A method for producing ethanol and lactic acid from organic waste

Abstract

This invention belongs to the field of organic waste fermentation treatment technology, specifically relating to a method for producing ethanol and lactic acid from organic waste. This method involves adding a certain amount of alcohol quorum sensing molecules to an anaerobic fermentation system of organic waste, which can promote the anaerobic fermentation synthesis of ethanol and lactic acid, increasing the yield of ethanol and / or lactic acid. The fermentation time is relatively short, making it highly valuable for widespread application and very suitable for industrial use.

Description

Technical Field

[0001] This invention belongs to the field of organic waste fermentation treatment technology. More specifically, it relates to a method for producing ethanol and lactic acid from organic waste. Background Technology

[0002] Food waste, a typical representative of organic waste, is a general term for restaurant waste and kitchen scraps. Its output is enormous, accounting for approximately 30% to 50% of urban household waste. Because its main components are protein, lipids, starch, and cellulose, it is rich in nutrients, highly perishable, emits foul odors, and spreads bacteria and viruses. With my country's rapid economic development, the output of food waste is increasing, and the resulting problems are becoming increasingly serious. How to effectively treat and dispose of food waste and other organic wastes is an urgent problem to be solved. Among these solutions, utilizing such organic waste to produce fuel ethanol and lactic acid is an effective way to achieve its high-value utilization.

[0003] Currently, among existing technologies for the resource utilization of such organic waste, Chinese patent application CN103484521A discloses a method for promoting the production of ethanol and lactic acid from kitchen waste through hydrothermal treatment. This method involves pre-treating the kitchen waste with hydrothermal heat before fermentation to ensure more thorough hydrolysis, followed by the sequential addition of ethanol and lactic acid fermentation bacteria for stepwise fermentation to obtain ethanol and lactic acid. However, this method requires hydrothermal treatment, resulting in high energy consumption and complex steps. Therefore, developing a highly efficient method for treating organic waste, increasing the yield of ethanol and / or lactic acid, and reducing treatment costs has become an urgent technical problem to be solved in this field. Summary of the Invention

[0004] The technical problem to be solved by the present invention is to overcome the defects and shortcomings of the prior art in the resource utilization of organic waste, which requires a lot of energy and involves complicated steps, and to provide a method for producing ethanol and lactic acid from organic waste.

[0005] Another objective of this invention is to provide the application of the method in the fermentation of organic waste.

[0006] The above-mentioned objective of this invention is achieved through the following technical solution:

[0007] A method for producing ethanol and lactic acid from organic waste involves adding alcohol quorum sensing molecules at a concentration of 1–500 μmol / L to an anaerobic fermentation system of organic waste.

[0008] The alcohol quorum sensing molecule is phenylethanol, tyrosol, or cromoglycan.

[0009] When faced with environmental stress, microorganisms activate quorum sensing systems to adapt to changes. Utilizing quorum sensing to regulate the physiological metabolism during anaerobic fermentation can promote the synthesis and transformation of microbial metabolites, thus optimizing anaerobic fermentation technology. Quorum sensing is a cell-density-dependent microbial communication mechanism that can be used to regulate cellular physiological metabolism. Quorum sensing can induce microorganisms to exhibit physiological characteristics not present in small numbers or individual microbial cells, such as biofluorescence, antibiotic synthesis, and biofilm formation. Regulating the cellular physiological characteristics of anaerobic fermenting microorganisms through quorum sensing and promoting the synthesis of cellular metabolites is a feasible measure to improve the efficiency of anaerobic fermentation of food waste and other organic waste.

[0010] The inventors unexpectedly discovered that during the fermentation of mixed raw material food waste, a specific concentration of quorum sensing molecules can increase the yield of ethanol and / or lactic acid. Based on this, the present invention provides a method for adding alcohol quorum sensing molecules to promote the production of ethanol and lactic acid from organic waste.

[0011] Furthermore, the organic waste includes kitchen waste, lignocellulosic waste, starchy waste, and other carbon-containing organic waste.

[0012] Preferably, when the organic waste is kitchen waste, the alcohol quorum sensing molecule is phenylethanol at 50±15μmol / L or 300±50μmol / L, or tyrosol at 25±15μmol / L, 50±15μmol / L or 300±50μmol / L, or chromool at 50±15μmol / L or 300±50μmol / L.

[0013] Preferably, when the organic waste is lignocellulosic waste, the alcohol quorum sensing molecule is phenylethanol at 50±15μmol / L or 200±50μmol / L, or tyrosol at 25±15μmol / L, 50±15μmol / L, 100±50μmol / L, or 300±50μmol / L, or chromool at 25±15μmol / L, 100±50μmol / L, 200±50μmol / L, or 300±50μmol / L.

[0014] Preferably, when the organic waste is starch-based waste, the alcohol quorum sensing molecule is 25±15μmol / L phenylethanol or 50±15μmol / L tyrosol.

[0015] Preferably, the microbial strains in the anaerobic fermentation system include bacteria and fungi.

[0016] Furthermore, the bacteria are two or more species selected from the genera *Limosilactobacillus*, *Weissella*, *Pediococcus*, *Clostridiaceae*, *Bacillus*, *Proteiniphilum*, and *Aminobacterium*. Using species from any of these genera will achieve the desired effect.

