Agarwood ferment and method for preparing the same

Abstract

This invention belongs to the field of deep processing of traditional Chinese medicine, specifically relating to an agarwood fermentation product and its preparation method. The agarwood fermentation product is obtained by fermenting agarwood with enzymes; the enzymes are cellulase, β-glucanase, lignin peroxidase, and neutral protease; the cellulase and β-glucanase are added to the agarwood at the beginning of fermentation; the lignin peroxidase is added to the agarwood after one day of fermentation, and the neutral protease is added to the agarwood after three days of fermentation. This invention is based on a three-stage enzymatic regulation of the natural fermentation of agarwood monomers to obtain an agarwood fermentation product. Testing shows that it has a high content of active ingredients and a rich aroma, making it suitable for use as a traditional Chinese medicine to enhance efficacy, and also as an excellent fragrance to improve quality of life, possessing practical application value.

Description

Technical Field

[0001] This invention belongs to the field of deep processing of traditional Chinese medicine, specifically relating to a fermented agarwood product and its preparation method. Background Technology

[0002] Agarwood is a precious complex substance formed from the resin secreted by trees of the genus *Aquilaria* in the Thymelaeaceae family after external trauma, as a defense against fungal infection, and which matures naturally over many years. Its quality is typically classified into three grades based on resin content and density: sinking-grade, semi-sinking, and yellow-ripe agarwood. Since ancient times, agarwood has not only been regarded as a top-grade spice and traditional Chinese medicine, possessing properties such as promoting qi circulation and relieving pain, warming the stomach and stopping vomiting, and calming asthma, but also carries profound cultural significance. Today, its value has expanded to multiple fields, including the modern perfume industry, high-end crafts, and biomedical research and development.

[0003] Agarwood fermentation products typically refer to products obtained by treating agarwood raw materials with microorganisms (such as fungi and bacteria) or enzyme preparations, thereby altering their physicochemical properties and flavor characteristics through fermentation. This process aims to simulate natural aging to enhance aroma purity, increase medicinal active ingredients, or improve taste. CN103320232A discloses a method for fermenting agarwood using mixed enzyme preparations. This method primarily aims to increase the yield of agarwood essential oil but does not address the quality of the agarwood fermentation products. Summary of the Invention

[0004] To improve the sensory quality and active ingredient content of agarwood, thereby comprehensively enhancing its overall quality, this invention provides an agarwood ferment, which is produced by the natural fermentation of agarwood with added enzymes; the enzymes are cellulase, β-glucanase, lignin peroxidase, and neutral protease. The cellulase and β-glucanase were added to the agarwood at the start of fermentation; lignin peroxidase was added to the agarwood one day after fermentation, and neutral protease was added to the agarwood three days after fermentation.

[0005] Furthermore, an enzyme is added after water is added to the agarwood; the mass ratio of water to agarwood is 100:40~60.

[0006] Furthermore, the mass ratio of water to agarwood is 100:52.

[0007] Further, the mass percentage of cellulase or β-glucanase to agarwood is 0.5%-1.5%, the mass percentage of lignin peroxidase to agarwood is 0.2%-0.8%, and the mass percentage of neutral protease to agarwood is 0.3%-1.0%.

[0008] Furthermore, the cellulase or β-glucanase has a mass percentage of 0.5% with agarwood, the lignin peroxidase has a mass percentage of 0.2% with agarwood, and the neutral protease has a mass percentage of 0.3% with agarwood.

[0009] Furthermore, the fermentation temperature is 30~40℃, the humidity is 60~70%, and the time is 6~8 days.

[0010] Furthermore, the fermentation temperature is 30°C, the humidity is 60%, and the time is 6 days.

[0011] This invention also provides a method for fermenting agarwood, comprising the following steps: Take agarwood powder, add water and mix well, then spray with enzyme solution, stir evenly and let it ferment naturally to obtain the product; The enzyme solutions are respectively cellulase, β-glucanase, lignin peroxidase and neutral protease; The cellulase and β-glucanase were sprayed into the agarwood at the start of fermentation; lignin peroxidase was sprayed into the agarwood one day after fermentation, and neutral protease was sprayed into the agarwood three days after fermentation.

