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Starch-deficient mutant strain of Scenedesmus apicula and its application

A technology for synthesizing starch and defective type, which is used in the preparation of mutants, unicellular algae, biofuels, etc., to achieve the effect of improving oil conversion rate, strong operability, and fast growth rate

Active Publication Date: 2021-01-19
GUOTOU BIO TECH INVESTMENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are relatively few domestic studies on the use of microalgae to produce oil, especially the studies on mutants with deficient starch synthesis.

Method used

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  • Starch-deficient mutant strain of Scenedesmus apicula and its application
  • Starch-deficient mutant strain of Scenedesmus apicula and its application
  • Starch-deficient mutant strain of Scenedesmus apicula and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0064] Embodiment 1, the screening of mutant strain

[0065] 1. Determine the germination rate and semi-lethal rate of Scenedesmus under the solid plate of heterotrophic medium:

[0066] Determination of the concentration of ethyl formate (EMS): inoculate the GT-2 single clone grown on the solid Endol medium into a 50ml Erlenmeyer flask containing 20ml Endol liquid medium, cultivate in the dark at 30°C, 150rpm / min for 5- 6 days, when the number of cells reaches 7 x 10 7 ind / ml, transferred to a 250ml Erlenmeyer flask containing 100ml liquid medium, the inoculum size was 7×10 4 ind / ml. When the number of cells grows to the logarithmic phase, it is about 7×10 7 ind / ml. Take 100 μl of algae liquid GT-2 in the logarithmic growth phase and treat it with different concentrations of ethylmethanesulfonate (EMS) solutions. The concentrations were 0, 0.5%, 1%, 1.5%, 2%, 2.5% (W / V), respectively.

[0067] The concentration of EMS is 0%, the treatment time is 1h, 2h and 4h, the survi...

Embodiment 2

[0081] Example 2. Determination of dry weight curve, starch and oil content under the induction of high light and low nitrogen.

[0082] (1) Preparations before induction:

[0083] First, through normal heterotrophic and autotrophic conditions, find out the fast-growing, biomass-accumulated starch synthesis-deficient mutants and wild-type (hereinafter referred to as GT-2) single clones for activation:

[0084] Heterotrophic culture: Pick the target single clone and culture it in BG11 liquid medium under light, and the light intensity at 26°C is 50 μmol.m -2 .s -1 , 150rpm / min, cultured for one week, if it can grow normally after one week, it is a candidate starch synthesis defective mutant. Pick candidate starch synthesis deficient mutant strains and GT-2 and culture them in 20ml Endol liquid medium, keep away from light at 30°C, 150rpm / min, and cultivate them for 5 days. The same concentration (9.25×10 4 ind / ml) reactivation. Adjust the algae solution in the reactivated ...

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Abstract

The invention provides the starch-deficient mutant strains MSA1 and MSA5 of Scenedesmus apicula obtained after mutation screening; their preservation numbers are respectively CGMCC No.13864 and CGMCC No.13866, and are preserved in the General Microbiology Center of China Microbiological Culture Collection Management Committee . The present invention screens out starch synthesis deficient mutants that do not synthesize starch or have a very small amount of starch synthesis, and find out varieties that can be both heterotrophic and photoautotrophic from these alternative mutants, and finally select the mutants that can be produced under heterotrophic culture. A starch synthesis-deficient mutant with fast growth rate, high biomass, and high neutral lipid and triacylglycerol synthesis under photoautotrophic conditions. The finally obtained mutant strains MSA1 and MSA5 can quickly accumulate total fatty acid content under heterotrophic fermentation conditions, and the starch yield is greatly reduced. In the final fermentation result, the total fatty acid content is increased by 5 times to 10 times compared with the wild type. The conversion rate of carbon source into oil is greatly improved, and it can be used as industrial-grade "oil-producing microalgae" and widely used in the preparation of biodiesel.

Description

technical field [0001] The invention relates to the field of screening mutant strains of dominant algal strains of energy microalgae, in particular to a starch-deficient mutant strain of Scenedesmus cuspidatum, a screening method and an application thereof. Background technique [0002] At present, the utilization rate of fossil fuels such as coal and oil is unsustainable. At the same time, the burning of fossil fuels will cause a large amount of greenhouse gas emissions, which is the main factor leading to global warming. This is contrary to the sustainable green development advocated by the CPC Central Committee. Therefore, it is urgent to find a renewable green alternative. Microalgae emerged as the times require to become an energy carrier for the sustainable development of the tertiary industry that maintains the economy and the environment. [0003] Scenedesmus apicula belongs to Chlorophyta, Chlorophyceae, Chlorococcuses, Scenedesmusceae, and Scenedesmus genus. Sce...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N15/01C12N1/12C12P7/64C12Q1/04C12R1/89
CPCC12N1/12C12N15/01C12P7/649C12Q1/04C12N1/125C12R2001/89Y02E50/10
Inventor 韩丹翔孙文超吴明灿胡强
Owner GUOTOU BIO TECH INVESTMENT CO LTD
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