[0017] Furthermore, the *Lactobacillus* genus is selected from one or more of *Lactobacillus delbrueckii*, *Lactobacillus plantarum*, *Lactobacillus helveticus*, *Lactobacillus acidophilus*, and *Lactobacillus caudatus*; the *Weissella* genus is selected from one or more of *Weissella fusionis*, *Weissella esculenta*, *Weissella entericae*, *Weissella viride*, *Weissella univar*, *Weissella cannii*, *Weissella oryzae*, *Weissella henryi*, *Weissella microphylla*, *Weissella staphylococcus*, and *Weissella serrata*; the *Pediococcus* genus is selected from one or more of *Pediococcus brevis*, *Pediococcus lactis*, *Pediococcus pentosaceus*, and *Pediococcus halophilus*; the *Clostridium* genus is selected from one or more of *Clostridium butyricum* and *Clostridium acetobutanol*; the *Bacillus* genus is selected from one or more of *Bacillus polymyxa*, *Bacillus subtilis*, *Bacillus cereus*, *Bacillus licheniformis*, and *Bacillus brevis*; the *Proteophile* genus is selected from one or more of *Acetobacter acetophilus* and *Akkermansia muciniphilus*; and the *Aminobacter* genus is selected from one or more of *Aminobacterium hydrophilum* and *Aminobacterium aminophilum*.

[0018] Preferably, the abundance ratio of the species of *Limosilactobacillus*, *Weissella*, *Pediococcus*, *Clostridiaceae*, *Bacillus*, *Proteiniphilum*, and *Aminobacterium* is: (30–65):(1–45):(0.5–15):(0.01–27):(0.1–20):(0.01–5):(0.01–15).

[0019] Furthermore, the fungus is two or more species selected from the genera *Saccharomyces*, *Aspergillus*, *Fusarium*, *Cyberlindnera*, *Cladosporium*, *Mortierella*, *Alternaria*, *Penicillium*, *Filobasidium*, *Trichoderma*, and *Rhizopus*. Using species from any of these genera will achieve the effects described in this application.

[0020] Furthermore, the yeast genus is selected from one or more of *Saccharomyces cerevisiae*, *Schizosaccharomyces*, *Hansenula*, *Pichia pastoris*, *Candida*, *Gnaphalium*, and *Rhodotorula*; the *Aspergillus* genus is selected from one or more of *Aspergillus awamori*, *Aspergillus niger*, *Aspergillus ochre*, *Aspergillus oryzae*, *Aspergillus sacchari*, *Aspergillus styracifolius ... and *Aspergillus stigmata*; the *Fusarium* genus is selected from one or more of *Fusarium*, *Fusarium*, *Fusarium*, and *Fusarium*; the *Cyclocarya* genus is selected from one or more of *Saccharomyces cerevisiae* and *Saccharomyces cerevisiae*; the *Cladosporium* genus is selected from *Cladosporium xanthosporium*, *Cladosporium multiflorum*, *Cladosporium lantana*, and *Cladosporium trifolium*. One or more of the following: * *Morchella* is selected from *Morchella longisporum*, *Morchella alpineensis*, *Morchella microsporum*, *Morchella ramanense*, *Morchella scabranchii*, and *Morchella longibranchii*; * *Alternaria* is selected from one or more of *Alternaria longipetalum*, *Alternaria scabranchii*, *Alternaria scabranchii*, and *Alternaria microsporum*; * *Penicillium* is selected from one or more of *Penicillium reskeletoni*, *Penicillium heckii*, *Penicillium chrysogenum*, and *Penicillium salicum*; * *Filamentosa* is selected from one or more of rust fungi, smut fungi, lamellar fungi, and gastromycetes; * *Trichoderma* is selected from one or more of *Trichoderma viride*, *Trichoderma cornigerum*, *Trichoderma echinosporum*, *Trichoderma harzianum*, and *Trichoderma longibranchii*; * *Rhizopus* is selected from one or more of *Rhizopus nigricans*, *Rhizopus huassifolia*, and *Rhizopus oryzae*.

[0021] Preferably, the abundance ratio of the genera *Saccharomyces*, *Aspergillus*, *Fusarium*, *Cyberlindnera*, *Cladosporium*, *Mortierella*, *Alternaria*, *Penicillium*, *Filobasidium*, *Trichoderma*, and *Rhizopus* is: (1–80):(0.001–45):(0.001–1):(0.0001–0.1):(0.001–1):(0.001–1):(0.001–5):(0.001–1):(0.001–3):(0.001–3):(0.001–25).

[0022] Preferably, the concentration of the microbial strains in the anaerobic fermentation system is 1×10⁻⁶. 6 ~1.5×10 9 cfu / mL.

[0023] Preferably, the inoculum size of the anaerobic fermentation system is 5% to 30% of the volume of the organic waste.

[0024] Preferably, the fermentation time is 3 to 30 days.

[0025] Preferably, the temperature of the anaerobic fermentation system is 25–65°C.

[0026] In addition, the present invention also provides the application of the method in the fermentation of organic waste.

[0027] The present invention has the following beneficial effects: The present invention provides a method for producing ethanol and lactic acid from organic waste, namely, adding a certain amount of alcohol quorum sensing molecules to an organic waste fermentation system inoculated with mixed microorganisms, which can promote the anaerobic fermentation of organic waste to synthesize ethanol and lactic acid, improve the yield of ethanol and / or lactic acid, and shorten the fermentation time. It has great value for promotion and application and is very suitable for industrial application. Attached Figure Description

[0028] Figure 1 The effect of phenylethanol on ethanol production from anaerobic fermentation of kitchen waste;

[0029] Figure 2 The effect of phenylethanol on lactic acid production from anaerobic fermentation of kitchen waste;

[0030] Figure 3 The effect of tyrosol on ethanol production from anaerobic fermentation of food waste;

[0031] Figure 4 The effect of tyrosol on lactic acid production from anaerobic fermentation of food waste;

