WO2023155503A1

WO2023155503A1 - Photosynthetic green algae-based production apparatus for carbon sink algae liquid

Info

Publication number: WO2023155503A1
Application number: PCT/CN2022/131687
Authority: WO
Inventors: 李世峰; 王康杰; 卢群伟; 胡蓓娟; 王雅凡; 秦康曦; 王祎; 梁盼
Original assignee: 九江地福来农业科技发展有限公司
Priority date: 2022-02-21
Filing date: 2022-11-14
Publication date: 2023-08-24
Also published as: CN114686359A

Abstract

Disclosed are a photosynthetic green algae-based production apparatus for carbon sink algae liquid, a preparation method, and use. The preparation method comprises the following steps: feeding an algae carbon capture tank with purified air and purified water; adding photosynthetic green algae liquid to the algae carbon capture tank, mixing citric acid, CaCl 2, MgSO 4·7H2O, KH 2PO 4, CuSO 4·5H2O, EDTA-Fe, and purified water, and feeding the algae carbon capture tank with the mixture; mixing a pinellia ternate agglutinin protein with purified water, and feeding the algae carbon capture tank with the mixture; setting a preset illumination intensity by means of an illumination apparatus and keeping the preset illumination intensity, and setting a preset temperature by means of a temperature monitor and keeping the preset temperature; performing continuous culture, monitoring a cell concentration of photosynthetic green algae every day by means of an algae concentration monitor, and when the cell concentration reaches a preset peak value, discharging the carbon sink algae liquid in the algae carbon capture tank into a high-concentration algae liquid storage tank; and discharing the carbon sink algae liquid in the high-concentration algae liquid storage tank into a filling apparatus for filling to form a carbon sink product so as to be suitable for the space outside the forest, so that larger carbon sink intensity can be generated.

Description

Production equipment, preparation method and application of carbon sequestration algae liquid based on photosynthetic green algae

This application claims the priority of the Chinese patent application submitted to the China Patent Office on February 21, 2022, with the application number 202210158236.2, and the title of the invention is "Carbon sink algae liquid production equipment, preparation method and application based on photosynthetic green algae". The entire contents are incorporated by reference in this application.

technical field

The invention relates to the technical field of carbon sequestration algae liquid production, in particular to a carbon sink algae liquid production equipment, preparation method and application based on photosynthetic green algae.

Background technique

Climate change is a major crisis and severe challenge faced by all mankind, and has become a common concern in the fields of international politics, diplomacy, economy and ecology. To deal with climate change, we should pay equal attention to both mitigation and adaptation. Climate change mitigation is a long-term and arduous task, while adaptation to climate change is a more realistic and urgent task. Forestry is one of the areas most affected by climate change, and it is also one of the key areas for adapting to climate change identified by my country. Doing a good job in forestry adaptation to climate change is of great significance to enhancing the overall adaptability of the country and maintaining ecological security and climate security.

However, my country's forest resource endowment is insufficient. my country's forest coverage rate is far lower than the global average of 31%. The per capita forest area is only 1/4 of the world's average, and the per capita forest stock is only 1/7 of the world's average. The wetland rate is lower than the global average of 8.6% % of the average level, the per capita wetland area is only 1/5 of the world's per capita, the pressure of wetland protection is great, and the restoration is difficult; the haze is frequent, sandstorms occur frequently, and the task of sand prevention and control is heavy; landscape fragmentation and endangered species are intensified , Biodiversity protection is very urgent. Ecological fragility is still the basic national condition of our country, and the shortage of ecological products is still a prominent shortcoming. Forests, wetlands and desert ecosystems are relatively sensitive to climate change, and climate risks are relatively high. At the same time, the foundation for forestry adaptation to climate change is weak. The awareness of adapting to climate change in the forestry field is generally low, the capacity is relatively weak, the work system is not perfect, and the talent team is in short supply. All work needs to be strengthened urgently.

