Extracellular polysaccharide complex of Chlorella protothecoides with immunity regulation activity, and preparation method and application thereof

An immunomodulatory activity and extracellular polysaccharide technology, which is applied in the directions of organic active ingredients, medical preparations containing active ingredients, and applications, and can solve the application of less researched and no Chlorella protothecoides exopolysaccharides and their complexes. Value and other issues, to achieve the effect of simple preparation, reduction of inflammatory factors ROS, and cost reduction

Active Publication Date: 2020-02-04
XIANGTAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Chlorella contains polysaccharides and their complexes that are rich in anti-tumor activity, anti-pathogenic bacteria, anti-viral infection and enhancing immunity, but there are few studies on the polysaccharides of Chlorella protothecoides, only Mario et al. Analyzed its cell wall polysaccharide composition (Arch.Mikrobiol.,1973,92,227-233), Qiao Dairong et al. optimized the extraction process of its intracellular polysaccharide (Chin.J.Appl.Environ.Biol.,2014,20(4):615 -620), so far there is no relevant report on the application value of Chlorella protothecoides exopolysaccharide and its complex

Method used

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  • Extracellular polysaccharide complex of Chlorella protothecoides with immunity regulation activity, and preparation method and application thereof
  • Extracellular polysaccharide complex of Chlorella protothecoides with immunity regulation activity, and preparation method and application thereof
  • Extracellular polysaccharide complex of Chlorella protothecoides with immunity regulation activity, and preparation method and application thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Preparation of Chlorella exopolysaccharide complex with immunomodulatory activity

[0039] (1) Inoculate Chlorella protothecoides CS-41 on Basal solid medium, and culture it under 28°C and 2000Lux light conditions for 5 days to rejuvenate the algae; pick the rejuvenated Chlorella and transfer it to common In the Basal liquid culture medium, 2000Lux (12h light: 12h darkness is carried out alternately), 28 ℃, 160rpm concurrent culture 5 days, obtains first-class seed liquid; 5% first-class seed liquid is inoculated in the fresh liquid medium, isotropic Obtain secondary seed liquid after cultivating for 5 days;

[0040] (2) Get 1% secondary seed liquid to expand and cultivate in the Basal liquid medium that contains 10‰NaCl salinity (about 2.784g / L), the light intensity is 2000Lux, and the light cycle is 8h light: 16h dark (light- Darkness is carried out alternately) under the condition of 28 ℃, 160rpm concurrent culture culture 7 days, obtain the chlorella fermented liquid...

Embodiment 2

[0045] Composition, Purity and Molecular Weight Determination of Chlorella exopolysaccharide complexes with immunomodulatory activity

[0046] (1) Determination of sugar content

[0047] Take 0, 0.05mL, 0.1mL, 0.15mL, 0.2mL and 0.25mL glucose standard solution (100μg / mL) respectively, make up to 0.25mL with distilled water, and add 1mL of anthrone (mass percentage: 0.2%)-sulfuric acid solution, mix well; after boiling water bath for 10 minutes, quickly stop the reaction in ice bath, and then equilibrate to room temperature. Detect its absorbance value at 620nm by a spectrophotometer, and take the glucose concentration as the abscissa, and use the OD 620 As the ordinate, make a standard curve. At the same time, take 2 μL of the aqueous solution (10 mg / mL) of the chlorella exopolysaccharide complex and make up to 0.25 mL with distilled water. The other steps are the same as above, and the sugar content is calculated according to the standard curve.

[0048] (2) Determination ...

Embodiment 3

[0053] Monosaccharide composition and fatty acid composition analysis of exopolysaccharide complexes of Chlorella algae with immunomodulatory activity

[0054] (1) Monosaccharide composition

[0055] Weigh 20mg of dried chlorella exopolysaccharide CPEPS-2, add 10mL trifluoroacetic acid (2mol / L), hydrolyze at 110°C for 6h; add 2mL methanol after drying the hydrolyzate, evaporate to dryness under reduced pressure, repeat 3-4 times , to remove trifluoroacetic acid; then the sample was mixed with 10 mg of hydroxylamine hydrochloride and 0.6 mL of pyridine, and reacted in a water bath at 90 ° C for 30 min; after cooling, 0.5 mL of acetic anhydride was added to continue the reaction at 90 ° C for 30 min; the reaction product was dried and dissolved in 1 mL of chloroform. GC-MS analysis. Chromatographic conditions: HP-5 chromatographic column, injection volume: 2 μL, inlet temperature: 250°C, flow rate 1mL / min; heating program: keep at 110°C for 2 minutes, increase temperature from ...

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Abstract

The invention discloses an extracellular polysaccharide complex of Chlorella protothecoides with immunity regulation activity as well as a preparation method and application thereof. The extracellularpolysaccharide complex of the Chlorella protothecoides disclosed by the invention is extracted from Chlorella protothecoides CS-41 fermentation broth, and is specifically prepared by the following steps: carrying out filtration, centrifuging, concentration and ethanol precipitation, and removing proteins by adopting a Sevage method; and then, carrying out separation and purification procedures, including anion exchange chromatography, gel exclusion chromatography and the like, so as to obtain the extracellular polysaccharide complex of the Chlorella protothecoides. The prepared extracellularpolysaccharide complex of the Chlorella protothecoides is white powder of which the average relative molecular weight is 170 kDa; and the extracellular polysaccharide complex of the Chlorella protothecoides is composed of lipid and polysaccharides, wherein the lipid contains palmitic acid and stearic acid, and the polysaccharides are composed of rhamnose, xylose, mannose and glucose. The extracellular polysaccharide complex of the Chlorella protothecoides disclosed by the invention is clear in compositions, and has the effect of immunity regulation; and moreover, the extracellular polysaccharide complex is capable of significantly reducing levels of inflammatory factors (ROS, iNOS, IL-6 and TNF-alpha). Thus, the extracellular polysaccharide complex of the Chlorella protothecoides has broadprospects of application in clinical as well as related medical and health-caring fields.

Description

technical field [0001] The invention relates to the field of functional health food or biomedicine, in particular to a chlorella exopolysaccharide complex with immunoregulatory activity and its preparation method and application. Background technique [0002] Microalgal biodiesel has become the third-generation sustainable biofuel recognized all over the world, and its development and application are of great significance to solve the current shortage of fossil fuels and air pollution. However, the microalgae energy industry is still facing the dilemma of high cost, and it is still in its infancy, making commercial application difficult. One of the effective solutions to reduce the cost of microalgae biodiesel is to increase the oil accumulation rate in algae cells and increase oil production. [0003] Focusing on the characteristics of algae cell growth and oil accumulation at home and abroad, by changing environmental conditions such as temperature, light, nutritional con...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/715A61P37/04A61P29/00A23L33/125A61K36/05
CPCA61K31/715A61P37/04A61P29/00A23L33/125A61K2236/19A61K36/05A23V2002/00A23V2200/324A23V2200/30A23V2250/51
Inventor 李玉芹雷铮宇周蓉唐裕芳祝佳惠李超贾淑婷
Owner XIANGTAN UNIV
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