[0012] Furthermore, the powder is a powder that has passed through an 80-mesh sieve.

[0013] Furthermore, the mass ratio of water to agarwood is 100:40~60.

[0014] Furthermore, the fermentation temperature is 30-40°C, the humidity is 60-70%, and the time is 6-8 days.

[0015] Furthermore, the cellulase or β-glucanase is 0.5%-1.5% of the mass of agarwood, the lignin peroxidase is 0.2%-0.8% of the mass of agarwood, and the neutral protease is 0.3%-1.0% of the mass of agarwood.

[0016] Furthermore, the cellulase or β-glucanase has a mass percentage of 0.5% with agarwood, the lignin peroxidase has a mass percentage of 0.2% with agarwood, and the neutral protease has a mass percentage of 0.3% with agarwood.

[0017] Furthermore, the enzyme solution is prepared by using an enzyme pH buffer solution.

[0018] Furthermore, the cellulase solution in the enzyme solution is a cellulase solution diluted with pH 4.8 citrate-sodium citrate buffer to an enzyme concentration of 10 mg / mL; β-glucanase solution is an enzyme solution in which β-glucanase is diluted with acetate-sodium acetate buffer at pH 5.0 to an enzyme concentration of 10 mg / mL. Lignin peroxidase solution is an enzyme solution in which lignin peroxidase is diluted with tartaric acid-sodium tartrate buffer at pH 3.5 to an enzyme concentration of 5 mg / mL. Neutral protease solution is an enzyme solution in which neutral protease is diluted with Tris-HCl buffer at pH 7.0 to an enzyme concentration of 8 mg / mL.

[0019] The natural fermentation described in this invention refers to a fermentation process that does not involve artificial inoculation with specific strains, but relies on the raw materials themselves and naturally occurring microorganisms in the environment to carry out the fermentation.

[0020] The agarwood fermentation product of this invention adopts a three-stage enzymatic regulation technology, that is, it is prepared by adding specific enzyme preparations at three different time points to drive the natural fermentation of agarwood monomers. The test results show that the fermentation product of this invention has a high content of active ingredients, a rich and fragrant aroma, and its overall quality is significantly better than that of similar fermentation products reported in the public. It can be used as a Chinese medicine to enhance the efficacy of medicines, and as a high-grade fragrance to improve the quality of life. It has a wide range of practical application value.

[0021] Obviously, based on the above description of the present invention, and according to common technical knowledge and conventional methods in the field, various other modifications, substitutions or alterations can be made without departing from the basic technical concept of the present invention.

[0022] The following detailed embodiments further illustrate the above-described content of the present invention. However, this should not be construed as limiting the scope of the present invention to the following embodiments. All technologies implemented based on the above-described content of the present invention fall within the scope of the present invention. Attached Figure Description

[0023] Figure 1 This is a schematic diagram of the three-stage enzymatic fermentation process for agarwood according to the present invention; Figure 2 A graph comparing the endpoints of active ingredients in agarwood ferment under different enzyme addition strategies; Figure 3 Heatmap of normalized performance and comprehensive score for each experimental group; Figure 4 A normalized comparison chart of quality and marker components. Detailed Implementation

[0024] The raw materials, reagents, and equipment used in the specific embodiments of this invention were all purchased commercially. The method for preparing the enzyme solution used is as follows: Cellulase solution: Dilute cellulase with pH 4.8 citrate-sodium citrate buffer to an enzyme concentration of 10 mg / mL; β-glucanase solution: Dilute β-glucanase with acetate-sodium acetate buffer at pH 5.0 to an enzyme concentration of 10 mg / mL; Lignin peroxidase solution: Lignin peroxidase was diluted with tartaric acid-sodium tartrate buffer at pH 3.5 to an enzyme concentration of 5 mg / mL; Neutral protease solution: Dilute neutral protease with Tris-HCl buffer at pH 7.0 to an enzyme concentration of 8 mg / mL.

