Aspergillus tubingensis, microbial agent, biomass fermentation product, and preparation and use thereof

Abstract

Provided are a strain of Aspergillus tubingensis AMCC 20011, a microbial agent containing the strain and a preparation method therefor, a fermentation culture product of the strain and a preparation method therefor, and use of the strain or the microbial agent. The strain is deposited in the China Center for Type Culture Collection (CCTCC) with a deposit number of CCTCC NO: M 20241203. The strain has the ability to produce cellulase, β-glucosidase, and xylanase, has the characteristic of saccharifying biomass, and has the characteristic of improving the protein content in the biomass by means of synergistic fermentation with yeast.

Description

A type of Aspergillus tabineus, inoculant, biomass fermentation product, its preparation and application

[0001] Related applications

[0002] This application claims priority to the earlier application No. 202411653202.6 filed with the China National Intellectual Property Administration on November 18, 2024, the entire contents of which are incorporated herein by reference. Technical Field

[0003] This invention belongs to the field of microbiology, specifically relating to Aspergillus tabineum, inoculants, biomass fermentation products, their preparation and application. Background Technology

[0004] Aspergillus tubingensis is a safe, traditionally used microorganism for fermenting foods, primarily listed in the "List of Traditional Fermented Food Microorganisms in China" for its application in tea fermentation. In the fermentation of Pu-erh tea and green brick tea, Aspergillus tubingensis can shorten the fermentation cycle while maintaining a similar taste to traditional fermentation methods. Furthermore, it can enzymatically degrade bitter substances in the fermentation of citrus plants and rutin, thereby improving bitterness. In addition, Aspergillus tubingensis is known for its ability to degrade plastics, possessing the capacity to produce pectinase, tanninase, phytase, and laccase; it also has the ability to degrade lignocellulose. However, current technologies for the biotransformation and utilization of plant biomass using Aspergillus tubingensis still have limitations.

[0005] Plant biomass refers to organisms formed through photosynthesis. Plant biomass includes straw, sugarcane bagasse, bran, distiller's grains, tea leaves, etc. Plant biomass has a high cellulose content, mainly containing cellulose, hemicellulose, and lignin. However, it has a low content of nitrogen-free extracts (starch, sugars). Furthermore, most strains lack the ability to produce cellulase, hemicellulase, and ligninase, and most strains rely heavily on glucose or starch during fermentation using carbon sources. Insufficient utilization of pentoses leads to carbon metabolism repression, resulting in low saccharification and fermentation efficiency in most strains during plant biomass fermentation. Additionally, although some strains have the ability to produce cellulase, the cellobiose and glucose produced during the decomposition of lignocellulose by cellulase alone can have a feedback inhibitory effect on the cellulase.

[0006] In addition, during the pretreatment of plant biomass using physical or chemical methods, organic inhibitors such as weak acids, aldehydes, and phenols that inhibit the growth of bacterial strains are generated, which can reduce fermentation efficiency to some extent.

[0007] Therefore, in the process of high-value utilization of plant biomass, by developing a food-grade Aspergillus fermentation method for plant biomass, we can achieve deep processing, transformation, and utilization of plant biomass to produce high-value-added products, improve the utilization efficiency and value of plant biomass, maximize resource utilization, and meet the needs in the fields of medicine, food, and agriculture. Summary of the Invention

[0008] The problems with the existing technology are that most strains used for fermenting plant biomass have poor ability to produce cellulase, hemicellulase and ligninase. At the same time, most strains used for fermenting plant biomass rely heavily on glucose or starch during the fermentation process and do not make sufficient use of pentose sugars, resulting in carbon metabolism inhibition. This leads to low saccharification efficiency and low fermentation efficiency when fermenting plant biomass.

[0009] In view of the problems existing in the prior art mentioned above, the present invention provides Aspergillus tabineus, inoculum, biomass fermentation product, and its preparation and application.

[0010] Specifically, the present invention provides the following technical solution:

[0011] In a first aspect, the present invention provides a *Aspergillus tubingensis*, specifically *Aspergillus tubingensis* AMCC 20011, which is deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCCNO: M 20241203.

[0012] Preferably, the ITS gene sequence of the Aspergillus tabingensis strain is shown in SEQ ID NO.3.

[0013] Preferably, the BenA gene sequence of Aspergillus tabineum AMCC 20011 is shown in SEQ ID NO.6.

[0014] Preferably, the Aspergillus tabingensis strain has the characteristics of producing cellulase, and / or producing β-glucosidase, and / or producing xylanase, and / or saccharifying biomass, and / or co-fermenting with yeast to increase the protein content in biomass.

[0015] Preferably, the filter paper enzyme activity of cellulase produced by Aspergillus tabineis is 15-17 U / mL, wherein the amount of enzyme required to degrade filter paper and release 1 μmol of glucose per minute per milliliter of crude enzyme solution of Aspergillus tabineis is defined as one unit of enzyme activity of filter paper enzyme.

[0016] Preferably, the β-glucosidase produced by Aspergillus tabineis has an enzyme activity of 1.5-1.55 U / mL. One unit of β-glucosidase activity is defined as the amount of enzyme required to produce 1 μmol of p-nitrophenol per minute by enzymatically hydrolyzing a p-nitrophenyl-β-D-glucopyranoside solution with a concentration of 5 mmol / L and a pH of 5.0 per milliliter of crude enzyme solution from Aspergillus tabineis.

[0017] Preferably, the xylanase produced by Aspergillus tabineum has an enzyme activity of 437-445 U / mL, wherein one unit of xylanase activity is defined as the amount of enzyme required to release 1 μmol of xylose per minute from a xylan solution with a concentration of 10 mg / mL per milliliter of crude enzyme solution from Aspergillus tabineum.

[0018] Secondly, the present invention provides a tabine aspergillus agent containing the aforementioned tabine aspergillus.

[0019] Thirdly, the present invention provides a method for preparing the aforementioned Aspergillus tabinea agent, comprising the following steps: amplifying and culturing the aforementioned Aspergillus tabinea.

[0020] Preferably, the amplification culture temperature is 10-37℃.

[0021] Fourthly, the present invention provides a feed containing the aforementioned Aspergillus tabineus strain, or the aforementioned Aspergillus tabineus agent, or the Aspergillus tabineus agent prepared by the aforementioned preparation method.

[0022] Fifthly, the present invention provides a fermented tea containing the aforementioned Aspergillus tabineus strain, or the aforementioned Aspergillus tabineus agent, or the Aspergillus tabineus agent prepared by the aforementioned preparation method.

[0023] In a sixth aspect, the present invention provides a biomass fermentation product containing the aforementioned Aspergillus tabineus strain, or the aforementioned Aspergillus tabineus agent, or the Aspergillus tabineus agent prepared by the aforementioned preparation method.

[0024] In a seventh aspect, the present invention provides a method for preparing the biomass fermentation product, comprising the following steps: culturing the Aspergillus tabineus strain, or the Aspergillus tabineus agent, or the Aspergillus tabineus agent prepared by the preparation method.

[0025] Preferably, the preparation method includes the following steps:

[0026] (1) The Aspergillus tabineus was amplified and cultured to obtain a suspension of Aspergillus tabineus spores;

[0027] (2) The Aspergillus tabingensis spore suspension obtained in step (1) is inoculated into a fermentation medium containing plant biomass for fermentation culture to obtain biomass fermentation product.