Forests are the largest carbon pool in terrestrial ecosystems, and expanding forest coverage is an important measure to mitigate and respond to climate change that is economically feasible and low-cost in the next 30 to 50 years. Forests have a huge function of sequestering carbon dioxide, which is called "carbon sink". At the same time, deforestation, forest land degradation, forest fires, pests and diseases, etc., release the stored carbon dioxide into the atmosphere and become a "carbon source". Taking measures to strengthen the carbon storage and sink functions of forest ecosystems and reduce carbon emissions from forests has become a global consensus and action to deal with climate change, and it has also become one of the goals of China's two commitments to voluntarily reduce emissions. However, the existing carbon sink products are generally only used in forests, and the carbon sink intensity produced is not high.

Contents of the invention

Based on this, the object of the present invention is to provide a carbon sink algae liquid production equipment, preparation method and application based on photosynthetic green algae, so as to improve the carbon sink intensity.

In a first aspect, the present invention provides a carbon sequestration algae liquid production equipment based on photosynthetic green algae, including a device cavity, and an algae carbon capture tank disposed in the device cavity;

The algae carbon capture tank is connected to the air treatment equipment through the air inlet pipe, connected to the water treatment equipment through the water inlet pipe, and connected to the high-concentration algae liquid storage tank through the algae liquid outlet pipe;

The algae carbon capture tank is provided with an algae concentration monitor, a temperature monitor and an algae liquid inlet pipe, and the algae liquid inlet pipe is used to add photosynthetic green algae liquid;

The top of the equipment cavity is provided with lighting equipment, and the high-concentration algae liquid storage tank is connected with the filling equipment.

Further, the volume of the algae carbon capture tank is not less than 1m ³ .

In a first aspect, the present invention provides a method for preparing carbon sequestration algae liquid based on photosynthetic green algae, comprising the following steps:

Step S10, sending the purified air treated by the air treatment equipment into the algae carbon capture tank, and the purified water treated by the water treatment equipment into the algae carbon capture tank;

Step S11, after adding photosynthetic green algae liquid into the algae carbon capture tank, mixing citric acid, CaCl ₂ , MgSO ₄ 7H ₂ O, KH ₂ PO ₄ , CuSO ₄ 5H ₂ O, EDTA-Fe and purified water , into the algae carbon capture tank;

Step S12, mixing the pinellia lectin protein after multi-step purification with purified water, and sending it into the algae carbon capture tank;

Step S13, setting and maintaining the preset light intensity through the lighting equipment, and setting and maintaining the preset temperature by the temperature monitor;

Step S14, carry out continuous cultivation, and maintain the cell concentration of photosynthetic green algae every day through the algae concentration monitor. When the cell concentration reaches the preset peak value, discharge the carbon sequestration algae liquid in the algae carbon capture tank into the high concentration algae liquid storage tank middle;

Step S15, discharge the carbon sequestration algae liquid in the high-concentration algae liquid storage tank into the filling equipment for filling to form carbon sequestration products.

Further, the step S12 specifically includes:

Step S121, cloning the cloned pinellia lectin gene into a high-expression plasmid with a His tag, and transforming it into a bacterial culture for preliminary culture, and then inducing it with lactose analogue IPTG for secondary culture;

Step S122, after high-speed centrifugation of the recultivated bacterial culture to form a cell pellet, the cell pellet is lysed with a lysis buffer of a mixture of Tris, NaCl and protease inhibitors to form a cell lysate;

In step S123, the cell lysate is sequentially purified by nickel affinity chromatography, ion exchange chromatography column and gel filtration column to obtain Pinellia lectin protein with a preset purity.

Further, the step S123 specifically includes:

After the cell lysate was purified by nickel affinity chromatography, the protein was eluted with 500 mM imidazole to form a preliminary eluate;

Inject the protein peak collection solution in the primary eluent into the ion exchange chromatography column, and use different concentrations of NaCl for gradient purification based on the surface ionic charge to form a second eluent;

Inject the eluent again into a gel filtration column for separation and purification to obtain Pinellia lectin protein with preset purity.

Further, in the step S11, the concentration of citric acid is 20 mg/L, the concentration of CaCl ₂ is 2 g/L, the concentration of MgSO ₄ ·7H ₂ O is 2 g/L, and the concentration of KH ₂ PO ₄ is 2 g/L. L, the concentration of CuSO ₄ ·5H ₂ O is 2 mg/L, and the concentration of EDTA-Fe is 20 mg/L.