[0025] Cellulase was purchased from Shanghai Maclean Biotechnology Co., Ltd., with an enzyme activity ≥200,000 U / g; β-glucanase was purchased from Shanghai Maclean Biotechnology Co., Ltd., with an enzyme activity ≥30,000 U / g; lignin peroxidase was purchased from Shanghai Maclean Biotechnology Co., Ltd., with an enzyme activity ≥100 U / g; and neutral protease was purchased from BASF Biotechnology Co., Ltd. in Hefei, with an enzyme activity ≥50,000 U / g. The preparation method of agarwood powder is as follows: Take the resin-containing wood of Aquilaria sinensis (Lour.) Gilg, a plant of the Thymelaeaceae family, crush it and pass it through an 80-mesh sieve. The resulting agarwood powder is agarwood powder.

[0026] Example 1 Preparation of Agarwood Fermentation Product Take agarwood powder that has passed through an 80-mesh sieve, add water at a ratio of 100:52 (water to agarwood powder by weight), mix thoroughly, and then ferment at 30°C and 60% humidity. During fermentation, at the beginning of fermentation, cellulase solution and β-glucanase solution are sprayed and stirred evenly, so that the mass percentage of the two enzymes and agarwood powder reaches 0.5% respectively; after 1 day of fermentation, lignin peroxidase solution is sprayed and stirred evenly, so that the mass percentage of this enzyme and agarwood powder is 0.2%; after 3 days of fermentation, neutral protease solution is sprayed and stirred evenly, so that the mass percentage of this enzyme and agarwood powder is 0.3%; fermentation continues until the 6th day, and the fermented agarwood product is obtained.

[0027] Example 2 Preparation of Agarwood Fermentation Product Take agarwood powder that has passed through an 80-mesh sieve, add water at a ratio of 100:50 (water to agarwood powder by weight), mix thoroughly, and then ferment at a temperature of 40℃ and a humidity of 70%. During fermentation, at the beginning of fermentation, cellulase solution and β-glucanase solution are sprayed and stirred evenly, so that the mass percentage of the two enzymes and agarwood powder reaches 1.5% respectively; after 1 day of fermentation, lignin peroxidase solution is sprayed and stirred evenly, so that the mass percentage of the enzyme and agarwood powder is 0.8%; after 3 days of fermentation, neutral protease solution is sprayed and stirred evenly, so that the mass percentage of the enzyme and agarwood powder is 1.0%; fermentation continues until the 8th day, and the fermented agarwood product is obtained.

[0028] Example 3 Preparation of Agarwood Fermentation Product Take agarwood powder that has passed through an 80-mesh sieve, add water at a ratio of 100:60 (water to agarwood powder by weight), mix thoroughly, and then ferment at a temperature of 40℃ and a humidity of 70%. During fermentation, at the beginning of fermentation, cellulase solution and β-glucanase solution are sprayed and stirred evenly, so that the mass percentage of the two enzymes and agarwood powder reaches 1.2% respectively; after 1 day of fermentation, lignin peroxidase solution is sprayed and stirred evenly, so that the mass percentage of this enzyme and agarwood powder is 0.6%; after 3 days of fermentation, neutral protease solution is sprayed and stirred evenly, so that the mass percentage of this enzyme and agarwood powder is 0.5%; fermentation continues until the 8th day, and the fermented agarwood product is obtained.

[0029] Example 4 Preparation of Agarwood Fermentation Product Take agarwood powder that has passed through an 80-mesh sieve, add water at a ratio of 100:40 (water to agarwood powder by weight), mix thoroughly, and then ferment at 35°C and 65% humidity. During fermentation, at the beginning of fermentation, cellulase solution and β-glucanase solution are sprayed and stirred evenly, so that the mass percentage of the two enzymes and agarwood powder reaches 1.5% respectively; after 1 day of fermentation, lignin peroxidase solution is sprayed and stirred evenly, so that the mass percentage of this enzyme and agarwood powder is 0.8%; after 3 days of fermentation, neutral protease solution is sprayed and stirred evenly, so that the mass percentage of this enzyme and agarwood powder is 1.0%; fermentation continues until the 7th day, and the fermented agarwood product is obtained.

[0030] The following experimental examples illustrate the beneficial effects of the present invention.

