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Sorokin chlorella synthetic starch defect type mutant strain and application thereof

A technology for synthesizing starch and chlorella, which is used in the preparation of mutants, unicellular algae, biochemical equipment and methods, etc. Effect

Active Publication Date: 2017-09-01
GUOTOU BIO TECH INVESTMENT CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These screening methods that directly target the lipid content of cells require specialized large-scale equipment and dedicated personnel to operate, and are difficult to be widely used

Method used

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  • Sorokin chlorella synthetic starch defect type mutant strain and application thereof
  • Sorokin chlorella synthetic starch defect type mutant strain and application thereof
  • Sorokin chlorella synthetic starch defect type mutant strain and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0056] Embodiment 1: Mutagenesis screening of mutant strains

[0057] 1. Determine the germination rate and semi-lethal rate of Chlorella under the solid plate of heterotrophic medium

[0058] Determination of germination rate: smear 100 μL of algae liquid on each Endo solid medium, the number of cells per 100 μL is 1000, 500, and 200, and each cell concentration gradient is made in three parallels, and then cultured in a 37°C incubator for 5 days , to determine the cell germination rate according to the number of single colonies grown from each plate.

[0059] The formula of Endo heterotrophic medium is as follows:

[0060] Glucose 20g / L; KNO 3 2g / L; KH 2 PO 4 1.2g / L; MgSO 4 ·7H 2 O 1.2g / L; Trisodium Citrate 0.2g / L; CaCl 2 2H 2 O mother liquor (1000×) 1ml; FeSO 4 ·7H 2 O and EDTA·2Na mother solution (1000×) 1mL; trace element mother solution (1000×) 1mL.

[0061] in:

[0062] CaCl 2 Preparation of mother liquor: 105g CaCl 2 2H 2 O dissolved in 1000ml H 2 O; ...

Embodiment 2

[0078] Example 2, under the induction of high light and low nitrogen, the dry weight curve, starch and oil content of mutant strains were measured, and the best mutant strains were selected

[0079] (1) The medium formula of low-nitrogen (1 / 16N) autotrophic medium BG11 is as follows:

[0080]

[0081]

[0082] The ingredients of Trace metal mix A5 are:

[0083] Element Mother liquor concentration (g / L) h 3 BO 3

2.86 MnCl 2 .4H 2 o

1.86 ZnSO 4 .7H 2 o

0.22 Na 2 MoO 4 .2H 2 o

0.39 CuSO 4 .5H 2 o

0.08 Co(NO3) 2 .6H 2 o

0.05

[0084] K 2 HPO 4 It is sterilized separately from Ferric ammonium citrate mother solution, and added after other solutions are sterilized.

[0085] The configuration method of A5 (you need to add each ingredient to the water in turn, stir until dissolved and then add another ingredient).

[0086] Finally, adjust the pH of the prepared BG11 medium to 7.1 with 1...

Embodiment 3

[0139] Example 3: Determination of the dry weight curve, cell growth curve, oil content and TAG content of the SLM2 mutant strain with the highest oil production under heterotrophic fermentation culture

[0140] The mutant strain SLM2 with low starch content and high oil content was selected and cultured at high density in a fermenter to study its dry weight, cell growth and oil content under full medium.

[0141] Proceed as follows:

[0142] Select monoclonal algae strain SLM2 and wild type to activate in 20ml Endo liquid medium for 2 days (primary culture); when entering the logarithmic phase of growth, adjust monoclonal algae strain SLM2 and wild type to the same cell concentration and transfer to In a 250ml Erlenmeyer flask, the final volume of the algae solution is 100ml. The temperature is 37°C, 150rpm (secondary culture); on the third day, transfer 40ml of algae liquid to a 1000ml Erlenmeyer flask, add fresh Endo medium to 400ml (tertiary culture) and cultivate for 3.5...

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Abstract

The invention provides sorokin chlorella synthetic starch defect type mutant strains GT-1-SLM1, GT-1-SLM2 and GT-1-SLM3 through mutation screening. The preservation numbers of the strains are respectively CGMCC No.13861, CGMCC No.13862 and CGMCC No.13863, and the strains are preserved in the China General Microbiological Culture Collection Center. As related channels or a small part of the channels for synthesizing starch through the strains are blocked, the content of starch in cells is remarkably reduced, the grease synthesis velocity is remarkably increased, and the highest grease content in the cells is remarkably increased. By adopting the mutant strains provided by the invention, the microalgae oil generation period can be shortened, so that the risk of disease infection in the culture period can be reduced, and meanwhile as biomasses have relatively high content of grease, later grease extraction and preparation processes can be simplified, and high-quality germplasm resources can be provided for industrialization of microalgae oil production.

Description

technical field [0001] The invention relates to the field of mutant screening of dominant algal strains of energy microalgae, in particular to a starch-deficient mutant strain of Chlorella sorokinus, a screening method and an application thereof. Background technique [0002] With the increase of world population and the continuous depletion of non-renewable energy sources, the crisis of shortage of fossil energy is inevitable, and attention to renewable energy has become a hot spot. Algae have the characteristics of wide distribution, high oil content, short production cycle, strong environmental adaptability, and high yield, among which microalgal biomass is considered to be the most promising biofuel raw material. The use of microalgae can not only produce biodiesel and other energy sources, but also carry out biological carbon sequestration, which is conducive to curbing global warming and has broad development prospects. [0003] At present, most of the microalgae used...

Claims

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Application Information

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IPC IPC(8): C12N1/12C12N15/01C12P7/64C12Q1/04C12R1/89
CPCC12N1/12C12N15/01C12P7/649C12Q1/04C12N1/125C12R2001/89Y02E50/10
Inventor 韩丹翔吴明灿孙文超胡强
Owner GUOTOU BIO TECH INVESTMENT CO LTD
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