A strain of Xanthophyllum and its cultivation and application
A technology of yellow hair algae and algae strains, applied in the direction of single-cell algae, microorganisms, microorganisms, etc., can solve the problems of slow growth, few research records, slow oil accumulation of yellow hair algae, etc., and achieve the effect of fast growth
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Embodiment 1
[0033] Example 1 Screening and molecular identification of Xanthophyllum ENN177
[0034] 1. Screening of Xanthophylla ENN177 (Tribonema sp.)
[0035] Soil samples were taken in the northeast of Inner Mongolia in autumn. In the laboratory, BG11 medium diluted 1 / 10 was added to the water samples, enriched and cultured for 2-3 days under natural light and room temperature, and the samples were treated with sterile deionized water. Dilute, take a single algae silk and carry out purification culture in a 24-well plate, the medium is BG11 medium, the culture temperature is 25°C, and the light is natural light. The purified algae filaments in the orifice plate were transferred to the Erlenmeyer flask for expansion culture, and finally the pure algae strain of ENN177 was obtained.
[0036] Table 1BG11 medium composition
[0037] Substance name Dosage per liter of deionized water NaNO 3 (sodium nitrate)
1.5g K 2 HPO 4 ·3H 2 O(dipotassium hydrogen phosp...
Embodiment 2
[0060] Indoor cultivation of embodiment 2ENN177 algal strain
[0061] In a column reactor with an inner diameter of 5cm and a height of 60cm, use sterile BG11 medium for cultivation, the cultivation temperature is 25°C, the initial inoculum concentration is about 1g / L, and the mixture of filtered air and carbon dioxide is introduced. Gas, wherein the content of carbon dioxide is 1-2% of the total amount of mixed gas. The initial light intensity is 120umol / m 2 / s, according to the growth of biomass in the later stage, the light intensity is increased to 200umol / m 2 / s. Samples were taken every day to determine the biomass and fatty acid content in the cells, and the cells were cultured for 4 days in total.
[0062] Biomass measurement method: Accurately measure 50mL of algae liquid, filter it with a glass fiber membrane (pore size of 0.45um) with a known initial weight after drying, rinse with 150mL of deionized water three times to remove ash, and then remove the algae cell...
Embodiment 3
[0064] The oil production of embodiment 3ENN177 strain
[0065] Insert the ENN177 algae at the end of the exponential growth stage into a column reactor with an inner diameter of 5 cm and a height of 60 cm. The initial inoculation amount of algal cells is about 1.5 g / L. The medium used is BG11 medium lacking nitrogen and phosphorus. The temperature is 25°C, and the initial light intensity is 150umol / m 2 / s, the subsequent light intensity is gradually increased to 200umol / m 2 / s, other parameters are the same as in Embodiment 2. Samples were taken every day to measure the fatty acid content in the cells, and the biomass was measured every two days for a total of 3 days of culture. The results were as follows: Figure 4 shown.
[0066] Fatty acid composition and content determination method: weigh 25mg of freeze-dried sample, add 2mL 2% H 2 SO 4 Anhydrous methanol-toluene solution. After filling with nitrogen, heat at 80°C for 2 hours. Afterwards, 1 mL of pure water and n...
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