Thraustochytrid and fatty acid composition and preparation method and application thereof
A thraustochytrium and fatty acid technology, applied in biochemical equipment and methods, microbial-based methods, drug combinations, etc., can solve problems such as unsatisfactory levels of omega-6 fatty acids and low productivity
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Embodiment 1
[0174] Identification of an isolated Thraustochytrium deposited under ATCC Accession No. PTA-9695 for classification.
[0175] Samples were collected from intertidal habitats during low tide. Place water, sediment, live plant matter and spoiled plant / animal debris into 50ml sterile tubes. Each sample component was plated together with water on a solid agar plate of separation medium. The separation medium consists of the following: 500ml artificial seawater, 500ml distilled water, 1g glucose, 1g glycerol, 13g agar, 1g glutamate, 0.5g yeast extract, 0.5g casein hydrolyzate, 1ml vitamin solution (100mg / L thiamine 0.5mg / L biotin, 0.5mg B12), 1ml trace mineral solution (PII metal, containing: 6.0g FeCl per liter 3 6H 2 O,6.84g H 3 BO 3 ,0.86g MnCl 2 4H 2 O, 0.06g ZnCl 2 ,0.026 CoCl 2 6H 2 O, 0.052g NiSO 4 h 2 O, 0.002g CuSO 4 5H 2 O and 0.005g Na 2 MoO 4 2H 2 0) and 500 mg each of penicillin G and streptomycin sulfate. Incubate the agar plate in the dark at 20-25...
Embodiment 2
[0196] An isolated Thraustochytrium deposited under ATCC Accession No. PTA-9695 produced high levels of cell growth under different culture conditions as described below. Typical media and culture conditions are shown in Table 1. High levels of fatty acids and DNA were also observed (ie greater than 50% by weight of dry cell weight was fatty acid and greater than 50% by weight fatty acid was DHA).
[0197] In a carbon and nitrogen fed culture with 8200 ppm Cl- at 22.5°C, 20% dissolved oxygen at pH 7.0, the isolate produced a dry cell weight of 140 g / L after 7 days of culture, with a fatty acid content of 70% by weight . A closed loop ammonia feed was used and the pH was maintained at 7.0. Omega-3 production under these conditions was 8.92 g / (L*day) over 7 days with 4.7 g / L EPA (5% by weight fatty acids) and 56.3 g / L DHA (57% by weight fatty acids).
[0198] In a carbon and nitrogen fed culture with 3640 ppm Cl- at 22.5°C, 20% dissolved oxygen at pH 7.0, the isolate produced...
Embodiment 3
[0201] Oil was extracted from a biomass sample of isolated Thraustochytrium deposited under ATCC Accession No. PTA-9695 (Sample A). 980ppm Cl at 22.5°C- The biomass samples were produced in a carbon and nitrogen fed culture with 20% dissolved oxygen at pH 7.0. Oil was extracted from biomass sample A by the hexane extraction method to produce microbial oil sample A1. Briefly, dry biomass was ground with hexane for about 2 h, using stainless steel tubes and stainless steel ball bearings. Vacuum filter the slurry and collect the filtrate. Hexane was removed using a rotary evaporator. also use Method (GEA Westfalia Separator UK Ltd., Milton Keynes, UK) oil was extracted from biomass sample A to produce microbial oil sample A2. Individual lipids were separated from microbial oil samples A1 and A2 by low-pressure flash chromatography, and the weight percentages of each category were determined. The distribution of fatty acids as fatty acid methyl esters (FAMEs) in each class w...
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