[0032] Figure 5 The effect of ascorbol on ethanol production from anaerobic fermentation of kitchen waste;

[0033] Figure 6 The effect of ascorbol on lactic acid production from anaerobic fermentation of kitchen waste;

[0034] Figure 7 The effect of phenylethanol on ethanol production from anaerobic fermentation of Chinese cabbage culture medium;

[0035] Figure 8 The effect of phenylethanol on lactic acid production during anaerobic fermentation of Chinese cabbage culture medium;

[0036] Figure 9 The effect of tyrosol on ethanol production from anaerobic fermentation of Chinese cabbage culture medium;

[0037] Figure 10 The effect of tyrosol on lactic acid production during anaerobic fermentation of Chinese cabbage culture medium;

[0038] Figure 11 The effect of ascorbol on ethanol production from anaerobic fermentation of Chinese cabbage culture medium;

[0039] Figure 12 The effect of ascorbol on lactic acid production during anaerobic fermentation of Chinese cabbage culture medium;

[0040] Figure 13 The effect of phenylethanol on ethanol production from anaerobic fermentation of rice culture medium;

[0041] Figure 14 The effect of phenylethanol on lactic acid production during anaerobic fermentation of rice culture medium;

[0042] Figure 15 The effect of tyrosol on ethanol production from anaerobic fermentation of rice culture medium;

[0043] Figure 16 The effect of tyrosol on lactic acid production during anaerobic fermentation of rice culture medium;

[0044] Figure 17 The effect of ascorbol on ethanol production from anaerobic fermentation of rice culture medium;

[0045] Figure 18 The effect of chromol on lactic acid production during anaerobic fermentation of rice culture medium.

[0046] In the examples, the concentration values ​​of the quorum sensing molecules added are represented in the figure as “letter [P(phenylethanol), T(tyrosol), I(chromosol)] + concentration value”, such as 100, 200, 300 μmol / L; in the figure, they are respectively recorded as P100 (or T100, I100), P200 (or T200, I200), P300 (or T300, I300), and the blank experimental group without any quorum sensing molecules added is recorded as K. Detailed Implementation

[0047] The present invention will be further described below with reference to the accompanying drawings and specific embodiments, but the embodiments do not limit the present invention in any way. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in this technical field.

[0048] Unless otherwise specified, all reagents and materials used in the following examples are commercially available.

[0049] Because the fermentation experiments with high concentrations (100–300 μmol / L) and low concentrations (25–50 μmol / L) of quorum sensing molecules were not conducted in the same batch, the fermentation substrate components collected were inconsistent, resulting in different concentrations of ethanol and lactic acid in the control group (K). The experimental results for high and low concentrations of quorum sensing molecules were plotted separately in this example. Furthermore, the basis for determining the effect of quorum sensing molecules on ethanol and lactic acid fermentation in this embodiment was: examining the difference between the daily product measurement and the control value. When the difference was positive, and the product measurement on any one day was greater than the maximum value of the control group, it indicated that the quorum sensing molecules promoted product production. Conversely, it indicated that the quorum sensing molecules inhibited product production. When the difference was the maximum value within 7 days, it indicated that the optimal effect of promoting product yield was achieved on that day. When the ethanol or lactic acid yield peaked earlier than the control group within seven days, and this peak value was greater than the yield value of the control group on the same day but less than the maximum value of the control group, it indicated that although the quorum sensing molecules did not increase product yield, they could rapidly stimulate microorganisms to produce products, shortening the product production time.

[0050] Example 1: Effect of adding phenylethanol on the production of ethanol and lactic acid in food waste as a fermentation substrate. 1. Experimental method

[0051] S1. Perform preliminary sorting on the collected kitchen waste to remove impurities that are not easily fermented;

[0052] S2. Add bacterial flora to make the abundance ratio of Lactobacillus delbrueckii, Weissella oryzae, Pediococcus lactis, Clostridium acetobutyricum, Bacillus subtilis, acetophilic proteobacterium, and aminoaminotrophic bacteria 30:7.9:15:27:0.1:5:15.

[0053] The abundance ratio of the fungi *Saccharomyces cerevisiae*, *Aspergillus niger*, *Fusarium graminearum*, *Giardia lamblia*, *Cladosporium xanthosporium*, *Morchella alpina*, *Alternaria microsporum*, *Penicillium reskeletoni*, *Ustilago maydis*, *Trichoderma corniglanense*, and *Rhizopus oryzae* was 80:0.001:1:0.1:1:1:5:1:3:3:4.899.

[0054] S3. Add 25 μmol / L, 50 μmol / L, 100 μmol / L, 200 μmol / L, and 300 μmol / L phenylethanol (these concentration values ​​are the concentrations in the anaerobic fermentation system);

[0055] S4. Anaerobic fermentation was carried out at 30℃ for 7 days. After fermentation, the mixture was centrifuged and the supernatant was purified into ethanol and lactic acid using conventional methods.

[0056] The bacteria are added at a rate of 8% (by volume), and the fungi are added at a rate of 8% (by volume).

[0057] 2. Experimental Results

[0058] The effect of phenylethanol on ethanol production from anaerobic fermentation of kitchen waste was as follows: Figure 1 As shown, in the anaerobic fermentation system using food waste as a substrate, high concentrations of phenylethanol generally inhibited ethanol synthesis, and only in the experimental group with a phenylethanol concentration of 50 μmol / L (P50) was ethanol synthesis promoted in the anaerobic fermentation system. The same promoting effect was obtained at concentrations near this value.

