Construction method and application of engineered Trichoderma reesei for producing lycopene

Abstract

The present invention discloses a method for constructing and applying an engineered Trichoderma reesei strain capable of producing lycopene, belonging to the field of bioengineering. This construction method uses the filamentous fungus Trichoderma reesei, which does not produce lycopene, or a derivative thereof, as a starting strain, and introduces exogenous genes encoding a single-function mutant of phytoene synthase and a phytoene dehydrogenase to obtain an engineered strain. Through genetic modification, the present invention obtains an engineered Trichoderma reesei strain capable of producing lycopene. The engineered strain uses glucose as a carbon source for direct fermentation to produce lycopene; it also uses lactose and cellulose as carbon sources for simultaneous fermentation to produce lycopene and cellulase. This engineered strain achieves dual production of enzymes and lycopene in a single fermentation process, increasing product profits and reducing production costs. This method also provides a new method for microbial production of lycopene, which can be applied to the industrial production of lycopene and cellulase.

Description

Technical Field

[0001] The present invention relates to the field of bioengineering, and in particular to a construction method and application of a Trichoderma reesei engineering bacterium for producing lycopene. Background Art

[0002] Lycopene is widely found in tomatoes, tomato products, and fruits such as watermelon and grapefruit. It is the primary pigment in ripe tomatoes. Lycopene's long-chain polyunsaturated olefin molecular structure gives it strong free radical scavenging and antioxidant properties. Current research on its biological effects focuses on antioxidant activity, reducing cardiovascular disease risk, reducing genetic damage, and inhibiting tumor development. Furthermore, the acidity of lycopene-rich tomato products lowers the pH of meat, inhibiting the growth of spoilage microorganisms to a certain extent. Therefore, lycopene can be used as a preservative and colorant in meat products, partially replacing nitrite. Consequently, lycopene is not only widely used as a natural pigment but is also increasingly being incorporated into functional foods, pharmaceuticals, and cosmetics.

[0003] Mammals cannot synthesize lycopene on their own and must obtain it from their diet. With increasing interest in dietary health, lycopene is becoming increasingly popular. Furthermore, it's also attracting attention as a feed additive. Studies have shown that lycopene, when used in aquaculture, can brighten the color of aquatic animals and improve their quality. It can also protect phagocytes from oxidative damage and promote lymphocyte proliferation, thereby enhancing the animal's immune system. This can effectively prevent and suppress disease in animals, replace or reduce the use of antibiotics, and mitigate food safety issues caused by feed safety concerns.

[0004] Currently, lycopene production through microbial fermentation offers a short production cycle, is not restricted by location or season, and uses mostly inexpensive food crops, resulting in low production costs and safety. With the deepening of research and the emergence of innovative products in various fields, lycopene has promising application prospects.

[0005] Advantages of Trichoderma reesei: As an important industrial production strain, a GRAS (Generally Regarded as Safe) strain, Trichoderma reesei is well-suited for industrial scale-up and has been widely used in the fermentation industry to produce various enzyme preparations needed in the food, feed, and other industries. However, since Trichoderma reesei strains cannot produce lycopene, the present invention uses genetic modification technology to construct an engineered Trichoderma reesei strain that can produce lycopene while fermenting enzyme preparations. Summary of the Invention

[0006] The purpose of the present invention is to provide a method for constructing and applying an engineered bacterium of Trichoderma reesei for producing lycopene, so as to solve the problems existing in the above-mentioned prior art. The engineered bacterium can simultaneously ferment and produce lycopene and cellulase, thereby realizing the dual production of enzyme and lycopene in a single fermentation process, thereby increasing product profits and reducing production costs.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The present invention provides a method for constructing an engineered Trichoderma reesei strain for producing lycopene. The method uses the filamentous fungus Trichoderma reesei, which does not produce lycopene, or a derivative of Trichoderma reesei as a starting strain, and introduces exogenous genes encoding a single-function mutant of phytoene synthase and a phytoene dehydrogenase to obtain an engineered strain.

[0009] Furthermore, the starting strains include Trichoderma reesei QM6a, QM9414, Rut-C30, RL-P37, NG14 and PC-3-7.

[0010] Furthermore, the amino acid sequence of the single-functional mutant of the phytoene synthase is shown in SEQ ID NO: 5, and the gene sequence encoding the single-functional mutant of the phytoene synthase is shown in SEQ ID NO. 6; the amino acid sequence of the phytoene dehydrogenase is shown in SEQ ID NO. 7, and the gene sequence encoding the phytoene dehydrogenase is shown in SEQ ID NO. 8.

