Method for recycling biomass waste residues

Abstract

The application discloses a biomass waste residue recycling method. The biomass waste residue recycling method comprises the following steps: mixing filamentous fungi and yeast with biomass waste residue, carrying out fermentation culture, carrying out solid-liquid separation, and extracting biomass oil from the separated solid. The filamentous fungi comprise one or a combination of two selected from Trichoderma reesei and Aspergillus niger; the yeast comprises one or a combination of two selected from Lipomyces starkeyi and Rhodotorula. The biomass waste residue recycling method can treat fruit residue, coffee residue or medicine residue to produce biomass oil.

Description

Technical Field

[0001] This invention relates to a method for reusing biomass waste, particularly a method for reusing biomass waste (including fruit pomace, coffee grounds, or medicinal residues without pectin) to produce biomass oil. Background Technology

[0002] Biomass waste such as fruit pomace, coffee grounds, and medicinal residues can cause environmental pollution if not handled properly, such as producing leachate and breeding bacteria. Fruit pomace is the solid residue remaining after extracting useful components (such as fructose, oils, and juice) from fruits, and pectin is also removed through acid extraction. Coffee grounds are the solid residue after coffee extraction. Medicinal residues are the solid residue remaining after extracting active ingredients from traditional Chinese medicine. The main component of these wastes is lignocellulose, which does not contain pectin or sugars. Current treatment methods mostly involve landfilling, incineration for power generation, or composting for use as natural fertilizer. However, some wastes are prone to spoilage, require pH adjustment, or contain hazardous substances, making them unsuitable for these methods. They also suffer from low utilization rates and low added value. While some fermentation methods exist, they often require complex pretreatment of the waste, such as enzymatic hydrolysis, or multiple fermentation steps, resulting in high processing costs.

[0003] The information disclosed in the background section is only intended to enhance the understanding of the background of this application, and therefore may include information that does not constitute prior art known to those skilled in the art. Summary of the Invention

[0004] This invention provides a method for recycling biomass waste, which can process fruit pomace, coffee grounds or medicinal residues to produce biomass oil. At the same time, it does not require complicated enzymatic pretreatment and can be processed in one step by fermentation, resulting in low processing costs.

[0005] The present invention adopts the following technical solution:

[0006] A method for reusing biomass waste includes: mixing filamentous fungi and yeast with biomass waste, fermenting and culturing it, then performing solid-liquid separation, and extracting biomass oil from the separated solid.

[0007] The filamentous fungi include one or a combination of two selected from Trichoderma reesei and Aspergillus niger;

[0008] The yeast includes one or more combinations selected from Saccharomyces stevidae and Rhodotorula rubrum.

[0009] In some preferred embodiments, the biomass waste includes fruit pomace, coffee grounds, or Chinese medicine residue.

[0010] In some preferred embodiments, the fruit pomace, coffee grounds, or Chinese medicine residue is waste residue from which sugars, oils, or pectin have been extracted, and the mass percentage of cellulose, lignin, and hemicellulose in the waste residue is greater than 90%.

[0011] In some preferred embodiments, the biomass waste residue is mixed with the filamentous fungi and yeast for fermentation without pretreatment or pH adjustment to 4.5-6.

[0012] In some preferred embodiments, the red yeast includes one or more combinations selected from Rhodotorula glutinis, Rhodotorula natans, and Rhodotorula spp.

[0013] In some preferred embodiments, the filamentous fungus is 0.3 × 10 7 ~1.5×10 7 The yeast was added to the first culture medium at an inoculum size of 0.3 × 10⁻⁶ cells / mL for pre-culturing. 7 ~1.5×10 7 An inoculum of 100 cells / mL was added to the second culture medium for pre-culture, and the two pre-culture products were mixed with biomass waste.