Further, in the step S13, the light intensity is 2800 lux, and the temperature is 28°C.

Further, in the step S14, the preset peak value is 20 million/ml. Before reaching the preset peak value, the carbon sink strength of the carbon sequestration algae liquid increases with the increase of culture time. After reaching the preset peak value, continue to Cultivation will reduce the carbon sink intensity of the carbon sink algae liquid, and the best time to discharge the carbon sink algae liquid and repeat the culture step is 90 days.

In the third aspect, in the present invention, the application of the carbon sink algae liquid prepared by the above-mentioned carbon sink algae liquid based on photosynthetic green algae preparation method, the carbon sink algae liquid is evenly sprayed on the leaf surface of Quercus liaotungensis, and the carbon sink The higher the concentration of algae sink liquid, the stronger the photosynthesis of Quercus liaotungensis and the increase of carbon sink.

In the fourth aspect, in the present invention, the application of the carbon sink algae liquid prepared by the above-mentioned method for preparing the carbon sink algae liquid based on photosynthetic green algae is to spray the carbon sink algae liquid evenly on the leaf surface of the hawthorn tree, and the carbon sink The higher the concentration of algae sink, the stronger the photosynthesis of hawthorn, the increase of carbon sink, the sugar content of mature hawthorn and the content of unsaturated fatty acid increased.

Compared with the prior art, the present invention has the following advantages:

First, the equipment occupies a small area, simple structure and low cost;

Second, the carbon sequestration products after filling can be applied to the space outside the forest, so as to generate greater carbon sequestration intensity;

Third, the carbon dioxide captured by the carbon sink algae liquid becomes the nutrients needed for the life activities of photosynthetic green algae through photosynthesis, and the high concentration of photosynthetic green algae in the carbon sink algae liquid can be applied to the leaves of various trees, making trees The photosynthesis of leaves is improved, thereby increasing the carbon sink of the entire forest.

Instructions attached

Fig. 1 is a schematic block diagram of the carbon sink algae liquid production equipment based on photosynthetic green algae in the present invention.

The following specific embodiments will further illustrate the present invention in conjunction with the above-mentioned drawings.

Detailed ways

In order to facilitate the understanding of the present invention, the present invention will be described more fully below with reference to the associated drawings. Several embodiments of the invention are shown in the drawings. However, the present invention can be embodied in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided so that the disclosure of the present invention will be thorough and complete.

Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the technical field of the invention. The terms used herein in the description of the present invention are for the purpose of describing specific embodiments only, and are not intended to limit the present invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.

Please refer to Fig. 1, a kind of carbon sequestration algae liquid production equipment based on photosynthetic green algae provided in an embodiment of the present invention, comprises equipment cavity, and the algae carbon capture tank that is located in the equipment cavity;

It should be noted that, in the present invention, the volume of the algae carbon capture tank is not less than 1 m ³ , which is convenient for operation, storage and transportation.

Among the present invention, a kind of preparation method of the carbon sequestration algae liquid based on photosynthetic green algae, comprises the following steps:

(1), the air treatment equipment purifies the air inhaled from the outside of the equipment room, while maintaining the inhaled carbon dioxide concentration at 33%, nitrogen at 50%, ventilation rate at 200ml/min, and air pressure at 0.6Mpa;

(2), the water treatment equipment purifies the water, the pH value of the water is controlled at 7.5, and the TDS value is 20, and the purified water is added to the algae carbon capture tank through the water inlet pipe;

(3), 10L of photosynthetic green algae liquid is added to the algae carbon capture tank through the algae liquid inlet pipe;

(4), mix 20mg/L citric acid, 2g/L CaCl ₂ , 2g/L MgSO ₄ 7H ₂ O, 2g/L KH ₂ PO ₄ , 2mg/LCuSO ₄ 5H ₂ O, 20mg/LEDTA-Fe The water purified by the water treatment equipment is added to the algae carbon capture tank through the algae liquid pipe to serve as the nutrients needed for the growth of photosynthetic green algae, each component is 5g;