[0028] Preferably, the concentration of the Aspergillus tabineus spore suspension obtained in step (1) is adjusted to 6 × 10⁻⁶. 6 -6×10 8 After obtaining cfu / mL, the mixture is inoculated into a fermentation medium containing plant biomass for fermentation to obtain biomass fermentation product; more preferably, the fermentation temperature is 10-37℃ and / or the fermentation time is 15-30h.

[0029] Preferably, the fermentation medium containing plant biomass comprises: plant biomass and water, and preferably, the weight ratio of plant biomass to water is 1-1.5:1.

[0030] Preferably, in step (2), the inoculation amount of Aspergillus tabineus spore suspension is: 5-10 mL of Aspergillus tabineus spore suspension per 100 g of fermentation medium containing plant biomass.

[0031] Eighthly, the present invention provides a compound biomass fermentation product containing the aforementioned Aspergillus tabineus strain, or the aforementioned Aspergillus tabineus agent, or the Aspergillus tabineus agent prepared by the aforementioned preparation method, and yeast.

[0032] Preferably, the yeast is from the genera *Saccharomyces*, *Kluyveromyces*, *Pichia*, *Cyberlindnera*, *Arxiozyma*, *Cylindricascospora*, *Cyniclomyces*, *Eremothecium*, *Grigorovia*, *Hagleromyces*, *Henningerozyma*, *Huiozyma*, *Jamesozyma*, or *Kazachstania*. One or more of the following genera: *Cea*, *Maudiozyma*, *Monosporozyma*, *Naumovozyma*, *Nakaseomyces*, *Oligophagozyma*, *Savitreea*, *Sungouiozyma*, *Stenotrophomyces*, *Tetrapisispora*, *Torulaspora*, *Vanderwaltozyma*, *Yueomyces*, *Zygosaccharomyces*, and *Zygotorulaspora*.

[0033] Preferably, the yeast includes one or more species from the group consisting of the genera *Saccharomyces*, *Kluyveromyces*, *Pichia*, and *Cyberlindnera*.

[0034] More preferably, the yeast genus (Saccharomyces) is Saccharomyces cerevisiae;

[0035] Most preferably, the brewing yeast is brewing yeast (Saccharomyces cerevisiae) d5.12, with the preservation number CCTCC NO: M 2016460.

[0036] Preferably, the crude protein content is 30% or more based on the weight of the compound biomass fermentation product, and more preferably, the crude protein content is 30-36%.

[0037] In a ninth aspect, the present invention provides a method for preparing the aforementioned composite biomass fermentation product, characterized by comprising the following steps: culturing the aforementioned Aspergillus tabineus strain, or the aforementioned Aspergillus tabineus agent, or the Aspergillus tabineus agent prepared by the aforementioned preparation method, and yeast.

[0038] Preferably, the preparation method includes the following steps:

[0039] (1) The Aspergillus tabineus was amplified and cultured to obtain a suspension of Aspergillus tabineus spores;

[0040] (2) The Aspergillus tabineus spore suspension obtained in step (1) is fermented in a fermentation medium containing plant biomass to obtain biomass fermentation product;

[0041] (3) The yeast was amplified and cultured to obtain yeast seed culture;

[0042] (4) The biomass fermentation product obtained in step (2) is mixed with the yeast seed liquid obtained in step (3) and fermented and cultured. After drying, the compound biomass fermentation product is obtained.

[0043] More preferably, in step (2), the concentration of the Aspergillus tabineus spore suspension obtained in step (1) is adjusted to 6 × 10⁻⁶. 6 -6×10 8 After obtaining cfu / mL, the sample is inoculated into a fermentation medium containing plant biomass for fermentation culture. Preferably, the fermentation culture temperature is 10-37℃ and / or the fermentation culture time is 15-30h.

[0044] More preferably, in step (3), the yeast is amplified to obtain a yeast seed culture, and the concentration of the yeast seed culture is adjusted to 5 × 10⁻⁶. 8 -8×10 8 cfu / mL.

[0045] More preferably, in step (4), the biomass fermentation product obtained in step (2) is mixed with the yeast seed liquid obtained in step (3) and then fermented to obtain a composite biomass fermentation product. Preferably, the fermentation temperature is 10-37℃ and / or the fermentation time is 3-7 days.

[0046] More preferably, in step (3), the inoculation amount of yeast seed liquid is: 5-10 mL of yeast seed liquid per 100 g of biomass fermentation product.

[0047] Preferably, the biomass is plant biomass; plant biomass includes one or more of the group consisting of carbohydrate biomass, starch biomass and lignocellulosic biomass; more preferably, lignocellulosic biomass includes one or a combination of two of straw biomass and lees biomass.

[0048] The preferred type of straw-based biomass includes crop straw and bran.

[0049] The preferred type of biomass includes one or more of the following groups of materials: Chinese medicinal residues, tea residues, distiller's grains, and sugarcane bagasse.

[0050] In a tenth aspect, the present invention provides the application of the described Aspergillus tabineus strain, or the described Aspergillus tabineus agent, or the Aspergillus tabineus agent prepared by the described preparation method, in the preparation of feed, fermented tea, biomass fermentation products, or compound biomass fermentation products.

[0051] Beneficial effects of this invention:

[0052] The Aspergillus tabineum provided by this invention possesses a cellulose complex enzyme system, enabling it to degrade cellulose, produce β-glucosidase, and exhibit biomass potential. Simultaneously, it has a large reducing sugar release capacity, improving biomass saccharification efficiency. Furthermore, it can co-ferment plant biomass with yeast, increasing the protein content in the fermented plant biomass, thereby enhancing the nutritional value of plant biomass and facilitating its resource utilization. Attached Figure Description

[0053] Figure 1 shows the colony morphology of Aspergillus tabingensis AMCC 20011.

[0054] Figure 2 shows the enzyme-producing phenotype of Aspergillus tabingensis AMCC 20011.

[0055] Figure 3 shows the crude protein content in the solid-state fermentation product co-fermented by Aspergillus tabbinensis AMCC 20011 and Saccharomyces cerevisiae d5.12, as well as the determination results of the crude protein content in the solid-state fermentation product co-fermented by Aspergillus niger 60B-3DW and Saccharomyces cerevisiae d5.12.

[0056] Microbial strain preservation information

[0057] The Aspergillus tubingensis AMCC 20011 provided by this invention was deposited at the China Center for Type Culture Collection on June 13, 2024, with accession number CCTCCNO: M 20241203. The deposit address is: Wuhan University, Wuhan, China, Postcode: 430072; Telephone: 027-68754052.

[0058] The *Saccharomyces cerevisiae* d5.12 used in this invention was deposited on September 5, 2016, at the China Center for Type Culture Collection (CCTCC), accession number: CCTCC NO: M2016460, address: Wuhan University, Wuhan, China, postcode: 430072; telephone: 027-68754052. This strain has been described in Chinese Patent Publication No. CN108200999A. Detailed Implementation

[0059] To better understand the above technical solutions, the technical solutions of the present invention will be clearly and completely explained below in conjunction with specific embodiments. It should be noted that the content of the specific embodiments is only a specific implementation and explanation of the technical solutions of the present invention, and should not be construed as a limitation on the scope of protection of the present invention.