[0014] In some embodiments, the amount of filamentous fungi added is 0.3 × 10⁻⁶. 7 ~1.5×10 7 The number of filamentous fungi added can be 0.3 × 10⁻⁶ per mL. 7 cells / mL, 0.4×10 7 cells / mL, 0.5×10 7 cells / mL, 0.6×10 7 cells / mL, 0.7×10 7 cells / mL, 0.8×10 7 cells / mL, 0.9×10 7 cells / mL, 1.0×10 7 cells / mL, 1.1×10 7 cells / mL, 1.2×10 7 cells / mL, 1.3×10 7 cells / mL, 1.4 × 10 7 cells / mL or 1.5 × 10⁻⁶ 7 / mL. In a more preferred embodiment, the amount of filamentous fungi added is 0.3 × 10⁻⁶. 7 cells / mL ~1.2×10 7 / mL. In a more preferred embodiment, the amount of filamentous fungi added is 0.3 × 10⁻⁶. 7 Cells / mL ~ 1×10 7 / mL. In a specific and preferred embodiment, the amount of filamentous fungi added is 0.4 × 10⁻⁶. 7per mL.

[0015] In some embodiments, the amount of yeast added is 0.3 × 10⁻⁶. 7 ~1.5×10 7 The yeast count can be 0.3 × 10⁶ cells / mL. 7 cells / mL, 0.4×10 7 cells / mL, 0.5×10 7 cells / mL, 0.6×10 7 cells / mL, 0.7×10 7 cells / mL, 0.8×10 7 cells / mL, 0.9×10 7 cells / mL, 1.0×10 7 cells / mL, 1.1×10 7 cells / mL, 1.2×10 7 cells / mL, 1.3×10 7 cells / mL, 1.4 × 10 7 cells / mL or 1.5 × 10⁻⁶ 7 / mL. In a more preferred embodiment, the amount of yeast added is 0.3 × 10⁻⁶. 7 cells / mL ~1.2×10 7 / mL. In a more preferred embodiment, the amount of yeast added is 0.3 × 10⁻⁶. 7 Cells / mL ~ 1×10 7 / mL. In a specific and preferred embodiment, the amount of yeast added is 0.4 × 10⁻⁶. 7 per mL.

[0016] In some embodiments, the amount of biomass waste added is 10-80 g / L. Preferably, the amount of biomass waste added is 10-50 g / L; more preferably, the amount of biomass waste added is 20-30 g / L.

[0017] In some preferred embodiments, the first culture medium includes a carbon source, a nitrogen source, a yeast extract, and an inorganic salt containing metal cations, wherein the inorganic salt containing metal cations is composed of potassium hydrogen phosphate 0.45-0.65 g / L, magnesium sulfate heptahydrate 0.45-0.65 g / L, zinc sulfate heptahydrate 0.04-0.07 g / L, and ferric chloride hexahydrate 0.005-0.011 g / L; the second culture medium includes YPD (yeast extract-tryptone-glucose medium) medium.

[0018] In some more preferred embodiments, the pre-culture time of the filamentous fungus is 12-48 hours, and the pre-culture time of the yeast is 12-48 hours.

[0019] In some preferred embodiments, filamentous fungi and yeasts are mixed with biomass waste and fermented for 48-120 hours.

[0020] In some preferred embodiments, the separated solids are washed to retain the precipitate, subjected to cell wall disruption treatment, and extracted with an organic solvent to obtain biomass oil.

[0021] In some specific and preferred embodiments, the method for reusing biomass waste is implemented as follows:

[0022] Step A: Take 0.3×10 7 ~1.5×10 7 Add 1 / mL of *Saccharomyces stearothermii* or *Rhodotorula rubra* to YPD medium and pre-culture at 25-35℃ and 80-120 rpm for 20-36 h.

