Breeding method of high-concentration CO2-tolerant microalgae species

A technology of CO2 and microalgae, which is applied in the direction of microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of single-gene knockout mutants unable to grow normally, achieve increased carotenoid content, high efficiency, The effect of less work

Inactive Publication Date: 2016-07-20
SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The single-gene knockout mutant just constructed couldn't even grow normally

Method used

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  • Breeding method of high-concentration CO2-tolerant microalgae species
  • Breeding method of high-concentration CO2-tolerant microalgae species
  • Breeding method of high-concentration CO2-tolerant microalgae species

Examples

Experimental program
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Effect test

Embodiment 1

[0032] A strain of Chlorella (Chlorella sorokiniana) was isolated and screened from the natural environment of Chongming Island, Shanghai. According to the identification of molecular biology, the homologous similarity between the algae and the algae in the known database is 99%. The algal strain was placed in 10% CO 2 conditions for adaptive evolution. The culture medium is BG11 medium.

[0033] Adaptive evolution culture conditions: the initial inoculation density of microalgae is 0.02g / L, the temperature is 28°C; the apparent ventilation rate is 0.26cm / s; the light intensity is about 95μmol photons (photons) / m 2 / s, continuous light (that is, the light-dark cycle ratio is 24 hours / 0 hours), and the culture system is 300ml. In order to obtain enough Chlorella as algal species for the next evolution cycle, the culture time of the first cycle was set to 7 days, and then the evolution time of each cycle was set to 3 days. Three parallel experiments were set up for each grou...

Embodiment 2

[0044] Unevolved Chlorella in 20% CO 2 Conditions for adaptive evolution (other conditions are the same as in Example 1). Using 20% ​​CO 2 The evolutionary strains obtained after evolution under the conditions of 10% and 20% CO respectively 2 and 30% after 11 days of cultivation, its dry weight can reach 3.0g / L (such as Figure 5 shown). Compared with the dry cell weight of unevolved algal strains (such as figure 1 shown), at 10% and 20% CO 2 Conditioned growth was enhanced, with 85% and 141% increases in dry cell weight, respectively.

[0045] Alternatively, with 20% CO 2 The evolutionary strains obtained by adapting to evolution under the conditions of 10%, 20% and 30% CO 2 (volume ratio) condition and cultivated for 11 days, its carotenoid content all can reach more than 24mg / L (such as Figure 6 shown).

Embodiment 3

[0047] 1) After inoculating Chlamydomonas reinhardti in TAP medium with a cell density of 0.2g / L, culture it for 10 days under the following conditions as an adaptive evolution culture cycle;

[0048] Conditions: 40°C, 40% CO 2 and 500 μmol photonsm -2 the s -1 , The light-dark cycle is 16 hours / 8 hours, and the apparent ventilation rate is 1.0cm / s;

[0049] 2) Dilute the Chlamydomonas reinhardtii cultivated in step 1) to a cell density of 0.2 g / L, and perform a new cycle of cultivation for 10 days under the same conditions as in the above 1);

[0050] 3) Take step 1) as a culture cycle, enter the next culture cycle according to the method of step 2), repeat the above process for 50 culture cycles, so that the growth rate tends to be stable, so as to obtain CO tolerant 2 species of Chlamydomonas reinhardtii.

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Abstract

The invention discloses a breeding method of a high-concentration CO2-tolerant microalgae species. The breeding method comprises the following steps: 1) cultivating inoculated microalgae with the presence of less than 40% of CO2 and 20-500 [micro]moles of photons m-2s-1 at 15-40 DEG C under the conditions of a light-dark cycle of (1-24h)/(0-23h) and an apparent ventilation rate of 0.01-1.0cm/s for 2-10 days as an adaptive evolution culture cycle; 2) inoculating the microalgae which is cultivated in the step 1) once again, and cultivating by a new cycle under conditions same with that of the step 1) for 2-10 days; and 3) turning to a next culture cycle as methods prescribed within the step 2) by taking operations prescribed within the step 1) as a culture cycle, and repeating the process by 10-200 culture cycles, so that the CO2-tolerant microalgae species is obtained. The breeding method disclosed by the invention is applicable to the breeding of microalgae bio-fuel species; and meanwhile, the breeding method can also provide a basis for microalgae biological studies.

Description

technical field [0001] The invention relates to a method for breeding microalgae species, in particular to a method for tolerant high concentration CO 2 Breeding method of microalgae species. Background technique [0002] With the rapid development of my country's economy, energy demand is increasing year by year. In my country, energy is mainly based on fossil energy. According to the 2013 Statistical Bulletin on National Economic and Social Development, my country produced 209 million tons of crude oil and 3.68 billion tons of raw coal in 2013. At the same time, 282 million tons of crude oil and 327 million tons of coal were imported. The problem of greenhouse gas emissions in the use of fossil energy is inseparable from the climate anomalies in recent years. In 2009, my country formally promised that by 2020, China's carbon dioxide emissions per unit of GDP would be reduced by 40-45% compared with 2005, and this would be incorporated into the medium and long-term plan...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12R1/89
Inventor 李登进孙予罕赵权宇汪靓
Owner SHANGHAI ADVANCED RES INST CHINESE ACADEMY OF SCI
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