[0059] The effect of phenylethanol on lactic acid production from anaerobic fermentation of kitchen waste was as follows: Figure 2 As shown, in an anaerobic fermentation system using food waste as a substrate, 300 μmol / L phenylethanol can promote lactic acid fermentation. The same promoting effect can be obtained at concentrations near this value.

[0060] Example 2: Effect of adding tyrosol on the production of ethanol and lactic acid in food waste as a fermentation substrate. 1. Experimental method

[0061] The main steps of this embodiment are the same as those of Example 1, except that in step S3, the added quorum sensing molecule is tyrosol.

[0062] 2. Experimental Results

[0063] The effect of tyrosol on ethanol production from anaerobic fermentation of food waste was as follows: Figure 3 As shown, in an anaerobic fermentation system using food waste as a substrate, high concentrations of tyrosol are detrimental to ethanol production, while low concentrations of tyrosol (T25, T50) can promote ethanol synthesis, and the lower the tyrosol concentration, the more significant the promoting effect. The tyrosol concentration of 25 μmol / L showed the best effect in promoting ethanol synthesis.

[0064] The effect of tyrosol on lactic acid production from anaerobic fermentation of food waste was as follows: Figure 4 As shown, in an anaerobic fermentation system using food waste as a substrate, tyrosol at concentrations of 25 μmol / L and 300 μmol / L promoted lactic acid production. Similar promoting effects were observed at concentrations near these values.

[0065] Example 3: Effect of Adding Chrysodium on Ethanol and Lactic Acid Production in Food Waste Fermentation Substrate 1. Experimental Methods

[0066] The main steps of this embodiment are the same as those of Example 1, except that in step S3, the added quorum sensing molecule is chromool.

[0067] 2. Experimental Results

[0068] The effect of trypsinol on ethanol production from anaerobic fermentation of food waste was as follows: Figure 5 As shown, in an anaerobic fermentation system using food waste as a substrate, high concentrations of tryptophan have no promoting effect on ethanol production; however, when the tryptophan concentration is 50 μmol / L, tryptophan has a positive effect on ethanol production. The same promoting effect can be obtained at concentrations near this value.

[0069] The effect of trypsinol on lactic acid production from anaerobic fermentation of food waste was as follows: Figure 6 As shown, in an anaerobic fermentation system using food waste as a substrate, 50 μmol / L and 300 μmol / L phenylethanol can increase the lactic acid production during the anaerobic fermentation of food waste. Similar promoting effects can be obtained at concentrations near these values.

[0070] Example 4: Effect of adding phenylethanol on the production of ethanol and lactic acid in leafy greens as a fermentation substrate.

[0071] 1. Experimental Methods

[0072] The main steps of this embodiment are the same as those of Example 1, except that the substrate for fermentation is green vegetables.

[0073] 2. Experimental Results

[0074] The effect of phenylethanol on ethanol production from anaerobic fermentation of Chinese cabbage culture medium was as follows: Figure 7 As shown, in the anaerobic fermentation system using leafy greens as a substrate, exogenous phenylethanol has an inhibitory effect on ethanol production. However, the inhibitory effect is not significant in the low-concentration phenylethanol test group.

[0075] The effect of phenylethanol on lactic acid production during anaerobic fermentation of Chinese cabbage culture medium was as follows: Figure 8 As shown, in an anaerobic fermentation system using leafy greens as a substrate, 50 μmol / L and 200 μmol / L phenylethanol can increase lactic acid production. Similar promoting effects can be obtained at concentrations near these values.

[0076] Example 5: Effect of adding tyrosol on the production of ethanol and lactic acid in green leafy vegetables as a fermentation substrate.

[0077] 1. Experimental Methods

[0078] The main steps of this embodiment are the same as those of Example 1, except that the substrate for fermentation is green vegetables and the added quorum sensing molecule is tyrosol.

[0079] 2. Experimental Results

[0080] The effect of tyrosol on ethanol production from anaerobic fermentation of Chinese cabbage culture medium was as follows: Figure 9 As shown, in an anaerobic fermentation system using leafy greens as a substrate, a tyrosol concentration of 50 μmol / L promotes ethanol synthesis. The same promoting effect is obtained at concentrations near this value.

[0081] The effect of tyrosol on lactic acid production during anaerobic fermentation of leafy greens was as follows: Figure 10 As shown, in the anaerobic fermentation system using Chinese cabbage as a substrate, high concentrations of tyrosol (100 μmol / L and 300 μmol / L) and low concentrations of tyrosol (25 μmol / L and 50 μmol / L) can all promote the fermentation of Chinese cabbage to produce lactic acid.

[0082] Example 6: Effect of adding croterol on the production of ethanol and lactic acid in green leafy vegetables as a fermentation substrate

[0083] 1. Experimental Methods

[0084] The main steps of this embodiment are the same as those of Example 1, except that the substrate for fermentation is green vegetables and the added quorum sensing molecule is croterol.

[0085] 2. Experimental Results

[0086] The effect of tryptophan on ethanol production from anaerobic fermentation of Chinese cabbage culture medium was as follows: Figure 11 As shown, in the anaerobic fermentation system using leafy greens as a substrate, high concentrations of croterol inhibited ethanol production; however, a croterol concentration of 50 μmol / L was beneficial for increasing ethanol content. The same promoting effect was achieved at concentrations near this value.

[0087] The effect of tryptophan on lactic acid production during anaerobic fermentation of Chinese cabbage culture medium was as follows: Figure 12 As shown, in an anaerobic fermentation system using leafy greens as a substrate, high concentrations (100, 200, 300 μmol / L) and 25 μmol / L of croterol can increase lactic acid synthesis yield.