[0023] Step B, take 0.3×10 7 ~1.5×10 7 Add Trichoderma reesei per mL to the culture medium and pre-culture at 25-35℃ and shaking speed of 80-120 rpm for 20-36 h;

[0024] Step C: Place fruit pomace, coffee grounds, or medicinal residue with a pH adjusted to 4.5-6 in a shaker flask;

[0025] Step D: Add the two pre-cultured systems from Step A and Step B into a shake flask and incubate for 60-84 hours at a temperature of 25-35℃ and a shaking speed of 60-100 rpm.

[0026] Step E: Centrifuge and collect the solid; wash the solid with PBS and retain the precipitate; add hydrochloric acid and mix the precipitate, then incubate in a water bath for 1-2 hours; add chloroform-methanol mixture and extract at 25-35°C and 80-120 rpm; then centrifuge, remove the chloroform phase at the bottom, and evaporate or blow dry.

[0027] The present invention adopts the above solution and has the following advantages:

[0028] The method proposed in this invention utilizes a combination of specific filamentous fungi and specific yeasts to perform mixed fermentation on biomass waste residue that is essentially free of sugars, oils, and pectin, thereby producing oil. This improves the utilization rate of the biomass waste residue and results in a product with high added value. Specifically, the fermentation residue produced by mixing the two types of fungi with the biomass waste residue in a one-step process can be easily extracted to obtain biomass oil. Furthermore, the biomass waste residue does not require a complex pretreatment process, making the entire reuse process simple and suitable for industrial application. Attached Figure Description

[0029] To more clearly illustrate the technical solution of the present invention, the accompanying drawings used in the description of the embodiments will be briefly introduced below. Obviously, the accompanying drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0030] Figure 1 Photographs of the fruit pomace used in Examples 1 to 3 in a fermentation tank.

[0031] Figure 2 This is a photograph of the material in the shake flask after fermentation in Example 1.

[0032] Figure 3 This is a microscope image of the fermented system in Example 1.

[0033] Figure 4 A photograph of the oil collected in Example 1.

[0034] Figure 5 This is a microscope image of the fermented system in Example 2.

[0035] Figure 6 A photograph of the oil collected in Example 2.

[0036] Figure 7 A photograph of the oil collected in Example 3.

[0037] Figure 8 A photograph of the oil collected in Example 4.

[0038] Figure 9 A photograph of the oil collected in Example 5. Detailed Implementation

[0039] The preferred embodiments of the present invention will now be described in detail with reference to the accompanying drawings, so that the advantages and features of the present invention can be more readily understood by those skilled in the art. It should be noted that the description of these embodiments is for the purpose of aiding understanding the present invention, but does not constitute a limitation thereof.

[0040] Difficult-to-process biomass waste mainly consists of pectin-extracted fruit pomace (from juice factories), medicinal residues (such as dried tangerine peel and astragalus from pharmaceutical factories), or coffee grounds. Fruit pomace, coffee grounds, or medicinal residues are waste from which sugars, oils, or pectin have already been extracted, with cellulose, lignin, and hemicellulose accounting for more than 90% of the residue by mass. Fruit pomace after pectin extraction is even more nutrient-poor, requiring adjustment of microorganisms for oil production. Medicinal residues contain antibacterial metabolites, resulting in low fermentation efficiency. This implementation method aims to reuse these types of biomass waste to produce biomass oil. Using a mixture of specific filamentous fungi and specific yeasts, a one-step mixed fermentation process eliminates the need for complex pretreatment of the aforementioned wastes (such as enzymatic hydrolysis). The resulting microbial residue can be directly fermented to extract biomass oil. The filamentous fungi selected are *Trichoderma reesei* or *Aspergillus niger*, and the yeasts selected are *Saccharomyces cerevisiae*, *Rhodotorula rubrum*, or *Kluyveromyces martensii*. The *Rhodotorula rubrum* is selected from *Rhodotorula glutinis*, *Rhodotorula glutinis*, or *Rhodotorula spp.* The inventors discovered that, compared to other filamentous fungi and yeasts, this combination of fungi (Trichoderma reesei / Aspergillus niger + Saccharomyces stevidae / Rhodotorula rubrum) can more effectively ferment and treat the above-mentioned biomass waste residue, and the oil yield is also higher; in particular, the combination of Trichoderma reesei and Rhodotorula rubrum or Rhodotorula rubrum has a higher oil yield and the product has a higher proportion of oleic acid.

