Method for transforming vegetable oil into very-long-chain highly unsaturated fatty acid enriched bio-oil
A technology of unsaturated fatty acids and vegetable oils, which is applied in the field of bio-oils rich in ultra-long-chain highly unsaturated fatty acids, can solve the problems of limited fish oil resources, low yield, and complicated purification process, so as to promote industrial utilization potential and increase growth Effect of Velocity and Velocity of Grease Accumulation
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Embodiment 1
[0031] 1. Strain purification
[0032] Thaustochytrium was placed on a solid medium plate under peptone, yeast extract powder, and glucose medium, streaked, and colonies with fast growth rates were selected for purification. The formula of 100ml medium: glucose 1%, peptone 0.15%, yeast extract 0.01%, agar 1%. The culture temperature was 20°C and the culture was static.
[0033] 2. Activation culture
[0034] Add 1 small drop of vegetable oil to the selected area on the above plate, suck it with a pipette gun, and transfer it to a 100ml Erlenmeyer flask. Each 100ml culture solution contains 1% glucose, 0.15% peptone, 0.01% yeast extract, and 1-1% vegetable oil. 2%; culture temperature is 15-20°C, shake culture, culture time is 7 days.
[0035] 3. Rapid cultivation
[0036] After the above culture solution was left to stand for 30 minutes, absorb a layer of liquid on the surface, approximately 30ml of the inoculum solution, take several bottles, and add the inoculum solution...
Embodiment 2
[0043] 1. Collect the biomass of the Thraustochytrium obtained in Implementation 1. The medium used has the following four types: YP, GYP, GYPO, YPO (G refers to glucose, Y refers to yeast extract, P refers to peptone, O refers to vegetable oil), wherein YPO20 refers to adding 20mL vegetable oil to 500mL YPO medium, and so on.
[0044] 2. The extraction method of bio-oil adopts the Bligh-Dyer method. Weigh 1g of the sample to be tested, add 10ml of methanol, add a small amount of acid-washed sand to grind, homogenize for 1 minute, then add 20ml of chloroform, continue homogenizing for 2 minutes, filter, and filter the residue Add 30ml of chloroform-methanol mixture (2:1v / v) to it, grind it, and filter. First wash the filter residue with 20ml chloroform and 10ml methanol, combine the filtrate, then add water with 1 / 4 of the volume of the whole filtrate, vibrate, leave to stand for stratification, remove the solids at the upper layer and the interface, and then add methanol-wate...
Embodiment 3
[0057] 1. According to the analysis results obtained in Example 2, it was found that the Thraustochytrium C16 fatty acid cultured with vegetable oil did not decrease compared with the Thraustochytrium without vegetable oil, but the ratio of C18 fatty acid increased to some extent. Exogenous vegetable oil interfered, so Thraustochytrium cultured under GYP and GYPO and pure vegetable oil were selected for lipid analysis.
[0058] 2. Sample Preparation for HPLC Total Lipid Analysis
[0059] Thraustochytrium samples cultured in GYP and GYPO medium and vegetable oil were subjected to total lipid extraction by Bligh-Dyer method, and analyzed by UPLC-Q-TOF-MS.
[0060] 3. Chromatography and mass spectrometry conditions of LCMS
[0061] Chromatographic conditions: the total lipid extracts of Thraustochytrium under different culture medium conditions were analyzed by WatersAcquityUltraPerformanceLC-(UPLC)-Q-TOFPremier ultra-high performance liquid chromatography-quadrupole-high resolu...
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