[0031] Experimental Example 1: Study on Enzymatic Fermentation Process of Agarwood To address the issue that enzymatic hydrolysis of agarwood can easily alter its original composition, affect the content of functional components, and influence its aroma characteristics, the research team, after referring to relevant reports and attempting to improve quality by adding different types of enzymes simultaneously without success, systematically explored and finally established a three-stage enzyme addition strategy—that is, adding cellulase, β-glucanase, lignin peroxidase, and neutral protease at three specific fermentation stages. This strategy can effectively increase the content of representative active ingredients in the final product of agarwood fermentation and improve the overall sensory quality. Figure 1 To further optimize this process, the key process parameters of the three-stage enzyme-added fermentation of agarwood are optimized in order to continuously improve the overall quality of the fermented agarwood.

[0032] I. Methods Using total polysaccharides, total triterpenes, total phenols, and aroma scores as comprehensive evaluation indicators, moisture content, enzyme levels, fermentation temperature, relative humidity, and fermentation time were progressively screened. During the screening process, the remaining steps and conditions were the same as in Example 1, only the factors to be investigated were changed.

[0033] Total triterpenoid content: determined by vanillin-perchloric acid colorimetric method. Specifically: accurately weigh 0.5 g of sample powder, add 50 mL of anhydrous ethanol, and extract ultrasonically for 30 min (power 200 W, temperature 40 ℃). Filter and dilute to 50 mL. Accurately measure 1 mL of the extract, add 0.2 mL of 5% vanillin-glacial acetic acid solution and 0.8 mL of perchloric acid, heat in a 60 ℃ water bath for 15 min, cool, add 5 mL of glacial acetic acid, and measure the absorbance at 548 nm. Plot a standard curve using oleanolic acid as a reference, and calculate the total triterpenoid content.

[0034] Total polysaccharide content: determined by the sulfuric acid-anthrone colorimetric method. Specifically: Accurately weigh 0.5 g of sample powder, place it in an Erlenmeyer flask, add 25 mL of water, extract in a boiling water bath for 2.5 h, cool, make up the weight loss, shake well, and filter. Take an appropriate amount of the filtrate, evaporate to dryness, dissolve the residue in 2 mL of hot water, add 30 mL of anhydrous ethanol, place at 4 ℃ for 12 h, centrifuge at 10000 r / min for 10 min, discard the supernatant, dissolve the precipitate in hot water, and dilute to 10 mL. Accurately measure 1 mL of the test solution, add 6 mL of sulfuric acid-anthrone solution, develop color, and measure the absorbance at 625 nm. Plot a standard curve using glucose as a reference, and calculate the total polysaccharide content.

[0035] Total phenol content: determined using the Folin-Ciocalteu colorimetric method. Specifically: accurately weigh 0.2 g of sample, add 30 mL of 70% ethanol, extract in an 80 ℃ water bath for 40 min, filter, and dilute to 50 mL. Accurately measure 1 mL of the extract, add 0.5 mL of Folin-Ciocalteu reagent, shake well, add 2 mL of 10% sodium carbonate solution, and dilute to 10 mL with water. Incubate at room temperature for 60 min, and measure the absorbance at 765 nm. Plot a standard curve using gallic acid as a reference standard, and calculate the total phenol content.

[0036] Aroma Evaluation: Sensory evaluation was adopted. An evaluation team of 5 professional aroma evaluators with more than 3 years of industry experience from the agarwood company was formed. Blind evaluation was conducted in a standard aroma evaluation environment (temperature 25 ℃, humidity 50%, no odor, and sufficient light). The evaluation indicators included aroma intensity (0-3 points), aroma purity (0-3 points), aroma persistence (0-2 points), and characteristic aroma richness (0-2 points), with a total score of 10 points. The evaluation process was as follows: 5 g of sample was placed in an aroma evaluation cup, 95 ℃ hot water was added, the cup was covered and steeped for 3 minutes, and then the cup was opened and the samples were smelled and scored in turn. The average score of the 5 people was taken as the final aroma score.