[0011] The present invention also provides an engineered strain of Trichoderma reesei obtained according to the construction method.

[0012] The present invention also provides a method for producing lycopene, which comprises fermenting the engineered Trichoderma reesei, collecting mycelium, and obtaining lycopene.

[0013] Furthermore, the fermentation conditions are: the inoculation amount is 10 8 spores / 50 mL culture medium, the fermentation temperature was 25-30℃, and the rotation speed was 220 rpm.

[0014] Furthermore, the culture medium includes the following concentration components: carbon source 60-100 g / L, peptone 7-10 g / L, yeast powder 0.5-1 g / L, corn steep liquor 10-12 g / L, KH2PO4 6 g / L, (NH4)2SO4 5 g / L, CaCl2·2H2O 0.5 g / L, MgSO4·7H2O 1 g / L and 1 mL / L trace element solution.

[0015] Furthermore, the carbon source includes any one of glucose, lactose and cellulose; the trace element solution includes the following components by weight: 1.6 g MnSO4·4H2O, 5 g FeSO4·7H2O, 2 g CoCl2·6H2O and 1.4 g ZnSO4·7H2O, which are dissolved in water and diluted to 1 L.

[0016] The present invention also provides the use of the engineered Trichoderma reesei in producing lycopene and / or cellulase. The engineered Trichoderma reesei is fermented in the culture medium, and mycelium and fermentation liquid are collected to obtain lycopene and cellulase.

[0017] Furthermore, when the carbon source of the culture medium is glucose, lactose or cellulose, the mycelium is collected to obtain lycopene; when the carbon source of the culture medium is lactose or cellulose, the fermentation liquid is collected to obtain cellulase.

[0018] The present invention discloses the following technical effects:

[0019] The present invention uses the non-lycopene-producing filamentous fungus Trichoderma reesei as a starting strain. After genetic modification, exogenous genes were successfully introduced, enabling the modified strain to express a single-function mutant of phytoene synthase and phytoene dehydrogenase, thereby acquiring the ability to produce lycopene. Experiments have demonstrated that the modified engineered Trichoderma reesei can directly ferment lycopene using glucose as a carbon source, and simultaneously produce two products: lycopene and cellulase using lactose and cellulose as carbon sources. Lycopene accumulates within the mycelium, while the cellulase is secreted into the fermentation broth. Therefore, the two products can be separated through simple operations such as filtration and centrifugation. This engineered strain can achieve dual production of enzymes and lycopene during a single fermentation, increasing product profits and reducing production costs.

[0020] Furthermore, Trichoderma reesei is a strain that meets food safety standards, and its lycopene-containing mycelium can be added to animal feed as a raw material, leveraging the biological functions of lycopene and improving animal nutrition and disease resistance. This invention provides a new method for microbial lycopene production, applicable to the feed industry. It also provides a new companion product for the fermentation of enzyme preparations such as cellulase, potentially enabling the industrial production of lycopene and cellulase. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0022] Figure 1 Flowchart for the construction of plasmids and expression vectors in the present invention; A: Construction diagram of P1.0 and EP1.0 plasmids; B: Construction diagram of expression vectors PcarRPm and PcarB-carRPm;

[0023] Figure 2 The engineered strains of the present invention produce lycopene by fermenting glucose as a carbon source; A: mycelium of the engineered strain; B: yield of lycopene produced by different engineered strains by fermenting glucose;

[0024] Figure 3 The engineered strain of the present invention produces cellulase and lycopene by fermentation using lactose and cellulase as carbon sources; A: cellulase yield; B: lycopene yield. DETAILED DESCRIPTION

[0025] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but rather as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0026] It should be understood that the terms described herein are intended only to describe particular embodiments and are not intended to limit the present invention. In addition, for numerical ranges herein, it should be understood that each intermediate value between the upper and lower limits of the range is also specifically disclosed. Each smaller range between any intermediate value within a stated value or stated range and any other stated value or intermediate value within the stated range is also encompassed by the present invention. The upper and lower limits of these smaller ranges may be independently included or excluded within the scope.

[0027] Unless otherwise indicated, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art. Although only preferred methods and materials are described herein, any methods and materials similar or equivalent to those described herein may also be used in the practice or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of any conflict with any incorporated document, the contents of this specification shall prevail.

