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Method for culturing chlorella with high-density and high-quality

A technology of chlorella and common chlorella, applied in the biological field, can solve the problems of easy contamination of miscellaneous bacteria, difficulty in amplification, inconvenient cleaning, etc., and achieve the effect of high-density and high-quality cultivation

Active Publication Date: 2006-09-27
JIAXING ZEYUAN BIOLOGICAL PROD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, from the perspective of large-scale industrial production, the cultivation system and cultivation process established by Ogbonna et al. have the following three problems that are difficult to solve: 1) When the U-shaped tubular photobioreactor is large in scale, it is fragile, inconvenient to clean, Oxygen decomposition is slow and other problems, it is difficult to enlarge; 2) In the process of heterotrophic-photoautotrophic series culture Aseptic operation cannot be achieved during cultivation; 3) The continuous cultivation process is very easy to contaminate miscellaneous bacteria, which is basically not used in industrial production, and more batch-fed cultivation techniques are adopted
[0005] The present inventor firstly carried out exploratory experiments on Chlorella vulgaris using the culture medium and culture method used by Ogbonna et al. The results showed that the cell density of Chlorella vulgaris was only 4.68g / L at the end of the cultivation, and the algae protein and chlorophyll were only 4.68g / L respectively. 23.3% and 1.2%, cannot achieve high-density and high-quality cultivation of Chlorella vulgaris
This shows that the culture medium and culture method used by Ogbonna et al. cannot be generally used for high-density and high-quality culture of other species of Chlorella.

Method used

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  • Method for culturing chlorella with high-density and high-quality
  • Method for culturing chlorella with high-density and high-quality
  • Method for culturing chlorella with high-density and high-quality

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Add the following heterotrophic culture medium into a 5L bioreactor, add tap water to 3.4L and then sterilize, then insert Chlorella vulgaris into 6% of the working volume when the temperature drops to 30°C, and start heterotrophic culture . Heterotrophic culture conditions: the temperature is 30°C, the rotation speed is gradually adjusted from 200r / min at the time of inoculation to 400r / min at the end of the culture, the air flow rate is 1vvm, the pH is less than 8.5, and the dissolved oxygen is controlled above 15%. 30h after inoculation, feed solution (20g / L glucose and 3.33g / LKNO 3 mixed solution), and then added once every 5 to 8 hours, for a total of 4 times. After culturing for 56.34h, the dry cell weight (□) reached 47.91g / L, and the algal protein (□) and chlorophyll (□) contents were 33.42% and 15.65mg / gDcw respectively (see figure 1 ), the heterotrophic culture period ended.

[0064] KNO 3 : 9.25 Glucose: 30

[0065] K H 2 PO 4 : 0.7 Na 2 HPO 4 12H2 O...

Embodiment 2

[0083] Add the culture medium adopted by heterotroph in Example 1 into a 5L bioreactor, and adopt the heterotrophic culture conditions and culture techniques used in Example 1 to cultivate Chlorella vulgaris. After culturing for 60.08 hours, the dry cell weight (□) reached 51.15g / L, and the contents of algae protein (○) and chlorophyll (□) were 33.65% and 12.80mg / gDcw respectively (see figure 2 ), the heterotrophic culture period ended.

[0084] Use the following dilutions of the medium used, where KNO 3 The contents are 2.60g / L, 4.95g / L and 7.36g / L respectively, and the high-density algal liquid obtained from heterotrophic culture is diluted to 11.45g / L (A-10) and 21.89g / L (A-20) respectively and 32.56g / L (A-30) were transferred to a 1L flat photobioreactor for photoautotrophic culture. The photoautotrophic culture conditions were the same as in Example 1. Continue to cultivate until 87.08h, when the initial dry cell weight is 11.45g / L after transferring to photoautotroph...

Embodiment 3

[0094] Add the medium used in heterotrophy in Example 1 to a 50L bioreactor, add tap water to 34L and then sterilize it, then insert Chlorella vulgaris into 6% of the working volume when the temperature drops to 30°C, and start heterotrophy to cultivate. The heterotrophic culture conditions and culture techniques used in Example 1 were used to cultivate Chlorella vulgaris, wherein the feeding times were increased to 5 times. Cultivate to 58.20h dry cell weight reaches 54.52g / L, algal body protein and chlorophyll content are respectively 32.01% and 12.70mg / gDcw (see image 3 ), the heterotrophic culture period ended.

[0095] The high-density algae cell that 50L bioreactor heterotrophic culture obtains adopts the substratum (wherein KNO 3 Content is 2.60g / L) is diluted to 14.88g / L, then transfers to 30L plate type photobioreactor and carries out photoautotrophic culture. Photoautotrophic culture conditions: the temperature is 30°C, the liquid volume is 24L, the air flow rate...

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Abstract

The invention discloses a small ball algae cultivation method with high density and high quality, which comprises the following phases: high-density heterotrophy cultivation, high-density algae liquid dilution and photo-autotroph cultivation. The invention makes the small ball algae reach high-density level within shorter cultivation period, which improves the small ball algae quality.

Description

technical field [0001] The invention belongs to the field of biotechnology and relates to a method suitable for high-density and high-quality cultivation of chlorella. Background technique [0002] People have a long history of research on chlorella, among which Chlorella vulgaris and Chlorella pyrenoidosa are the algal species that have been studied more, and are also commonly used in commercial production. [0003] At present, there are two commonly used culture methods for Chlorella: open outdoor large pond photoautotrophic culture and fermenter heterotrophic culture. Open outdoor large pool culture is a common method for commercial mass production at present. Although its cost is low, its development is greatly restricted due to the disadvantages of low cell density, easy pollution, and large footprint. Heterotrophic culture is currently basically in the stage of laboratory research and development. Although it can greatly increase the density of algae cells, compared w...

Claims

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Application Information

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IPC IPC(8): C12N1/12A23K1/16A23L1/337A23K10/30A23L17/60
Inventor 李元广李兴武沈国敏钱峰慧安洋魏鸿刚王伟
Owner JIAXING ZEYUAN BIOLOGICAL PROD
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