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Culture method for efficiently inducing lipid accumulation in Botryococcus braunii

A technology of botrytis brannicola and culture medium, which is applied in the field of high-efficiency induction of fat accumulation of botrytis brawny, can solve the problems of slow growth of botrytis and high-density cultivation technology of botrytis, and achieve environmental protection, The effect of promoting cell division

Inactive Publication Date: 2012-12-12
北京芳能科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But at present, the high-density cultivation technology of botrytis has not been well solved, and it is still in the stage of laboratory exploration. This situation is largely caused by the slow growth of botrytis.

Method used

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  • Culture method for efficiently inducing lipid accumulation in Botryococcus braunii
  • Culture method for efficiently inducing lipid accumulation in Botryococcus braunii
  • Culture method for efficiently inducing lipid accumulation in Botryococcus braunii

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] The formula of FN16 medium is as follows:

[0047] Potassium nitrate 0.4g / L Dipotassium hydrogen phosphate 0.08g / L

[0048] Sodium fluoride is 0.05, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4mg / L respectively

[0049] Magnesium sulfate heptahydrate 0.2g / L Calcium chloride dihydrate 0.107g / L

[0050] Ferrous sulfate 0.02g / L Citric acid 0.1g / L

[0051] Sodium chloride 1.75g / L Trace elements 1ml / L

[0052] The configuration method of trace elements is: 0.02mg of cobalt chloride, 0.44mg of zinc sulfate heptahydrate, 5.72mg of boron, 0.16mg of copper sulfate pentahydrate, 3.62mg of manganese chloride tetrahydrate, 0.084mg of sodium molybdate and distilled water to 1L.

[0053] Dissolve the above substances, dilute to 1 L with distilled water, and autoclave at 121°C for 20 minutes. Add 100ml of FN16 culture medium to 250ml shake flask respectively, wherein the initial concentration of sodium fluoride is respectively 0.05, 0.2, 0.4, 0.6, 0.8, 1.0, 1.2, 1.4mg / L, and the inoculum con...

Embodiment 2

[0056] 1. The formula of FN16 medium is as follows:

[0057] Potassium nitrate 0.4g / L Dipotassium hydrogen phosphate 0.08g / L

[0058] Magnesium sulfate heptahydrate 0.2g / L Calcium chloride dihydrate 0.107g / L

[0059] Ferrous sulfate 0.02g / L Citric acid 0.1g / L

[0060] Sodium chloride 1.75g / L Sodium fluoride 0.6mg / L

[0061] Trace elements 1ml / L

[0062] The configuration method of trace elements is: 0.02mg of cobalt chloride, 0.44mg of zinc sulfate heptahydrate, 5.72mg of boron, 0.16mg of copper sulfate pentahydrate, 3.62mg of manganese chloride tetrahydrate, 0.084mg of sodium molybdate and distilled water to 1L.

[0063] Dissolve the above substances, dilute to 1 L with distilled water, and autoclave at 121°C for 20 minutes. Add 100ml of FN16 culture medium to a 250ml shake flask, the amount of solution A added every day is 0.5, 1.0, 1.5, 2.0ml / L respectively, and the inoculation concentration is 3.25×10 5 cells / ml, cultured at a temperature of 26°C, light:dark=14:10, li...

Embodiment 3

[0074] 1. Multi-conditional induction by combining ultrasound and culture conditions

[0075] In the 300L small-scale photoreactor system, add FN16 medium, the ratio of Staphylococcus branici-X09 to the medium volume ratio is 1:10, inoculate Staphylococcus branici-X09 into the medium, and the inoculation concentration is 2.24×10 5 The temperature is 26-28°C, the light is 15000-18000 lux, and the light time is 14 hours / day. Add solution A to the microalgae culture solution every day at 1ml / L, and start to induce fat accumulation after 13 days of culture; the ultrasonic frequency is 20kHz during the fat-induced accumulation stage The frequency is 5w, the treatment time is 25s, and the treatment is performed twice, and then the concentration of 6% CO is injected at a flow rate of 1.2L / min. 2 , Supplement solution A at 15μl / L every day, add sodium chloride 5.85g / L at one time, and add 150mg / L ferrous sulfate at the same time, after induction culture for 2 days, then use 12kHz freq...

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Abstract

The invention discloses a culture method for efficiently inducing lipid accumulation in Botryococcus braunii, which comprises the following steps of: preparing a culture medium and daily added nutrient solution, culturing the Botryococcus braunii for 10 to 18 days of biomass accumulation, introducing CO2 into the Botryococcus braunii culture solution, regulating the concentration of the added nutrient solution, performing ultrasonic treatment on the Botryococcus braunii culture solution for 1 to 3 times by using an ultrasonic instrument at a certain frequency, spacing 10 seconds each time, and treating 5 to 60 seconds each time to inducing the lipid accumulation in the Botryococcus braunii. For the Botryococcus braunii cultured by the method, the biomass is improved by 1 to 1.5 times, thetotal fat content is improved by 30 to 55 percent, and the growth cycle is shortened by 2 to 3 days. A carbon source is supplemented in a mode of introducing CO2; and in the process of producing the lipid accumulation in the Botryococcus braunii on a large scale, a great amount of CO2 waste gas generated in the industrial production process is absorbed, and the method has significance for saving energy, reducing emission and protecting environment.

Description

Technical field: [0001] The invention relates to a method for efficiently increasing the fat content of the botrytis brachyphyllum-X09 and cultivating the botrytis brachyphylla-X09 in an open mode at high density. Background technique [0002] The global energy shortage and environmental pollution make the development and utilization of renewable and pollution-free bio-energy an important research direction. The use of microalgae to produce bioenergy has the following unique advantages: Microalgae can effectively use solar energy and fix CO through photosynthesis 2 , to convert inorganic substances into energy substances such as highly unsaturated alkanes and oils; microalgae bioenergy is renewable. Botryococcus (Botryococcus.sp), also known as Conglococcus, belongs to Chlorophyta, Chlorophyceae, and Botrytisaceae. It is a single-celled green algae distributed worldwide in freshwater or brackish water. The algae has attracted widespread attention as a non-polluting and ren...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/12C12N13/00C12R1/89
Inventor 单东杰苏娇娇
Owner 北京芳能科技有限公司
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