[0088] Example 7: Effect of adding phenylethanol on the production of ethanol and lactic acid in rice-based fermentation substrates

[0089] 1. Experimental Methods

[0090] The main steps of this embodiment are the same as those of Example 1, except that the substrate for fermentation is rice.

[0091] 2. Experimental Results

[0092] The effect of phenylethanol on ethanol production from anaerobic fermentation of rice culture medium was as follows: Figure 13 As shown, in the anaerobic fermentation system using rice as a substrate, the experimental group (P25) with a phenylethanol concentration of 25 μmol / L promoted ethanol synthesis in the rice anaerobic fermentation system. Similar promoting effects were observed at concentrations near this value.

[0093] The effect of phenylethanol on lactic acid production from anaerobic fermentation of rice culture medium was shown in the following results. Figure 14 As shown, in the anaerobic fermentation system using rice as the substrate, the maximum lactic acid concentration occurred in the control group on day 7, indicating that exogenous low-concentration phenylethanol did not increase the yield of lactic acid synthesis in rice. However, compared to the control group reaching its maximum lactic acid concentration on day 7, the experimental groups consistently produced higher lactic acid yields in the first 6 days. Furthermore, the P25 group reached its peak on day 4, with a peak value greater than that of the control group on the same day. The P50 group reached its first peak on day 3, earlier than the control group, and its peak value was also greater than that of the control group on the same day. This demonstrates that exogenous low-concentration phenylethanol can rapidly stimulate microbial lactic acid production and shorten the lactic acid production time.

[0094] Based on previous experimental results, it was found that high concentrations of quorum sensing molecules generally inhibited ethanol production. Therefore, in the anaerobic fermentation system using rice as a substrate, only experiments with low concentrations of quorum sensing molecules were conducted. Example 8: Effect of adding tyrosol on ethanol and lactic acid production in rice-based fermentation systems.

[0095] 1. Experimental Methods

[0096] The main steps of this embodiment are the same as those of Example 1, except that the substrate for fermentation is rice and the added quorum sensing molecule is tyrosol.

[0097] 2. Experimental Results

[0098] The effect of tyrosol on ethanol production from anaerobic fermentation of rice culture medium was as follows: Figure 15 As shown, in the anaerobic fermentation system using rice as a substrate, when the tyrosol concentration was 50 μmol / L, the ethanol concentration showed a good growth trend. The same promoting effect was also obtained at concentrations near this value.

[0099] The effect of tyrosol on lactic acid production during anaerobic fermentation of rice culture medium was as follows: Figure 16As shown, in the anaerobic fermentation system using rice as a substrate, based on the maximum lactic acid concentration, exogenous low-concentration tyrosol did not increase the yield of lactic acid synthesis in rice. However, compared to the control group, which reached its maximum lactic acid concentration on the 7th day, the experimental group's lactic acid production was generally higher than that of the control group in the first 6 days. The P25 group showed peak values ​​on the 3rd day, earlier than the control group, and the peak values ​​were greater than those of the control group on the corresponding days. The P50 group showed peak values ​​on the 3rd and 6th days, earlier than the control group, and the peak values ​​were greater than those of the control group on the corresponding days. This indicates that exogenous low-concentration tyrosol can rapidly stimulate microorganisms to produce lactic acid and shorten the lactic acid production time.

[0100] Example 9: Effect of adding colorol on the production of ethanol and lactic acid in rice-based fermentation substrate

[0101] 1. Experimental Methods

[0102] The main steps of this embodiment are the same as those of Example 1, except that the substrate for fermentation is rice and the added quorum sensing molecule is chromol.

[0103] 2. Experimental Results

[0104] The effect of tryptophan on ethanol production from anaerobic fermentation of rice culture medium was shown in the following results. Figure 17 As shown, in the anaerobic fermentation system using rice as a substrate, group I25 reached its peak value on the second day, earlier than the control group, and the peak value was greater than that of the control group on the same day. This indicates that exogenous low-concentration tryptol at a concentration of 25 μmol / L can rapidly stimulate microorganisms to produce ethanol and shorten the ethanol production time. Low-concentration tryptol at 50 μmol / L did not show a promoting effect on the anaerobic fermentation synthesis of ethanol in rice culture medium.

[0105] The effect of tryptophan on lactic acid production during anaerobic fermentation of rice culture medium was as follows: Figure 18 As shown, in the anaerobic fermentation system using rice as the substrate, based on the maximum lactic acid concentration, exogenous low-concentration tryptol did not increase the yield of lactic acid synthesis in rice. However, compared to the control group, which reached its maximum lactic acid concentration on the 7th day, the experimental groups consistently produced higher lactic acid in the first 6 days. Furthermore, the I25 group reached its peak on the 3rd and 5th days, and the I50 group reached its peak on the 2nd and 6th days, earlier than the control group, and all peak values ​​were higher than those of the control group on the corresponding days. This demonstrates that exogenous low-concentration tryptol can rapidly stimulate microbial lactic acid production and shorten the lactic acid production time.

[0106] Example 10: Effect of adding phenylethanol on ethanol production in food waste as a fermentation substrate

[0107] 1. Experimental Methods

[0108] S1. Perform preliminary sorting on the collected kitchen waste to remove impurities that are not easily fermented;

[0109] S2. Add bacterial flora to make the abundance ratio of Lactobacillus plantarum, Clostridium butyricum, Bacillus licheniformis, and Bacillus amine-degrading bacteria 50:20:20:10;

[0110] The abundance ratio of the fungal species *Schizosaccharomyces*, *Aspergillus charcoalii*, and *Saccharomyces cerevisiae* was 75:25:0.005.

