Method for producing main components of sustainable aviation fuel using photolyase decarboxylation reaction
By constructing and immobilizing a Chlorella fatty acid photodecarboxylase mutant, the problem of low catalytic efficiency of fatty acid photodecarboxylase was solved, achieving efficient conversion of long-chain fatty acids into hydrocarbons and producing high-purity alkane products suitable for industrial applications in sustainable aviation fuels.
Patent Information
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- TANGSHAN JINLIHAI BIODIESEL
- Filing Date
- 2026-03-10
- Publication Date
- 2026-07-14
AI Technical Summary
The low photocatalytic efficiency of existing fatty acid photodecarboxylases limits their large-scale industrial application in sustainable aviation fuel production.
By constructing a fatty acid photodecarboxylase mutant, screening for the best Chlorella, extracting the Chlorella fatty acid photodecarboxylase gene, constructing engineered bacteria, immobilizing the active fatty acid photodecarboxylase, and using the immobilized enzyme to convert long-chain fatty acids into a mixture of hydrocarbons, and finally separating and purifying them to generate alkane products.
It improves the operational stability and catalytic efficiency of enzymes, reduces production costs, meets the quality requirements of industrial applications for sustainable aviation fuel components, and is suitable for the conversion of various raw materials.
Smart Images
Figure CN122382149A_ABST
Abstract
Description
Technical Field
[0001] This invention belongs to the field of biocatalysis and green chemistry, and relates to the production of sustainable aviation fuel (SAF) and fine chemicals, especially a method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction. Background Technology
[0002] With the increasing global demand for carbon emission reduction, especially in the aviation industry, the development of carbon-neutral fuels has become crucial. Traditional aviation fuels have significant carbon emissions, and sustainable aviation fuel (SAF) is considered the most promising alternative to reduce these emissions. Current major SAF production technologies, such as hydrotreated fatty acids and esters (HEFA), while having a relatively mature technological foundation, still suffer from high energy consumption and low selectivity, resulting in significantly higher costs than traditional aviation fuels. Fatty acid photocatalytic decarboxylase technology, as an emerging photocatalytic method, has been found to decarboxylate long-chain fatty acids into alkane fuel molecules under mild conditions, thus providing another potential solution for the green production of SAF. However, the low photocatalytic efficiency of current fatty acid photocatalytic decarboxylases limits their large-scale industrial application. There is an urgent need for a method with high catalytic efficiency that can be applied on a large scale industrially to convert long-chain fatty acids into carbon-neutral liquid fuels and hydrocarbons via photocatalytic decarboxylation reactions, producing sustainable aviation fuels that meet demand. Summary of the Invention
[0003] To address the problem of low photocatalytic efficiency of fatty acid photodecarboxylases, a method is proposed that can efficiently convert long-chain fatty acids into carbon-neutral liquid fuels and hydrocarbons through photoenzymatic decarboxylation.
[0004] To achieve the above objectives, the present invention adopts the following technical solution: A method for producing key components of sustainable aviation fuel using photoenzymatic decarboxylation includes the following steps: S1: Construct a fatty acid photodecarboxylase mutant; screen for the optimal Chlorella strain; S2: Extract the Chlorella fatty acid photodecarboxylase gene from the selected optimal Chlorella species; S3: Construct engineered bacteria; S4: Extract active fatty acid photodecarboxylase from the engineered bacteria produced in step S3; S5: Immobilize the active fatty acid photodecarboxylase to obtain immobilized fatty acid photodecarboxylase; S6: Using immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into mixed products containing hydrocarbons; S7: Separate and purify the mixed products containing hydrocarbons to generate alkane products that can be used as components of aviation fuel mixtures.
[0005] Preferably, in step S1, constructing the fatty acid photodecarboxylase mutant specifically involves designing primers and using PCR to amplify the Chlorella fatty acid photodecarboxylase gene. The PCR amplification conditions are as follows: Pre-denaturation at 98℃ for 2 minutes; denaturation at 98℃ for 30 seconds, annealing at 55℃ for 30 seconds, extension at 72℃ for 1 minute, for a total of 30 cycles; final extension at 72℃ for 5 minutes.