(5) Pinellia lectin was purified to promote cell division of photosynthetic green algae, and the Pinellia lectin gene was cloned into a high-expression plasmid with a His tag. Bacterial cultures transformed with a plasmid carrying the gene of interest were grown at 37°C to OD 0.4, induced with 0.2 mM lactose analogue IPTG, and then cultured overnight at 24°C. The next day, the bacterial culture was centrifuged at high speed, and the cell pellet was lysed with lysis buffer containing 20 mM Tris-pH 7.5, 150 mM NaCl, and protease inhibitor cocktail. Cell lysates will first be purified using nickel affinity chromatography and proteins will be eluted with 500 mM imidazole. The protein peak pool is then injected into an ion-exchange column to further purify the protein based on its surface ionic charge. During this step, proteins will be eluted from the column using a 100mM to 1M NaCl gradient. Finally, the eluate from the ion exchange column will be injected into the gel filtration column, which will further separate and purify the protein according to the molecular size. Finally, through this three-step purification scheme, high-purity pinellia lectin protein can be obtained. Mix the Pinellia lectin protein into the water purified by the water treatment equipment, and add the algae carbon capture tank through the algae liquid inlet pipe;

(6), the lighting equipment keeps the light intensity at 2800lux;

(7), the temperature monitor keeps the temperature at 28°C;

(8), the algae concentration detector detects the cell concentration of photosynthetic green algae every day;

(9), the whole process lasts for 20-100 days, and the algae liquid is discharged into the high-concentration algae liquid storage tank through the algae liquid outlet pipe;

(10), discharging the algae liquid in the high-concentration algae liquid storage tank into the solution tank of the filling equipment, and performing the filling operation to form a carbon sink product;

(11), repeat step (1) to step (10);

(12), measure step (1) to step (11), the maximum fixed carbon dioxide rate of photosynthetic green algae is 0.9-2g/(L.d).

What needs to be further clarified is that the present invention is to manufacture equipment with larger carbon sinks similar to forest carbon sinks in a narrower space. Wide application can increase my country's "plain forests", and the carbon dioxide captured at the same time will increase Through photosynthesis, it becomes the nutrients needed for the life activities of algae, and the high-concentration photosynthetic green algae produced by the carbon sink can be used in the forest to increase the photosynthesis of the leaves of the forest, thereby increasing the carbon sink of the forest.

The invention will be described in detail below with specific embodiments.

Example 1:

The overall process steps are: the air treatment equipment purifies the air inhaled from the outside of the equipment room, while maintaining the inhaled carbon dioxide concentration at 33%, nitrogen at 50%, ventilation rate at 200mL/min, and air pressure at 0.6Mpa; water treatment equipment purifies water , the water pH value is controlled at 7.5, and the TDS value is at 20; the purified water is added to the algae carbon capture tank through the water inlet pipe; 10L of photosynthetic green algae liquid is added to the algae carbon capture tank through the algae liquid inlet pipe; , 2g/L CaCl ₂ , 2g/L MgSO ₄ 7H ₂ O, 2g/L KH ₂ PO ₄ , 2mg/L CuSO ₄ 5H ₂ O, 20mg/L EDTA-Fe mixed into the purified water of the water treatment equipment, Add the algae carbon capture tank through the algae liquid tube; the pinellia lectin is purified, and the Pinellia lectin gene is cloned into a high-expression plasmid with a His tag. Bacterial cultures transformed with a plasmid carrying the gene of interest were grown at 37°C to OD 0.4, induced with 0.2 mM lactose analogue IPTG, and then grown overnight at 24°C. The next day, the bacterial culture was high-speed centrifuged and the cell pellet was lysed with lysis buffer containing 20 mM Tris-pH 7.5, 150 mM NaCl and protease inhibitor cocktail. Cell lysates will first be purified using nickel affinity chromatography and proteins will be eluted with 500 mM imidazole. The protein peak pool is then injected into an ion-exchange column to further purify the protein based on its surface ionic charge. In this step, the protein will be eluted from the column using a 100 mM to 1 M NaCl gradient. Finally, the eluate from the ion exchange column will be injected into the gel filtration column, which will further separate and purify the protein according to the molecular size. Finally, through this three-step purification scheme, high-purity pinellia lectin protein can be obtained. Mix the pinellia lectin protein into the water purified by the water treatment equipment, and add the algae carbon capture tank through the algae liquid pipe; the lighting equipment keeps the light The intensity is 2800lux; the temperature control equipment keeps the temperature at 28 degrees Celsius; the algae concentration detector detects the concentration of photosynthetic green algae cells every day; The algae liquid in the concentration algae liquid storage tank is discharged into the solution tank of the filling equipment, and the filling operation is carried out to form a carbon sink product.

