Lipase mutant, gene thereof and application thereof

Abstract

The application discloses a lipase mutant and a gene and application thereof, relates to the technical field of genetic engineering and enzyme engineering, and provides a lipase mutant TLL2AZL.The amino acid sequence of the lipase mutant TLL2AZL is shown as SEQ ID NO:2.The lipase mutant TLL2AZL has significantly improved thermal stability, can maintain more than 80% enzyme activity at 85 DEG C for 3 min, and has more than 50% stability after being treated at pH 7.0-10.0 for 1 h.The lipase mutant TLL2AZL has an optimal pH of 9.0, an optimal temperature of 60 DEG C, good stability, and a wide application prospect.

Description

Technical Field

[0001] This invention relates to the fields of genetic engineering and enzyme engineering technology, specifically to a lipase mutant, its gene, and its applications. Background Technology

[0002] Lipases are widely found in animals, plants, and microorganisms. Their function in organisms is primarily involved in lipid metabolism. In animals, lipases are mainly distributed in the pancreas and adipose tissue of higher animals; however, animal-derived lipases often contain other components such as hormones or viruses, making them unsuitable for food processing. Plant-derived lipases are extracted from seeds, but due to their low content and difficulty in extraction and separation, large-scale production is not feasible. Lipases are widely distributed in microorganisms. Because microorganisms have characteristics such as rapid growth, ease of modification, high yield of metabolites, and convenient separation, microbial-derived lipases are widely used in industrial production.

[0003] Thermomyces lanuginosus is a type of fungus with a high upper limit for growth temperature and thermophilic properties, widely distributed in nature. Its lipases also exhibit relatively good temperature tolerance and are widely used in various industrial sectors. In the feed industry, it degrades anti-nutritional factors and improves feed utilization; in the paper industry, it optimizes pre-bleaching processes and reduces pollution; in brewing and fermentation, it enhances flavor compound generation and raw material conversion; in the bioenergy and oil industries, it efficiently converts biomass and utilizes oils for high value; and in environmental remediation, it accelerates the decomposition of organic waste. Its thermal stability and multi-enzyme synergistic properties make it a core microbial resource for promoting green industrial upgrading, resource recycling, and sustainable development.

[0004] Researchers are currently using genetic engineering and protein engineering to modify lipases, optimizing their structure and performance to improve their catalytic efficiency, stability, and substrate specificity. These improvements not only help enhance the efficiency and product quality of existing industrial processes but also open up possibilities for developing new biotransformation technologies and products.

[0005] Chinese patent CN119144581A discloses a lipase mutant. Compared with wild-type lipase RL, the lipase mutant containing the P313L single-point mutation provided by this invention has significantly improved heat resistance. After treatment at 80℃ for 3 minutes, the enzyme activity residual rate is as high as 75.77%, but its thermal stability is still relatively poor.

[0006] Therefore, further improving lipases through genetic engineering, protein engineering, and other means, and optimizing their structure and performance to enhance their catalytic efficiency, stability, and substrate specificity is the current research focus. Summary of the Invention

[0007] The purpose of this invention is to provide a lipase mutant, its gene, and its applications.

[0008] To achieve the above-mentioned objectives, the technical solution of the present invention is as follows:

[0009] In a first aspect, the present invention provides a lipase mutant TLL2AZL, the amino acid sequence of which is shown in SEQ ID NO:2.

[0010] Specifically, the lipase mutant TLL2AZL is obtained by mutation based on the sequence shown in SEQ ID NO:1.

[0011] Furthermore, the mutation is to mutate glycine at position 109 to tryptophan (G109W), serine at position 116 to leucine (S116L), and aspartic acid at position 167 to tyrosine (D167Y) in the sequence shown in SEQ ID NO:1.

[0012] SEQ ID NO:1:

[0013] EVSQDLFNQFNLFAQYSAAAYCGKNNDAPAGTNITCTGNACPEVEKADATFLYSFEDSGVGDVTGFLALDNTNKLIVLSFRGSRSIENWIANLNFDLKEINDICSGCRGHDGFTSSWRSVADTLRQKVEDAVREH PDYRVVFTGHSLGALATVAGADLRGNGYDIDVFSYGAPRVGNRAFAEFLTVQTGGTLYRITHTNDIVPRLPPREFGYSHSSPEYWIKSGTLVPVTRNDIVKIEGIDATGGNNQPNIPDIPAHLWYFGLIGTCL.