[0060] This invention provides a strain of Aspergillus tubingensis, AMCC 20011, which was deposited on June 13, 2024, at the China Center for Type Culture Collection (CCTCC), accession number CCTCC NO: M 20241203, address: Wuhan University, Wuhan, China, postcode: 430072; telephone: 027-68754052.

[0061] The colonies of this fungus have a velvety texture with radial grooves. The mycelium is white, and the colonies are initially white, gradually turning black from the center outwards. The reverse side of the colony is yellowish-brown. Microscopically, the conidiophores are spherical; the conidiophores have smooth walls; and the apical sacs are spherical, 50-60 μm in diameter. The ITS gene sequence of *Aspergillus tabinei* AMCC 20011 is shown in SEQ ID NO.3. The BenA gene sequence of *Aspergillus tabinei* AMCC 20011 is shown in SEQ ID NO.6.

[0062] To better understand the technical solution of the present invention, the technical solution of the present invention will be described in detail below with reference to specific embodiments.

[0063] Unless otherwise stated, all reagents / instruments used in the embodiments of this invention are conventional commercially available products. The sources of the experimental reagents used in this invention are shown in Table 1, and the sources of information on the experimental instruments are shown in Table 2.

[0064] Table 1. Experimental Reagent Information

[0065] Table 2. Experimental Instrument Information Sheet

[0066] I. The composition of the culture medium involved in the examples is as follows:

[0067] 1. The preparation method of PDA solid culture medium is as follows:

[0068] (1) Wash and peel the potatoes, cut them into small pieces, add 200g to 1L of distilled water and boil for 20-30 minutes until they can be crushed with a glass rod. Filter the mixture while it is still hot through 8 layers of gauze and retain the filtrate.

[0069] (2) Add 20g of agar to the potato filtrate obtained in step (1), continue heating and mixing. After the agar is completely dissolved, turn off the heat and add 20g of glucose and stir evenly. After cooling slightly, add water to 1L, dispense, stopper and wrap, sterilize at 115℃ for 20min, take out and prepare slant and plate, and the preparation of PDA solid culture medium is completed.

[0070] 2. Preparation method of β-glucosidase screening medium: Based on the weight of PDA solid medium, add 0.1% by weight of aescin and 0.25% by weight of ferric ammonium citrate to PDA solid medium, and sterilize at 115℃ for 20 min to obtain β-glucosidase screening medium.

[0071] 3. Cellulase screening medium: Mix 10g sodium carboxymethyl cellulose, 2.5g dipotassium hydrogen phosphate, 1.25g magnesium sulfate, 20g agar and 1000mL distilled water, and sterilize at 115℃ for 20min to obtain cellulase screening medium.

[0072] 4. Enzyme-producing fermentation medium: Mix 4g of distiller's grains, 0.5g of ammonium sulfate, 0.3g of potassium dihydrogen phosphate, 0.05g of magnesium sulfate, 0.1mL of Tween 80 and 1000mL of distilled water, and sterilize at 115℃ for 20min to obtain the enzyme-producing fermentation medium.

[0073] 5. Solid-state fermentation medium: Mix the lees and water at a weight ratio of 1.5:1, dispense into sterilization bags, and sterilize at 115℃ for 20 minutes to obtain the solid-state fermentation medium.

[0074] II. The reagent preparation methods used in the examples are as follows:

[0075] 1. Buffer solution with pH 5.5: Dissolve 5.39g of citric acid (C6H8O7·H2O) and 21.9g of trisodium citrate (Na3C6H5O7·2H2O) in 900mL of distilled water, determine the pH, adjust the pH to 5.50 with 3mol / L hydrochloric acid solution or 3mol / L sodium hydroxide solution, and bring the volume to 1000mL to obtain a buffer solution with pH 5.5.

[0076] 2. Xylan solution with a concentration of 10 mg / mL: Weigh 1.0000 g of anhydrous xylan and place it in 40 mL of 0.1 mol / L sodium hydroxide solution. Heat and stir magnetically at 70 °C until completely dissolved. Cool to room temperature, add buffer solution to make a volume of 90 mL, adjust the pH to 5.50 with 3 M hydrochloric acid solution, and then make up to 100 mL with pH 5.5 buffer solution to obtain a 10 mg / mL xylan solution.

[0077] 3. Standard xylose solution with a concentration of 10 mg / mL: Weigh 1.0000 g of anhydrous xylose and dissolve it in 80 mL of distilled water by stirring. After it is completely dissolved, transfer it to a volumetric flask and make up to 100 mL to obtain a standard xylose solution with a concentration of 10 mg / mL.

[0078] 4. Sodium acetate buffer solution with pH 5.0: Dissolve 4.1015g of sodium acetate in distilled water, add 2mL of acetic acid, adjust the pH to 5.0, and bring the volume to 1L.

[0079] 5. 5 mmol / L, pH 5.0 p-nitrophenyl-β-D-glucopyranoside (pNPG) solution: Accurately weigh 0.1506 g of p-nitrophenyl-β-D-glucopyranoside (pNPG) into 80 mL of pH 5.0 acetate-sodium acetate buffer. After complete dissolution, transfer to a volumetric flask and bring the volume to 100 mL with acetate-sodium acetate buffer.

[0080] 6. A 1 mg / mL p-nitrophenol solution: Weigh 0.1000 g of p-nitrophenol and dissolve it in 80 mL of distilled water by stirring. After complete dissolution, transfer the solution to a volumetric flask and bring the volume to 100 mL to obtain a 1 mg / mL standard p-nitrophenol solution.

[0081] 7. 1 mol / L sodium carbonate: Weigh 10.599 g of sodium carbonate and dissolve it in 80 mL of distilled water by stirring. After it is completely dissolved, transfer it to a volumetric flask and make up to 100 mL to obtain a 1 mol / L sodium carbonate solution.

[0082] 8. Glucose solution with a concentration of 1 mg / mL: Weigh 0.1000 g of glucose and dissolve it in 80 mL of distilled water by stirring. After it is completely dissolved, transfer it to a volumetric flask and make up to 100 mL to obtain a standard glucose solution with a concentration of 1 mg / mL.

[0083] 9. 0.1% Congo Red: Weigh 0.1g of Congo Red into 100mL of distilled water, and obtain a 0.1% Congo Red solution after it is completely dissolved.

[0084] 10. 0.1M Sodium Chloride: Weigh 5.844g of sodium chloride and dissolve it in 80mL of distilled water by stirring. After it is completely dissolved, transfer it to a volumetric flask and make up to 100mL to obtain a 0.1M sodium chloride solution.

[0085] Example 1: Isolation and identification of Aspergillus tabineus AMCC 20011 strain

[0086] (1) Strains were isolated: Fermented tea samples obtained in Yichang City, Hubei Province (collected during the fermentation process of black tea preparation) were serially diluted 10-fold to prepare 10 strains. -5 10 -6 The bacterial suspension was spread onto PDA solid medium and cultured at 28°C for 72 hours to obtain single colonies.