[0006] Preferably, in step S3, the process of constructing engineered bacteria is as follows: The Chlorella fatty acid photodecarboxylase gene and a His-tagged expression plasmid were ligated, and the recombinant plasmid was then transformed into E. coli. High-efficiency expression was driven by the T7 promoter. Protein expression was induced by 0.5-1.0 mM isopropyl-β-D-thiogalactoside. LB medium containing 50 μg / mL kanamycin was used as the culture medium. The initial culture temperature was 37℃. When the OD600 reached 0.6-0.8, the temperature was lowered to 16-20℃ for induction. The stirring speed was 200 rpm, and the induction time was 4-16 hours to promote proper folding and improve the activity of Chlorella fatty acid photodecarboxylase.
[0007] Preferably, in step S4, the process of extracting active fatty acid photodecarboxylase is as follows: SS1: The engineered bacterial cells are lysed using ultrasound to release their contents and form a cell suspension; SS2: The lysed cell suspension was centrifuged to remove cell debris and precipitate, and a supernatant containing fatty acid photodecarboxylase was obtained. SS3: Gradually add ammonium sulfate to the supernatant until it reaches 30-60% saturation. After stirring evenly, let it stand at 4°C for 1 hour to allow fatty acid photodecarboxylase to precipitate. SS4: Centrifuge the fatty acid photodecarboxylase precipitate again, collect the fatty acid photodecarboxylase precipitate and dissolve it in phosphate buffer solution; SS5: Using Ni²⁺-NTA resin to bind the His tag of fatty acid photodecarboxylase, the bound fatty acid photodecarboxylase solution is added to a pre-packed affinity column and operated at 4°C to ensure the activity of fatty acid photodecarboxylase. SS6: Elute the enzyme solution with buffer solution and collect the eluent; SS7: DEAE-Sepharose is used as the anion exchange carrier to remove excess salt and buffer components; SS8: High-purity liquid active fatty acid photodecarboxylase was obtained by concentrating the eluent using an ultrafiltration membrane with a molecular weight cutoff of 30 kDa.
[0008] Preferably, in the process of extracting active fatty acid photodecarboxylase, the ultrasonic lysis parameters in step SS1 are a frequency of 20 kHz, a power of 300-400 W, a treatment time of 30 seconds, an interval of 30 seconds, and repeated 5-10 times to avoid protein denaturation. The pH was maintained at 7.5 using 50 mM phosphate buffer, and 1 mM protease inhibitor PMSF was added to protect fatty acid photodecarboxylase from degradation. In step SS2, the centrifugation speed was 12,000 rpm, the temperature was 4℃, and the time was 20 minutes; In step SS4, the centrifugation speed was 12,000 rpm, the temperature was 4℃, the time was 20 minutes, and the phosphate buffer solution was 50 mM. In step SS6, the buffer solution is an imidazole buffer containing 20-250 mM.
[0009] Preferably, the process of immobilizing the active fatty acid photodecarboxylase in step S5 is as follows: SS1: Amination of the carrier; SS2: A cross-linking agent is coated on the surface of the amination-treated carrier, and then an active fatty acid decarboxylase is added, so that the active fatty acid photodecarboxylase is covalently bonded to it to generate a solidified fatty acid decarboxylase.
[0010] Preferably, in the process of immobilizing the active fatty acid photodecarboxylase, the carrier amination process in step SS1 is as follows: the carrier is immersed in dimethyl sulfoxide containing cyanuric acid chloride and stirred at room temperature for 2-4 hours to ensure the binding of the activator to the carrier; The carrier was washed with anhydrous DMSO to remove unreacted activator. The activated carrier is immersed in a 5-10% ethylenediamine solution, maintaining a pH of 8-9, and the reaction is carried out with gentle stirring at room temperature or 30°C for 4-12 hours. After the reaction is complete, thoroughly wash the gel carrier with distilled water to remove any unreacted ethylenediamine residue; Treat the support surface with Ninhydrin solution and observe whether a blue-violet reaction occurs, indicating the presence of amine groups; The carrier is a polyacrylamide gel.