[0014] SEQ ID NO:2:

[0015] EVSQDLFNQFNLFAQYSAAAYCGKNNDAPAGTNITCTGNACPEVEKADATFLYSFEDSGVGDVTGFLALDNTNKLIVLSFRGSRSIENWIANLNFDLKEINDICSGCRWHDGFTSLWRSVADTLRQKVEDAVREH PDYRVVFTGHSLGALATVAGADLRGNGYDIYVFSYGAPRVGNRAFAEFLTVQTGGTLYRITHTNDIVPRLPPREFGYSHSSPEYWIKSGTLVPVTRNDIVKIEGIDATGGNNQPNIPDIPAHLWYFGLIGTCL.

[0016] Secondly, the present invention provides a gene encoding the above-mentioned lipase mutant.

[0017] Specifically, the gene is tll2azl Gene, tll2azl The nucleotide sequence of the gene is shown in SEQ ID NO:3.

[0018] SEQ ID NO:3:

[0019] .

[0020] Thirdly, the present invention provides a gene engineering vector containing the above-mentioned genes.

[0021] Specifically, the genetic engineering vector is constructed by fusing the aforementioned gene with an expression vector.

[0022] Preferably, the expression vector is selected from one or more of pPIC9K, pET-28a, pEZZ18, pTA1529, pINIII-ompA, pUB110, pE194, pUCX05-bgaB, pHT304, pMK3, pPIC9, pHIL-S1, pPICZα, pYAM75P, PNZ8149-usp45, pQE2, pQE9, pQE30, pQE31, pQE32, pQE40, pQE70, pQE80, pRSET-A, pRSET-B, pRSET-C, pGEX-5X-1, pBV220, pBV221, pBV222, pTrc99A, pTwin1, pEZZ18, and pKK232-8.

[0023] Furthermore, the expression vector is pPIC9K.

[0024] According to some embodiments of the present invention, the gene engineering vector contains tll2azl Genes pPIC9K carrier, for pPIC9K - tll2azl .

[0025] According to some embodiments of the present invention, the method for preparing the gene engineering vector is as follows:

[0026] expression carrier pPIC9K Double enzyme digestion ( EcoR I + Not I ), and simultaneously encode the lipase mutant TLL2AZL tll2azl Gene (SEQ ID NO.3) double enzyme digestion ( EcoR I + Not I ),Will tll2azl Genes and expression vectors pPIC9K Connect, to obtain the content tll2azl Genes pPIC9K Carrier.

[0027] Fourthly, the present invention provides genetically engineered cells containing the above-mentioned genetic engineering vector.

[0028] Specifically, the genetically engineered cells are obtained by converting a genetically engineered vector into a host cell.

[0029] Preferably, the host cell includes Pichia pastoris cells, brewer's yeast cells, or polymorphonuclear yeast cells.

[0030] More preferably, the host cell is Pichia pastoris.

[0031] More preferably, the host cell is Pichia pastoris. GS115 .

[0032] Fifthly, the present invention provides a method for preparing a lipase mutant, the method comprising: culturing the above-mentioned genetically engineered cells.

[0033] Specifically, the culture method is as follows: the above-mentioned genetically engineered cells are seeded in BMGY culture medium and cultured at 30°C and 200 rpm for 48 hours with shaking.

[0034] In a sixth aspect, the present invention provides a genetically engineered preparation comprising one or more of the following: a culture of genetically engineered cells, a culture extract, cell fragments, bacterial cells, fermentation broth, fermentation broth precipitate, and lyophilized powder.

[0035] In a seventh aspect, the present invention provides the application of the above-mentioned lipase mutants, genes, genetic engineering vectors, genetic engineering cells, and genetic engineering preparations in food production, feed production, oil processing, pulping and papermaking, or textiles.

[0036] Specifically, when applied to feed production, it can degrade anti-nutritional factors and improve feed utilization.

[0037] Specifically, when applied to pulp and paper making or textiles, it can optimize the pre-bleaching process and reduce pollution.

[0038] Specifically, it is applied in food production, enhancing the generation of flavor compounds and the transformation of raw materials in the brewing and fermentation field.

[0039] Specifically, when applied to oil processing, it can efficiently convert biomass and utilize oils at high value.

