Method for extracting microalgae lipid through biological enzyme catalysis wall breaking

Abstract

The invention discloses a method for extracting microalgae lipid through biological enzyme catalysis wall breaking. The method includes the steps that A, a phosphate buffer solution and a biological enzyme catalyst are sequentially added into wet algae mud, or algae slurry, or dry powder or fermentation liquor for culturing harvested lipid-generating microalgae, and the mixture is stirred and heated; B, after the enzyme catalysis degradation reaction is finished, the pH value of a reaction system is adjusted, and the temperature of enzyme catalysis degradation liquid is raised and kept; C, after the reaction is finished, the temperature of the enzyme catalysis degradation system is lowered, an extraction agent is added into the reaction system for lipid extraction, and then demulsifier is added to carry out layering through centrifugation; D, reduced pressure distillation or thin film evaporation concentration is carried out on the oil phase obtained through separation to obtain the microalgae lipid. The method is easy to implement and convenient to operate; due to efficient wall-breaking lipid extraction for the lipid-generating microalgae particularly photoautotroph lipid-generating microalgae, energy input in the microalgae lipid extraction process is reduced, the energy efficiency ratio of microalgae energy is increased, and then comprehensive production cost of microalgae biomass energy is greatly reduced.

Description

technical field

[0001] The invention relates to the fields of microalgae biochemical industry, microalgae biomass energy, microalgae high value-added natural product chemistry and the like, in particular to a cell wall breaking and oil extraction method of oil-producing microalgae. Background technique

[0002] With the rising price of fossil fuels and the recognition of the non-renewability of fossil fuels, people's demand for alternative new energy sources is also expanding. Microalgal photosynthetic autotrophy can reduce CO under normal temperature and pressure 2 Perform efficient absorption, convert light energy into fat or starch, store it in cells, and release oxygen. Microalgae have extremely high photosynthetic efficiency, efficient concentration and fixation of CO 2 The ability and the ability to use non-food crop land such as oceans, tidal flats, and deserts as breeding sites are considered to be a very promising bulk raw material for biodiesel production in the ...

Examples

Embodiment 1

[0024] A novel bio-enzyme-catalyzed method for extracting oil from microalgae by breaking walls, the steps of which are:

[0025] A. Weigh 10g of chlorella freeze-dried algal powder obtained from heterotrophic fermentation culture and place it in a stoppered conical flask, add 50ml of 0.05M phosphate buffer solution (PH7.2) to mix well with the algae, then add 150mg of chitin The compound enzyme (1:1:1, m / m / m) composed of enzyme, cellulase and lipase was shaken well, and placed in a water bath at 50°C for 4 hours.

[0026] B. Further, adjust the pH of the system to 7.5 or 8.5 or 9 with 0.1M sodium hydroxide solution and raise the temperature of the water bath to 70 or 75 or 80° C. for 2 hours. After the reaction is completed, cool the flask to below 40°C, then add 10ml of a mixed solvent of n-hexane and ether (1:1, v / v) and transfer it to a centrifuge tube for vigorous shaking for extraction, then add 20ml of methanol and continue shaking extraction.

[0027] C. Further plac...

Embodiment 2

[0029] A novel bio-enzyme-catalyzed method for extracting oil from microalgae by breaking walls, the steps of which are:

[0030] A. Take 500g of fresh algae mud of photoautotrophic Chlorella, which has a solid content of 21.0%, and put it in a three-necked bottle, add 400ml of 0.05M phosphate buffer solution (PH7.2) and stir well with the algae mud, then add 2.0 g of compound enzyme (1:1:1, m / m / m) composed of chitinase, pectinase and alkaline protease was stirred evenly, and placed in a 50°C water bath for 4 hours.

[0031] B. Further, adjust the pH of the system to 7.8 or 8.3 or 9 with 0.1M sodium hydroxide solution and raise the temperature of the water bath to 70 or 73 or 76 or 80° C. for 2 hours. After the reaction is complete, cool the flask to below 40°C, then add 150ml of a mixed solvent of n-hexane and ether (1:1, v / v) to the three-neck flask and shake vigorously for extraction, then add 200ml of methanol and continue shaking for 5 minutes.

[0032] C. Further transfer...

Embodiment 3

[0035] A novel bio-enzyme-catalyzed method for extracting oil from microalgae by breaking walls, the steps of which are:

[0036] A, get heterotrophic inferior chytrid chytrid fermented liquid 20L and place in the glass extraction kettle, the solid content of this fermented liquid is 108g / L, add potassium dihydrogen phosphate 4.8g and 36 grams of dipotassium hydrogen phosphate successively and stir and dissolve evenly, then Add 32g of compound enzyme composed of chitinase and alkaline protease (1:1, m / m), stir evenly, and then heat to 45°C for 3h reaction.