Example 2:

Example 3:

The overall process steps are: the air treatment equipment purifies the air inhaled from the outside of the equipment room, while maintaining the inhaled carbon dioxide concentration at 33%, nitrogen at 50%, ventilation rate at 200mL/min, and air pressure at 0.6Mpa; water treatment equipment purifies water , the water pH value is controlled at 7.5, and the TDS value is at 20; the purified water is added to the algae carbon capture tank through the water inlet pipe; 10L of photosynthetic green algae liquid is added to the algae carbon capture tank through the algae liquid inlet pipe; , 2g/L CaCl ₂ , 2g/L MgSO ₄ 7H ₂ O, 2g/L KH ₂ PO ₄ , 2mg/L CuSO ₄ 5H ₂ O, 20mg/L EDTA-Fe mixed into the purified water of the water treatment equipment, Add the algae carbon capture tank through the algae liquid tube; the pinellia lectin is purified, and the Pinellia lectin gene is cloned into a high-expression plasmid with a His tag. Bacterial cultures transformed with a plasmid carrying the gene of interest were grown at 37°C to OD 0.4, induced with 0.2 mM lactose analogue IPTG, and then grown overnight at 24°C. The next day, the bacterial culture was high-speed centrifuged and the cell pellet was lysed with lysis buffer containing 20 mM Tris-pH 7.5, 150 mM NaCl and protease inhibitor cocktail. Cell lysates will first be purified using nickel affinity chromatography and proteins will be eluted with 500 mM imidazole. The protein peak pool is then injected into an ion-exchange column to further purify the protein based on its surface ionic charge. In this step, the protein will be eluted from the column using a 100 mM to 1 M NaCl gradient. Finally, the eluate from the ion exchange column will be injected into the gel filtration column, which will further separate and purify the protein according to the molecular size. Finally, through this three-step purification scheme, high-purity pinellia lectin protein can be obtained. Mix the pinellia lectin protein into the water purified by the water treatment equipment, and add the algae carbon capture tank through the algae liquid pipe; the lighting equipment keeps the light The intensity is 2800lux; the temperature control equipment keeps the temperature at 28 degrees Celsius; the algae concentration detector detects the concentration of photosynthetic green algae cells every day; The algae liquid in the concentration algae liquid storage tank is discharged into the solution tank of the filling equipment for filling operation to form a carbon sink product.

Example 4:

The basic overall process steps are: the air treatment equipment purifies the air inhaled from the outside of the equipment room, while maintaining the inhaled carbon dioxide concentration at 33%, nitrogen at 50%, ventilation rate at 200mL/min, and air pressure at 0.6Mpa; water treatment equipment purifies Water, the pH value of the water is controlled at 7.5, and the TDS value is 20; the purified water is added to the algae carbon capture tank through the water inlet pipe; 10L of photosynthetic green algae liquid is added to the algae carbon capture tank through the algae liquid inlet pipe; Acid, 2g/L CaCl ₂ , 2g/L MgSO ₄ 7H ₂ O, 2g/L KH ₂ PO ₄ , 2mg/LCuSO ₄ 5H ₂ O, 20mg/L EDTA-Fe are mixed into the purified water of the water treatment equipment, Add the algae carbon capture tank through the algae liquid tube; the pinellia lectin is purified, and the pinellia lectin gene is cloned into a high-expression plasmid with a His tag. Bacterial cultures transformed with a plasmid carrying the gene of interest were grown at 37°C to OD 0.4, induced with 0.2 mM lactose analogue IPTG, and then cultured overnight at 24°C. The next day, the bacterial culture was high-speed centrifuged and the cell pellet was lysed with lysis buffer containing 20 mM Tris-pH 7.5, 150 mM NaCl and protease inhibitor cocktail. Cell lysates will first be purified using nickel affinity chromatography and proteins will be eluted with 500 mM imidazole. The protein peak pool is then injected into an ion-exchange column to further purify the protein based on its surface ionic charge. In this step, the protein will be eluted from the column using a 100 mM to 1 M NaCl gradient. Finally, the eluate from the ion exchange column will be injected into the gel filtration column, which will further separate and purify the protein according to the molecular size. Finally, through this three-step purification scheme, high-purity pinellia lectin protein can be obtained. Mix the pinellia lectin protein into the water purified by the water treatment equipment, and add the algae carbon capture tank through the algae liquid pipe; the lighting equipment keeps the light The intensity is 2800lux; the temperature control equipment keeps the temperature at 28 degrees Celsius; the algae concentration detector detects the concentration of photosynthetic green algae cells every day; The algae liquid in the concentration algae liquid storage tank is discharged into the solution tank of the filling equipment for filling operation to form a carbon sink product.

