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A method of extracting a microalga intracellular metabolite sample

A metabolite and microalgae technology, applied in the field of microalgae intracellular metabolite sample extraction, to achieve the effect of fewer operation steps

Inactive Publication Date: 2017-08-15
DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the centrifugation time at 4°C is only a few minutes, the cells still have some viability during the process

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] 1) Isochrysis zhangjiangensis was cultured in seawater supplemented with F / 2 medium, and the final concentration of the main component of the F / 2 nutrient salt used was NaNO 3 75mg / L, NaH 2 PO 4 2H 2 O 5mg / L, Vitamin B 12 0.5μg / L, Biotin 0.5μg / L, vitamin B 1 0.1mg / L, FeCl 3 ·6H 2 O 3.16mg / L, Na 2 EDTA·2H 2 O 4.36mg / L, CuSO 4 ·5H 2 O9.8μg / L, Na 2 MoO 4 2H 2 O 6.3 μg / L, ZnSO 4 ·7H 2 O 22 μg / L, CoCl 2 ·6H 2 O 12μg / L, MnCl 2 4H 2 O 0.18mg / L. They were cultured in a 500mL photobioreactor for six days, and 7mL samples were taken on days 3, 4, 5 and 6 (corresponding to biomass of 3.4, 5.5, 7.5 and 7.2 mg). One day in advance, take 7mL of seawater sterilized by filtration with a 0.22 micron membrane, put it into a 15mL centrifuge tube with a cover, place it horizontally in a -80°C refrigerator, and prepare an ice slope at 45° to the horizontal plane. After sampling, directly add to the 15mL centrifuge tube prepared with ice slope, close the lid, shake vigoro...

Embodiment 2

[0022] Embodiment 2: The same operation as in Example 1, except that the difference from Example 1 is that during the above operation, isotope-labeled lactic acid, malic acid and succinic acid with a final concentration of 0.125 μg / mL were added to the aqueous methanol solution, It is used for subsequent quantification of small organic molecules in the aqueous phase by chromatography-mass spectrometry.

Embodiment 3

[0023] Embodiment 3: After the operation process of the above-mentioned embodiment 1, the organic phase is taken out, and weighed after purging the solvent to a constant weight with N2 gas, to obtain the total fat-soluble substance amount determined by gravimetric method. Alternatively, at the same time, the organic phase can be diluted to 1 mg / mL (using acetonitrile:isopropanol solution with a volume ratio of 65:35), and the lipids in it can be determined by mass spectrometry.

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Abstract

A method of extracting a microalga intracellular metabolite sample is disclosed. The method includes rapidly deactivating intracellular enzymes by adopting an ice bath manner, performing ultrasonic disintegration with dichloromethane or chloroform, rapidly separating polar compounds, non-polar compounds, insoluble saccharides and protein, for subsequent metabonomical analysis, by adopting a dichloromethane (or chloroform)-methanol-water three-phase system. The method has characteristics of few operation steps, and capability of simultaneously measuring polar and non-polar intracellular components.

Description

technical field [0001] The invention relates to a method for extracting microalgae intracellular metabolite samples. Background technique [0002] Microalgae are autotrophic microorganisms, also known as microalgae plants because of the photosynthesis system in their cells. Microalgae have the ability to efficiently absorb solar energy and fix carbon dioxide to produce biomass (that is, the ability to photosynthesize); at the same time, as a kind of microorganism, microalgae proliferates faster. As single-celled organisms, microalgae are mainly composed of protein, fat, carbohydrates and nucleic acids. At present, microalgae has become an important breeding bait and widely used by farming enterprises. At the same time, microalgae has also become a source of health food such as carotenoids, proteins, and polysaccharides. As the prospect of fossil energy depletion intensifies, the potential application of microalgae in energy products and fine chemicals has also received gre...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N1/28
Inventor 薛松曹旭鹏
Owner DALIAN INST OF CHEM PHYSICS CHINESE ACAD OF SCI
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