[0041] The amount of filamentous fungi added was 0.3 × 10⁻⁶. 7 ~1.5×10 7 Cells / mL, preferably 0.3 × 10⁻⁶. 7 Cells / mL ~ 1×10 7 The yeast count was 0.3 × 10⁶ cells / mL. 7 ~1.5×10 7 Cells / mL, preferably 0.3 × 10⁻⁶. 7 Cells / mL ~ 1×10 7 The amount of biomass waste added is 10~80 g / L, preferably 10~50 g / L, and more preferably 20~30 g / L.

[0042] In one specific implementation, 0.3 × 10 7 ~1.5×10 7 0.3 × 10⁶ cells / mL of *Saccharomyces cerevisiae* and *Rhodotorula rubrum* were added to YPD medium and pre-cultured at 25–35°C and a shaking speed of 80–120 rpm for 20–36 h. In one specific embodiment, 0.3 × 10⁶ cells / mL of *Saccharomyces cerevisiae* and *Rhodotorula rubrum* were 7 ~1.5×10 7Trichoderma reesei was added at a concentration of 1 / mL to the first culture medium and pre-cultured at 25–35°C and a shaking speed of 80–120 rpm for 20–36 h. The first culture medium comprised a carbon source, a nitrogen source, yeast extract, and an inorganic salt containing metal cations. The inorganic salt containing metal cations consisted of 0.45–0.65 g / L potassium hydrogen phosphate, 0.45–0.65 g / L magnesium sulfate heptahydrate, 0.04–0.07 g / L zinc sulfate heptahydrate, and 0.005–0.011 g / L ferric chloride hexahydrate. The first culture medium used in the following examples was prepared according to the formulation disclosed in Example 1 of patent document CN116622518A. PDB (potato dextrose broth) medium can also be used as the first culture medium.

[0043] In one specific embodiment, fruit pomace, coffee grounds, or medicinal residue, with the pH adjusted to 4.5-6 (preferably 5.5-6), is placed in a shake flask at a concentration of 20-30 g / L. In another specific embodiment, the two pre-cultured systems are added to the shake flask and incubated at 25-35°C and a shaking speed of 60-100 rpm for 60-84 h. In yet another specific embodiment, the solid is collected by centrifugation; the solid is washed with PBS, retaining the precipitate; hydrochloric acid is added to mix the precipitate, and the mixture is incubated in a water bath for 1-2 h; a chloroform-methanol mixture is added, and extraction is performed at 25-35°C and a shaking speed of 80-120 rpm; then, the mixture is centrifuged, the lower chloroform phase is collected, and the mixture is evaporated or dried to extract the biomass oil.

[0044] raw material:

[0045] The fruit pomace, sourced from Yantai Northern Andeli Juice Co., Ltd., consists of 10 g / L apple pomace and 10 g / L lemon pomace, both of which have had pectin removed by acid extraction and pH adjusted to 5.5-6.

[0046] Coffee grounds: waste from coffee shops.

[0047] Medicinal residue: Derived from the tangerine peel and astragalus residue from pharmaceutical factories after the extraction of active ingredients. Medicinal residue contains antibacterial metabolites, and conventional fermentation methods have low efficiency.

[0048] Trichoderma reesei: ATCC 56765.

[0049] Strychnospermum oryzae: JCM3767.

[0050] Red yeast rice: ATCC 9449.

[0051] Red syringa yeast: ATCC 10657.