[0111] S3. Add 50 μmol / L phenylethanol (these concentration values ​​are for the anaerobic fermentation system);

[0112] S4. Anaerobic fermentation was carried out at 30℃ for 7 days. After fermentation, the mixture was centrifuged and the supernatant was purified into ethanol using conventional methods.

[0113] The bacteria are added at a rate of 8% (by volume), and the fungi are added at a rate of 8% (by volume).

[0114] 2. Experimental Results

[0115] In food waste with a phenylethanol concentration of 50 μmol / L, after adding the above-mentioned microbial community, the ethanol concentration reached its highest value on the first day of fermentation, approximately 1.2 times higher than the maximum ethanol concentration in the control group (without phenylethanol). Example 11: Effect of adding phenylethanol on lactic acid production in food waste as a fermentation substrate.

[0116] 1. Experimental Methods

[0117] S1. Perform preliminary sorting on the collected kitchen waste to remove impurities that are not easily fermented;

[0118] S2. Add bacterial strains to make the abundance ratio of Lactobacillus acidophilus and Clostridium butyricum in the bacteria 65:27;

[0119] The abundance ratio of Candida albicans and Aspergillus oryzae in the fungi was 80:20.

[0120] S3. Add 300 μmol / L phenylethanol (these concentration values ​​are for the anaerobic fermentation system);

[0121] S4. Anaerobic fermentation was carried out at 30℃ for 7 days. After fermentation, the mixture was centrifuged and the supernatant was purified into lactic acid using conventional methods.

[0122] The bacteria are added at a rate of 8% (by volume), and the fungi are added at a rate of 8% (by volume).

[0123] 2. Experimental Results

[0124] When the above-mentioned microbial community was added to kitchen waste with a phenylethanol concentration of 300 μmol / L, the lactic acid concentration reached its highest value on the first day of fermentation, which was about 1.8 times higher than the maximum lactic acid concentration value of the control group (without phenylethanol).

[0125] In summary, when the yield of ethanol or lactic acid peaks within seven days and exceeds the maximum value of the control group, it indicates that the quorum sensing molecules at that concentration promote product production, and have the same promoting effect at concentrations near this value. When the yield of ethanol or lactic acid peaks earlier than that of the control group within seven days, and this peak value is greater than the yield value of the control group on the same day but less than the maximum value of the control group, it indicates that although the quorum sensing molecules do not increase product yield, they can quickly stimulate microorganisms to produce products and shorten the product production time.

[0126] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A method for producing ethanol from organic waste, characterized in that, Add alcohol quorum sensing molecules at a concentration of 1~500 μmol / L to the anaerobic fermentation system of organic waste; The organic waste is either lignocellulosic waste or starch waste; When the organic waste is lignocellulose waste, the alcohol group sensing molecule is 50±15 μmol / L of tyrosol. When the organic waste is starch-based waste, the alcohol quorum sensing molecule is 25±15 μmol / L phenylethanol; The anaerobic fermentation system contains bacteria and fungi. The bacteria consist of *Limosilactobacillus*, *Weissella*, *Pediococcus*, *Clostridiaceae*, *Bacillus*, *Proteiniphilum*, and *Aminobacterium* in a species abundance ratio of (30–65):(1–45):(0.5–15):(0.01–27):(0.1–20):(0.01–5):(0.01–15). The *Lactobacillus* genus is selected from one or more of *Lactobacillus delbrueckii*, *Lactobacillus plantarum*, *Lactobacillus helveticus*, *Lactobacillus acidophilus*, and *Lactobacillus caudatus*; the *Weissella* genus is selected from one or more of *Weissella fusionis*, *Weissella esculenta*, *Weissella entericae*, *Weissella viride*, *Weissella univar*, *Weissella cannii*, *Weissella oryzae*, *Weissella henryi*, *Weissella microphylla*, *Weissella staphylococcus*, and *Weissella serrata*; the *Pediococcus* genus is selected from one or more of *Pediococcus brevis*, *Pediococcus lactis*, *Pediococcus pentosaceus*, and *Pediococcus halophilus*; the *Clostridium* genus is selected from one or more of *Clostridium butyricum* and *Clostridium acetobutanol*; the *Bacillus* genus is selected from one or more of *Bacillus polymyxa*, *Bacillus subtilis*, *Bacillus cereus*, *Bacillus licheniformis*, and *Bacillus brevis*; the *Proteophile* genus is selected from one or more of *Acetobacter acetophilus* and *Akkermansia muciniphilus*; the *Aminomyxobacter* genus is selected from one or more of *Bacillus amyloliquefaciens* and *Aminomyxobacterium taurinae*. The fungi consist of the genera *Saccharomyces*, *Aspergillus*, *Fusarium*, *Cyberlindnera*, *Cladosporium*, *Mortierella*, *Alternaria*, *Penicillium*, *Filobasidium*, *Trichoderma*, and *Rhizopus*, in a species abundance ratio of (1~80):(0.001~45):(0.001~1):(0.0001~0.1):(0.001~1):(0.001~1):(0.001~5):(0.001~1):(0.001~3):(0.001~3):(0.001~25). The yeast genus is selected from one or more of *Saccharomyces cerevisiae*, *Schizosaccharomyces*, *Hansenula polymorpha*, *Pichia pastoris*, *Candida*, *Gnaphalium*, and *Rhodotorula*. The *Aspergillus* genus is selected from one or more of *Aspergillus awamori*, *Aspergillus niger*, *Aspergillus ochre*, *Aspergillus oryzae*, *Aspergillus sacchari*, *Aspergillus styracifolius ... tabinei*, *Aspergillus umbellatus*, *Aspergillus urinaria*, and *Aspergillus fibrosum*. The *Fusarium* genus is selected from one or more of *Fusarium*, *Fusarium*, *Fusarium*, and *Fusarium*. The *Cyclocarya* genus is selected from one or more of *Saccharomyces cerevisiae* and *Saccharomyces cerevisiae*. The *Cladosporium* genus is selected from one or more of *Cladosporium xanthosporium*, *Cladosporium multiflorum*, *Cladosporium lantana*, and *Cladosporium trifolium*. The fungi are selected from one or more of the following: *Morchella spp.*, *Morchella alpineensis*, *Morchella microsporum*, *Morchella ramanense*, *Morchella scabranchii*, and *Morchella longiflora*; *Alternaria* is selected from one or more of the following: *Alternaria longiflora*, *Alternaria scabranchii*, *Alternaria scabranchii*, and *Alternaria microsporum*; *Penicillium* is selected from one or more of the following: *Penicillium reskeletoni*, *Penicillium heckii*, *Penicillium chrysogenum*, and *Penicillium salicum*; *Basidia* is selected from one or more of the following: rust fungi, smut fungi, lamellar fungi, and gastromycetes; *Trichoderma* is selected from one or more of the following: *Trichoderma viride*, *Trichoderma cornigerum*, *Trichoderma echinosporum*, *Trichoderma harzianum*, and *Trichoderma longiflora*; *Rhizopus* is selected from one or more of the following: *Rhizopus nigricans*, *Rhizopus huassifolia*, and *Rhizopus oryzae*.