[0028] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments described herein without departing from the scope or spirit of the invention. Other embodiments will be apparent to those skilled in the art from the description of the invention. The description and examples are intended to be exemplary only.

[0029] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0030] The strains involved in the following examples are: Trichoderma reesei strain QM6a (purchased from American Type Culture Collection ATCC13631), QM9414 (purchased from American Type Culture Collection ATCC 26921), Rut-C30 (purchased from American Type Culture Collection ATCC 56765), RL-P37 (Agricultural Research Institute Type Culture Collection NRRL 15709), NG14 (purchased from American Type Culture Collection ATCC 56767), PC-3-7 (purchased from American Type Culture Collection ATCC 66589).

[0031] Example 1 Fermentation production of lycopene using genetically engineered strains

[0032] The genetically engineered bacteria were inoculated into 50 mL of a culture medium containing glucose, lactose, and cellulose as carbon sources (formula: carbon source 60 g / L, peptone 10 g / L, yeast powder 0.5 g / L, corn steep liquor 10 g / L, KH2PO4 6 g / L, (NH4)2SO4 5 g / L, CaCl2·2H2O 0.5 g / L, MgSO4·7H2O 1 g / L, 1 mL / L trace element solution. The formula of the trace element solution (1.6 g MnSO4·4H2O, 5 g FeSO4·7H2O, 2 g CoCl2·6H2O, 1.4 g ZnSO4·7H2O, dissolved in water and fixed to 1 L) in a 250 mL Erlenmeyer flask. The inoculation size was 10 8 The culture medium was incubated at 28°C and 220 rpm for 10 spores / 50 mL. Samples were taken on the 7th day to determine the lycopene content.

[0033] Example 2 Method for Determining Lycopene Content

[0034] 5 mL of the fermentation broth from Example 1 was centrifuged at 14,000 rpm for approximately 10 minutes. The supernatant was completely removed and the mycelium was collected. The supernatant contained secreted cellulase, which was used to measure cellulase activity. The enzyme activity measurement protocol can be found in previously published literature (Chen et al. Mn 2+ Lycopene is insoluble in water and cannot be secreted outside the cell. Instead, it accumulates in the mycelium, causing it to turn red.

[0035] An appropriate amount of ceramic beads was added to the mycelium, followed by 5 mL of acetone. The mycelium was then placed in a freeze grinder (JXFSTPRP-CL, Shanghai Jingxin Industrial Development Co., Ltd.) and thoroughly ground 4-6 times at 55 Hz for 300 s until the visible pigment was completely extracted. The mycelium was then centrifuged at 14,000 rpm for approximately 10 minutes to pellet the mycelial debris. The supernatant was used for lycopene quantification.

[0036] The treated samples were analyzed for lycopene content using high-performance liquid chromatography (HPLC). The mobile phase consisted of acetonitrile:methanol:isopropanol (5:3:2) at a flow rate of 1 mL / min. The column was a Wondasil C18 (4.6 mm × 150 mm, 5 μm particle size, GLSciences, Japan). The injection volume was 10 μL, the column temperature was set at 40°C, and the detection wavelength was 472 nm.

[0037] Example 3 Construction of expression plasmid EP1.0

[0038] 1. Use primers Ppdc-F and Ppdc-R to amplify the Ppdc sequence using the Trichoderma reesei genome as a template.

[0039] Ppdc-F: 5'-ACTAGTGAGTCATTTATGAAAGGAGGGAGCATTCTTCGA-3';

[0040] Ppdc-R: 5'-AAAGCCATTTAAATCATGATTGTGCTGTAGCTGCGC-3'.

[0041] Amplification reaction system: 10× PCR Buffer for KOD-Plus-Neo 5 μL; 2 mM dNTPs 5 μL; 25 mM MgSO4 3 μL; primers (10 μM each) 1.5 μL; genomic template (20 ng) 1 μL; KOD-Plus-Neo (1 U / μL) 1 μL.

[0042] Reaction program: 94°C for 2 min; 98°C for 10 sec, 58°C for 30 sec, 68°C for 45 sec, for 30 cycles; 68°C for 5 min.

[0043] 2. Using primers Tcbh2-1 and Tcbh2-2, amplify the Tcbh2 sequence using the Trichoderma reesei genome as a template;

[0044] Tcbh2-1: 5'-ATCATGATTTAAATGGCTTTCGTGACCGGGCTT-3';

[0045] Tcbh2-2: 5'-AGTGCCAAGCTTATTTTGGGTATGGTTTCCACGTGCA-3'.