[0040] Specifically, it can be applied to environmental governance, accelerate the decomposition of organic waste, and its thermal stability and multi-enzyme synergistic properties make it a core microbial resource for promoting green industrial upgrading, resource recycling and sustainable development.

[0041] The beneficial effects of this invention are as follows:

[0042] This invention obtains the lipase mutant TLL2AZL by mutating the sequence shown in SEQ ID NO.1 with G109W, S116L, and D167Y. The lipase mutant TLL2AZL exhibits significantly improved thermostability, maintaining over 80% enzyme activity after 3 minutes at 85°C; and maintaining over 50% stability after 1 hour of treatment at pH 7.0-10.0. The lipase mutant TLL2AZL of this invention possesses the following properties: optimal pH 9.0, optimal temperature 60°C, good stability, and broad application prospects. This invention not only promotes the optimization and upgrading of existing industrial processes but also opens up broad avenues for exploring novel biotransformation technologies. Attached Figure Description

[0043] Figure 1 The optimal pH for the recombinant lipase mutant.

[0044] Figure 2 pH stability of the recombinant lipase mutant.

[0045] Figure 3 The optimal temperature for the recombinant lipase mutant.

[0046] Figure 4 The thermostability of the recombinant lipase mutant. Detailed Implementation

[0047] To make the technical means, creative features, and achieved objectives and effects of this invention easier to understand, the invention is further illustrated below with specific embodiments. However, the following embodiments are merely preferred embodiments of this invention and not all embodiments. Other embodiments obtained by those skilled in the art based on the embodiments described herein without creative effort are all within the protection scope of this invention. Unless otherwise specified, the operating methods and equipment used in the following embodiments are conventional operating methods, and the materials and equipment used in each embodiment are the same.

[0048] Experimental materials and reagents:

[0049] 1. Strains and vectors: The lipase gene of this invention tll2azl The lipase genes tll2az and tll2azlh were synthesized by Beijing Ruiboxingke Biotechnology Co., Ltd., and were obtained by point mutation. The Pichia pastoris expression vector pPIC9K and strain GS115 were purchased from Invitrogen.

[0050] 2. Enzymes and other biochemical reagents: Endonuclease was purchased from TaKaRa, ligase was purchased from Invitrogen, and the rest were domestically produced reagents that can be purchased from ordinary biochemical reagent companies.

[0051] 3. Culture medium:

[0052] (1) Yeast culture medium YPD: 1% peptone, 0.5% yeast extract, 1% glucose, 2% agar, pH 7.0.

[0053] (2) Escherichia coli culture medium LB: 1% peptone, 0.5% yeast extract, 1% NaCl, pH 7.0.

[0054] (3) BMGY medium: 1% yeast extract, 2% peptone, 1.34% YNB, 0.00004% Biotin, 1% glycerol.

[0055] (4) BMMY medium: 1% yeast extract, 2% peptone, 1.34% YNB, 0.00004% Biotin, 0.5% methanol.

[0056] Note: Molecular biology experimental methods not specifically described in the following examples were performed in accordance with the specific methods listed in J. Sambrook's "Molecular Cloning: A Laboratory Manual" (3rd Edition), or according to the kit and product instructions.

[0057] Example 1: *Thermophilic filamentosa* (a type of filamentous fungus) Thermomyces lanuginosus lipase-encoding gene tll2azl Synthesis

[0058] This invention utilizes lipases derived from *Thermophilus spp.* tll2az Using the gene as a reference, its sequence (SEQ ID NO.1) was mutated into G109W, S116L, and D167Y, and insertions were added to the 5' and 3' ends of the mutated sequence, respectively. EcoR I and Not I Restriction enzyme sites were determined, and the sequence was sent to Beijing Ruiboxingke Biotechnology Co., Ltd. for artificial gene synthesis. The amino acid sequence of the artificially synthesized lipase mutant TLL2AZL is shown in SEQ ID NO.2. tll2azl The gene nucleotide sequence is shown in SEQ ID NO.3.

