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Method for improving microalgae cultivation condition by using metabonomics so as to improve oil-producing capacity

A technology of microalgae culture and metabolomics, which can be used in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., and can solve problems such as unclear metabolic mechanisms.

Active Publication Date: 2014-04-09
TIANJIN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The second-generation bioenergy is mainly ethanol produced from waste cellulose resources such as straw. The source of cellulose is extensive and abundant, but a large amount of toxic inhibitory substances produced during the pretreatment process is a major limiting factor.
[0005] At present, a large number of laboratory cultures and pilot tests have been used in the field of microalgae to produce diesel oil. Existing studies have shown that under the conditions of non-autotrophic, nitrogen deficiency or high density, it is helpful for the accumulation of intracellular oil in microalgae, but for The underlying metabolic mechanisms are still poorly understood, and this is a key limiting factor for the large-scale industrial use of microalgae for biodiesel production.

Method used

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  • Method for improving microalgae cultivation condition by using metabonomics so as to improve oil-producing capacity
  • Method for improving microalgae cultivation condition by using metabonomics so as to improve oil-producing capacity
  • Method for improving microalgae cultivation condition by using metabonomics so as to improve oil-producing capacity

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Experimental program
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Effect test

Embodiment 1

[0081] A method utilizing metabolomics to improve microalgae culture conditions to increase oil production capacity of microalgae, comprising the steps of:

[0082] (1) Determination of intracellular metabolites in Chlorella sorokiniana:

[0083] ① Chlorella sorokiniana cell collection and quenching:

[0084] The microalgae were cultured at different inoculation densities, and the initial inoculation density was set to 1×10 4 , 1×10 5 , 1×10 6 and 1×10 7 cells mL -1 , when the cells were cultured to the stationary stage, 5 samples of algae liquid were taken out for each inoculation density, 100 mL each, centrifuged at 3000 rpm at 4 °C for 3 min, the cells in the lower layer were collected, and quenched with 4 mL of metabolic termination solution at -40 °C for 5 minutes, terminate the metabolic reaction, and freeze-dry at -80°C for 4 hours to obtain freeze-dried cells;

[0085] The metabolic termination solution contains 1500 mg·L -1 NaNO 3 , 36mg·L -1 CaCl 2 2H 2 O,...

Embodiment 2

[0149] A method utilizing metabolomics to improve microalgae culture conditions to increase oil production capacity of microalgae, comprising the steps of:

[0150] (1) Determination of intracellular metabolites in Chlorella sorokiniana:

[0151] ① Chlorella sorokiniana cell collection and quenching:

[0152] The microalgae were cultured at different inoculation densities, and the initial inoculation density was set to 1×10 4 , 1×10 5 , 1×10 6 and 1×10 7 cells mL -1 , when the cells are cultured to the late exponential and stable phases, take 4 algae liquid samples at each time point in each inoculation density, each 120mL, centrifuge at 5000rpm at 4°C for 3min, collect the cells in the lower layer, and use 3mL of metabolic stop solution 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;

[0153] Metabolism termination liquid is the same as embodiment 1;

[0154] ②Extract intracellular met...

Embodiment 3

[0187] A method utilizing metabolomics to improve microalgae culture conditions to increase oil production capacity of microalgae, comprising the steps of:

[0188] (1) Determination of intracellular metabolites in Chlorella sorokiniana:

[0189] ① Chlorella sorokiniana cell collection and quenching:

[0190] The microalgae were cultured at different inoculation densities, and the initial inoculation density was set to 1×10 4 , 1×10 5 , 1×10 6 and 1×10 7 cells mL -1 , when the cells are cultured to the mid-exponential, late-exponential and stable phases, take 3 algae liquid samples at each time point under each inoculation density culture, each 150mL, centrifuge at 3000rpm at 4°C for 4min, and collect the cells in the lower layer. Use 5mL of metabolic termination solution to 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;

[0191] Metabolism termination liquid is the same as embodiment ...

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Abstract

The invention discloses a method for improving a microalgae cultivation condition by using metabonomics so as to improve the oil-producing capacity. The method comprises the following steps of: (1) detecting intracellular metabolites in microalgae; (2) analyzing the oil-producing capacity of the microalgae; (3) performing partial least squares (PLS) analysis; and (4) process analysis. The oil-producing process of the microalgae, an intracellular metabolism change rule and important metabolites in the oil-producing process can be researched wholly by the method, and the change rule of the metabolism levels lays a base for the internal mechanism of the oil-producing process, so a theoretical base is laid for further optimizing a microalgae cultivation process and improving the grease yield. A new concept and a new method are provided for the research on other oil-producing microorganism cultivation processes.

Description

field of invention [0001] The invention belongs to the field of biofuels, and relates to a method for improving microalgae culture conditions by using metabolomics to increase oil production capacity. Background technique [0002] With the depletion of fossil energy and people's increasing attention to environmental protection, the development of sustainable green new energy has been put on the agenda and attracted more and more attention. The earliest bioenergy was derived from food crops, but due to land competition with grain, it caused a serious food crisis, and now it is gradually replaced by the second generation of bioenergy. The second-generation bioenergy is mainly ethanol produced from waste cellulose resources such as straw. The source of cellulose is extensive and abundant, but a large amount of toxic inhibitory substances produced during the pretreatment process is a major limiting factor. And ethanol is more corrosive and less potent than gasoline or diesel. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P7/64C12R1/89
Inventor 元英进陆姝欢杨洁牛艳红
Owner TIANJIN UNIV
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