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Method of increasing the biomass and DHA productivity of isochrysis galbana parke

A technology of isochrysis globosa and biomass, applied in the field of algae, can solve the problems of inability to use algae oil, insufficient DHA content, unsatisfactory biomass and DHA yield, etc., so as to reduce the cost of producing DHA and improve the yield of DHA. Effect

Active Publication Date: 2018-12-18
杭州园泰生物科技有限公司
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Problems solved by technology

[0004] Isochrysis globosa is a single-celled microalgae belonging to the order Isodinoflagellates, Isoflagellates, and Isoflagellates. It is photoautotrophic, with simple culture methods, low cost, and fast proliferation. Rich in unsaturated fatty acids, but the DHA content is not high enough, only about 10%, and cannot be used as high-quality algae oil
Existing technology has carried out many researches on DHA produced by algae. Document 1 "Xiao Shang, Effect of Compound Carbon Sources on DHA Accumulation in Cryptidinosa, Journal of Henan Normal University (Natural Science Edition) 2013" mainly studies single carbon The effects of different carbon sources and compound carbon sources on the accumulation of DHA in Cryptidinosa show that glucose is the best single carbon source in batch fermentation, glucose and glycerol are the best compound carbon sources, and the DHA yield of compound carbon sources is higher than that of single glucose carbon sources More than 20%, when fed batch fermentation for many times, glucose and glycerol are the best single carbon source, the best composite carbon source is the composite carbon source of glucose and sucrose, and the final DHA yield is higher than that of single carbon source. Batch fermentation is 23.6% higher; however, the production process is still in the stage of laboratory culture bottles, and it needs industrialized large-scale production to test its specific effect
Document 2 "Wang Jufang, Journal of Zhanjiang Ocean University, 2001, Effects of Several Inorganic Salts on Crypthecodinium cohnii ATCC30556 and DHA Production". Show that: Cryptidoflagellate can grow in the culture medium that NaCl is the sole inorganic salt; The mass concentration of NaCl in the medium is 6g / L, and this moment Cryptidoflagellate has maximum biomass and DHA production; But biomass and DHA production are both It is not ideal enough and needs to be further improved to realize industrialized production

Method used

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  • Method of increasing the biomass and DHA productivity of isochrysis galbana parke
  • Method of increasing the biomass and DHA productivity of isochrysis galbana parke
  • Method of increasing the biomass and DHA productivity of isochrysis galbana parke

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Embodiment 1

[0031] The method for improving Isochrysis globosa biomass and DHA output, it comprises the steps:

[0032] Inoculate the Isochrysis globosa that has been cultivated to the logarithmic growth phase into the seed tank containing the seed medium, and the initial inoculation density is 1 × 10 5 seeds / ml, light intensity 5000lux, culture at 22°C, light-to-dark ratio 18:6, ventilation rate 0.2vvm, culture for 48 hours, and collect seed solution;

[0033] The components of the seed culture medium are: sodium nitrate 1.5g / L, sodium silicate 0.5g / L, sodium chloride 0.3g / L, ammonium sulfate 0.2g / L, potassium dihydrogen phosphate 0.1g / L, VB 1 0.5mg / L, VB 12 0.1mg / L;

[0034] Inoculate the seed liquid into the reaction tank containing the fermentation medium according to the inoculum amount of 3%, and ferment and cultivate at 22°C with an aeration rate of 0.5vvm; when the culture reaches 48 hours, add arachidonic acid and gallic acid propionate to the fermentation medium Esters, so th...

Embodiment 2

[0037] The method for improving Isochrysis globosa biomass and DHA output, it comprises the steps:

[0038] Inoculate the Isochrysis globosa that has been cultivated to the logarithmic growth phase into the seed tank containing the seed medium, and the initial inoculation density is 2×10 5 seeds / ml, light intensity 5000lux, culture at 25°C, light-to-dark ratio 18:6, ventilation rate 0.3vvm, culture for 48 hours, and collect seed solution;

[0039] The components of the seed culture medium are: sodium nitrate 1.5g / L, sodium silicate 0.5g / L, sodium chloride 0.3g / L, ammonium sulfate 0.2g / L, potassium dihydrogen phosphate 0.1g / L, VB 1 0.5mg / L, VB 12 0.1mg / L;

[0040] Inoculate the seed solution into the reaction tank containing the fermentation medium according to the inoculum amount of 5%, and ferment and cultivate at 25°C with an aeration rate of 0.5-1vvm; when the culture reaches 48 hours, add arachidonic acid and gallic acid to the fermentation medium Propyl ester, contro...

Embodiment 3

[0043] Determination of algae cell density and DHA content:

[0044] Determination of algae cell density: use the OD method to measure the absorbance in the culture system at 680nm, and use the formula "cell density (×10 4 cells / mL)=(OD680×1250-90.125)×dilution factor” to calculate the corresponding cell density and draw the growth curve.

[0045] Lipid composition analysis:

[0046] Take the algae solution, centrifuge at 4000r / min for 8min, wash the precipitate with distilled water for 3 times, dry it in vacuum at 50°C, crush the algae cells, and then add it to the mixed solution of chloroform and methanol (the volume ratio of chloroform and methanol is 2:1). For 1g powder: 3ml chloroform-methanol mixed solution, microwave extraction, microwave power 150W, extraction time 60min, extraction temperature 50°C, and then ultrasonic extraction, extraction temperature 60°C, ultrasonic power 300W, extraction time 60min, Then centrifuge, collect the chloroform phase, place it in nit...

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Abstract

The invention belongs to the technical field of algae and discloses a method of increasing the biomass and DHA productivity of isochrysis galbana parke. The method includes: S1) inoculating the isochrysis galbana parke, which has been cultured to logarithmic phase, on a seed pot filled with a seed culture medium, and culturing the isochrysis galbana parke for 48 h and collecting a seed liquid; S2)inoculating the seed liquid on a reaction tank filled with a fermenting culture liquid, performing fermenting culture at 22-25 DEG C, when the cultivation reaches 48 h, adding arachidonic acid and propyl gallate to the fermenting culture liquid and continuously culturing the seed liquid for 72-96 h through fermentation, and collecting algae cells. The method can increase the biomass and DHA productivity of the isochrysis galbana parke.

Description

technical field [0001] The invention belongs to the technical field of algae, and in particular relates to a method for increasing the biomass of dinoflagellates and the production of DHA. Background technique [0002] DHA, docosahexaenoic acid, commonly known as brain gold, is an unsaturated fatty acid that is very important to the human body and is an important member of the omega-3 unsaturated fatty acid family. Animals and humans cannot synthesize DHA by themselves and must be ingested from the outside world. DHA has important physiological functions: 1) DHA is a main component of the growth and maintenance of nervous system cells, and an important component of the brain and retina. Its content in the human cerebral cortex is as high as 20%, and it accounts for the largest proportion in the eye retina. , accounting for about 50%. Therefore, it is very important for the intelligence and vision development of fetuses and babies. For a long time, most people have supplemen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12P7/64C12R1/89
CPCC12N1/12C12P7/6427Y02E50/30
Inventor 应金良应金元夏瀚吴善青
Owner 杭州园泰生物科技有限公司
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