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Method for analyzing microalgae metabolome

A technology of metabolome and microalgae, applied in the direction of analysis materials, material separation, measurement devices, etc., can solve problems such as blanks and difficulties in microalgae wall breaking, and achieve the effect of promoting metabolism research

Active Publication Date: 2012-06-13
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] However, the oil production of microalgae is still in its infancy, and there is still a lot of technical knowledge to be accumulated before it can be adapted to large-scale industrial production. Research on the metabolic mechanism of microalgae cultivation is one of the key directions. It is difficult to break the wall, and the metabolomics research methods related to microalgae are almost blank

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  • Method for analyzing microalgae metabolome
  • Method for analyzing microalgae metabolome
  • Method for analyzing microalgae metabolome

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] A method for analyzing the metabolome of microalgae, comprising the steps of:

[0036] (1) Chlorella sorokiniana cell collection and quenching:

[0037] Cultivate Chlorella sorokiniana. When the culture reaches OD560 of 0.8, take 5 samples of algae liquid, 100 mL each, centrifuge at 3000 rpm at 4°C for 3 minutes, collect the cells in the lower layer, and use 4 mL of metabolic stop solution in - Quench at 40°C for 5 minutes to terminate the metabolic reaction, and freeze-dry at -80°C for 4 hours to obtain freeze-dried cells;

[0038] The metabolic termination solution contains 1500 mg·L -1 NaNO 3 , 36mg·L -1 CaCl 2 2H 2 O, 75mg·L -1 MgSO 4 ·7H 2 O and 40mg·L -1 K 2 HPO 4 ·3H 2 O in methanol aqueous solution, the volume ratio of methanol and water in the methanol aqueous solution is 1:2;

[0039] (2) Extract intracellular metabolites:

[0040] Take 5 parts of freeze-dried cells obtained in step (1), 15 mg each, and place them in centrifuge tubes, add 1 mL-40...

Embodiment 2

[0065] A method for analyzing the metabolome of microalgae, comprising the steps of:

[0066] (1) Chlorella sorokiniana cell collection and quenching:

[0067] Cultivate Chlorella sorokiniana. When the culture reaches OD560 of 1.0, take out 4 samples of algae liquid, 120 mL each, centrifuge at 5000 rpm at 4°C for 3 minutes, collect the cells in the lower layer, and use 3 mL of metabolic stop solution in - Quench at 40°C for 7 minutes to terminate the metabolic reaction, and freeze-dry at -80°C for 6 hours to obtain freeze-dried cells;

[0068] Metabolism termination liquid is the same as embodiment 1;

[0069] (2) Extract intracellular metabolites:

[0070] Take 4 parts of freeze-dried cells obtained in step (1), put 25 mg of each into centrifuge tubes, add 2 mL of metabolic extraction solution at -40 °C to each tube, mix well, cover tightly and put into liquid nitrogen, and freeze and thaw repeatedly for 5 The second time, centrifuge at 6000rpm at -20°C for 1min, collect t...

Embodiment 3

[0080] A method for analyzing the metabolome of microalgae, comprising the steps of:

[0081] (1) Chlorella sorokiniana cell collection and quenching:

[0082] Cultivate Chlorella sorokiniana. When the OD560 reaches 1.5, take 3 samples of algae liquid, each 150 mL, centrifuge at 3000 rpm at 4°C for 4 min, collect the cells in the lower layer, and use 5 mL of metabolic stop solution in the Quench at -40°C for 10 minutes to terminate the metabolic reaction, and freeze-dry at -80°C for 6 hours to obtain freeze-dried cells;

[0083] Metabolism termination liquid is the same as embodiment 1;

[0084] (2) Extract intracellular metabolites:

[0085] Take 3 parts of freeze-dried cells obtained in step (1), 20 mg each, and place them in centrifuge tubes, add 2 mL-40 °C metabolic extract to each tube, mix well, cover tightly and put into liquid nitrogen, freeze-thaw repeatedly for 5 The second time, centrifuge at 6000rpm at -20°C for 2min, collect the supernatant, add 1mL of metaboli...

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Abstract

The invention discloses a method for analyzing a microalgae metabolome, comprising the following steps: collecting and quenching microalgae cells, extracting metabolites in the cells, carrying out derivatization on the metabolites, and finally analyzing a sample through GC-MS to obtain a metabolome result. According to the invention, the method relates to the termination of metabolic reaction, the extraction of the metabolites, and the analysis of the metabolites, thus qualitative and quantitative results of the microalgae metabolites under different cultural conditions are acquired. The method has important meanings for promoting the research of microalgae metabolism.

Description

technical field [0001] The invention belongs to the field of biofuels and relates to a method for analyzing microalgae metabolites. Background technique [0002] With the continuous development of human society, the demand for energy is increasing day by day, but the current reserve of fossil energy is limited. With the consumption of fossil energy, the price of oil continues to rise, and international relations are becoming increasingly tense. In addition, the use of fossil energy causes carbon dioxide It is the main source of the greenhouse effect. Therefore, the development of sustainable green new energy has become a hot spot of concern in various countries. [0003] At present, the commonly used bioenergy is mainly bioethanol and biodiesel, among which bioethanol mainly comes from cellulose resources in agricultural waste, but the use of bioethanol needs to redesign the distribution infrastructure and engine, which is poorly compatible with existing technologies, and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N30/02G01N30/06
Inventor 元英进陆姝欢丁明珠牛艳红
Owner TIANJIN UNIV
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