[0037] B. Further, adjust the pH of the system to 7.5 or 7.8 or 8 with 0.1M sodium hydroxide solution, and raise the temperature of the water bath to 70 or 74 or 78 or 80°C for 1 hour. After the reaction was completed, 5 L of ethanol and 3 L of n-hexane were added to the above system and stirred for 30 min.

[0038] C. Further use a three-phase disc separator to separate the oil phase, and the disc centrifugal speed is 60...

technical field

[0001] The invention relates to the fields of microalgae biochemical industry, microalgae biomass energy, microalgae high value-added natural product chemistry and the like, in particular to a cell wall breaking and oil extraction method of oil-producing microalgae. Background technique

[0002] With the rising price of fossil fuels and the recognition of the non-renewability of fossil fuels, people's demand for alternative new energy sources is also expanding. Microalgal photosynthetic autotrophy can reduce CO under normal temperature and pressure 2 Perform efficient absorption, convert light energy into fat or starch, store it in cells, and release oxygen. Microalgae have extremely high photosynthetic efficiency, efficient concentration and fixation of CO 2 The ability and the ability to use non-food crop land such as oceans, tidal flats, and deserts as breeding sites are considered to be a very promising bulk raw material for biodiesel production in the ...

Examples

Embodiment 1

[0024] A novel bio-enzyme-catalyzed method for extracting oil from microalgae by breaking walls, the steps of which are:

[0025] A. Weigh 10g of chlorella freeze-dried algal powder obtained from heterotrophic fermentation culture and place it in a stoppered conical flask, add 50ml of 0.05M phosphate buffer solution (PH7.2) to mix well with the algae, then add 150mg of chitin The compound enzyme (1:1:1, m / m / m) composed of enzyme, cellulase and lipase was shaken well, and placed in a water bath at 50°C for 4 hours.

[0026] B. Further, adjust the pH of the system to 7.5 or 8.5 or 9 with 0.1M sodium hydroxide solution and raise the temperature of the water bath to 70 or 75 or 80° C. for 2 hours. After the reaction is completed, cool the flask to below 40°C, then add 10ml of a mixed solvent of n-hexane and ether (1:1, v / v) and transfer it to a centrifuge tube for vigorous shaking for extraction, then add 20ml of methanol and continue shaking extraction.

[0027] C. Further plac...

Embodiment 2

[0029] A novel bio-enzyme-catalyzed method for extracting oil from microalgae by breaking walls, the steps of which are:

[0030] A. Take 500g of fresh algae mud of photoautotrophic Chlorella, which has a solid content of 21.0%, and put it in a three-necked bottle, add 400ml of 0.05M phosphate buffer solution (PH7.2) and stir well with the algae mud, then add 2.0 g of compound enzyme (1:1:1, m / m / m) composed of chitinase, pectinase and alkaline protease was stirred evenly, and placed in a 50°C water bath for 4 hours.

[0031] B. Further, adjust the pH of the system to 7.8 or 8.3 or 9 with 0.1M sodium hydroxide solution and raise the temperature of the water bath to 70 or 73 or 76 or 80° C. for 2 hours. After the reaction is complete, cool the flask to below 40°C, then add 150ml of a mixed solvent of n-hexane and ether (1:1, v / v) to the three-neck flask and shake vigorously for extraction, then add 200ml of methanol and continue shaking for 5 minutes.

[0032] C. Further transfer...

Embodiment 3

[0035] A novel bio-enzyme-catalyzed method for extracting oil from microalgae by breaking walls, the steps of which are:

[0036] A, get heterotrophic inferior chytrid chytrid fermented liquid 20L and place in the glass extraction kettle, the solid content of this fermented liquid is 108g / L, add potassium dihydrogen phosphate 4.8g and 36 grams of dipotassium hydrogen phosphate successively and stir and dissolve evenly, then Add 32g of compound enzyme composed of chitinase and alkaline protease (1:1, m / m), stir evenly, and then heat to 45°C for 3h reaction.

[0037] B. Further, adjust the pH of the system to 7.5 or 7.8 or 8 with 0.1M sodium hydroxide solution, and raise the temperature of the water bath to 70 or 74 or 78 or 80°C for 1 hour. After the reaction was completed, 5 L of ethanol and 3 L of n-hexane were added to the above system and stirred for 30 min.

[0038] C. Further use a three-phase disc separator to separate the oil phase, and the disc centrifugal speed is 60...