[0059] SEQ ID NO:1:

[0060] EVSQDLFNQFNLFAQYSAAAYCGKNNDAPAGTNITCTGNACPEVEKADATFLYSFEDSGVGDVTGFLALDNTNKLIVLSFRGSRSIENWIANLNFDLKEINDICSGCRGHDGFTSSWRSVADTLRQKVEDAVREH PDYRVVFTGHSLGALATVAGADLRGNGYDIDVFSYGAPRVGNRAFAEFLTVQTGGTLYRITHTNDIVPRLPPREFGYSHSSPEYWIKSGTLVPVTRNDIVKIEGIDATGGNNQPNIPDIPAHLWYFGLIGTCL.

[0061] SEQ ID NO:2:

[0062] EVSQDLFNQFNLFAQYSAAAYCGKNNDAPAGTNITCTGNACPEVEKADATFLYSFEDSGVGDVTGFLALDNTNKLIVLSFRGSRSIENWIANLNFDLKEINDICSGCRWHDGFTSLWRSVADTLRQKVEDAVREHPDYRVVFTGHSLGGALATVAGADLRGNGYDIYVFSYGAPRVGNRAFAEFLTVQTGGTLYRITHTNDIVPRLPPREFGYSHSSPEYWIKSGTLVPVTRNDIVKIEGIDATGGNNQPNIPDIPAHLWYFGLIGTCL。

[0063] SEQ ID NO:3:

[0064] GAAGTGTCACAAGATCTGTTTAACCAGTTTAATCTATTTGCCCAATATTCCGCTGCCGCATATTGTGGAAAGAACAACGACGCTCCCGCAGGCACCAACATTACATGTACAGGTAACGCATGTCCTGAAGTTGAGAAAGCTGATGCTACTTTCCTTTATTCTTTCGAAGATTCTGGAGTTGGTGATGTAACAGGCTTCCTTGCTCTGGACAACACTAACAAATTGATAGTCCTGTCTTTTCGTGGTTCTAGAAGTATCGAGAACTGGATTGCAAATCTAAACTTCGATTTGAAGGAAATCAACGACATTTGTTCCGGATGCAGATGGCATGATGGTTTCACTTCCCTGTGGAGATCCGTTGCAGATACATTGCGTCAAAAGGTTGAGGATGCTGTCAGAGAACATCCTGACTACCGTGTGGTTTTCACAGGTCATTCCCTAGGTGGAGCTTTGGCTACCGTGGCTGGTGCTGACTTAAGAGGCAACGGCTACGATATTTACGTCTTTTCATACGGTGCCCCCAGAGTCGGAAACCGTGCATTTGCCGAGTTTTTGACTGTGCAGACAGGAGGTACCTTGTATCGTATAACTCACACGAATGATATCGTCCCTAGACTACCACCTAGAGAGTTTGGCTACTCCCATTCATCTCCAGAATATTGGATTAAGTCCGGAACCTTGGTTCCTGTCACCAGAAACGACATCGTTAAGATTGAGGGTATAGATGCCACCGGAGGAAACAACCAGCCAAACATTCCTGACATCCCAGCCCACTTGTGGTACTTCGGTCTGATTGGAACTTGTCTT。

[0065] Example 2 Lipase gene tll2azl Cloning

[0066] Digest the expression vector pPIC9K with double restriction enzymes ( EcoR I + Not I ), and at the same time digest the tll2azl gene encoding lipase mutant TLL2AZL (SEQ ID NO.3) with double restriction enzymes ( EcoR I+ Not I ),Will tll2azl Genes and expression vectors pPIC9K Connect, to obtain the content tll2azl Genes pPIC9K The vector was preserved as a puncture bacteria. The puncture bacteria were picked up with a sterile toothpick in a clean bench and placed in an LB shaker containing antibiotic Kan (working concentration: 100 μg / ml). The culture was carried out overnight at 37°C and 220 rpm. The next day, the gene-containing vector was extracted according to the instructions of the Kangwei Century Plasmid Extraction Kit PurePlasmid Mini Kit (CW0500).

[0067] Based on the lipase gene sequence, the following primers were designed and synthesized:

[0068] P1 (SEQ ID NO.4):

[0069] CCGGAATTCCGGGAAGTTTCTCAAGATTTGTTTA;

[0070] P2 (SEQ ID NO.5):

[0071] TTATAGCGGCCGCATATTTTTACAAACAAGTACCAATCAAACCAAA.

