene and may be because more squalene and IPP accumulated when the sterol pathway was blocked.…”

Section: Resultsmentioning

confidence: 99%

The role of fluconazole in the regulation of fatty acid and unsaponifiable matter biosynthesis in Schizochytrium sp. MYA 1381

Zhou

Pan

et al. 2019

BMC Microbiol

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BackgroundSchizochytrium has been widely used in industry for synthesizing polyunsaturated fatty acids (PUFAs), especially docosahexaenoic acid (DHA). However, unclear biosynthesis pathway of PUFAs inhibits further production of the Schizochytrium. Unsaponifiable matter (UM) from mevalonate pathway is crucial to cell growth and intracellular metabolism in all higher eukaryotes and microalgae. Therefore, regulation of UM biosynthesis in Schizochytrium may have important effects on fatty acids synthesis. Moreover, it is well known that UMs, such as squalene and β-carotene, are of great commercial value. Thus, regulating UM biosynthesis may also allow for an increased valuation of Schizochytrium.ResultsTo investigate the correlation of UM biosynthesis with fatty acids accumulation in Schizochytrium, fluconazole was used to block the sterols pathway. The addition of 60 mg/L fluconazole at 48 h increased the total lipids (TLs) at 96 h by 16% without affecting cell growth, which was accompanied by remarkable changes in UMs and NADPH. Cholesterol content was reduced by 8%, and the squalene content improved by 45% at 72 h, which demonstrated fluconazole’s role in inhibiting squalene flow to cholesterol. As another typical UM with antioxidant capacity, the β-carotene production was increased by 53% at 96 h. The increase of squalene and β-carotene could boost intracellular oxidation resistance to protect fatty acids from oxidation. The NADPH was found to be 33% higher than that of the control at 96 h, which meant that the cells had more reducing power for fatty acid synthesis. Metabolic analysis further confirmed that regulation of sterols was closely related to glucose absorption, pigment biosynthesis and fatty acid production in Schizochytrium.ConclusionThis work first reported the effect of UM biosynthesis on fatty acid accumulation in Schizochytrium. The UM was found to affect fatty acid biosynthesis by changing cell membrane function, intracellular antioxidation and reducing power. We believe that this work provides valuable insights in improving PUFA and other valuable matters in microalgae.

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“…There are some similar studies on oleagineous model systems including thraustochytrids [29][30][31][32]44,45] but none covering with quantitative methods the whole central carbon metabolism, redox-cofactors and lipid composition as completely as presented here. Those studies have mostly applied untargeted LC-MS and GC-MS profiling, few internal standards and usually only normalized to the total peak area.…”

Section: Methodsmentioning

confidence: 99%

“…Those studies have mostly applied untargeted LC-MS and GC-MS profiling, few internal standards and usually only normalized to the total peak area. Geng and coworkers used qualitative GC-MS analysis and reported 4-aminobutyric acid, proline and glutamine as potential biomarkers associated with growth and lipid accumulation in an engineered Schizochytrium strain [29]. Perez and coworkers used both LC-MS and GC-MS operated in nontarget qualitative mode to profile the metabolome of acetate and glucose grown Aurantiochytrium limacinum SR21 cells, and especially TCA metabolites were upregulated on acetate [30].…”

Section: Methodsmentioning

confidence: 99%

“…Establishment of genetic tools for thraustochytrid strain engineering has been challenging, but several successful attempts have been reported [24][25][26][27][28]. Most thraustochytrid studies are reported with cultivation data, total lipids and DHA-production, while only scarce information is available about intracellular metabolite levels [13,[29][30][31][32]. Such information is requested in ME projects, both to generate biological knowledge as basis for selection of targets to manipulate, but, equally important, to characterize the phenotype of the resulting mutants.…”

Section: Introductionmentioning

confidence: 99%

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Combined Metabolome and Lipidome Analyses for In-Depth Characterization of Lipid Accumulation in the DHA Producing Aurantiochytrium sp. T66

et al. 2021

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Thraustochytrids are marine heterotrophic microorganisms known for their potential to accumulate docosahexaenoic acid (DHA)-enriched lipids. There have been many attempts to improve thraustochytrid DHA bioprocesses, especially through traditional optimization of cultivation and media conditions. Nevertheless, thraustochytrid-based bioprocesses are still not commercially competitive for high volume-low cost production of DHA. Thus, it is realized that genetic and metabolic engineering strategies are needed for the development of commercially competitive thraustochytrid DHA cell factories. Here, we present an analytical workflow for high resolution phenotyping at metabolite and lipid levels to generate deeper insight into the thraustochytrid physiology, with particular focus on central carbon and redox metabolism. We use time-series sampling during unlimited growth and nitrogen depleted triggering of DHA synthesis and lipid accumulation (LA) to show-case our methodology. The mass spectrometric absolute quantitative metabolite profiling covered glycolytic, pentose phosphate pathway (PPP) and tricarboxylic acid cycle (TCA) metabolites, amino acids, complete (deoxy)nucleoside phosphate pools, CoA and NAD metabolites, while semiquantitative high-resolution supercritical fluid chromatography MS/MS was applied for the lipid profiling. Interestingly, trace amounts of a triacylglycerols (TG) with DHA incorporated in all three acyl positions was detected, while TGs 16:0_16:0_22:6 and 16:0_22:6_22:6 were among the dominant lipid species. The metabolite profiling data indicated that lipid accumulation is not limited by availability of the acyl chain carbon precursor acetyl-CoA nor reducing power (NADPH) but rather points to the TG head group precursor glycerol-3-phosphate as the potential cause at the metabolite level for the gradual decline in lipid production throughout the cultivation. This high-resolution phenotyping provides new knowledge of changes in the central metabolism during growth and LA in thraustochytrids and will guide target selection for metabolic engineering needed for further improvements of this DHA cell factory.

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Fermentation performance and metabolomic analysis of an engineered high-yield PUFA-producing strain of Schizochytrium sp.

Cited by 25 publications

References 37 publications

Supplementation with Chlorella vulgaris, Chlorella protothecoides, and Schizochytrium sp. increases propionate‐producing bacteria in in vitro human gut fermentation

Supplementation with Chlorella vulgaris, Chlorella protothecoides, and Schizochytrium sp. increases propionate‐producing bacteria in in vitro human gut fermentation

The role of fluconazole in the regulation of fatty acid and unsaponifiable matter biosynthesis in Schizochytrium sp. MYA 1381

Combined Metabolome and Lipidome Analyses for In-Depth Characterization of Lipid Accumulation in the DHA Producing Aurantiochytrium sp. T66

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