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Two-stage microalgae cultivation method with low nitrogen and low phosphorus stress

A culture method and two-stage technology, applied in the field of microalgae culture, can solve the problems of microalgae stress culture, damage repairing active substances, etc., and achieve the effects of reducing cell death rate, improving production efficiency, and shortening culture period

Pending Publication Date: 2020-05-19
NINGBO UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no public method of using low N and P nutrients to stress culture microalgae, and then make them produce and accumulate damage repair active substances in large quantities.

Method used

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  • Two-stage microalgae cultivation method with low nitrogen and low phosphorus stress
  • Two-stage microalgae cultivation method with low nitrogen and low phosphorus stress
  • Two-stage microalgae cultivation method with low nitrogen and low phosphorus stress

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Using Chlamydomonas (sp.cw15, purchased from the U.S. Duke University algae bank) cells as algae species to carry out a two-stage low-nitrogen and low-phosphorus stress microalgae cell culture method, the specific steps are as follows:

[0023] 1. Cultivate and obtain a logarithmic phase microalgae cell preculture solution. Prepare 6L of TAP medium, the specific formula is shown in Table 1 below, inoculate Chlamydomonas cells into it after sterilization, place the inoculated culture medium at 25°C, and the intensity of sunlight light is 100 μmol / (m 2 s) in a shaker light incubator with a rotating speed of 20 rpm, culture for 7 days to obtain a pre-culture solution of Chlamydomonas cells in the logarithmic phase.

[0024] Table 1 TAP medium formula

[0025]

[0026]

[0027] 2. Detect the concentration of Chlamydomonas cells in the logarithmic phase pre-culture solution. Shake the Chlamydomonas cell preculture solution and count under the microscope using a hemoc...

Embodiment 2

[0037] Chlorella (sp.dh8248, preserved in the laboratory, collected in East Lake, Wuhan) cells was used as a two-stage low-nitrogen and low-phosphorus stress microalgae cell culture method, and the specific steps were as follows:

[0038] 1. Cultivate and obtain a logarithmic phase microalgae cell preculture solution. Prepare 6L of TAP medium. The specific formula is shown in Table 1 below. After sterilization, inoculate the small ball cells into it, and place the inoculated culture medium at 25°C, and the intensity of sunlight light is 100 μmol / (m 2 s) in a shaker light incubator with a rotating speed of 20rpm, culture for 7 days to obtain a pre-culture solution of bead cells in the logarithmic phase.

[0039] Table 4BG11 medium formula

[0040] compound Final concentration NaNO 3

1.5g / L K 2 HPO 4

0.04g / L MgSO 4 ·7H 2 o

0.075g / L CaCl 2 2H 2 o

0.036g / L Na 2 CO 3

0.02g / L citric acid 0.006g / L ...

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Abstract

The invention discloses a two-stage microalgae cultivation method with low nitrogen and low phosphorus stress. The cultivation method is characterized in that the method includes the following steps:(1) obtaining a logarithmic phase microalgae cell pre-culture solution through cultivation under the optimal culture conditions; (2) performing centrifugation on the logarithmic phase microalgae cellpre-culture solution, and obtaining microalgae cells through collection; (3) performing amplification culture on the collected microalgae cells again in a culture medium with low nitrogen and low phosphorus stress; and (4) performing light ventilation culture at 25 DEG C for 10-14 d. Thus, the microalgae cells whose accumulation of polyunsaturated fatty acids (PUFAs) is significantly increased canbe obtained.

Description

technical field [0001] The invention belongs to the technical field of microalgae cultivation, and in particular relates to a two-stage method for culturing microalgae cells under low-nitrogen and low-phosphorus stress. Background technique [0002] Microalgae is a general term for small algae in algae. As a pioneer species on the earth, it is an autotrophic organism that can use light energy, carbon dioxide and water to synthesize various organic substances in its cells, and has a strong adaptability to the environment. This extremely strong adaptability is closely related to the efficient metabolic pathways formed by algae during the long-term evolution process and the massive production and accumulation of various damage repair active substances. A large number of research reports have pointed out that under the condition of nutrient deficiency stress, microalgae will produce and accumulate various damage repair active substances in the cells, among which polyunsaturated ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/12C12N1/38C12R1/89
CPCC12N1/12C12N1/38
Inventor 高翔周维成张弛王尧尧王高鸿李登峰
Owner NINGBO UNIV
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