[0052] Red yeast rice: ATCC 204091. Example 1

[0053] Place the fruit pomace in a 2L fermentation shaker (500mL fermentation scale). Specifically, add 500mL of a mixture of apple and lemon pomace to the fermentation shaker. This mixture is formed by mixing 10 g / L dry weight apple pomace and 10 g / L dry weight lemon pomace in a 1:1 volume ratio. Figure 1 As shown, the large particles in the flask are now very noticeable when shaken.

[0054] 1×10 7 Add one unit of Rhodotorula glutinis to 25 mL of YPD medium and incubate at 28°C and 100 rpm for 24 h.

[0055] 1×10 7 Add one Trichoderma reesei spore to 25 mL of the first culture medium and incubate at 28°C and 100 rpm for 24 hours.

[0056] The two pre-culture products were then simultaneously inoculated into the aforementioned 2L fermentation shake flasks, and incubated at 28°C and 75 rpm for a total of 72 hours. The fermentation results are as follows: Figure 2 and Figure 3 As shown, see Figure 2 Large solid particles are rarely found in the material system after shake-flask fermentation; see also Figure 3 After co-fermentation, Rhodotorula glutinis and Trichoderma reesei grew in large quantities in the material system.

[0057] After fermentation, solid-liquid separation was performed by centrifugation at 12000 rpm, and the solid was collected. The solid was added to one volume of PBS and washed twice at 5000 rpm, retaining the precipitate as the final separated sample. An appropriate amount of 4M hydrochloric acid was added to mix and disrupt the cell wall, and the mixture was placed in a 60℃ water bath for 1.5 h, inverting and mixing every 0.5 h. After the water bath, a 1:1 volume of methanol-chloroform (1:1) was added, and the mixture was incubated at 30℃ and 100 rpm for 1 h. Then, it was centrifuged at 5000 rpm for 10 min at room temperature. The lower chloroform phase was aspirated in a fume hood and then dried by blowing the chloroform. The resulting sample is shown below. Figure 4 The lipids extracted using the small extraction method were obtained as yellow-brown solid lipids. The average oil yield from the three parallel experiments was 0.260g.

[0058] Lipid analysis results showed that oleic acid accounted for the largest proportion at 61.7%, palmitic acid for the second largest at 13.9%, linoleic acid for the third largest at 11%, and stearic acid for the fourth largest at 5.37%. Other tested components were present in small proportions or were not detected. The oils were tested by a third-party company, PONY, and the results are shown in Table 1 below.

[0059]

[0060] Example 2

[0061] Place the fruit pomace in a 2L fermentation shaker (500mL fermentation scale). Specifically, add 500mL of a mixture of apple pomace and lemon pomace to the fermentation shaker. The mixture is formed by mixing apple pomace (10 g / L dry weight) and lemon pomace (10 g / L dry weight) in a 1:1 volume ratio.

[0062] 1×10 7 Add one spore of *Evodia rutaecarpa* to 25 mL of YPD medium and incubate at 28°C and 100 rpm for 24 h.

[0063] 1×10 7 Add one Trichoderma reesei spore to 25 mL of the first culture medium and incubate at 28°C and 100 rpm for 24 hours.

[0064] The two pre-culture products were then simultaneously inoculated into the aforementioned 2L fermentation shake flasks and incubated at 28°C and 75 rpm for a total of 72 hours. (See also...) Figure 5 After co-fermentation, a large number of *Trichoderma spp.* and *Trichoderma reesei* grew in the material system.

[0065] After fermentation, solid-liquid separation was performed by centrifugation at 12000 rpm, and the solid was collected. The solid was added to one volume of PBS and washed twice at 5000 rpm, retaining the precipitate as the final separated sample. An appropriate amount of 4M hydrochloric acid was added to mix and disrupt the cell wall, and the mixture was placed in a 60℃ water bath for 1.5 h, inverting and mixing every 0.5 h. After the water bath, a 1:1 volume of methanol-chloroform (1:1) was added, and the mixture was incubated at 30℃ and 100 rpm for 1 h. Then, it was centrifuged at 5000 rpm for 10 min at room temperature. The lower chloroform phase was aspirated in a fume hood and then dried by blowing the chloroform. The resulting sample is shown below. Figure 6 The lipids were extracted using a small extraction method, and the resulting solid lipids were yellowish-brown. The average oil yield from the three parallel experiments was 0.247g.