2. A method for producing lactic acid from organic waste, characterized in that, Add alcohol quorum sensing molecules at a concentration of 1~500 μmol / L to the anaerobic fermentation system of organic waste; The alcohol quorum sensing molecule is phenylethanol, tyrosol, or cromoglycine; The organic waste is kitchen waste or lignocellulosic waste; When the organic waste is kitchen waste, the alcohol quorum sensing molecule is 300±50 μmol / L phenylethanol or 300±50 μmol / L tyrosol or 50±15 μmol / L and 300±50 μmol / L color alcohol. When the organic waste is lignocellulosic waste, the alcohol quorum sensing molecule is 200±50 μmol / L phenethyl alcohol or 300±50 μmol / L tyrosol or 25±15 μmol / L, 200±50 μmol / L, or 300±50 μmol / L color alcohol. The anaerobic fermentation system contains bacteria and fungi. The bacteria consist of *Limosilactobacillus*, *Weissella*, *Pediococcus*, *Clostridiaceae*, *Bacillus*, *Proteiniphilum*, and *Aminobacterium* in a species abundance ratio of (30–65):(1–45):(0.5–15):(0.01–27):(0.1–20):(0.01–5):(0.01–15). The *Lactobacillus* genus is selected from one or more of *Lactobacillus delbrueckii*, *Lactobacillus plantarum*, *Lactobacillus helveticus*, *Lactobacillus acidophilus*, and *Lactobacillus caudatus*; the *Weissella* genus is selected from one or more of *Weissella fusionis*, *Weissella esculenta*, *Weissella entericae*, *Weissella viride*, *Weissella univar*, *Weissella cannii*, *Weissella oryzae*, *Weissella henryi*, *Weissella microphylla*, *Weissella staphylococcus*, and *Weissella serrata*; the *Pediococcus* genus is selected from one or more of *Pediococcus brevis*, *Pediococcus lactis*, *Pediococcus pentosaceus*, and *Pediococcus halophilus*; the *Clostridium* genus is selected from one or more of *Clostridium butyricum* and *Clostridium acetobutanol*; the *Bacillus* genus is selected from one or more of *Bacillus polymyxa*, *Bacillus subtilis*, *Bacillus cereus*, *Bacillus licheniformis*, and *Bacillus brevis*; the *Proteophile* genus is selected from one or more of *Acetobacter acetophilus* and *Akkermansia muciniphilus*; the *Aminomyxobacter* genus is selected from one or more of *Bacillus amyloliquefaciens* and *Aminomyxobacterium taurinae*. The fungi consist of the genera *Saccharomyces*, *Aspergillus*, *Fusarium*, *Cyberlindnera*, *Cladosporium*, *Mortierella*, *Alternaria*, *Penicillium*, *Filobasidium*, *Trichoderma*, and *Rhizopus*, in a species abundance ratio of (1~80):(0.001~45):(0.001~1):(0.0001~0.1):(0.001~1):(0.001~1):(0.001~5):(0.001~1):(0.001~3):(0.001~3):(0.001~25). The yeast genus is selected from one or more of *Saccharomyces cerevisiae*, *Schizosaccharomyces*, *Hansenula polymorpha*, *Pichia pastoris*, *Candida*, *Gnaphalium*, and *Rhodotorula*. The *Aspergillus* genus is selected from one or more of *Aspergillus awamori*, *Aspergillus niger*, *Aspergillus ochre*, *Aspergillus oryzae*, *Aspergillus sacchari*, *Aspergillus styracifolius ... tabinei*, *Aspergillus umbellatus*, *Aspergillus urinaria*, and *Aspergillus fibrosum*. The *Fusarium* genus is selected from one or more of *Fusarium*, *Fusarium*, *Fusarium*, and *Fusarium*. The *Cyclocarya* genus is selected from one or more of *Saccharomyces cerevisiae* and *Saccharomyces cerevisiae*. The *Cladosporium* genus is selected from one or more of *Cladosporium xanthosporium*, *Cladosporium multiflorum*, *Cladosporium lantana*, and *Cladosporium trifolium*. The fungi are selected from one or more of the following: *Morchella spp.*, *Morchella alpineensis*, *Morchella microsporum*, *Morchella ramanense*, *Morchella scabranchii*, and *Morchella longiflora*; *Alternaria* is selected from one or more of the following: *Alternaria longiflora*, *Alternaria scabranchii*, *Alternaria scabranchii*, and *Alternaria microsporum*; *Penicillium* is selected from one or more of the following: *Penicillium reskeletoni*, *Penicillium heckii*, *Penicillium chrysogenum*, and *Penicillium salicum*; *Basidia* is selected from one or more of the following: rust fungi, smut fungi, lamellar fungi, and gastromycetes; *Trichoderma* is selected from one or more of the following: *Trichoderma viride*, *Trichoderma cornigerum*, *Trichoderma echinosporum*, *Trichoderma harzianum*, and *Trichoderma longiflora*; *Rhizopus* is selected from one or more of the following: *Rhizopus nigricans*, *Rhizopus huassifolia*, and *Rhizopus oryzae*.