[0011] Preferably, step S6, which utilizes an immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, is as follows; SS1: Dissolve long-chain fatty acids in a mixture of water and ethanol, and use phosphate buffer to ensure a stable environment for enzyme activity; add immobilized fatty acid photodecarboxylase to carry out the conversion reaction; SS2: The reaction process is monitored online using a UV-Vis spectrometer or gas chromatography to track the consumption of long-chain fatty acids and the formation of products in real time; the light source is turned off after the reaction is completed to terminate the photocatalytic activity of the enzyme.
[0012] SS3: The product of centrifugation, which is separated into an organic phase and an aqueous phase; SS4: The organic solvent in the organic phase is recovered by rotary evaporator, and the hydrocarbons in the organic solvent are concentrated into crude product; SS5: Use silica gel column chromatography to purify hydrocarbons to produce high-purity alkane products.
[0013] Preferably, in step SS1, which uses an immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, the volume ratio of water to ethanol is 50:50, and the concentration is 10-50 mM. The phosphate buffer solution is 50 mM, pH 7.0; The concentration of immobilized fatty acid photodecarboxylase was controlled at 0.5-2.0 mg / mL; In step SS 3, the products are separated by centrifugation at 4,000 rpm for 10 minutes.
[0014] Preferably, the reaction conditions in step SS1, which uses immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, are as follows: Blue light with a wavelength of 450-480 nm is used as the excitation source; The light intensity was controlled within the range of 500-1000 μmol / m²·s; The concentration of substrate solutions containing long-chain fatty acids is 10-50 mM; The pH environment conditions are 6-8; Maintain the temperature between 20-30℃; The reaction time is 12-24 hours.
[0015] The beneficial effects of this invention: Directed evolutionary genetic engineering was used to obtain Chlorella fatty acid photodecarboxylases with higher photostability and a broader substrate spectrum. These photodecarboxylases maintained high catalytic activity under high substrate concentrations, and active fatty acid photodecarboxylases were extracted from engineered bacteria after PCR amplification. By immobilizing these active fatty acid photodecarboxylases, immobilized fatty acid photodecarboxylases were produced, improving the enzyme's operational stability, reusability, and catalytic efficiency under industrial reaction conditions.
[0016] The technology of this invention has the advantages of mild process, high carbon conversion rate and low production cost. The produced alkane products have high purity, which meets the quality requirements of sustainable aviation fuel components in industrial applications. It is suitable for the conversion of various raw materials such as waste lipids and industrial by-product fatty acids. Attached Figure Description
[0017] Figure 1 This is a process flow diagram of the present invention. Detailed Implementation
[0018] To facilitate understanding of the present invention, a more comprehensive description will be provided below. The present invention can be implemented in many different forms and is not limited to the embodiments described herein. Rather, these embodiments are provided to provide a thorough and complete understanding of the disclosure of the present invention. Unless otherwise defined, all technical and scientific terms used in this invention have the same meaning as commonly understood by one of ordinary skill in the art to which this invention pertains. The terminology used in this specification is for the purpose of describing particular embodiments only and is not intended to limit the invention.
[0019] This invention utilizes fatty acid photodecarboxylases to convert long-chain fatty acids (such as C16-C18) into hydrocarbons under mild conditions, forming components suitable for sustainable aviation fuels. Directed evolution technology was used to obtain a Chlorella fatty acid photodecarboxylase mutant with higher photostability and a broader substrate spectrum, maintaining high catalytic activity under high substrate concentrations. Active fatty acid photodecarboxylase was extracted from PCR-amplified engineered bacteria. Immobilization improved the operational stability, reusability, and catalytic efficiency of the active fatty acid photodecarboxylase under industrial reaction conditions. The activity of the immobilized fatty acid photodecarboxylase remained above 80% even after multiple uses.