[0066] Example 3

[0067] Place the fruit pomace in a 2L fermentation shaker (500mL fermentation scale). Specifically, add 500mL of a mixture of apple pomace and lemon pomace to the fermentation shaker. The mixture is formed by mixing apple pomace (10 g / L dry weight) and lemon pomace (10 g / L dry weight) in a 1:1 volume ratio.

[0068] 1×10 7 One spore of Rhodotorula glutinis was added to 25 mL of YPD medium and incubated at 28°C and 100 rpm for 24 h.

[0069] 1×10 7Add one Trichoderma reesei spore to 25 mL of the first culture medium and incubate at 28°C and 100 rpm for 24 hours.

[0070] Then, the two pre-culture products were simultaneously inoculated into the above 2L fermentation shake flask, and the culture temperature was 28 degrees Celsius, 75 rpm, for a total of 72 hours.

[0071] After fermentation, solid-liquid separation was performed by centrifugation at 12000 rpm, and the solid was collected. The solid was added to one volume of PBS and washed twice at 5000 rpm, retaining the precipitate as the final separated sample. An appropriate amount of 4M hydrochloric acid was added to mix and disrupt the cell wall, and the mixture was placed in a 60℃ water bath for 1.5 h, inverting and mixing every 0.5 h. After the water bath, a 1:1 volume of methanol-chloroform (1:1) was added, and the mixture was incubated at 30℃ and 100 rpm for 1 h. Then, it was centrifuged at 5000 rpm for 10 min at room temperature. The lower chloroform phase was aspirated in a fume hood and then dried by blowing the chloroform. The resulting sample is shown below. Figure 7 The lipids were extracted using a small extraction method, and the resulting solid lipids were yellowish-brown. The average oil yield in parallel experiments was 0.35g.

[0072] Example 4

[0073] The raw material was replaced with medicinal residue with a dry weight of 20 g / L, and the dosage was 500 mL. Other procedures were the same as in Example 3. The resulting sample was as follows: Figure 8 The average oil yield in the parallel experiments was 0.34g.

[0074] Example 5

[0075] The raw material was replaced with coffee grounds with a dry weight of 25 g / L, and the volume was 500 mL. All other steps were the same as in Example 3. The resulting sample was as follows: Figure 9 The average oil yield in the parallel experiments was 0.31g.

[0076] Examples 3 to 5 show that the method of the present invention can reuse different single biomass wastes (fruit pomace, medicinal residue, coffee grounds) to produce biomass oil.

[0077] As indicated in this specification and claims, the terms "comprising" and "including" only indicate the inclusion of explicitly identified steps and elements, and these steps and elements do not constitute an exclusive list, and the method or apparatus may also include other steps or elements.

[0078] It can be further understood that in this disclosure, "multiple" refers to two or more, and other quantifiers are similar.

[0079] It is further understood that the terms "first," "second," etc., are used to describe various types of information, but this information should not be limited to these terms. These terms are only used to distinguish information of the same type from one another, and do not indicate a specific order or degree of importance. In fact, the expressions "first," "second," etc., are completely interchangeable. For example, without departing from the scope of this disclosure, first information can also be referred to as second information, and similarly, second information can also be referred to as first information.

[0080] The above embodiments are only for illustrating the technical concept and features of the present invention, and are preferred embodiments. Their purpose is to enable those skilled in the art to understand the content of the present invention and implement it accordingly, and they cannot be used to limit the scope of protection of the present invention.