[0037] II. Results (1) Moisture content screening The study examined three water-to-agarwood powder ratios: 100:40, 100:52, and 100:60. The results are shown in Table 1. Table 1. Results of fermentation endpoint indicators under different moisture contents The results showed that the fermentation system with a water-to-agarwood powder ratio of 100:40 had a relatively high water content, and the fermented material was prone to becoming wet and sticky and locally sticky, which was not conducive to aeration and uniform fermentation. The fermentation system with a water-to-agarwood powder ratio of 100:60 had a relatively low water content, and the fermented material was relatively dry, which may limit the growth, metabolism and enzymatic transformation of microorganisms. In contrast, the fermentation state was more uniform under the 100:52 condition, and the overall performance of total polysaccharides, total triterpenes and total phenols was the best. Therefore, the preferred water-to-agarwood powder ratio was 100:52.

[0038] (2) Enzyme level screening Three enzyme levels were investigated: low, medium, and high. The low level consisted of 0.5% cellulase and 0.5% β-glucanase at the start of fermentation, 0.2% lignin peroxidase after one day of fermentation, and 0.3% neutral protease after three days of fermentation. The medium level consisted of 1.0% cellulase, 1.0% β-glucanase, 0.5% lignin peroxidase, and 0.7% neutral protease. The high level consisted of 1.5% cellulase, 1.5% β-glucanase, 0.8% lignin peroxidase, and 1.0% neutral protease. The results are shown in Table 2. Table 2. Fermentation endpoint results under different enzyme levels. The results showed that the low-level group performed best in the evaluation of the three indicators of total polysaccharides, total triterpenes, and total phenols at the endpoint. Although the medium and high levels could enhance the cell wall disruption in the early stage, the retention of total triterpenes decreased and the aroma coordination deteriorated in the later stage. Therefore, the amount of each enzyme added was determined to be 0.5%–1.5% for cellulase and β-glucanase, 0.2%–0.8% for lignin peroxidase, and 0.3%–1.0% for neutral protease, with the low-level group, namely 0.5% each of cellulase and β-glucanase, 0.2% for lignin peroxidase, and 0.3% for neutral protease, being the optimal concentration.

[0039] (3) Screening of fermentation temperature The study examined three temperature levels: 30℃, 35℃, and 40℃. The results are shown in Table 3. Table 3. Results of fermentation endpoint indicators under different fermentation temperatures. The results showed that the substrate layer was stable and the aroma purity was good at 30℃, with the best overall performance in terms of total polysaccharides, total triterpenes, and total phenols. Conversion could also be completed at 35℃, but the final indicators were slightly lower. While 40℃ accelerated the initial reaction, the aroma became more mixed in the later stages, and the overall color and quality were inferior to the 30℃ group. Therefore, the fermentation temperature range was determined to be 30–40℃, with 30℃ being the optimal temperature.

[0040] (4) Relative humidity screening The study examined three levels: 60%, 65%, and 70%. The results are shown in Table 4. Table 4. Results of fermentation endpoint indicators under different relative humidity conditions The results showed that 60%–70% relative humidity was suitable for the three-stage enzymatic fermentation of agarwood. At 60%, the material state was stable and reproducibility was good. While fermentation could be maintained at 65% and 70%, the moisture content of the material layer fluctuated more, resulting in slightly lower final indicators. Therefore, the relative humidity range was determined to be 60%–70%, with 60% being optimal.

[0041] (5) Screening of fermentation time The study examined three levels: 6 days, 7 days, and 8 days. The results are shown in Table 5. Table 5 Results of fermentation endpoint indicators under different fermentation time conditions The results showed that the release of active ingredients and aroma formation reached a good balance by day 6 of fermentation; further extending the fermentation to 7-8 days resulted in limited gains in some indicators, and the retention of total triterpenes showed a decreasing trend. Therefore, the fermentation time range was determined to be 6-8 days, with 6 days being the optimal time.

[0042] Based on the above results, the optimal three-stage enzyme fermentation process was determined as follows: Agarwood powder passing through an 80-mesh sieve was mixed with water to a water-to-agarwood powder mass ratio of 100:52, and fermented for 6 days at 30℃ and 60% relative humidity. At the start of fermentation, 0.5% each of cellulase and β-glucanase were added; after 1 day of fermentation, 0.2% lignin peroxidase was added; and after 3 days of fermentation, 0.3% neutral protease was added. This process ensures the release of active ingredients while also contributing to stable and balanced agarwood fermentation quality.