[0020] The gene sequence of the Chlorella fatty acid photodecarboxylase mutant described in this invention is as follows:
[0021] The present invention will be further described in detail below with reference to specific embodiments: A method for producing key components of sustainable aviation fuel using photoenzymatic decarboxylation includes the following steps: S1: Construct a fatty acid photodecarboxylase mutant; screen for the optimal Chlorella strain; S2: Extract the Chlorella fatty acid photodecarboxylase gene from the selected optimal Chlorella species; S3: Construct engineered bacteria; S4: Extract active fatty acid photodecarboxylase from the engineered bacteria produced in step S3; S5: Immobilize the active fatty acid photodecarboxylase to obtain immobilized fatty acid photodecarboxylase; S6: Using immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into mixed products containing hydrocarbons; S7: Separate and purify the mixed products containing hydrocarbons to generate alkane products that can be used as components of aviation fuel mixtures.
[0022] In step S1, the construction of the fatty acid photodecarboxylase mutant specifically involves designing primers and using PCR to amplify the Chlorella fatty acid photodecarboxylase gene. The PCR amplification conditions are as follows: Pre-denaturation at 98℃ for 2 minutes; denaturation at 98℃ for 30 seconds, annealing at 55℃ for 30 seconds, extension at 72℃ for 1 minute, for a total of 30 cycles; final extension at 72℃ for 5 minutes.
[0023] In S3, the process of constructing the engineered bacteria was as follows: the Chlorella fatty acid photodecarboxylase gene and the expression plasmid containing the His tag were ligated, and then the recombinant plasmid was transformed into Escherichia coli, and high-efficiency expression was driven by the T7 promoter; protein expression was induced by 0.5-1.0 mM isopropyl-β-D-thiogalactoside (IPTG); LB medium containing 50 μg / mL kanamycin was used in the culture medium, the initial culture temperature was 37℃, and when the OD600 reached 0.6-0.8, the temperature was reduced to 16-20℃ for induction expression, the stirring speed was 200 rpm, and the induction time was 4-16 hours to promote correct folding and improve the activity of Chlorella fatty acid photodecarboxylase.
[0024] In step S4, the process of extracting active fatty acid photodecarboxylase is as follows: SS1: The engineered bacterial cells are lysed using ultrasound to release their contents and form a cell suspension; SS2: The lysed cell suspension was centrifuged to remove cell debris and precipitate, and a supernatant containing fatty acid photodecarboxylase was obtained. SS3: Gradually add ammonium sulfate to the supernatant until it reaches 30-60% saturation. After stirring evenly, let it stand at 4°C for 1 hour to allow fatty acid photodecarboxylase to precipitate. SS4: Centrifuge the fatty acid photodecarboxylase precipitate again, collect the fatty acid photodecarboxylase precipitate and dissolve it in phosphate buffer solution; SS5: Using Ni²⁺-NTA resin to bind the His tag of fatty acid photodecarboxylase, the bound fatty acid photodecarboxylase solution is added to a pre-packed affinity column and operated at 4°C to ensure the activity of fatty acid photodecarboxylase. SS6: Elute the enzyme solution with buffer solution and collect the eluent; SS7: DEAE-Sepharose is used as the anion exchange carrier to remove excess salt and buffer components; SS8: High-purity liquid active fatty acid photodecarboxylase was obtained by concentrating the eluent using an ultrafiltration membrane with a molecular weight cutoff of 30 kDa.
[0025] In the extraction of active fatty acid photodecarboxylase, the ultrasonic lysis parameters in step SS1 are: a frequency of 20 kHz, a power of 300-400 W, 30 seconds per treatment, a 30-second interval, and repeated 5-10 times to avoid protein denaturation; phosphate buffer (50 mM) is used to maintain the pH environment at 7.5, and 1 mM protease inhibitor PMSF is added to protect the fatty acid photodecarboxylase from degradation; in step SS2, the centrifugation speed is 12,000 rpm, the temperature is 4℃, and the time is 20 minutes; in step SS4, the centrifugation speed is 12,000 rpm, the temperature is 4℃, and the time is 20 minutes, with 50 mM phosphate buffer; in step SS6, the buffer is an imidazole (gradient elution) buffer containing 20-250 mM (50 mM phosphate buffer, pH 7.5).