[0087] (2) Purification of the strain: After a single colony has grown to the sporulation stage, spores are streaked onto a PDA plate and cultured at 28°C for 5 days to complete one purification. This process is repeated three times.

[0088] (3) Preservation of strains: The strains that have been purified three times were inoculated onto PDA slant medium and preserved at 4°C to obtain strain preservation slant.

[0089] (4) Identification of strains: Take one strain of the strain preservation slant, add 5 mL of sterile physiological saline and shake to mix, then filter through sterile gauze to obtain a spore suspension containing Aspergillus tabineum AMCC20011. Then, use an inoculation loop to dip the spore suspension and streak it on PDA solid medium. Incubate at 28°C for 7 days to observe the colony morphology.

[0090] A strain of fungus was obtained. The colony texture of this fungus is velvety with radial grooves, and the mycelium is white.

[0091] The colonies are initially white, gradually turning black from the center outwards, with a yellowish-brown underside. Microscopically, the conidiophores are spherical; the conidiophores have smooth walls; and the apical sacs are spherical, 50-60 μm in diameter. The genome of this bacterium was extracted. Using ITS1 (5'-TCCGTAGGTGAACCTGCGG-3', SEQ ID NO.1) and ITS4 (5'-TCCTCCGCTTATTGATATGC-3', SEQ ID NO.2) as primers, a PCR program of 94℃ pre-denaturation for 5 min, 94℃ denaturation for 30 s, 55℃ annealing for 45 s, and 72℃ extension for 90 s, for 30 cycles, followed by a final 72℃ extension for 10 min, was used to amplify the ITS gene sequence. After 1% gel electrophoresis and sequencing, the sequence was compared with sequences in GenBank using BLAST analysis. The sequence similarity was greater than 99%, indicating the same species. The determined ITS gene sequence of this strain is shown in SEQ ID NO.3.

[0092] (5) Identification of the BenA gene sequence of this strain: The genome of this strain was extracted, and the PCR program was performed using Bt2a (5'-GGTAACCAAATCGGTGCTGCTTTC-3', i.e. SEQ ID NO.4) and Bt2b (5'-ACCCTCAGTGTAGTGACCCTTGGC-3', i.e. SEQ ID NO.5) as primers. The PCR program was as follows: 94℃ pre-denaturation for 5 min, 94℃ denaturation for 30 s, 55℃ annealing for 45 s, 72℃ extension for 90 s, 30 cycles, and a final extension at 72℃ for 10 min to amplify the ITS gene sequence. After detection by 1% gel electrophoresis and sequencing, the sequence was compared with the sequence in GenBank by BLAST analysis. The sequence similarity was greater than 99% and the strain belonged to the same species. The BenA gene sequence of this strain was determined, as shown in SEQ ID NO.6:

[0093] Based on morphological analysis and molecular identification, the bacterium was identified as *Aspergillus tubingensis* AMCC 20011, which was deposited at the China Center for Type Culture Collection (CCTCC) on June 13, 2024, with accession number CCTCC NO: M 20241203. Figure 1 shows the colony morphology of *Aspergillus tubingensis* AMCC 20011.

[0094] Example 2: Determination of the ability of Aspergillus tabingii AMCC 20011 to produce β-glucosidase and cellulase

[0095] Take one specimen of *Aspergillus tabineus* AMCC20011 preservation slant, add 5 mL of sterile physiological saline, shake to mix, and then filter through sterile gauze to obtain a spore suspension of *Aspergillus tabineus* AMCC20011. Take 20 μL of the spore suspension and spot it at the center of both the β-glucosidase and cellulase selection media, and incubate statically at 28℃. Observe the colony phenotype daily and record the morphological characteristics of the earliest enzyme-producing phenotype in the β-glucosidase selection medium. The results are shown in the left column of Figure 2. Simultaneously, on the third day of static incubation at 28℃, add 0.1% Congo red staining solution to the cellulase selection medium for 60 min, followed by destaining with 1M sodium chloride for 30 min. Record the enzyme-producing phenotype in the cellulase selection medium. The results are shown in the right column of Figure 2.

[0096] As shown in the left column of Figure 2, the left column of Figure 2 represents the morphological characteristics of the enzyme-producing phenotypes in the β-glucosidase selection medium. The upper part of the left column in Figure 2 shows the morphological characteristics of the enzyme-producing phenotypes on the front side of the β-glucosidase selection medium; the lower part shows the morphological characteristics of the enzyme-producing phenotypes on the back side of the β-glucosidase selection medium. The results indicate that after culturing *Aspergillus tabingensis* AMCC20011 on the β-glucosidase selection medium for 1 day, a black hydrolysis zone appeared on the β-glucosidase selection medium. At this time, the colony diameter d = 1 cm, the diameter of the black hydrolysis zone D = 4 cm, and D / d = 4. Because esculin and ferric ammonium citrate were added to the β-glucosidase screening medium, when *Aspergillus tabinei* decomposes esculin into aescin, aescin reacts with the ferrous ions in ferric ammonium citrate to form a black compound, turning the medium black. Aesculin and cellobiose have similar β-D-glucopyranose structures, and enzymes in enzymatic reactions typically have specific recognition capabilities for substrate structures. β-glucosidase, which can decompose cellobiose, can also recognize and bind to esculin. Therefore, *Aspergillus tabinei* AMCC20011 possesses the ability to produce β-glucosidase.

[0097] As shown in the right column of Figure 2, the right column of Figure 2 represents the morphological characteristics of enzyme-producing phenotypes in the cellulase selection medium. The upper part of the right column in Figure 2 shows the morphological characteristics of enzyme-producing phenotypes on the front side of the cellulase selection medium, while the lower part shows the morphological characteristics of enzyme-producing phenotypes on the back side. The results indicate that on the third day of cultivation on the cellulase selection medium, *Aspergillus tabinei* AMCC20011, after staining and destaining, exhibited light-colored clear zones around the colonies. Because Congo red can form a red complex with carboxymethyl cellulose in the cellulase selection medium, while non-polysaccharide substances such as cellobiose and glucose cannot be stained by Congo red, when cellulose is decomposed by cellulase, the red complex between cellulose and Congo red cannot form, resulting in clear zones centered on cellulose-decomposing bacteria in the cellulase selection medium. Therefore, *Aspergillus tabinei* AMCC20011 possesses the ability to produce cellulase.

[0098] Example 3: Determination of the enzyme activities of filter paper cellulase, β-glucosidase, and xylanase produced by Aspergillus tabingensis AMCC20011.

[0099] (1) Preparation of spore suspension of Aspergillus tabineum AMCC20011: The specific preparation method is as follows: Take one slant of Aspergillus tabineum strain AMCC20011, add 5 mL of sterile physiological saline and shake to mix. Then filter through sterile gauze to obtain the spore suspension of Aspergillus tabineum AMCC20011. Then use an inoculation loop to dip the spore suspension into the plate and streak it on PDA solid medium. Incubate at 28℃ for 7 days to complete the activation culture. Then add 5 mL of sterile physiological saline to the activated culture plate to slowly wet it so that Aspergillus tabineum AMCC 20011 is completely immersed in the sterile physiological saline. Filter through sterile gauze to obtain the spore suspension of Aspergillus tabineum AMCC20011.