[0043] Experimental Example 2: Comparison of Different Enzyme Addition Strategies To verify the superiority of the present invention, the optimized three-stage enzyme fermentation process, the adjusted three-stage enzyme fermentation process, and the commonly used one-time addition of compound enzymes for agarwood enzymatic hydrolysis fermentation were systematically compared.

[0044] I. Methods Agarwood was fermented according to the experimental groups in Table 6, with 3 parallel samples in each group. The total polysaccharides, total triterpenes and total phenols of the fermentation end products were detected, and aroma scores, comprehensive evaluations and determination of characteristic components such as ash, agaritol and isoagaritol were performed.

[0045] The detection methods for total polysaccharides, total triterpenes, and total phenols are the same as in Experimental Example 1; Ash content was determined by constant weight ashing at 550–600 °C.

[0046] Agaricone and iso-agaricone were determined by HPLC using the external standard method. Specific chromatographic conditions were as follows: C18 column (250 mm × 4.6 mm, 5 μm); mobile phase: acetonitrile (A)-water (B); gradient elution: 0–10 min, A 18%; 10–25 min, A 18%–25%; 25–35 min, A 25%–30%; 35–45 min, A 30%; flow rate 1.0 mL / min; column temperature 30 ℃; detection wavelength 252 nm; injection volume 10 μL. Reference solutions: Agaricone and iso-agaricone reference standards were accurately weighed, dissolved in methanol, and prepared into a series of standard solutions. Test solution: Approximately 0.5 g of sample powder was accurately weighed, added to 25 mL of methanol, and extracted ultrasonically for 30 min. After cooling, the weight loss was adjusted, filtered, and then filtered through a 0.22 μm microporous membrane before injection for determination. The contents of agaritol and isoaraitol were calculated using the external standard method.

[0047] Table 6 Experimental Groups and Process Settings II. Results 1. The results of the main active ingredient detection are shown in Table 7. Table 7. Results of endpoint active ingredient detection Note: p The values ​​are obtained by comparing each control group with the invention group.

[0048] From Table 7 and Figure 2 As can be seen, the present invention group is at or near the best level in all three representative indicators. Among them, the polysaccharide content is statistically significantly different from that of T1 holoenzyme, T3 holoenzyme, reverse enzyme addition and literature process control; the total phenol content also shows a significant increase compared with T0 holoenzyme, T1 holoenzyme and reverse enzyme addition, indicating that the three-stage enzyme addition strategy of the present invention is more conducive to the staged release and transformation of agarwood matrix.

[0049] 2. Aroma Score and Overall Evaluation Based on the active ingredient data, five professional aroma evaluators from the agarwood company, each with over three years of industry experience, conducted a blind evaluation under a standard aroma evaluation environment. They assessed the aroma intensity, purity, persistence, and richness of characteristic aromas in each sample group. The four factors were directly summed to obtain a 10-point aroma score. The 100-point comprehensive score is not a newly added testing item but rather an auxiliary evaluation value used for process screening and ranking. It is calculated by weighting the total polysaccharide, total triterpenes, total phenols, and aroma score after dimensionless evaluation. The formula is: Comprehensive Score = 30 × (Total Polysaccharide / Maximum Value in this Group) + 30 × (Total Triterpenes / Maximum Value in this Group) + 20 × (Total Phenols / Maximum Value in this Group) + 20 × (Aroma Score / 10).

[0050] Table 8 Aroma Score and Overall Performance From Table 8 and Figure 3 It is evident that the invention group achieved the highest aroma score of 8.9 / 10, and significantly outperformed other control groups in the overall score.

[0051] 3. Determination of quality and marker components The actual test results of ash content and marker components in each group are shown in Table 9 and 2019. Figure 4 .

[0052] Table 9 Actual test results of ash content and marker components in each group Note: Data are expressed as mean ± standard deviation (n=3). Different lowercase letters in the same column indicate significant differences (p<0.05), while those containing the same letter indicate no significant differences.

