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Method for high-efficiency and high-density culture of biological bait

A technology of high-density cultivation and biological bait, which is applied in the field of high-efficiency and high-density cultivation of biological bait, can solve the problems of high harvesting cost, low density of algae cells, and unbalanced nutrients, so as to reduce production costs, simplify the process flow, and ensure effect of homogeneity

Inactive Publication Date: 2018-03-23
KUSN INNOVATION INST OF NANJING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although low-cost open culture is a large-scale culture method commonly used for microalgae cells, it has the following disadvantages: (1) The density of algae cells is low, the area is large, and it is easy to be polluted by protozoa, bacteria, fungi, and miscellaneous algae (2) Since the open culture is greatly affected by the climate conditions, the quality of the bait microalgae is uneven, the nutrient content is unbalanced, and the bait potency is low; (3) The bait supply is not timely, and it is often out of touch with the seedling raising progress, which is difficult to meet The supply and demand of bait in aquaculture seedlings leads to low survival rate of seedlings, poor disease resistance of seedlings, and high cost of seedlings
However, the heterotrophic culture of bait microalgae still has the following defects: (1) the protein content and chlorophyll content of algal cells are low; (2) the algae cell culture process is inhibited by high concentration of substrate, and the cell density is low
Although the two-stage culture method combines the high-density culture characteristics of closed reactors and the high-efficiency induction characteristics of open bioreactors, it also has the disadvantages of both.
This cultivation method still cannot solve the problems of high harvesting cost, uneven product quality, and contamination by harmful organisms.
Therefore, how to achieve high-density and efficient cultivation of biological bait is still a technical problem hindering its industrial production

Method used

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  • Method for high-efficiency and high-density culture of biological bait
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  • Method for high-efficiency and high-density culture of biological bait

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0069] Add 250ml of basal medium into a 500ml shake flask (recipe is shown in Table 1), insert the algae cells that have been checked for sterility and normal appearance into the shake flask for culture, the culture conditions are: completely dark culture, and the culture temperature is 30°C , the initial speed is 150rpm. The experimental design was divided into 3 groups:

[0070] Experiment 1 (batch culture process): Batch culture was used to culture without supplementing the basal medium concentrate.

[0071] Experiment 2 (feed-fed culture process): Firstly, the batch culture method was used to culture, and the basal medium concentrate was supplemented every 12 hours from the 3rd day of culture.

[0072] Experiment 3 (feed-starvation cycle culture process): Firstly, the batch culture method was used to culture, and the basal medium concentrate was supplemented every 24 hours from the 3rd day of culture.

[0073] Samples were taken at regular intervals to determine cell dry...

Embodiment 2

[0081] Add 250ml of basal medium into a 500ml shake flask (recipe is shown in Table 1), insert the algae cells that have been checked for sterility and normal appearance into the shake flask for starvation culture for 16 hours, and the culture conditions are: completely dark culture, culture temperature is 30°C, initial rotation speed 150rpm. The C / N ratio is set to: 69, 52, 34, 23, 17. The C / P ratio is set to: ∞, 606, 302, 151, 75. Samples were taken at regular intervals to determine cell dry weight, protein content, glucose content, nitrate content and phosphate content.

[0082] Phosphate content was determined by molybdenum acid spectrophotometry (GB / T 11893-1989 Determination of total phosphorus in water quality by ammonium molybdate spectrophotometry).

[0083] Analysis of results: by figure 2 It can be seen that too low nitrogen concentration affects the accumulation of intracellular protein in Chlorella cells, mainly because there is a certain threshold for the act...

Embodiment 3

[0085] Add basal culture medium (recipe is shown in Table 2) in the bioreactor, and the filling factor of the reactor is 70%, inserts the algae cell that checks sterility and normal appearance state into the bioreactor to cultivate, and the culture condition is: culture temperature The temperature is 30°C, the gas-liquid ratio is 1.0vvm, and the initial rotation speed is 200rpm. When Do<20%, the rotation speed increases by 50rpm, and the maximum rotation speed is 1000rpm. The experimental design is divided into 2 groups:

[0086] Experiment 1 (feed-feeding process): First, batch culture was used to cultivate Chlorella, and when the cell density reached a certain value (ie, glucose<20g / L), fed-feed medium was added (see Table 3 for the formula).

[0087] Experiment 2 (feed-starvation cycle culture process): Firstly, the batch culture method was used to cultivate Chlorella, and when the cell density reached a certain value (i.e. glucose <20g / L), fed-batch feeding medium (recipe...

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Abstract

The invention relates to the technical fields of fermentation engineering and aquaculture, and especially relates to a method for high-efficiency and high-density culture of a biological bait. The method comprises the following steps: 1, performing batch culture; 2, performing feeding-hunger cycle culture: feeding a medium when the cell density reaches 80-100 g / L until the pH value is 6-8, stopping the feeding for 2-8 h to adjust the cell metabolism to a hungry state, afresh recovering the feeding of the medium to recover the cell metabolism to a nutrition metabolism state, and repeating aboveprocesses; and 3, inducing the target product: going to an induction stage when the cell density reaches 100 g / L or above, and reducing the feeding rate or removing at least one substrate nutrient component from the medium to reduce the cell growth rate and induce the synthesis of the target product, wherein the substrate nutrient component can be one or more of a carbon source, a nitrogen source, a phosphorus source, a sulfur source, vitamins and heavy metal ions. The method has the advantages of high culture efficiency, one-step realization of protein accumulation, and low harvesting cost.

Description

technical field [0001] The invention relates to the technical fields of fermentation engineering and aquaculture, in particular to a method for cultivating biological bait with high efficiency and high density. Background technique [0002] Biological bait refers to high-quality bait organisms that have been artificially screened, can be artificially cultivated, and are suitable for aquaculture. The protein content of microalgae can reach more than 40%, and the content of essential amino acids is equivalent to or even better than that of fish meal. A large number of nutritional and toxicological studies have proved that microalgae are a suitable biological bait. There are three main types of microalgae bait used in aquaculture: (1) As a living biological bait used in aquaculture, in addition to being directly ingested by aquatic animals, its other role is also reflected in the improvement of aquaculture water quality, Regulate the balance of water body micro-ecological env...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12N1/20
CPCC12N1/12C12N1/20
Inventor 杨欢张京奎刘士涛潘志龙邹志刚吴聪萍刘建国
Owner KUSN INNOVATION INST OF NANJING UNIV
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