[0072] PCR amplification was performed using the extracted vector as a template. The PCR reaction parameters were: denaturation at 94℃ for 5 min; followed by denaturation at 94℃ for 30 sec, annealing at 55℃ for 30 sec, extension at 72℃ for 1 min, and 32 cycles, followed by incubation at 72℃ for 10 min. A fragment of approximately 807 bp was obtained. This fragment was recovered, ligated into the pMD19 vector, and sent to Beijing Ruiboxingke Biotechnology Co., Ltd. for sequencing. The predicted protein molecular weight was 29.54 kDa.

[0073] Based on the nucleotide sequences obtained from sequencing, the DNAMan software was used to compare the obtained nucleotide sequences with... tll2azl The sequence (lipase sequence) was compared and confirmed to be correct.

[0074] Example 3 Preparation of recombinant lipase TLL2AZL

[0075] expression carrier pPIC9K Double enzyme digestion ( EcoR I + Not I ), and at the same time, the gene encoding the lipase mutant TLL2AZL tll2azl (SEQ ID NO.3) Double enzyme digestion ( EcoR I + Not I ), enzyme digestion to extract the gene fragment encoding lipase and expression vector pPIC9K Linkage to obtain a gene containing lipase. tll2azlrecombinant plasmid pPIC9K - tll2azl And transform Pichia pastoris GS115 Recombinant Pichia pastoris strain was obtained GS115 / TLL2AZL .

[0076] Take recombinant Pichia pastoris strain GS115 / TLL2AZL Compared with the control strain (i.e., the unmutated strain) GS115 / TLL2AZ The preparation method is the same as GS115 / TLL2AZL The cells were inoculated into 300 mL of BMGY medium and cultured at 30°C with shaking at 200 rpm for 48 h. The cells were then collected by centrifugation. The cells were then resuspended in 150 mL of BMGY medium and cultured at 30°C with shaking at 200 rpm. After induction for 72 h, the supernatant was collected by centrifugation to obtain recombinant lipase TLL2AZL and unmutated lipase TLL2AZ. Lipase activity was then measured.

[0077] Example 4 Activity analysis of recombinant lipase TLL2AZL

[0078] The method for determining lipase activity was as follows: 4 mL of olive oil emulsion was used as the substrate, 5 mL of Tris-HCl (pH=7.0, 50 mM) was used as the buffer, and 1 mL of appropriately diluted fermentation supernatant was added. The reaction was carried out at 40℃ for 15 min, then removed and placed on ice. 15 mL of 95% ethanol was added to terminate the reaction. 30 μL of phenolphthalein was added to each sample and control group as an indicator, and titrated with 50 mmol / L NaOH standard solution. The titration endpoint was determined when the reaction solution turned red. The lipase activity was calculated based on the volume difference of NaOH standard solution consumed by the sample and control groups. One unit of enzyme activity is defined as the amount of titratable fatty acid produced by hydrolyzing the substrate in 1 min under the above temperature and pH conditions through 1 mL of liquid enzyme.

[0079] The results showed that the expression level of recombinant lipase TLL2AZL was 2288 U / mL, while the expression level of non-mutated lipase TLL2AZ was 1480 U / mL.

[0080] Example 5: Determination of the optimal pH and pH stability of recombinant lipase TLL2AZL

[0081] The enzymatic properties of recombinant lipase TLL2AZL and unmutated lipase TLL2AZ were determined and compared. Simultaneously, a mutant recombinant lipase TLL2AZLH was added for enzymatic property comparison. Recombinant lipase TLL2AZLH was modified from SEQ ID NO. 1 by incorporating the G109A and D167P mutations. The specific preparation methods for the TLL2AZL mutant and recombinant enzyme TLL2AZLH are as described in the examples above.

[0082] 1. Determination of optimal pH

[0083] Purified unmutated lipase TLL2AZ, recombinant lipase TLL2AZL, and recombinant lipase TLL2AZLH were subjected to enzymatic reactions at different pH values ​​for 15 min to determine their optimal pH. The enzyme activities of the lipases were determined by alkaline titration at pH values ​​of 6.5–8.0 (phosphate buffer), 8.0–9.5 (Tris-HCl buffer), and 9.5–11.0 (glycine-sodium hydroxide buffer) to explore their optimal reaction pH.

