Method for improving micro algae growth rate and cell fat content employing high-salinity domestication

A technology of growth rate and oil content, applied in the directions of microorganism-based methods, methods of using microorganisms, biochemical equipment and methods, etc. The ability of cells to enrich oil, the effect of oil content increase

Inactive Publication Date: 2015-05-06
ZHEJIANG UNIV
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Problems solved by technology

Ben-Amotz et al. (Ben-Amotz et al., 1985) confirmed that some species of algae have high oil content under high salinity, and Cohen et al. (Cohen et al., 1988) also found that growth in high salinity The cells of Porphyridium Cruentum algae contain a higher p

Method used

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  • Method for improving micro algae growth rate and cell fat content employing high-salinity domestication

Examples

Experimental program
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Effect test

Embodiment 1

[0028] (1) According to the inoculum size, inoculate the Nitzschia liquid into a 1L Erlenmeyer flask (800mL culture medium), at a constant temperature of 20°C, with a light intensity of 3500lux, and under the conditions of 12h of light and 12h of darkness in turn every day, air is passed into it to amplify the culture 10 days.

[0029] (2) Get the algal species after the expanded culture and inoculate it into a 400mL cylindrical bottle with a capacity of 5% inoculum, the medium in the cylindrical bottle is 250mL, the initial salinity is 0%, feed the air of 70mL / min, at temperature 25°C, light intensity 8000lux, light-to-dark ratio 12h:12h, and initial pH value of 9. After culturing for 8 days, transfer it to a culture medium with a salinity of 0.2% at 5-10% for 8 days. sky.

[0030] The medium consists of: 0.15g NaHCO 3 , 0.02g KH 2 PO 4 , 0.027g VB 1 , 1.5×10 -6 f 12 , 0.2g Na 2 SiO 3 9H 2 O, 1.0g NaNO 3 , 0.0005g Biotin, 1mL trace elements and 980mL artificial sea...

Embodiment 2

[0034] (1) Inoculate the Chlorella liquid into a 1L Erlenmeyer flask (800mL culture medium) according to the inoculum size of 8%, and pass air into it to amplify under the conditions of constant temperature 23°C, light intensity 3750lux, and 12h of light and 12h of darkness every day. Cultured for 12 days.

[0035] (2) Get the algal species after the enlarged culture and inoculate it into a capacity of 400mL cylindrical bottle with 8% inoculum, the medium in the cylindrical bottle is 300mL, the initial salinity is 0%, feed the air of 80mL / min, at temperature After culturing for 10 days under the conditions of 25°C, light intensity 8000lux, light-dark ratio 12h:12h, and initial pH value of 8, it was transferred to a medium with a salinity of 0.3% for 10 days at an inoculum size of 8%.

[0036] The medium consists of: 0.15g NaHCO 3 , 0.02g KH 2 PO 4 , 0.027g VB 1 , 1.5×10 -6 f 12 , 0.2g Na 2 SiO 3 9H 2 O, 1.0g NaNO 3 , 0.0005g Biotin, 1mL trace elements and 980mL artif...

Embodiment 3

[0040] (1) Inoculate the Nannochloropsis liquid into a 1L Erlenmeyer flask (800mL culture medium) according to the inoculum size of 10%, and pass air under the conditions of a constant temperature of 25°C, a light intensity of 4000 lux, and 12 hours of light and 12 hours of darkness every day. Cultured for 15 days.

[0041] (2) Get the algal species after the expanded culture and inoculate it into a 400mL cylindrical bottle with a capacity of 10% inoculum, the medium in the cylindrical bottle is 350mL, the initial salinity is 0%, feed the air of 90mL / min, at temperature After 12 days of culture at 29°C, light intensity of 9000lux, light-to-dark ratio of 12h:12h, and initial pH value of 10, it was transferred to a medium with a salinity of 0.4% for 12 days at a 10% inoculum size.

[0042] The medium consists of: 0.15g NaHCO 3 , 0.02g KH 2 PO 4 , 0.027g VB 1 , 1.5×10 -6 f 12 , 0.2g Na 2 SiO 3 9H 2 O, 1.0g NaNO 3, 0.0005g Biotin, 1mL trace elements and 980mL artificial ...

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Abstract

The invention relates to a biomass energy utilization technology, and aims at providing a method for improving micro-algae growth rate and cell fat content employing high-salinity domestication. The method comprises the following steps: inoculating a micro algae liquid into a culture medium of a triangular flask and carrying out enlarge cultivation; and carrying out gradient increase according to 0.3% salinity until inoculating algae species into the culture medium of which the initial salinity is 2.9%-3.1%, and continuously cultivating for 3-5 generations, so as to obtain the high-fat content algae species capable of rapidly growing. By virtue of a high-salinity gradient domestication method, a carbon-nitrogen metabolism pathway of micro-algae cells and the synthesis pathway are reconstructed; the metabolic pathway change of proline, ABC transportprotein, proteasome and glutathione is relatively large; and the key biological enzyme expression quantities of acetyl-coa carboxylase (ACCase), beta-ketoacyl-ACP synthase I and II and the like are regulated upwards, so that the micro algae growth rate and the cell oil-enrichment capacity are effectively improved.

Description

technical field [0001] The invention relates to biomass energy utilization technology, in particular to a method for increasing microalgae growth rate and cell oil content by acclimating to high salinity. Background technique [0002] The use of marine microalgae to absorb solar energy to produce clean and renewable liquid fuels such as biodiesel is of great significance for solving the crisis and contradictions of the increasing shortage of fossil energy such as petroleum and serious combustion pollution. Microalgae grow very rapidly, have powerful physiological and metabolic functions, and have high photosynthesis efficiency. They use seawater resources and absorb a large amount of CO 2 Carry out photosynthesis and synthesize enriched oil through green and clean biochemical processes (Sheehan et al., 1998; Huntley et al., 2007), which can be converted into high-grade biodiesel with a calorific value of up to 41MJ / kg, so it can replace Petrochemical diesel is used in vehic...

Claims

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Application Information

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IPC IPC(8): C12N1/36C12N1/12C12N1/13C12P7/64C12R1/89
CPCC12N1/36C12P7/649C12N1/125C12R2001/89Y02E50/10
Inventor 程军岑可法刘建忠周俊虎张彦威周志军王智化黄镇宇杨卫娟
Owner ZHEJIANG UNIV
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