[0100] (2) The preparation method of the spore suspension of Aspergillus niger 60B-3DW is the same as the preparation method of the spore suspension of Aspergillus tabinea AMCC20011 in step (1). The Aspergillus niger 60B-3DW was purchased from the Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, and the Aspergillus niger 60B-3DW has been disclosed in Chinese patent with patent number CN 113403207 B.

[0101] (3) The number of spores in the spore suspension of *Aspergillus tabinei* AMCC20011 was calculated. The specific counting method was as follows: Prepare a Tommy hemocytometer, cover it with a coverslip, and take 20 μL of the spore suspension of *Aspergillus tabinei* AMCC20011. Slowly draw it in from the edge of the coverslip, let it stand for 5 minutes, and under a microscope with an objective magnification of 40× and an eyepiece magnification of 10×, count the spores in five squares (80 small squares) in a diagonal orientation: the upper left, lower left, upper right, lower right, and the center square. The spore count was then calculated, and the number of *Aspergillus tabinei* AMCC 20011 spores was found to be 6.40 × 10⁻¹⁰. 6 cfu / mL.

[0102] The method for calculating the number of spores in the spore suspension of Aspergillus niger 60B-3DW was the same as that for Aspergillus tabineum AMCC20011, yielding a spore count of 6.15 × 10⁻⁶ for Aspergillus niger 60B-3DW. 6 cfu / mL.

[0103] The number of spores per milliliter of spore suspension is calculated as follows: (total number of 5 square cells / 80) × 400 × 10000 × dilution factor.

[0104] (4) Cultivating the crude enzyme solution of Aspergillus tabineus AMCC20011, the specific cultivation method is as follows: dilute the number of spores in the spore suspension of Aspergillus tabineus AMCC20011 to 6×10⁻⁶. 6 The concentration of cfu / mL was inoculated into 300 mL of enzyme-producing fermentation medium at a volume of 0.5%. Fermentation was carried out at 28°C and 220 rpm for 5 days to obtain the enzyme-producing fermentation broth of *Aspergillus tabineum* AMCC20011. Then, 45 mL of the enzyme-producing fermentation broth of *Aspergillus tabineum* AMCC20011 was transferred to a 50 mL centrifuge tube and centrifuged at 8000 rpm for 10 min. The supernatant was retained as the crude enzyme solution of *Aspergillus tabineum* AMCC20011.

[0105] The method for culturing the crude enzyme solution of Aspergillus niger 60B-3DW is the same as that for culturing the crude enzyme solution of Aspergillus tabineum AMCC20011.

[0106] (5) Determination of cellulase activity on filter paper: The cellulase activity of *Aspergillus tabinei* AMCC20011 and *Aspergillus niger* 60B-3DW on filter paper was determined according to GB / T 23881-2009 (Determination of cellulase activity in feed—Filter paper method). Three replicates were performed for each crude enzyme solution of *Aspergillus tabinei* AMCC20011 and crude enzyme solution of *Aspergillus niger* 60B-3DW. The standard curve was plotted with glucose concentration on the x-axis and absorbance at OD540nm on the y-axis. The linear formula for the glucose standard curve was obtained by fitting the curve as follows: y = 0.003x + 0.0398, R0 2 =0.9905.

[0107] One unit of cellulase activity is defined as the amount of enzyme required to release 1 μmol of glucose per minute per milliliter of crude enzyme solution from filter paper under the conditions of 37°C, pH 5.5, and 60 min of reaction.

[0108] (6) Determination of β-glucosidase activity:

[0109] (6.1) Method for determining the β-glucosidase activity of crude enzyme solution of Aspergillus tabineus AMCC20011: A 5 mmol / L solution of p-nitrophenyl-β-D-glucopyranoside (pNPG) with pH 5.0 was used as the substrate. The p-nitrophenyl-β-D-glucopyranoside (pNPG) solution was prepared by passing it through 50 mM sodium acetate solution (pH 5.0).

[0110] 200 μL of the substrate was mixed thoroughly with 100 μL of crude enzyme solution of *Aspergillus tabineum* AMCC20011 and incubated at 50°C for 60 min. Then, 200 μL of 1 mol / L Na₂CO₃ was added to terminate the reaction. 300 μL of the supernatant was transferred to 1 mL of distilled water to obtain a diluent. 200 μL of this diluent was transferred to the wells of an ELISA plate, and the absorbance at 410 nm was measured using an ELISA reader. Standard curve: A standard curve for p-nitrophenol was obtained by mixing 0-100 μg / mL p-nitrophenol solution with 200 μL of 1 mol / L Na₂CO₃ and developing the color. The absorbance at 410 nm was used as the ordinate, and the linear formula was: y = 0.0532x + 0.098, R₀ = 0.0532x + 0.098. 2 =0.9945.

[0111] One unit of β-glucosidase activity is defined as the amount of enzyme required to produce 1 μmol of p-nitrophenol (p-NP) per minute per milliliter of crude enzyme solution at a concentration of 5 mmol / L and a pH of 5.0, under conditions of 50°C and pH 5.0.

[0112] (6.2) The method for determining the β-glucosidase activity of crude enzyme solution of Aspergillus niger 60B-3DW is the same as that for determining the β-glucosidase activity of crude enzyme solution of Aspergillus tabbinatus AMCC20011.

[0113] Each sample of crude enzyme solution from Aspergillus tabineus AMCC20011 and Aspergillus niger 60B-3DW was used in three replicate experiments. Simultaneously, the crude enzyme solutions of inactivated Aspergillus tabineus AMCC20011 and Aspergillus niger 60B-3DW were used as blank controls to eliminate the influence of reagents.

[0114] (7) Determine the enzyme activity of xylanase:

[0115] (7.1) Method for determining the xylanase activity of crude enzyme solution of *Aspergillus tabineum* AMCC20011: Take 4 test tubes and add 0.5 mL of 10 mg / mL xylan solution as substrate to each of the 4 test tubes. Preheat the solution of crude enzyme solution of *Aspergillus tabineum* AMCC20011 at 50℃ for 5 min. Then add 0.5 mL of crude enzyme solution of *Aspergillus tabineum* AMCC20011 diluted 100 times to the 1st, 2nd and 3rd test tubes and react at 50℃ for 15 min. After the reaction is complete, add 1.5 mL of DNS reagent to the 1st, 2nd, 3rd and 4th test tubes respectively. Add 0.5 mL of crude enzyme solution of *Aspergillus tabineum* AMCC20011 to the 4th test tube and shake well. Then react in a boiling water bath for 5 min. After rapidly cooling to room temperature, make up to 10 mL with distilled water. Using the fourth test tube as a control, the absorbance of the first, second, and third test tubes was measured at a wavelength of 540 nm.

[0116] Standard curve: Mix 1 mL of 0-500 μg / mL xylose solution with 1.5 mL of DNS reagent, boil for 5 min, cool and bring to a final volume of 10 mL. Plot the standard curve with absorbance at 540 nm as the ordinate and xylose concentration as the abscissa. The linear formula for the xylose standard curve is: y = 0.0039x - 0.2557, R0 2 =0.9905.

[0117] One unit of xylanase activity is defined as the amount of enzyme required to release 1 μmol of xylose per minute from a xylan solution with a concentration of 10 mg / mL, at 50°C and pH 5.5.

