Production of Lipids and Proteome Variation in a Chilean Thraustochytrium striatum Strain Cultured under Different Growth Conditions
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Total lipids and docosahexaenoic acid (DHA) production by a Chilean isolated thraustochytrid were evaluated under different growth conditions in shake flasks. The analyzed strain was identified as Thraustochytrium striatum according to an 18S rRNA gene sequence analysis. The strain (T. striatum AL16) showed negligible growth in media prepared with artificial seawater at concentrations lower than 50% v/v and pH lower than 5. Maltose and starch were better carbon sources for growth than glucose. DHA content of the biomass grown with maltose (60 g L−1) was doubled by increasing the agitation rate from 150 to 250 rpm. The DHA (0.8–6%) and eicosapentaenoic acid (0.2–21%) content in the total lipids varied depending on culture conditions and culture age. Lipid and DHA concentration increased (up to 5 g L−1 and 66 mg L−1, respectively) by regularly feeding the culture with a concentrated starch solution. Carotenoid accumulation was detected in cells grown with maltose or starch. Contrasting conditions of starch and glucose cultures were selected for comparative proteomics. Total protein extracts were separated by two-dimensional gel electrophoresis; 25 spots were identified using ESI-MS/MS. A protein database (143,006 entries) for proteomic interrogation was generated using de novo assembling of Thraustochytrium sp. LLF1b – MMETSP0199_2 transcriptome; 18 proteins differentially expressed were identified. Three ATP synthases were differentially accumulated in cultures with glucose, whereas malate dehydrogenase was more abundant in cells cultured with starch.
KeywordsCulture conditions Docosahexaenoic acid Polyunsaturated fatty acids Microbial lipids Thraustochytrium
This research was supported by the Centre for Biotechnology and Bioengineering (CeBiB) FB-0001 and partially supported by the supercomputing infrastructure of the NLHPC (ECM-02; Powered@NLHPC). Parts of the experiments (Galaxy server) were performed at the Bordeaux Bioinformatic Center (CBIB). The authors thank the Dirección de Investigación at Universidad de La Frontera for economic support provided through GAP.
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