De novo transcriptomic and metabolomic analysis of docosahexaenoic acid (DHA)-producing Crypthecodinium cohnii during fed-batch fermentation

Pei, Guangsheng; Li, Xingrui; Liu, Liangsen; Liu, Jing; Wang, Fangzhong; Chen, Lei; Zhang, Weiwen

doi:10.1016/j.algal.2017.07.031

Cited by 37 publications

(34 citation statements)

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“…2). It was also consistent with previous findings that the lipid content (% DCW, w/w) of C. cohnii was increased with fermentation progressing [9,40].…”

Section: Gc-ms Based Metabolomic Analysis Of the Repeated Fed-batch Csupporting

confidence: 93%

“…Briefly, 25 mg freeze-dried cells of C. cohnii was extracted by chloroform:methanol (2:1, v/v) adding 0.01% butylated hydroxytoluene for three times, and then was washed by 1 M KCl solution and ddH 2 O for one time, respectively. Fatty acids profile were measured according to the previous study [40,46]. Briefly, freeze-dried cells of C. cohnii adding with heptadecanoic acid methanol solution (an internal standard), methanol of 3% (v/v) sulfuric acid, and chloroform were conducted for reaction at 100 °C for 4 h. The chloroform phase was used to detect after stratification by ddH 2 O.…”

Section: Analysis Of Dcw Glucose Concentration Total Lipid Content mentioning

confidence: 99%

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Repeated fed-batch strategy and metabolomic analysis to achieve high docosahexaenoic acid productivity in Crypthecodinium cohnii

et al. 2020

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Background: Docosahexaenoic acid (DHA) is essential for human diet. However, high production cost of DHA using C. cohnii makes it currently less competitive commercially, which is mainly caused by low DHA productivity. In recent years, repeated fed-batch strategies have been evaluated for increasing the production of many fermentation products. The reduction in terms of stability of culture system was one of the major challenges for repeated fed-batch fermentation. However, the possible mechanisms responsible for the decreased stability of the culture system in the repeated fedbatch fermentation are so far less investigated, restricting the efforts to further improve the productivity. In this study, a repeated fed-batch strategy for DHA production using C. cohnii M-1-2 was evaluated to improve DHA productivity and reduce production cost, and then the underlying mechanisms related to the gradually decreased stability of the culture system in repeated fed-batch culture were explored through LC-and GC-MS metabolomic analyses. Results: It was discovered that glucose concentration at 15-27 g/L and 80% medium replacement ratio were suitable for the growth of C. cohnii M-1-2 during the repeated fed-batch culture. A four-cycle repeated fed-batch culture was successfully developed and assessed at the optimum cultivation parameters, resulting in increasing the total DHA productivity by 26.28% compared with the highest DHA productivity of 57.08 mg/L/h reported using C. cohnii, including the time required for preparing seed culture and fermentor. In addition, LC-and GC-MS metabolomics analyses showed that the gradually decreased nitrogen utilization capacity, and down-regulated glycolysis and TCA cycle were correlated with the decreased stability of the culture system during the long-time repeated fed-batch culture. At last, some biomarkers, such as Pyr, Cit, OXA, FUM, l-tryptophan, l-threonine, l-leucine, serotonin, and 4-guanidinobutyric acid, correlated with the stability of culture system of C. cohnii M-1-2 were identified. Conclusions: The study proved that repeated fed-batch cultivation was an efficient and energy-saving strategy for industrial production of DHA using C. cohnii, which could also be useful for cultivation of other microbes to improve productivity and reduce production cost. In addition, the mechanisms study at metabolite level can also be useful to further optimize production processes for C. cohnii and other microbes.

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“…2). It was also consistent with previous findings that the lipid content (% DCW, w/w) of C. cohnii was increased with fermentation progressing [9,40].…”

Section: Gc-ms Based Metabolomic Analysis Of the Repeated Fed-batch Csupporting

confidence: 93%

Section: Analysis Of Dcw Glucose Concentration Total Lipid Content mentioning

confidence: 99%

Repeated fed-batch strategy and metabolomic analysis to achieve high docosahexaenoic acid productivity in Crypthecodinium cohnii

et al. 2020

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“…The solvents were removed using a vacuum concentrator system (ZLS-1, Hunan, China). The lipid profile was analyzed using an Agilent 5975 MSD/7890 instrument (Agilent Corp., Santa Clara, CA, United States) according to previous publications ( Xiong et al, 2008 ; Pei et al, 2017 ).…”

Section: Methodsmentioning

confidence: 99%

“…Crypthecodinium cohnii , a flagellated marine microalga, has been considered as a prolific producer of DHA, which contains 25–60% DHA while less than 1% of other types of PUFAs in its total fatty acids (TFAs) ( Pei et al, 2017 ). However, up to now, no genome sequence is available for C. cohnii , and the DHA biosynthetic pathway in C. cohnii remains elusive ( de Swaaf et al, 2003 ; Mendes et al, 2008 ).…”

Section: Introductionmentioning

confidence: 99%

“…Previous studies suggested that C. cohnii might utilize the polyketide synthases (PKS) route for the biosynthesis and accumulation of DHA, which requires no oxygen or NADPH-dependent desaturases ( Ratledge, 2004 ). Meanwhile, Pei et al (2017) recently applied de novo transcriptome analysis to characterize central carbohydrate and fatty acid biosynthesis in C. cohnii , which suggested that C. cohnii might utilize a combination of PKS systems and desaturase steps for DHA biosynthesis. In addition, metabolomics analysis has also been employed to understand the possible mechanisms responsible for the increased lipid accumulation ( Li et al, 2015 ).…”

Section: Introductionmentioning

confidence: 99%

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13C Metabolic Flux Analysis of Enhanced Lipid Accumulation Modulated by Ethanolamine in Crypthecodinium cohnii

et al. 2018

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The heterotrophic microalga Crypthecodinium cohnii has attracted considerable attention due to its capability of accumulating lipids with a high fraction of docosahexaenoic acid (DHA). In our previous study, ethanolamine (ETA) was identified as an effective chemical modulator for lipid accumulation in C. cohnii. In this study, to gain a better understanding of the lipid metabolism and mechanism for the positive effects of modulator ETA, metabolic flux analysis was performed using 13C-labeled glucose with and without 1 mM ETA modulator. The analysis of flux distribution showed that with the addition of ETA, flux in glycolysis pathway and citrate pyruvate cycle was strengthened while flux in pentose phosphate pathway was decreased. In addition, flux in TCA cycle was slightly decreased compared with the control without ETA. The enzyme activity of malic enzyme (ME) was significantly increased, suggesting that NADP+-dependent ME might be the major source of NADPH for lipid accumulation. The flux information obtained by this study could be valuable for the further efforts in improving lipid accumulation and DHA production in C. cohnii.

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Research and Production of Microbial Polyunsaturated Fatty Acids

Christophe

Fontanille

Larroche

2019

Bioprocessing for Biomolecules Production

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De novo transcriptomic and metabolomic analysis of docosahexaenoic acid (DHA)-producing Crypthecodinium cohnii during fed-batch fermentation

Cited by 37 publications

References 63 publications

Repeated fed-batch strategy and metabolomic analysis to achieve high docosahexaenoic acid productivity in Crypthecodinium cohnii

Repeated fed-batch strategy and metabolomic analysis to achieve high docosahexaenoic acid productivity in Crypthecodinium cohnii

13C Metabolic Flux Analysis of Enhanced Lipid Accumulation Modulated by Ethanolamine in Crypthecodinium cohnii

Research and Production of Microbial Polyunsaturated Fatty Acids

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