[0053] From Table 9 and Figure 4 It is evident that the three-stage enzymatic fermentation process employed by the invention group resulted in the lowest ash content and the highest contents of agarwood fermentation products containing agarwood tetraol and iso-anagarwood tetraol. Compared to the control groups, the invention group demonstrated a better effect in reducing ash content, indicating that this process can more effectively control inorganic residues and impurity load during fermentation. Simultaneously, the invention group exhibited higher levels of both marker components, suggesting that this process is beneficial for the release and retention of agarwood's characteristic components. Combined with the significance analysis results, the invention group outperformed other control groups in most indicators, demonstrating that this process has significant advantages in improving product quality stability and marker component levels, achieving a dual improvement in purity and enrichment of medicinal substances, and reflecting a more comprehensive overall quality benefit.

[0054] In summary, the agarwood ferment obtained by the present invention through three-stage enzymatic fermentation has a high content of active ingredients and a rich and fragrant aroma. Its overall quality is significantly better than that of similar fermented products reported in the public. It can be used as a Chinese medicine to enhance efficacy and as a high-grade fragrance to improve quality of life, and has broad practical application value.

Claims

1. A fermented agarwood product, characterized in that: It is made by natural fermentation of agarwood with enzymes; the enzymes are cellulase, β-glucanase, lignin peroxidase and neutral protease; The cellulase and β-glucanase were added to the agarwood at the start of fermentation; lignin peroxidase was added to the agarwood one day after fermentation, and neutral protease was added to the agarwood three days after fermentation.

2. The agarwood fermentation product according to claim 1, characterized in that: Add water to agarwood and then add enzymes; the mass ratio of water to agarwood is 100:40~60.

3. The agarwood fermentation product according to claim 1, characterized in that: The cellulase or β-glucanase is present in a mass percentage of 0.5%-1.5% of agarwood, the lignin peroxidase is present in a mass percentage of 0.2%-0.8% of agarwood, and the neutral protease is present in a mass percentage of 0.3%-1.0% of agarwood.

4. The agarwood fermentation product according to claim 1, characterized in that: The fermentation temperature is 30~40℃, the humidity is 60~70%, and the time is 6~8 days.

5. A fermentation method for agarwood, characterized in that: Includes the following steps: Take agarwood powder, add water and mix well, then spray with enzyme solution, stir evenly and let it ferment naturally to obtain the product; The enzyme solutions are respectively cellulase, β-glucanase, lignin peroxidase and neutral protease; The cellulase and β-glucanase were sprayed into the agarwood at the start of fermentation; lignin peroxidase was sprayed into the agarwood one day after fermentation, and neutral protease was sprayed into the agarwood three days after fermentation.

6. The fermentation method according to claim 5, characterized in that: The fermentation temperature is 30-40°C, the humidity is 60-70%, and the time is 6-8 days.

7. The fermentation method according to claim 5, characterized in that: The mass ratio of water to agarwood is 100:40~60.

8. The fermentation method according to claim 5, characterized in that: The cellulase or β-glucanase is present in a mass percentage of 0.5%-1.5% of agarwood, the lignin peroxidase is present in a mass percentage of 0.2%-0.8% of agarwood, and the neutral protease is present in a mass percentage of 0.3%-1.0% of agarwood.

9. The fermentation method according to claim 5, characterized in that: The enzyme solution is prepared using an enzyme pH buffer solution.

10. The fermentation method according to claim 9, characterized in that: The cellulase solution in the enzyme solution is cellulase diluted with pH 4.8 citrate-sodium citrate buffer to an enzyme concentration of 10 mg / mL; β-glucanase solution is an enzyme solution in which β-glucanase is diluted with acetate-sodium acetate buffer at pH 5.0 to an enzyme concentration of 10 mg / mL. Lignin peroxidase solution is an enzyme solution in which lignin peroxidase is diluted with tartaric acid-sodium tartrate buffer at pH 3.5 to an enzyme concentration of 5 mg / mL. Neutral protease solution is an enzyme solution in which neutral protease is diluted with Tris-HCl buffer at pH 7.0 to an enzyme concentration of 8 mg / mL.

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