[0118] (7.2) The method for determining the xylanase activity of crude enzyme solution of Aspergillus niger 60B-3DW is the same as that for determining the xylanase activity of crude enzyme solution of Aspergillus tabbina AMCC20011.

[0119] Xylanase activity (U / mL) = ((equal amount of xylose) / 150 / 15 / 0.5) × n

[0120] In the formula: 150 represents xylose converted from micrograms to micromoles; 15 represents the reaction time between the crude enzyme solution and the substrate; 0.5 represents the amount of crude enzyme solution added to the reaction; n represents the dilution factor of the crude enzyme solution, wherein the dilution factor in this embodiment is 100.

[0121] Table 3. Enzyme activities of filter paper cellulase from Aspergillus tabingensis AMCC20011 and Aspergillus niger 60B-3DW

[0122] Unit: U / mL

[0123] Table 4. Enzyme activities of β-glucosidase in Aspergillus tabingensis AMCC20011 and Aspergillus niger 60B-3DW

[0124] Unit: U / mL

[0125] Table 5. Enzyme activities of xylanase in Aspergillus tabingensis AMCC20011 and Aspergillus niger 60B-3DW

[0126] Unit: U / mL

[0127] Table 6. Ratio of mean enzyme activities of Aspergillus tabineum AMCC20011 and Aspergillus niger 60B-3DW

[0128] As shown in Tables 3, 4, 5 and 6, the results indicate that, under the same spore inoculation, substrate and fermentation conditions, the enzyme activities of filter paper cellulase, β-glucosidase and xylanase in Aspergillus tabinea AMCC 20011 were significantly higher than those in Aspergillus niger 60B-3DW.

[0129] As shown in Tables 3 and 4, the results indicate that the filter paper cellulase activity of Aspergillus tabineum AMCC 20011 was 15.96 ± 0.80 U / mL, and its average value was 376% of that of Aspergillus niger 60B-3DW. Since the filter paper cellulase activity characterizes the total enzyme activity of the cellulose complex enzyme system in the crude enzyme solution, the above results show that the total enzyme activity of the cellulose complex enzyme system of Aspergillus tabineum AMCC 20011 is much higher than that of Aspergillus niger 60B-3DW, which proves that Aspergillus tabineum AMCC 20011 has a stronger ability to degrade cellulose.

[0130] As shown in Tables 3 and 5, the results indicate that the β-glucosidase activity of *Aspergillus tabineum* AMCC 20011 was 1.51 ± 0.01 U / mL, with an average activity 130% higher than that of *Aspergillus niger* 60B-3DW. Since β-glucosidase is the rate-limiting enzyme in cellulose degradation, primarily breaking down cellobiose, an important intermediate product in cellulose degradation, and one of the main functions of β-glucosidase is to break down cellobiose, through hydrolysis, it converts cellobiose into glucose molecules. This is a crucial step in the degradation of cellulose into usable sugar molecules. Therefore, the rate-limiting effect of β-glucosidase in cellulose degradation is largely related to its ability to break down cellobiose. Therefore, the above results show that the β-glucosidase activity of Aspergillus tabingensis strain AMCC20011 is 30% higher than that of Aspergillus niger 60B-3DW. This proves that Aspergillus tabingensis strain AMCC20011 can rapidly decompose cellobiose, thereby avoiding the feedback inhibition of cellulase activity caused by cellobiose accumulation to a certain extent. It has the potential to ferment plant biomass and is conducive to the complete decomposition and utilization of lignocellulose.

[0131] As shown in Tables 3 and 6, the results indicate that the xylanase activity of *Aspergillus tabinei* AMCC 20011 was 440.43 ± 3.01 U / mL, with an average value 220% higher than that of *Aspergillus niger* 60B-3DW. The xylanase activity characterizes the hemicellulose activity in the crude enzyme solution, demonstrating its ability to decompose xylan into xylose, a fermentable sugar. These results show that the xylanase activity of *Aspergillus tabinei* AMCC 20011 is significantly higher than that of *Aspergillus niger* 60B-3DW, proving that *Aspergillus tabinei* AMCC 20011 has a stronger hemicellulose degradation capacity. This further demonstrates that *Aspergillus tabinei* AMCC 20011 is suitable for fermenting lignocellulose to a certain extent and has the potential to ferment plant biomass.

[0132] Example 4: Application of Aspergillus tabine in high-value fermentation of distiller's grains

[0133] (1) Colonies from the preservation slant of *Saccharomyces cerevisiae* d5.12 were picked and cultured overnight in YPD liquid medium. Then, 1% by volume was inoculated into 100 ml of YPD medium and cultured for 20 h at 30 °C and 180 rpm to obtain the seed culture of *Saccharomyces cerevisiae* d5.12 (the cell concentration of the seed culture of *Saccharomyces cerevisiae* d5.12 was 5 × 10⁻⁶). 8 (cfu / mL).

[0134] (2) Dilute the spore suspensions of Aspergillus tabingensis AMCC20011 and Aspergillus niger 60B-3DW to a uniform concentration of 6×10⁻⁶. 6 CFU / mL, 10 mL to 100 g of solid-state fermentation medium (mixed with distiller's grains and water at a weight ratio of 1.5:1, dispensed into sterile bags, and sterilized at 115℃ for 20 min to obtain solid-state fermentation medium) were inoculated separately. After fermentation at 28℃ for 24 h, solid-state fermentation products of *Aspergillus tabinei* AMCC20011 and *Aspergillus niger* 60B-3DW were obtained. The reducing sugar content of *Aspergillus tabinei* AMCC20011 and *Aspergillus niger* 60B-3DW solid-state fermentation products was then determined, and the results are shown in Table 7.

[0135] (3) 10 mL of *Saccharomyces cerevisiae* seed culture was inoculated into 100 g of *Aspergillus tabineum* AMCC20011 solid-state fermentation product and 100 g of *Aspergillus niger* 60B-3DW solid-state fermentation product, respectively, to obtain solid-state fermentation products co-fermented by *Aspergillus tabineum* AMCC20011 and *Saccharomyces cerevisiae* d5.12, and co-fermented by *Aspergillus niger* 60B-3DW and *Saccharomyces cerevisiae* d5.12. Fermentation was carried out at 30℃ for 7 days. On days 0, 3, 5, and 7 of fermentation, the co-fermented solid-state fermentation products were dried at 65℃ to constant weight, and the crude protein content was determined. The results of the crude protein content determination are shown in Figure 3. The crude protein determination method was in accordance with GB / T 6432-2018.

[0136] The reducing sugar was determined using the 3,5-dinitrosalicylic acid (DNS) colorimetric method. Specifically, 1 g of fresh fermented sample was weighed into a 250 mL Erlenmeyer flask, a small amount of distilled water was added to make a paste, and then 50 mL of distilled water was added to make up the paste. The mixture was boiled at 100 °C for 5 min to leach out the reducing sugar. The leachate (containing the precipitate) was then transferred to a 50 mL centrifuge tube and centrifuged at 8000 r / min for 5 min. The precipitate was washed once with 20 mL of distilled water and centrifuged again. The supernatant was collected in a 100 mL volumetric flask and diluted to the mark with distilled water. The solution was mixed well to obtain the 100-fold diluted reducing sugar test solution.