[0046] Amplification reaction system: 10× PCR Buffer for KOD-Plus-Neo 5 μL; 2 mM dNTPs 5 μL; 25 mM MgSO4 3 μL; primers (10 μM each) 1.5 μL; genomic template (20 ng) 1 μL; KOD-Plus-Neo (1 U / μL) 1 μL.

[0047] Reaction program: 94°C for 2 min; 98°C for 10 sec, 58°C for 30 sec, 68°C for 15 sec, for 30 cycles; 68°C for 5 min.

[0048] 3. An expression vector was constructed using LML2.0a (Zhang et al. Light-inducible genetic engineering and control of non-homologous end-joining in industrial eukaryoticmicroorganisms: LML 3.0 and OFN 1.0. Scientific Reports. 2016, 6:20761) as the backbone. The existing plasmid LML2.0a was digested with restriction endonuclease SwaI and homologous recombination was performed using the Vazyme OneStep Clone Kit. The Ppdc and Tcbh2 sequences were ligated to obtain plasmid P1.0 ( Figure 1 Middle A).

[0049] Among them, the nucleotide sequence of Ppdc is shown as SEQ ID NO. 1, and the nucleotide sequence of Tcbh2 is shown as SEQ ID NO. 2.

[0050] 4. Use primers Peno-F and Peno-R to amplify the Peno sequence using the Trichoderma reesei genome as a template.

[0051] Peno-F: 5'-GATTACGAATTCTTAATGCCAACTCCTTGACGCCAA-3';

[0052] Peno-R: 5'-GGAGCTTTAATTAACATTTTGAAGCTATTTCAGGT-3'.

[0053] Amplification reaction system: 10× PCR Buffer for KOD-Plus-Neo 5 μL; 2 mM dNTPs 5 μL; 25 mM MgSO4 3 μL; primers (10 μM each) 1.5 μL; genomic template (20 ng) 1 μL; KOD-Plus-Neo (1 U / μL) 1 μL.

[0054] Reaction program: 94°C for 2 min; 98°C for 10 sec, 58°C for 30 sec, 68°C for 45 sec, for 30 cycles; 68°C for 5 min.

[0055] 5. Use primers Tcbh1-1 and Tcbh1-2 to amplify the Tcbh1 sequence using the Trichoderma reesei genome as a template.

[0056] Tcbh1-1: 5'-AAAATGTTAATTAAGCTCCCGTGGCGAAAGCC-3';

[0057] Tcbh1-2: 5'-CATTATACGAAGTTATTCTAGAATTTCCACTGTTGCTATTATGCTGT-3'.

[0058] Amplification reaction system: 10× PCR Buffer for KOD-Plus-Neo 5 μL; 2 mM dNTPs 5 μL; 25 mM MgSO4 3 μL; primers (10 μM each) 1.5 μL; genomic template (20 ng) 1 μL; KOD-Plus-Neo (1 U / μL) 1 μL.

[0059] Reaction program: 94°C for 2 min; 98°C for 10 sec, 58°C for 30 sec, 68°C for 15 sec, for 30 cycles; 68°C for 5 min.

[0060] 6. With P1.0 ( Figure 1 A) was used as the backbone to construct an expression vector. The plasmid P1.0 constructed above was double-digested with restriction endonucleases PacI / XbaI, and homologous recombination was performed using the Vazyme One Step Clone Kit. The Peno and Tcbh1 sequences were ligated to obtain plasmid EP1.0 ( Figure 1 A).

[0061] Among them, the nucleotide sequence of Peno is shown in SEQ ID NO.3, and the nucleotide sequence of Tcbh1 is shown in SEQ ID NO.4.

[0062] Example 4 Construction of an expression vector PcarB-carRPm for simultaneously expressing the single-function mutant encoding gene carRPm for phytoene synthase and the gene carB encoding phytoene dehydrogenase

[0063] 1. Using primers carRPm-1 and carRPm-2, a codon-optimized carRPm DNA fragment (synthesized by Shanghai Jierui Biotechnology Co., Ltd., the nucleotide sequence of which is shown in SEQ ID NO. 6) was used as a template to amplify the carRPm sequence.