[0026] The process of immobilizing the active fatty acid photodecarboxylase in step S5 is as follows: SS1: Amination of the carrier; SS2: A cross-linking agent is coated on the surface of the amination-treated carrier, and then an active fatty acid decarboxylase is added, so that the active fatty acid photodecarboxylase is covalently bonded to it to generate a solidified fatty acid decarboxylase.
[0027] In the process of immobilizing active fatty acid photodecarboxylase, the carrier amination process in step SS1 is as follows: the carrier is immersed in dimethyl sulfoxide containing cyanuric acid chloride and stirred at room temperature for 2-4 hours to ensure the binding of the activator to the carrier; the carrier is washed with anhydrous DMSO to remove unreacted activator; the activated carrier is immersed in 5-10% (v / v) ethylenediamine solution, maintaining a pH environment of pH 8-9, and gently stirred at room temperature or 30°C for 4-12 hours; after the reaction is completed, the gel carrier is thoroughly washed with distilled water to remove unreacted ethylenediamine residue; the surface of the carrier is treated with Ninhydrin solution, and the presence of a blue-purple reaction is observed, indicating the presence of amino groups; the carrier is a polyacrylamide gel.
[0028] Step S6, which utilizes immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, is as follows; SS1: Dissolve long-chain fatty acids in a mixture of water and ethanol, and use phosphate buffer to ensure a stable environment for enzyme activity; add immobilized fatty acid photodecarboxylase to carry out the conversion reaction; SS2: The reaction process is monitored online using a UV-Vis spectrometer or gas chromatography (GC) to track the consumption of long-chain fatty acids and the formation of products in real time; the light source is turned off after the reaction is completed to terminate the photocatalytic activity of the enzyme.
[0029] SS3: The product of centrifugation, which is separated into an organic phase and an aqueous phase; SS4: The organic solvent in the organic phase is recovered by rotary evaporator, and the hydrocarbons in the organic solvent are concentrated into crude product; SS5: Use silica gel column chromatography to purify hydrocarbons to produce high-purity alkane products.
[0030] In step SS1, which uses immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, the ratio of water to ethanol is 50:50 v / v, and the concentration is 10-50 mM; the phosphate buffer is 50 mM, pH 7.0; and the concentration of immobilized fatty acid photodecarboxylase is controlled at 0.5-2.0 mg / mL. In step SS3, the products are separated by centrifugation at 4,000 rpm for 10 minutes.
[0031] The reaction conditions in step SS1, which uses immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, are as follows: blue light with a wavelength of 450-480 nm is used as the excitation source; the light intensity is controlled within the range of 500-1000 μmol / m²·s; the concentration of the substrate solution containing long-chain fatty acids is 10-50 mM; NADH coenzyme may be added; the pH environment is 6-8; the temperature is maintained at 20-30℃; and the reaction time is 12-24 hours. As a preferred embodiment, the optimal reaction conditions in step SS1 are: blue light is set to a wavelength of 450 nm for excitation; the light intensity is controlled at 1000 μmol / m²·s; the concentration of the substrate solution containing long-chain fatty acids is 50 mM; no NADH coenzyme is added; the pH environment is 8; the temperature is 30℃; and the reaction time is 12 hours.
[0032] The reactor design for the reaction in which long-chain fatty acids are converted into a mixture containing hydrocarbons using immobilized fatty acid photodecarboxylase is as follows: A transparent quartz stirred photoreactor is used to ensure light transmission. A reflective surface and a light-diffusing plate are installed on the reactor exterior to improve light utilization and reduce light energy loss. The light source output is adjusted via a variable power supply, allowing for free setting of the light intensity. A temperature control device is installed in the reactor to set and maintain a constant temperature. Magnetic stirring ensures uniform distribution of the substrate and enzyme within the reactor, promoting effective contact.