[0137] Add 1 mL of the reducing sugar test solution to a 15 mL graduated test tube, add 2 mL of DNS reagent, boil in water for 2 minutes, cool, and then bring the volume to 15 mL with water. Measure the absorbance at 540 nm. Substitute the absorbance value into the standard curve to obtain the amount of glucose per milliliter of the reducing sugar test solution. Standard Curve: Take 0, 0.2, 0.4, 0.6, 0.8, and 1 mL of 1 mg / mL glucose standard solution into 15 mL test tubes, respectively, and bring the volume to 1 mL with distilled water. Add 2 mL of DNS reagent to each tube, heat in a boiling water bath for 2 minutes, cool under running water, and then bring the volume to 15 mL with water. Measure the absorbance at 540 nm to obtain the standard curve.

[0138] Table 7. Reducing sugar release 1 day after Aspergillus tabineus inoculation

[0139] As shown in Table 7, the results indicate that when Aspergillus tabineum AMCC 20011 fermented distiller's grains alone for one day, the reducing sugar release in the three replicate fermentation groups was 4.01%, 3.96%, and 4.06%, respectively. When Aspergillus niger 60B-3DW was inoculated for one day, the reducing sugar release in the three replicate fermentation groups was 3.13%, 3.17%, and 3.22%, respectively. The reducing sugar release in the Aspergillus tabineum AMCC 20011 fermentation group was on average 26.5% higher than that in the Aspergillus niger 60B-3DW group. Therefore, Aspergillus tabineum AMCC 20011 exhibits the characteristics of highly efficient saccharification of biomass.

[0140] It should be noted that the reducing sugar release of a strain represents its ability to break down or utilize sugars to produce reducing sugars under specific conditions. Reducing sugars are sugars with reducing properties, such as glucose, fructose, and maltose. These sugars typically possess one or more free aldehyde or ketone groups, thus exhibiting reducing properties. The reducing sugar release of a strain is often used as an indicator to assess its metabolic activity, sugar utilization capacity, and potential application value.

[0141] Saccharification refers to the process of converting polysaccharides (such as cellulose and starch) in biomass into monosaccharides (such as glucose). Monosaccharides are important raw materials in many biological processes and industrial applications, especially in biofermentation and bioenergy production. During the saccharification of biomass, bacterial strains produce enzymes that can break down polysaccharides in the biomass, thereby accelerating the saccharification process.

[0142] Figure 3 shows the crude protein content in the solid-state fermentation products of *Aspergillus tabineum* AMCC 20011 and *Saccharomyces cerevisiae* d5.12, and the crude protein content in the solid-state fermentation products of *Aspergillus niger* 60B-3DW and *Saccharomyces cerevisiae* d5.12. In Figure 3, 20011 represents the solid-state fermentation product of *Aspergillus tabineum* AMCC 20011 and *Saccharomyces cerevisiae* d5.12, and 60B-3DW represents the solid-state fermentation product of *Aspergillus niger* 60B-3DW and *Saccharomyces cerevisiae* d5.12. The results indicate that the highest crude protein content was obtained in the solid-state fermentation product of *Aspergillus tabineum* AMCC 20011 and *Saccharomyces cerevisiae* d5.12 on day 5 of fermentation. Furthermore, the crude protein content of the solid-state fermentation product co-fermented with *Aspergillus tabineum* AMCC20011 and *Saccharomyces cerevisiae* d5.12 was 35.90±0.09%, while the crude protein content of the solid-state fermentation product co-fermented with *Aspergillus niger* 60B-3DW and *Saccharomyces cerevisiae* d5.12 was 33.44±0.08%. Therefore, the crude protein content of the solid-state fermentation product co-fermented with *Aspergillus tabineum* AMCC20011 and *Saccharomyces cerevisiae* d5.12 was 7.36% higher than that of the solid-state fermentation product co-fermented with *Aspergillus niger* 60B-3DW and *Saccharomyces cerevisiae* d5.12.

[0143] The above results indicate that Aspergillus tabineum AMCC 20011 has a high reducing sugar release rate, and the solid-state fermentation product obtained by co-fermenting Aspergillus tabineum AMCC 20011 and Saccharomyces cerevisiae d5.12 has a high crude protein enhancement effect. This proves that Aspergillus tabineum AMCC 20011 is suitable for fermenting distiller's grains, and further demonstrates the potential of Aspergillus tabineum AMCC 20011 in the high-value fermentation and utilization of low-value biomass such as distiller's grains, tea, straw, sugarcane bagasse and bran, and has broad application prospects.

[0144] The above embodiments are only for further explanation and understanding of the technical solution of the present invention, and are not intended to limit the present invention. Any improvements made by those skilled in the art on this basis that do not highlight substantive features or make significant progress should fall within the protection scope of the present invention.

Claims

1. A type of Aspergillus tabineus, characterized in that, The Aspergillus tubingensis mentioned is AMCC 20011, which is deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC NO: M 20241203.

2. The Aspergillus tabineum according to claim 1, characterized in that, The ITS gene sequence of the Aspergillus tabingensis strain is shown in SEQ ID NO.

3.

3. The Aspergillus tabineum according to claim 1 or 2, characterized in that, The BenA gene sequence of Aspergillus tabingensis AMCC 20011 is shown in SEQ ID NO.

6.

4. Aspergillus tabineum according to any one of claims 1-3, characterized in that, The Aspergillus tabingensis strain has the characteristics of producing cellulase, and / or producing β-glucosidase, and / or producing xylanase, and / or saccharifying biomass, and / or co-fermenting with yeast to increase the protein content in biomass.

5. Aspergillus tabineum according to any one of claims 1-4, characterized in that, The filter paper enzyme activity of cellulase produced by Aspergillus tabineis is 15-17 U / mL. The amount of enzyme required to degrade filter paper and release 1 μmol of glucose per minute per milliliter of crude enzyme solution from Aspergillus tabineis is defined as one unit of filter paper enzyme activity.

6. Aspergillus tabineum according to any one of claims 1-5, characterized in that, The enzyme activity of β-glucosidase produced by Aspergillus tabineis is 1.5-1.55 U / mL. One unit of β-glucosidase activity is defined as the amount of enzyme required to produce 1 μmol of p-nitrophenol per minute by enzymatically hydrolyzing a p-nitrophenyl-β-D-glucopyranoside solution with a concentration of 5 mmol / L and a pH of 5.0 per milliliter of crude enzyme solution from Aspergillus tabineis.

7. Aspergillus tabineum according to any one of claims 1-6, characterized in that, The enzyme activity of xylanase produced by Aspergillus tabineis is 437-445 U / mL. One unit of xylanase activity is defined as the amount of enzyme required to release 1 μmol of xylose per minute from a xylan solution with a concentration of 10 mg / mL per milliliter of crude enzyme solution from Aspergillus tabineis.

8. A tabine aspergillus agent, characterized in that, Contains *Aspergillus tabineum* as described in any one of claims 1-7.

9. A method for preparing the Aspergillus tabine agent according to claim 8, characterized in that, The method includes the following steps: amplifying and culturing Aspergillus tabineum according to any one of claims 1-7.