Example 5:

Embodiment 6:

Embodiment 7:

The overall process steps are: the air treatment equipment purifies the air inhaled from the outside of the equipment room, while maintaining the inhaled carbon dioxide concentration at 33%, nitrogen at 50%, ventilation rate at 200mL/min, and air pressure at 0.6Mpa; water treatment equipment purifies water , the water pH value is controlled at 7.5, and the TDS value is at 20; the purified water is added to the algae carbon capture tank through the water inlet pipe; 10L of photosynthetic green algae liquid is added to the algae carbon capture tank through the algae liquid inlet pipe; , 2g/L CaCl ₂ , 2g/L MgSO ₄ 7H ₂ O, 2g/L KH ₂ PO ₄ , 2mg/L CuSO ₄ 5H ₂ O, 20mg/L EDTA-Fe mixed into the purified water of the water treatment equipment, Add the algae carbon capture tank through the algae liquid tube; the pinellia lectin is purified, and the Pinellia lectin gene is cloned into a high-expression plasmid with a His tag. Bacterial cultures transformed with a plasmid carrying the gene of interest were grown at 37°C to OD 0.4, induced with 0.2 mM lactose analogue IPTG, and then grown overnight at 24°C. The next day, the bacterial culture was high-speed centrifuged and the cell pellet was lysed with lysis buffer containing 20 mM Tris-pH 7.5, 150 mM NaCl and protease inhibitor cocktail. Cell lysates will first be purified using nickel affinity chromatography and proteins will be eluted with 500 mM imidazole. The protein peak pool is then injected into an ion-exchange column to further purify the protein based on its surface ionic charge. In this step, the protein will be eluted from the column using a 100 mM to 1 M NaCl gradient. Finally, the eluate from the ion exchange column will be injected into the gel filtration column, which will further separate and purify the protein according to the molecular size. Finally, through this three-step purification scheme, high-purity pinellia lectin protein can be obtained. Mix the pinellia lectin protein into the water purified by the water treatment equipment, and pass through the algal liquid inlet pipe

Add the algae carbon capture tank; the lighting equipment keeps the light intensity at 2800lux; the temperature control equipment keeps the temperature at 28 degrees Celsius; the algae concentration detector detects the concentration of photosynthetic green algae cells every day; the whole process lasts for 80 days, and the algae liquid is discharged through the algae liquid outlet pipe into the high-concentration algae liquid storage tank; the algae liquid in the high-concentration algae liquid storage tank is discharged into the solution tank of the filling equipment for filling operation to form a carbon sink product.

Embodiment 8:

Embodiment 9:

Test example 1:

By calculating the carbon fixation of photosynthetic green algae of embodiment 1-embodiment 9, the following data can be obtained:

It can be seen from the above table that due to the reproduction of the carbon sink algae cells themselves, the peak concentration is 20 million/ml.

Before reaching the peak, the carbon sink strength of algae increases with the increase of culture time. After reaching the peak, continuing to cultivate algae will reduce the carbon sink strength. The best time to remove algae liquid and repeat the culture step is 90 days.