[0064] carRPm-1: 5'-AGCTACAGCACAATCATGCTGCTGACCTACATGGAGG-3';

[0065] carRPm-2: 5'-CCGGTCACGAAAGCCTCAGATGGTGTTCAGGTTTCGC-3'.

[0066] Amplification reaction system: 10× PCR Buffer for KOD-Plus-Neo 5 μL; 2 mM dNTPs 5 μL; 25 mM MgSO4 3 μL; primers (10 μM each) 1.5 μL; synthetic plasmid template (10 ng) 1 μL; KOD-Plus-Neo (1 U / μL) 1 μL.

[0067] Reaction program: 94°C for 2 min; 98°C for 10 sec, 58°C for 30 sec, 68°C for 60 sec, for 30 cycles; 68°C for 5 min.

[0068] 2. Using the plasmid EP1.0 in Example 3 ( Figure 1 Middle B) was used as the backbone to construct an expression vector. The restriction endonuclease SwaI on the existing plasmid EP1.0 was digested, and homologous recombination was performed using the Vazyme One Step Clone Kit. The carRPm sequence was ligated to obtain the carRPm single gene expression vector PcarRPm ( Figure 1 Middle B).

[0069] Among them, the amino acid sequence of carRPm is shown as SEQ ID NO.5, and the nucleotide sequence of carRPm is shown as SEQ ID NO.6.

[0070] 3. Use primers carB-1 and carB-2 to amplify the carB sequence using a codon-optimized carB DNA fragment (synthesized by Shanghai Jierui Biotechnology Co., Ltd., the nucleotide sequence of which is shown in SEQ ID NO. 8) as a template.

[0071] carB-1: 5'-TGAAATAGCTTCAAAATGTCTAAGAAGCACATTGTTATCATTG-3';

[0072] carB-2: 5'-TTTCGCCACGGAGCTTCAGATGACGTTGGAGTTGTGG-3'.

[0073] Amplification reaction system: 10× PCR Buffer for KOD-Plus-Neo 5 μL; 2 mM dNTPs 5 μL; 25 mM MgSO4 3 μL; primers (10 μM each) 1.5 μL; synthetic plasmid template (10 ng) 1 μL; KOD-Plus-Neo (1 U / μL) 1 μL.

[0074] Reaction program: 94°C for 2 min; 98°C for 10 sec, 58°C for 30 sec, 68°C for 60 sec, for 30 cycles; 68°C for 5 min.

[0075] 4. With PcarRPm( Figure 1 Middle B) was used as the backbone to construct an expression vector. The restriction endonuclease PacI on the existing plasmid PcarRPm was digested, and homologous recombination was performed using the Vazyme One Step Clone Kit. The carB sequence was ligated to obtain the carB and carRPm dual gene expression vector PcarB-carRPm ( Figure 1 Middle B).

[0076] Among them, the amino acid sequence of carB is shown in SEQ ID NO.7, and the nucleotide sequence of carB is shown in SEQ ID NO.8.

[0077] Example 5 Construction of an engineered strain for lycopene production

[0078] The expression or heterologous expression involved in the present invention is Agrobacterium-mediated transformation of Trichoderma reesei and cloning screening, integrating the relevant genes into the Trichoderma reesei genome for expression. The transformation method involved in the present invention is Agrobacterium tumefaciens-mediated conjugative transfer.

[0079] 1. The expression vector PcarB-carRPm constructed in Example 4 was electroporated into Agrobacterium, and then the Agrobacterium containing the expression vector PcarB-carRPm was co-cultured with the Trichoderma reesei host strains QM6a (ATCC 13631), QM9414 (ATCC 26921), Rut-C30 (ATCC 56765), RL-P37 (NRRL 15709), NG14 (ATCC 56767), and PC-3-7 (ATCC 66589) on IM plates (Covert et al. Agrobacterium tumefaciens-mediated transformation of Fusarium circinatum. Mycol. Res. 105(3):259-264) to perform Agrobacterium tumefaciens-mediated conjugative transfer. After two days of co-cultivation, the transformants were transferred to culture medium containing cefotaxime (300 μg / mL) and hygromycin B (75 μg / mL) on PDA plates until transformants grew hyphae and spores, and then were screened and verified.

[0080] 2. The above-verified transformants were inoculated into 50 mL of culture medium with glucose as carbon source (see Example 1) in a 250 mL Erlenmeyer flask, with an inoculum size of 10 8 The culture medium was incubated at 28°C and 220 rpm with spores per 50 mL of culture medium. Samples were taken on the 7th day to determine the lycopene content.