10. The preparation method according to claim 9, characterized in that, The amplification culture temperature is 10-37℃.

11. A feed, characterized in that, The preparation contains the Aspergillus tabineum strain according to any one of claims 1-7, or the Aspergillus tabineum agent according to claim 8, or the Aspergillus tabineum agent prepared by the preparation method according to claim 9 or 10.

12. A fermented tea, characterized in that, The preparation contains the Aspergillus tabineum strain according to any one of claims 1-7, or the Aspergillus tabineum agent according to claim 8, or the Aspergillus tabineum agent prepared by the preparation method according to claim 9 or 10.

13. A biomass fermentation product, characterized in that, The preparation contains the Aspergillus tabineum strain according to any one of claims 1-7, or the Aspergillus tabineum agent according to claim 8, or the Aspergillus tabineum agent prepared by the preparation method according to claim 9 or 10.

14. A method for preparing the biomass fermentation product according to claim 13, characterized in that, The process includes the following steps: culturing the Aspergillus tabineum strain according to any one of claims 1-7, or the Aspergillus tabineum agent according to claim 8, or the Aspergillus tabineum agent prepared by the preparation method according to claim 9 or 10.

15. The preparation method according to claim 14, characterized in that, Includes the following steps: (1) Amplify and culture Aspergillus tabineum according to any one of claims 1-7 to obtain Aspergillus tabineum spore suspension; (2) The Aspergillus tabingensis spore suspension obtained in step (1) is inoculated into a fermentation medium containing plant biomass for fermentation culture to obtain biomass fermentation product.

16. The preparation method according to claim 15, characterized in that, The concentration of the obtained suspension of Aspergillus tubingensis spores from step (1) is adjusted to 6 x 10 6 -6 x 10 8 cfu / mL, and the suspension is inoculated into a fermentation medium containing plant biomass and is subjected to a fermentation culture, whereby a biomass fermentation product is obtained; preferably, the fermentation culture is carried out at a temperature of 10-37°C and / or for a period of 15-30 h.

17. The preparation method according to claim 15 or 16, characterized in that, The fermentation medium containing plant biomass comprises: plant biomass and water, preferably, the weight ratio of plant biomass to water is 1-1.5:

1.

18. The preparation method according to claim 15, characterized in that, In step (2), the inoculation amount of Aspergillus tabineus spore suspension is: 5-10 mL of Aspergillus tabineus spore suspension per 100 g of fermentation medium containing plant biomass.

19. A compound biomass fermentation product, characterized in that, The product contains the Aspergillus tabineum strain according to any one of claims 1-7, or the Aspergillus tabineum agent according to claim 8, or the Aspergillus tabineum agent prepared by the preparation method according to claim 9 or 10, and yeast.

20. The compound biomass fermentation product according to claim 19, characterized in that, Yeasts include genera such as *Saccharomyces*, *Kluyveromyces*, *Pichia*, *Cyberlindnera*, *Arxiozyma*, *Cylindricascospora*, *Cyniclomyces*, *Eremothecium*, *Grigorovia*, *Hagleromyces*, *Henningerozyma*, *Huiozyma*, *Jamesozyma*, and *Kazachstania*. The group consisting of one or more genera including *Maudiozyma*, *Monosporozyma*, *Naumovozyma*, *Nakaseomyces*, *Oligophagozyma*, *Savitreea*, *Sungouiozyma*, *Stenotrophomyces*, *Tetrapisispora*, *Torulaspora*, *Vanderwaltozyma*, *Yueomyces*, *Zygosaccharomyces*, and *Zygotorulaspora*. Preferably, the yeast includes one or more species from the group consisting of the genera *Saccharomyces*, *Kluyveromyces*, *Pichia*, and *Cyberlindnera*. More preferably, the yeast genus (Saccharomyces) is Saccharomyces cerevisiae; Most preferably, the brewing yeast is brewing yeast (Saccharomyces cerevisiae) d5.12, with the preservation number CCTCC NO: M 2016460.

21. The compound biomass fermentation product according to claim 19 or 20, characterized in that, The crude protein content is 30% or more by weight of the compound biomass fermentation product, preferably 30-36%.

22. A method for preparing a compound biomass fermentation product according to any one of claims 19-21, characterized in that, The process includes the following steps: culturing the Aspergillus tabineum strain according to any one of claims 1-7, or the Aspergillus tabineum agent according to claim 8, or the Aspergillus tabineum agent prepared by the preparation method according to claim 9 or 10, and yeast.

23. The preparation method according to claim 22, characterized in that, Includes the following steps: (1) Amplify and culture Aspergillus tabineum according to any one of claims 1-7 to obtain Aspergillus tabineum spore suspension; (2) The Aspergillus tabineus spore suspension obtained in step (1) is fermented in a fermentation medium containing plant biomass to obtain biomass fermentation product; (3) The yeast was amplified and cultured to obtain yeast seed culture; (4) The biomass fermentation product obtained in step (2) is mixed with the yeast seed liquid obtained in step (3) and fermented and cultured. After drying, the compound biomass fermentation product is obtained.

24. The preparation method according to claim 23, characterized in that, In step (2), the concentration of the Aspergillus tubigensis spore suspension obtained in step (1) is adjusted to 6 x 10 6 -6 x 10 8 Afterwards, the spore suspension is inoculated into a fermentation medium containing plant biomass and is subjected to a fermentation culture, preferably at a fermentation culture temperature of 10-37°C and / or for a fermentation culture time of 15-30 h.

25. The preparation method according to claim 23, characterized in that, In step (3), the yeast is amplified and cultured to obtain yeast seed culture, and the concentration of the yeast seed culture is adjusted to 5×10⁻⁶. 8 -8×10 8 cfu / mL.

26. The preparation method according to any one of claims 23-25, characterized in that, In step (4), the biomass fermentation product obtained in step (2) is mixed with the yeast seed liquid obtained in step (3) and then fermented to obtain a composite biomass fermentation product. Preferably, the fermentation temperature is 10-37℃ and / or the fermentation time is 3-7 days.

27. The preparation method according to any one of claims 22-26, characterized in that, In step (3), the inoculation amount of yeast seed liquid is 5-10 mL of yeast seed liquid per 100 g of biomass fermentation material.

28. The biomass fermentation product according to claim 13, or the biomass fermentation product prepared by the preparation method according to any one of claims 14-18, or the composite biomass fermentation product according to any one of claims 19-21, or the composite biomass fermentation product prepared by the preparation method according to any one of claims 22-27, characterized in that, The biomass is plant biomass; preferably, the plant biomass includes one or more of the group consisting of carbohydrate biomass, starch biomass and lignocellulosic biomass; more preferably, the lignocellulosic biomass includes one or a combination of two of straw biomass and lees biomass. The preferred type of straw-based biomass includes crop straw and bran. The preferred type of biomass includes one or more of the following groups of materials: Chinese medicinal residues, tea residues, distiller's grains, and sugarcane bagasse.

29. The Aspergillus tabineum strain according to any one of claims 1-7, or the Aspergillus tabineum agent according to claim 8, or the Aspergillus tabineum agent prepared by the preparation method according to claim 9 or 10, in the preparation of feed, fermented tea, biomass fermentation products or compound biomass fermentation products.

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