[0084] The measurement results are as follows Figure 1 As shown, the optimal pH for unmutated lipase TLL2AZ, recombinant lipase TLL2AZL, and recombinant lipase TLL2AZLH is 9.0. Among them, recombinant lipase TLL2AZL exhibits over 50% relative enzyme activity at pH 8.0-10.0, while unmutated lipase TLL2AZ and recombinant lipase TLL2AZLH show less than 50% activity at pH 8.0, indicating that the recombinant lipase TLL2AZL has a significantly wider pH range of application.

[0085] 2. Determination of pH stability

[0086] Purified unmutated lipase TLL2AZ, recombinant lipase TLL2AZL, and recombinant lipase TLL2AZLH were treated at 37°C for 1 h in pH 6.5-8.0 (phosphate buffer), pH 8.0-9.5 (Tris-HCl buffer), or pH 9.5-11.0 (glycine-sodium hydroxide buffer), and then the enzyme activity was measured at 60°C in a pH 9.0 buffer system to study the pH tolerance of the enzymes.

[0087] The results are as follows Figure 2 As shown, the recombinant lipase TLL2AZL is very stable between pH 7.0 and 10.0. After treatment within this pH range for 1 hour, the remaining enzyme activity is more than 50%, which is significantly higher than that of TLL2AZ and TLL2AZLH. This indicates that the recombinant enzyme has good pH stability over a wide pH range.

[0088] Example 6: Determination of the optimal temperature and thermal stability of recombinant lipase TLL2AZL

[0089] 1. Determination of optimal temperature

[0090] Purified unmutated lipase TLL2AZ, recombinant lipase TLL2AZL, and recombinant lipase TLL2AZLH were reacted at different temperatures for 15 min in a phosphate buffer system at pH 8.0, and then their enzyme activities were measured at 60 °C.

[0091] The results of the optimal temperature measurement are as follows:Figure 3 As shown, the results indicate that the optimal temperature for unmutated lipase TLL2AZ, recombinant lipase TLL2AZL, and recombinant lipase TLL2AZLH is 60℃, and recombinant lipase TLL2AZL maintains high enzyme activity at 50-70℃.

[0092] 2. Thermal stability determination

[0093] The temperature resistance was determined by treating purified unmutated lipase TLL2AZ, recombinant lipase TLL2AZL, and recombinant lipase TLL2AZLH at 85°C for 3 min in a phosphate buffer solution at pH 7.0, followed by enzyme activity determination at 60°C.

[0094] The results of the enzyme's thermostability test are as follows: Figure 4 As shown, the recombinant lipase TLL2AZL exhibits good thermostability, retaining over 80% of its activity after incubation at 85℃ for 3 minutes. In contrast, the unmutated TLL2AZ retains less than 60% of its activity, and the recombinant lipase TLL2AZLH retains only 68%. This demonstrates that TLL2AZL exhibits significantly improved thermostability and excellent heat resistance.

[0095] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the protection scope of the present invention.

Claims

1. A lipase mutant TLL2AZL, characterized in that, The amino acid sequence of the lipase mutant TLL2AZL is shown in SEQ ID NO:

2.

2. The gene encoding the lipase mutant of claim 1.

3. The gene according to claim 2, characterized in that, The nucleotide sequence of the gene is shown in SEQ ID NO:

3.

4. A gene engineering vector containing the gene described in any one of claims 2-3.

5. The gene engineering vector according to claim 4, characterized in that, The genetic engineering vector is constructed by fusing the gene described in any one of claims 2-3 with an expression vector.

6. Genetically engineered cells containing the gene engineering vector according to any one of claims 4-5, characterized in that, The genetically engineered cells are obtained by transforming a genetically engineered vector into a host cell, wherein the host cell includes Pichia pastoris cells, brewer's yeast cells, or polymorphonuclear yeast cells.

7. The method for preparing the lipase mutant TLL2AZL according to claim 1, characterized in that, The preparation method includes: culturing the genetically engineered cells as described in claim 6.

8. A genetically engineered preparation, characterized in that, The genetically engineered preparation comprises one or more of the following: cell fragments of the genetically engineered cells as described in claim 6, fermentation broth, fermentation broth precipitate, and lyophilized powder.

9. The use of the lipase mutant TLL2AZL of claim 1, the gene of any one of claims 2-3, the gene engineering vector of any one of claims 4-5, the gene engineering cell of claim 6, and the gene engineering preparation of claim 8 in food production, feed production, oil processing, pulp and paper making, or textiles.

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