Test example 2:

The carbon sequestration product high-concentration photosynthetic green algae obtained by embodiment 1-embodiment 9 is respectively applied to 1 mu of Quercus liaotungensis under the same management conditions, evenly sprayed on the leaves, and 1 liter is applied to each mu of land, and the annual carbon Mobvista, the following data can be obtained:

It can be seen from the above table that the higher the concentration of photosynthetic green algae, the more photosynthesis of trees can be increased, and the carbon sink of trees can be increased.

Test example 3

The high-concentration photosynthetic green algae of the carbon sink product obtained through Examples 1-9 are applied to Hawthorn. Select 2 identical areas under the same management conditions, each area is 1 mu, and one of the areas is fertilized normally (comparative example 1), and the other area is fertilized according to the conventional amount, and the leaves are sprayed with carbon sink products with high concentration of photosynthetic green algae 1L, after the hawthorn matured, the sugar content and unsaturated fatty acid content in the hawthorn in the two regions were measured respectively, and the average increase relative to Comparative Example 1 was calculated. The calculation results are shown in the following table:

It can be seen from the above table that the higher the concentration of photosynthetic green algae, the more the photosynthesis of hawthorn can be increased, the sugar content of hawthorn will be increased, and the effect of increasing unsaturated fatty acids will be more obvious.

In this specification, various embodiments are described in a progressive manner, each embodiment focuses on the difference from other embodiments, and the same or similar parts of various embodiments can be referred to each other. Moreover, the above-mentioned embodiments only express several implementation modes of the present invention, and the descriptions thereof are relatively specific and detailed, but should not be construed as limiting the patent scope of the present invention. It should be noted that, for those skilled in the art, several modifications and improvements can be made without departing from the concept of the present invention, and these all belong to the protection scope of the present invention. Therefore, the protection scope of the patent for the present invention should be based on the appended claims.

The above is only a preferred embodiment of the present invention, it should be pointed out that, for those of ordinary skill in the art, without departing from the principle of the present invention, some improvements and modifications can also be made, and these improvements and modifications can also be made. It should be regarded as the protection scope of the present invention.

Claims

A carbon sequestration algae liquid production device based on photosynthetic green algae, characterized in that: it includes a device cavity, and an algae carbon capture tank arranged in the device cavity;

The algae carbon capture tank is connected to the air treatment equipment through the air inlet pipe, connected to the water treatment equipment through the water inlet pipe, and connected to the high-concentration algae liquid storage tank through the algae liquid outlet pipe;

The algae carbon capture tank is provided with an algae concentration monitor, a temperature monitor and an algae liquid inlet pipe, and the algae liquid inlet pipe is used to add photosynthetic green algae liquid;

The top of the equipment cavity is provided with lighting equipment, and the high-concentration algae liquid storage tank is connected with the filling equipment.
The carbon sequestration algae liquid production equipment based on photosynthetic green algae according to claim 1, characterized in that the volume of the algae carbon capture tank is not less than 1 m 3 .
A method for preparing carbon sequestration algae liquid based on photosynthetic green algae, characterized in that it comprises the following steps:

Step S10, sending the purified air treated by the air treatment equipment into the algae carbon capture tank, and the purified water treated by the water treatment equipment into the algae carbon capture tank;

Step S11, after adding photosynthetic green algae liquid into the algae carbon capture tank, mixing citric acid, CaCl 2 , MgSO 4 7H 2 O, KH 2 PO 4 , CuSO 4 5H 2 O, EDTA-Fe and purified water , into the algae carbon capture tank;

Step S12, mixing the pinellia lectin protein after multi-step purification with purified water, and sending it into the algae carbon capture tank;

Step S13, setting and maintaining the preset light intensity through the lighting equipment, and setting and maintaining the preset temperature by the temperature monitor;

Step S14, carry out continuous cultivation, and detect the cell concentration of photosynthetic green algae every day through the algae concentration monitor. When the cell concentration reaches the preset peak value, discharge the carbon sequestration algae liquid in the algae carbon capture tank into the high concentration algae liquid storage tank middle;