3. A method for producing ethanol and lactic acid from organic waste, characterized in that, Add alcohol quorum sensing molecules at a concentration of 1~500 μmol / L to the anaerobic fermentation system of organic waste; The organic waste is kitchen waste; The alcohol quorum sensing molecule is tyrosol at a concentration of 25 ± 15 μmol / L; The anaerobic fermentation system contains bacteria and fungi. The bacteria consist of *Limosilactobacillus*, *Weissella*, *Pediococcus*, *Clostridiaceae*, *Bacillus*, *Proteiniphilum*, and *Aminobacterium* in a species abundance ratio of (30–65):(1–45):(0.5–15):(0.01–27):(0.1–20):(0.01–5):(0.01–15). The *Lactobacillus* genus is selected from one or more of *Lactobacillus delbrueckii*, *Lactobacillus plantarum*, *Lactobacillus helveticus*, *Lactobacillus acidophilus*, and *Lactobacillus caudatus*; the *Weissella* genus is selected from one or more of *Weissella fusionis*, *Weissella esculenta*, *Weissella entericae*, *Weissella viride*, *Weissella univar*, *Weissella cannii*, *Weissella oryzae*, *Weissella henryi*, *Weissella microphylla*, *Weissella staphylococcus*, and *Weissella serrata*; the *Pediococcus* genus is selected from one or more of *Pediococcus brevis*, *Pediococcus lactis*, *Pediococcus pentosaceus*, and *Pediococcus halophilus*; the *Clostridium* genus is selected from one or more of *Clostridium butyricum* and *Clostridium acetobutanol*; the *Bacillus* genus is selected from one or more of *Bacillus polymyxa*, *Bacillus subtilis*, *Bacillus cereus*, *Bacillus licheniformis*, and *Bacillus brevis*; the *Proteophile* genus is selected from one or more of *Acetobacter acetophilus* and *Akkermansia muciniphilus*; the *Aminomyxobacter* genus is selected from one or more of *Bacillus amyloliquefaciens* and *Aminomyxobacterium taurinae*. The fungi consist of the genera *Saccharomyces*, *Aspergillus*, *Fusarium*, *Cyberlindnera*, *Cladosporium*, *Mortierella*, *Alternaria*, *Penicillium*, *Filobasidium*, *Trichoderma*, and *Rhizopus*, in a species abundance ratio of (1~80):(0.001~45):(0.001~1):(0.0001~0.1):(0.001~1):(0.001~1):(0.001~5):(0.001~1):(0.001~3):(0.001~3):(0.001~25). The yeast genus is selected from one or more of *Saccharomyces cerevisiae*, *Schizosaccharomyces*, *Hansenula polymorpha*, *Pichia pastoris*, *Candida*, *Gnaphalium*, and *Rhodotorula*. The *Aspergillus* genus is selected from one or more of *Aspergillus awamori*, *Aspergillus niger*, *Aspergillus ochre*, *Aspergillus oryzae*, *Aspergillus sacchari*, *Aspergillus styracifolius ... tabinei*, *Aspergillus umbellatus*, *Aspergillus urinaria*, and *Aspergillus fibrosum*. The *Fusarium* genus is selected from one or more of *Fusarium*, *Fusarium*, *Fusarium*, and *Fusarium*. The *Cyclocarya* genus is selected from one or more of *Saccharomyces cerevisiae* and *Saccharomyces cerevisiae*. The *Cladosporium* genus is selected from one or more of *Cladosporium xanthosporium*, *Cladosporium multiflorum*, *Cladosporium lantana*, and *Cladosporium trifolium*. The fungi are selected from one or more of the following: *Morchella spp.*, *Morchella alpineensis*, *Morchella microsporum*, *Morchella ramanense*, *Morchella scabranchii*, and *Morchella longiflora*; *Alternaria* is selected from one or more of the following: *Alternaria longiflora*, *Alternaria scabranchii*, *Alternaria scabranchii*, and *Alternaria microsporum*; *Penicillium* is selected from one or more of the following: *Penicillium reskeletoni*, *Penicillium heckii*, *Penicillium chrysogenum*, and *Penicillium salicum*; *Basidia* is selected from one or more of the following: rust fungi, smut fungi, lamellar fungi, and gastromycetes; *Trichoderma* is selected from one or more of the following: *Trichoderma viride*, *Trichoderma cornigerum*, *Trichoderma echinosporum*, *Trichoderma harzianum*, and *Trichoderma longiflora*; *Rhizopus* is selected from one or more of the following: *Rhizopus nigricans*, *Rhizopus huassifolia*, and *Rhizopus oryzae*.

4. The application of the method according to any one of claims 1 to 3 in the fermentation of organic waste.

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