[0081] 3. The engineered strains obtained by transforming the expression vector PcarB-carRPm into Trichoderma reesei can significantly produce lycopene, making the mycelium red ( Figure 2 The strain with the highest yield was the QM6a engineered strain obtained by transforming the expression vector into QM6a. When glucose was used as the carbon source, the lycopene yield of the QM6a engineered strain reached 60 mg / L ( Figure 2 Middle B).

[0082] 4. Rut-C30 engineered strain and PC-3-7 engineered strain can ferment and produce cellulase with common carbon sources such as lactose and cellulose ( Figure 3 A), while producing lycopene ( Figure 3 (B) The cellulase production level of the engineered strain was comparable to that of the original strain, indicating that the lycopene production process of the engineered strain did not affect its ability to secrete the expressed enzyme preparation.

[0083] As can be seen from the results of the above examples, the present invention successfully fermented and produced lycopene after genetically modifying Trichoderma reesei. The research results of the present invention are the first to show that although the original strain of Trichoderma reesei cannot produce lycopene, it can be fermented and produced lycopene using common carbon sources such as glucose, lactose, and cellulose as substrates after genetic engineering. Experiments have also confirmed that: while the engineered strain of Trichoderma reesei can induce secretion and expression of cellulase, it can also produce lycopene within the mycelium; the engineered strain can achieve dual production of enzymes and lycopene in a single fermentation process, increasing product profits and reducing production costs. Trichoderma reesei is a strain that meets food safety standards, and the lycopene-containing mycelium can be added as a raw material to animal feed to exert the biological function of lycopene and increase the nutrition and disease resistance of animals.

[0084] The embodiments described above are merely descriptions of preferred embodiments of the present invention and are not intended to limit the scope of the present invention. Without departing from the spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by persons skilled in the art shall fall within the scope of protection defined by the claims of the present invention.

Claims

1. A method for producing lycopene and cellulase, characterized in that: The engineered Trichoderma reesei bacteria are fermented in a culture medium, and mycelium and fermentation liquid are collected to obtain lycopene and cellulase; the culture medium uses cellulose as a carbon source; The method for constructing the engineered Trichoderma reesei comprises: using the filamentous fungus Trichoderma reesei, which does not produce lycopene, as a starting strain, introducing exogenous genes encoding a single-function mutant of phytoene synthase and a phytoene dehydrogenase to obtain an engineered strain; The amino acid sequence of the single-function mutant of the phytoene synthase is shown in SEQ ID NO.5, and the gene sequence encoding the single-function mutant of the phytoene synthase is shown in SEQ ID NO.6; the amino acid sequence of the phytoene dehydrogenase is shown in SEQ ID NO.7, and the gene sequence encoding the phytoene dehydrogenase is shown in SEQ ID NO.

8.

2. The method according to claim 1, characterized in that The starting strains include Trichoderma reesei QM6a, QM9414, Rut-C30, RL-P37, NG14 and PC-3-7.

3. The method according to claim 1, characterized in that The fermentation conditions are: the inoculum size is 10 8 spores / 50mL culture medium, the fermentation temperature is 25-30℃, and the rotation speed is 220rpm.

4. The method according to claim 3, characterized in that The culture medium comprises the following components in concentrations: 60-100 g / L of cellulose, 7-10 g / L of peptone, 0.5-1 g / L of yeast powder, 10-12 g / L of corn steep liquor, 6 g / L of KH2PO4, 5 g / L of (NH4)2SO4, 0.5 g / L of CaCl2·2H2O, 1 g / L of MgSO4·7H2O and 1 mL / L of trace element solution.

5. The method according to claim 4, characterized in that The trace element solution includes the following components by weight: 1.6 g MnSO4·4H2O, 5 g FeSO4·7H2O, 2 g CoCl2·6H2O and 1.4 g ZnSO4·7H2O, which are dissolved in water and diluted to 1 L.

6. Use of the engineered Trichoderma reesei according to claim 1 in producing lycopene and / or cellulase, characterized in that: The engineered Trichoderma reesei is fermented in the culture medium described in claim 4, mycelium and fermentation liquid are collected, and lycopene and cellulase are obtained; When the carbon source of the culture medium is glucose, lactose or cellulose, the mycelia are collected to obtain lycopene; when the carbon source of the culture medium is lactose or cellulose, the fermentation liquid is collected to obtain cellulase.

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