Step S15, discharging the carbon sequestration algae liquid in the high-concentration algae liquid storage tank into the filling equipment for filling to form a carbon sink product.
The preparation method according to claim 3, characterized in that, in the step S10, the air treatment equipment purifies the air inhaled from the outside of the equipment room, while maintaining the inhaled carbon dioxide concentration at 33%, the nitrogen concentration at 50%, and ventilation The rate is 200mL/min, and the air pressure is 0.6Mpa.
The preparation method according to claim 3, characterized in that, in the step S10, the purified water treated by the water treatment equipment has a pH value of 7.5 and a TDS value of 20.
The preparation method according to claim 3, wherein said step S12 specifically comprises:

Step S121, cloning the cloned pinellia lectin gene into a high-expression plasmid with a His tag, and transforming it into a bacterial culture for preliminary culture, and then inducing it with lactose analogue IPTG for secondary culture;

Step S122, after high-speed centrifugation of the recultivated bacterial culture to form a cell pellet, the cell pellet is lysed with a lysis buffer of a mixture of Tris, NaCl and protease inhibitors to form a cell lysate;

In step S123, the cell lysate is sequentially purified by nickel affinity chromatography, ion exchange chromatography column and gel filtration column to obtain pinellia lectin protein with preset purity.
The preparation method according to claim 6, characterized in that, in the step S121, the temperature of the preliminary cultivation is 37°C, and the preliminary cultivation is carried out until the OD value is 0.4.
The preparation method according to claim 6, characterized in that, in the step S121, the concentration of the lactose analogue is 0.2 mM, and the re-cultivation is overnight at 24°C.
The preparation method according to claim 6, characterized in that, in the step S122, the pH value of Tris in the lysis buffer is 7.5, the concentration is 20 mM, and the concentration of NaCl is 150 mM.
The preparation method according to claim 6, wherein the step S123 specifically comprises:

After the cell lysate was purified by nickel affinity chromatography, the protein was eluted with 500 mM imidazole to form a preliminary eluate;

Inject the protein peak collection solution in the primary eluent into the ion exchange chromatography column, and use different concentrations of NaCl for gradient purification based on the surface ionic charge to form a second eluent;

Inject the eluate again into the gel filtration column for separation and purification to obtain Pinellia lectin protein with preset purity.
The preparation method according to claim 10, characterized in that the concentration range of the NaCl of different concentrations is 100mM～1M.
The preparation method according to claim 3, characterized in that, in the step S11, the concentration of citric acid is 20 mg/L, the concentration of CaCl 2 is 2 g/L, and the concentration of MgSO 4 ·7H 2 O is 2 g/L. L, the concentration of KH 2 PO 4 is 2g/L, the concentration of CuSO 4 ·5H 2 O is 2mg/L, and the concentration of EDTA-Fe is 20mg/L.
The preparation method according to claim 3, characterized in that, in the step S13, the light intensity is 2800 lux, and the temperature is 28°C.
The preparation method according to claim 3, characterized in that, in the step S14, the duration of the continuous culture is 20-100 days.
The preparation method according to claim 3, characterized in that, in the step S14, the preset peak value is 20 million/ml, and before reaching the preset peak value, the carbon sink intensity of the carbon sink algae liquid increases with the cultivation After reaching the preset peak value, continuing to cultivate will reduce the carbon sink strength of the carbon sink algae liquid, and the best time to discharge the carbon sink algae liquid and repeat the cultivation step is 90 days.
A carbon sink algae liquid prepared by the method for preparing photosynthetic green algae-based carbon sink algae liquid according to any one of claims 3 to 15.
An application of the carbon sequestration algae liquid as claimed in claim 16, characterized in that, the carbon sequestration algae liquid is evenly sprayed on the leaf surface of Quercus liaotungensis, the higher the concentration of the carbon sequestration algae liquid is, the higher the concentration of the carbon sequestration algae liquid is. The stronger the photosynthesis, the higher the carbon sink.
An application of the carbon sequestration algae liquid as claimed in claim 16, characterized in that the carbon sequestration algae liquid is evenly sprayed on the leaf surface of the hawthorn tree, the higher the concentration of the carbon sequestration algae liquid, the greater the yield of the hawthorn tree. The stronger the photosynthesis, the higher the carbon sink, the higher the sugar content and the unsaturated fatty acid content of mature hawthorn.