Claims

1. A method for recycling biomass waste, characterized in that, include: Yeast was cultured at a rate of 0.3 × 10⁻⁶. 7 ~1.5×10 7 The yeast pre-culture product was obtained by adding cells / mL to YPD medium and pre-culturing; Trichoderma reesei was added at a concentration of 0.3 × 10⁻⁶. 7 ~1.5×10 7 The pre-culture product of Trichoderma reesei was obtained by adding 1 / mL of Trichoderma reesei to the first culture medium or PDB medium; the pre-culture product of Trichoderma reesei and the pre-culture product of yeast were added to the biomass waste and mixed in a fermentation shaker, fermented for 60-84h, and then solid-liquid separation was performed to extract biomass oil from the separated solid. The material system in the fermentation shake flask before fermentation consists of the biomass waste residue, the pre-culture product of Trichoderma reesei, and the pre-culture product of yeast; the first culture medium includes a carbon source, a nitrogen source, yeast extract, and an inorganic salt containing metal cations, wherein the inorganic salt containing metal cations consists of potassium hydrogen phosphate 0.45-0.65 g / L, magnesium sulfate heptahydrate 0.45-0.65 g / L, zinc sulfate heptahydrate 0.04-0.07 g / L, and ferric chloride hexahydrate 0.005-0.011 g / L; The yeast includes one or more combinations selected from Rhodotorula glutinis and Rhodotorula natans. The biomass waste includes fruit pomace, coffee grounds, or traditional Chinese medicine residue from which sugars, oils, or pectin have been extracted, and the waste contains more than 90% by mass of cellulose, lignin, and hemicellulose. The biomass oil includes oleic acid, palmitic acid, and linoleic acid.

2. The method for recycling biomass waste according to claim 1, characterized in that, The biomass waste residue is mixed with Trichoderma reesei and yeast for fermentation without pretreatment or pH adjustment to 4.5-6.

3. The method for recycling biomass waste according to claim 1, characterized in that, The pre-culture time for Trichoderma reesei is 12-48 hours, and the pre-culture time for yeast is 12-48 hours.

4. The method for recycling biomass waste according to claim 1, characterized in that, 0.3×10 7 ~1.5×10 7 Trichoderma reesei was added at a concentration of 1 / mL to the first culture medium and pre-cultured at 25–35°C and 80–120 rpm for 20–36 h to obtain the pre-culture product of Trichoderma reesei; 0.3 × 10⁻⁶ cells / mL was then added to the medium. 7 ~1.5×10 7 The yeast cells per mL were added to YPD medium and pre-cultured at 25-35°C and 80-120 rpm for 20-36 hours to obtain the pre-culture product of yeast.

5. The method for recycling biomass waste according to claim 1, characterized in that, The separated solids were washed to retain the precipitate, which was then subjected to cell wall disruption treatment and extracted with an organic solvent to obtain biomass oil.

6. The method for recycling biomass waste according to claim 1, characterized in that, The specific implementation methods for the reuse of biomass waste are as follows: Step A: Take 0.3×10 7 ~1.5×10 7 Add yeast cells / mL to YPD medium and pre-culture at 25-35℃ and shaking speed of 80-120 rpm for 20-36 h; Step B, take 0.3×10 7 ~1.5×10 7 Add Trichoderma reesei per mL to the culture medium and pre-culture at 25-35℃ and shaking speed of 80-120 rpm for 20-36 h; Step C: Place fruit pomace, coffee grounds, or medicinal residue with a pH adjusted to 4.5-6 in a shaker flask; Step D: Add the two pre-cultured systems from Step A and Step B into a shake flask and incubate for 60-84 hours at a temperature of 25-35℃ and a shaking speed of 60-100 rpm. Step E: Centrifuge and collect the solid; wash the solid with PBS and retain the precipitate; Add hydrochloric acid, mix well to precipitate, and incubate in a water bath for 1-2 hours; add chloroform-methanol mixture and extract at 25-35°C and 80-120 rpm; then centrifuge, remove the chloroform phase at the bottom, and evaporate or blow dry.

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