Culture method for increasing yield of fucoxanthin contained in diatom
A medium fucoxanthin and culture method technology, applied in the field of diatom culture and fucoxanthin yield culture, can solve the problems of low fucoxanthin content and unsatisfactory fucoxanthin yield
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Embodiment 1
[0047] Cultivate Cyclotella as follows:
[0048] (1) Use a 250ml Erlenmeyer flask as the culture container, with a working volume of 120ml. The activated Cyclotella was cultured heterotrophically in a shake flask, and the Cyclotella seed solution in the logarithmic growth phase was obtained after 7 days;
[0049] (2) Inoculate 10% (v / v) inoculum into newly prepared medium I for heterotrophic culture in shake flasks, and the initial biomass concentration converted to dry weight is 0.2±0.01g / L. Place the Erlenmeyer flask in a dark temperature-controlled shaker, the shaker speed is 130rpm, the temperature is controlled at 27±1°C, and the culture time is 8 days (192 hours). Samples were collected on the 6th, 7th, and 8th day of heterotrophic culture, washed by centrifugation, freeze-dried, and the content of fucoxanthin in the algae powder was tested. Among them, the formula of medium Ⅰ is: 0.66g MgSO dissolved in 1L saline 4 ·7H 2 O, 0.3g urea, 50.5mg KH 2 PO 4 , 34mg H 3 ...
Embodiment 2~5
[0055] Embodiments 2 to 5 are basically the same as in Example 1, except that the glucose concentration in the medium I used in step 2 is different, and the glucose concentrations in Examples 2 to 5 are respectively 10g / L, 15g / L, and 20g / L and 25g / L.
[0056] Embodiment 1~5 culture effect analysis:
[0057] see image 3 , when Examples 1 to 5 carried out heterotrophic culture, it was found that the biomass increased with the prolongation of the culture time. After cultivating for 8 days, the biomass dry weight concentration of Examples 1 to 5 were respectively 1.12g / L and 1.71g / L , 1.96g / L, 1.77g / L and 1.76g / L, while Example 3 (that is, the concentration of glucose in medium I was 15g / L) had the highest biomass dry weight concentration, but in the early stage (cultivation time Figure 4 ), it can be seen that there is a negative correlation between the concentration of carbon source and the content of fucoxanthin, that is, the content of fucoxanthin decreases with the increas...
Embodiment 6
[0059] Cultivate Cyclotella as follows:
[0060] (1) The activated Cyclotella shake flask was cultured heterotrophically for 7 days to obtain the seed solution in the logarithmic growth phase;
[0061] (2) The fermenter was used as the culture vessel for batch culture, and a 5L glass fermenter was selected with a working volume of 3.5L. Inject the Chlorella seed liquid into the newly prepared medium Ⅰ according to the inoculum amount of 15% (v / v), the initial biomass concentration is converted to dry weight of 0.29g / L, control the dissolved oxygen saturation > 50%, and stir The speed is 130rpm, the pH is controlled at 7.5, the culture temperature is controlled at 27±0.5°C, and the fermentation period is 9 days (216 hours). Samples were taken every 24 hours to measure the dry weight. After 3 days of culture (>72h), samples were taken every 24 hours and freeze-dried, then the pigment was extracted, and the content of fucoxanthin was quantitatively determined by liquid chromatog...
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