[0033] In the embodiments of this application, it has been demonstrated that, under blue light (450 nm) irradiation, the fatty acid photodecarboxylase mutant can catalyze the decarboxylation of long-chain fatty acids to generate alkanes with one fewer carbon atom. For example, when palmitic acid is used as a substrate, pentadecane can be generated; when stearic acid is used as a substrate, heptadecane can be generated; when a mixture of fatty acids (palmitic acid, stearic acid, and oleic acid) is used as a substrate, the resulting alkanes are mainly distributed in the C15–C17 range, which can serve as an important component of sustainable aviation fuel.
[0034] Detailed data is shown in the table below: The embodiments described above are merely illustrative of several implementations of the present invention, and while the descriptions are relatively specific and detailed, they should not be construed as limiting the scope of the invention patent. It should be noted that those skilled in the art can make various modifications and improvements without departing from the concept of the present invention, and these all fall within the protection scope of the present invention. Therefore, the protection scope of this invention patent should be determined by the appended claims.
Claims
1. A method for producing the main components of sustainable aviation fuel using a photoenzymatic decarboxylation reaction, characterized in that, Includes the following steps: S1: Construct a fatty acid photodecarboxylase mutant; screen for the optimal Chlorella strain; S2: Extract the Chlorella fatty acid photodecarboxylase gene from the selected optimal Chlorella species; S3: Construct engineered bacteria; S4: Extract active fatty acid photodecarboxylase from the engineered bacteria produced in step S3; S5: Immobilize the active fatty acid photodecarboxylase to obtain immobilized fatty acid photodecarboxylase; S6: Using immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into mixed products containing hydrocarbons; S7: Separate and purify the mixed products containing hydrocarbons to generate alkane products that can be used as components of aviation fuel mixtures.
2. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 1, characterized in that, In step S1, constructing the fatty acid photodecarboxylase mutant specifically involves designing primers and using PCR to amplify the Chlorella fatty acid photodecarboxylase gene. The PCR amplification conditions are as follows: Pre-denaturation at 98℃ for 2 minutes; denaturation at 98℃ for 30 seconds, annealing at 55℃ for 30 seconds, extension at 72℃ for 1 minute, for a total of 30 cycles; final extension at 72℃ for 5 minutes.
3. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 1, characterized in that, In step S3, the process of constructing engineered bacteria is as follows: The Chlorella fatty acid photodecarboxylase gene and a His-tagged expression plasmid were ligated, and the recombinant plasmid was then transformed into E. coli. High-efficiency expression was driven by the T7 promoter. Protein expression was induced by 0.5-1.0 mM isopropyl-β-D-thiogalactoside. LB medium containing 50 μg / mL kanamycin was used as the culture medium. The initial culture temperature was 37℃. When the OD600 reached 0.6-0.8, the temperature was lowered to 16-20℃ for induction. The stirring speed was 200 rpm, and the induction time was 4-16 hours to promote proper folding and improve the activity of Chlorella fatty acid photodecarboxylase.
4. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 3, characterized in that, In step S4, the process of extracting active fatty acid photodecarboxylase is as follows: SS1: The engineered bacterial cells are lysed using ultrasound to release their contents and form a cell suspension; SS2: The lysed cell suspension was centrifuged to remove cell debris and precipitate, and a supernatant containing fatty acid photodecarboxylase was obtained. SS3: Gradually add ammonium sulfate to the supernatant until it reaches 30-60% saturation. After stirring evenly, let it stand at 4°C for 1 hour to allow fatty acid photodecarboxylase to precipitate. SS4: Centrifuge the fatty acid photodecarboxylase precipitate again, collect the fatty acid photodecarboxylase precipitate and dissolve it in phosphate buffer solution; SS5: Using Ni²⁺-NTA resin to bind the His tag of fatty acid photodecarboxylase, the bound fatty acid photodecarboxylase solution is added to a pre-packed affinity column and operated at 4°C to ensure the activity of fatty acid photodecarboxylase. SS6: Elute the enzyme solution with buffer solution and collect the eluent; SS7: DEAE-Sepharose is used as the anion exchange carrier to remove excess salt and buffer components; SS8: High-purity liquid active fatty acid photodecarboxylase was obtained by concentrating the eluent using an ultrafiltration membrane with a molecular weight cutoff of 30 kDa.
5. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 4, characterized in that, In the process of extracting active fatty acid photodecarboxylase, the ultrasonic lysis parameters in step SS1 are a frequency of 20 kHz, a power of 300-400 W, a treatment time of 30 seconds, an interval of 30 seconds, and repeated 5-10 times to avoid protein denaturation. The pH was maintained at 7.5 using 50 mM phosphate buffer, and 1 mM protease inhibitor PMSF was added to protect fatty acid photodecarboxylase from degradation. In step SS2, the centrifugation speed is 12,000 rpm, the temperature is 4℃, and the time is 20 minutes; In step SS4, the centrifugation speed was 12,000 rpm, the temperature was 4°C, the time was 20 minutes, and the phosphate buffer solution was 50 mM. In step SS6, the buffer solution is an imidazole buffer containing 20-250 mM.
6. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 1, characterized in that, The process of immobilizing the active fatty acid photodecarboxylase in step S5 is as follows: SS1: Amination of the carrier; SS2: A cross-linking agent is coated on the surface of the amination-treated carrier, and then an active fatty acid decarboxylase is added, so that the active fatty acid photodecarboxylase is covalently bonded to it to generate a solidified fatty acid decarboxylase.
7. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 6, characterized in that, In the process of immobilizing the active fatty acid photodecarboxylase, the carrier amination process in step SS1 is as follows: the carrier is immersed in dimethyl sulfoxide containing cyanuric acid chloride and stirred at room temperature for 2-4 hours to ensure the binding of the activator to the carrier. The carrier was washed with anhydrous DMSO to remove unreacted activator. The activated carrier is immersed in a 5-10% ethylenediamine solution, maintaining a pH of 8-9, and the reaction is carried out with gentle stirring at room temperature or 30°C for 4-12 hours. After the reaction is complete, thoroughly wash the gel carrier with distilled water to remove any unreacted ethylenediamine residue; Treat the support surface with Ninhydrin solution and observe whether a blue-violet reaction occurs, indicating the presence of amine groups; The carrier is a polyacrylamide gel.
8. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 1, characterized in that, The step S6, which utilizes immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, is as follows: SS1: Dissolve long-chain fatty acids in a mixture of water and ethanol, and use phosphate buffer to ensure a stable environment for enzyme activity; add immobilized fatty acid photodecarboxylase to carry out the conversion reaction; SS2: The reaction process was monitored online using a UV-Vis spectrometer or gas chromatography to track the consumption of long-chain fatty acids and the formation of products in real time; the light source was turned off after the reaction was completed to terminate the photocatalytic activity of the enzyme. SS3: The product of centrifugation, which is separated into an organic phase and an aqueous phase; SS4: The organic solvent in the organic phase is recovered by rotary evaporator, and the hydrocarbons in the organic solvent are concentrated into crude product; SS5: Use silica gel column chromatography to purify hydrocarbons to produce high-purity alkane products.
9. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 8, characterized in that, In step SS1, which uses immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, the volume ratio of water to ethanol is 50:50 and the concentration is 10-50 mM. The phosphate buffer solution is 50 mM, pH 7.0; The concentration of immobilized fatty acid photodecarboxylase was controlled at 0.5-2.0 mg / mL; In step SS 3, the products are separated by centrifugation at 4,000 rpm for 10 minutes.
10. The method for producing the main components of sustainable aviation fuel using photoenzymatic decarboxylation reaction according to claim 8, characterized in that, In step SS1, which utilizes immobilized fatty acid photodecarboxylase to convert long-chain fatty acids into a mixed product containing hydrocarbons, the reaction conditions are as follows: Blue light with a wavelength of 450-480 nm is used as the excitation source; The light intensity was controlled within the range of 500-1000 μmol / m²·s; The concentration of substrate solutions containing long-chain fatty acids is 10-50 mM; The pH environment conditions are 6-8; Maintain the temperature between 20-30℃; The reaction time is 12-24 hours.