Phyllanthus emblica fermentation liquor and preparation method thereof

Abstract

The invention relates to the technical field of deep processing of phyllanthus emblica, in particular to phyllanthus emblica fermentation liquor and a preparation method thereof. The preparation method comprises the following steps: uniformly mixing crushed phyllanthus emblica pulp with water; after sterilization, adding lactic acid bacteria mixed powder for fermentation treatment; cellulase is added into the fermentation liquor for hydrolysis, and hydrolysate is obtained after enzyme deactivation; the hydrolysate is subjected to extraction treatment and solid-liquid separation, and the phyllanthus emblica fermentation liquor is obtained. By adopting the method disclosed by the invention, 107.11 mg of gallic acid, 360.76 mg of polyphenol and 65.56 mg of flavone can be extracted from each gram of the phyllanthus emblica raw material respectively, and the contents of gallic acid, polyphenol and flavone are increased by about 2.1 times, 4.5 times and 1.7 times respectively compared with those of gallic acid, polyphenol and flavone which are not subjected to fermentation treatment; the phyllanthus emblica fermentation liquor has alpha-glucosidase inhibitory activity, xanthine oxidase inhibitory activity, pancreatic lipase inhibitory activity, tyrosinase inhibitory activity and antioxidant activity; the phyllanthus emblica fermentation liquor is good in taste and flavor, and has the functions of reducing blood sugar, blood fat and uric acid and resisting oxidation.

Description

Technical Field

[0001] The invention relates to the technical field of emblica emblica deep processing, and in particular to emblica emblica fermentation liquid and a preparation method thereof. Background Art

[0002] Traditional Chinese medicine fermentation mostly utilizes microorganisms in the natural environment, and through their growth and metabolism, changes the medicinal properties of Chinese medicinal materials, reduces toxicity, and improves the efficacy of Chinese medicine and the utilization rate of Chinese medicine resources. Among them, lactic acid bacteria are widely used in Chinese medicine fermentation due to their high safety and market recognition. The use of fermentation methods can combine the functions of lactic acid bacteria with Chinese medicine, promote the dissolution and transformation of effective ingredients of Chinese medicine, and change the functions of Chinese medicine. In addition, the intestinal flora regulation function of lactic acid bacteria can assist the function of Chinese medicine, and lactic acid bacteria can improve the bioavailability of Chinese medicine during the fermentation process, so that it can exert better medicinal functions.

[0003] Phyllanthus emblica is a plant of the Euphorbiaceae family. Phyllanthus emblica L.) are rich in amino acids, flavonoids, terpenes, volatile oils and other compounds. Emblica extract has multiple biological activities such as anti-microbial infection, anti-oxidation, anti-tumor, lowering blood lipids and blood sugar, lowering blood pressure, and tonic.

[0004] At present, the commonly used extraction methods of Phyllanthus emblica components include solvent method, ultrasonic assisted method, maceration method, enzyme assisted method, subcritical water extraction method, microwave assisted method, reflux method and decompression method, etc. The current extraction method is single, and the extraction rate and effective ingredient content are low. Therefore, the existing extraction process of Phyllanthus emblica needs to be further improved. Summary of the invention

[0005] In order to solve the deficiencies in the prior art mentioned in the above background technology, the present invention provides a method for preparing a fermented liquid of Phyllanthus emblica, and its technical scheme is as follows: The method for preparing the emblica emblica fermented liquid comprises the following steps: Raw material pretreatment: crushing the emblica pulp, sieving, taking the sieved material to obtain the emblica pulp powder, mixing the crushed emblica pulp powder with water, sterilizing, and obtaining a slurry; Fermentation: adding lactic acid bacteria powder and a carbon source to the slurry for fermentation to obtain a fermentation liquid; the lactic acid bacteria powder is one or more of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum; Hydrolysis: adding cellulase to the fermentation broth for hydrolysis to obtain a hydrolyzate; Extraction: extracting the hydrolyzate at a certain temperature for a certain time to obtain an extract; The extract is subjected to solid-liquid separation, and the supernatant is taken for suction filtration to obtain the emblica fermentation liquid.

[0006] In one embodiment, in the raw material pretreatment step, the aperture of the sieve when the emblica flesh is crushed and sieved is 0.05-0.85 mm; the mass ratio of the emblica flesh powder to water is 1:(7.5-30), and during the sterilization process, the sterilization temperature is 85-95°C, and the sterilization time is 20-30 min.

[0007] When the emblica pulp is ground and sieved with a sieve aperture of 0.05 to 0.85 mm, the effective ingredients in the emblica pulp powder are more easily dissolved.

[0008] In one embodiment, in the fermentation step, the lactic acid bacteria powder is added in an amount of 0.005% to 0.03% of the mass of water.

[0009] In one embodiment, the Lactobacillus paracasei is Lactobacillus paracasei YYS-K1, whose preservation number is CGMCC No. 26405; the Lactobacillus plantarum is Lactobacillus plantarum BXM2, whose preservation number is CGMCC No. 16436; and the Lactobacillus fermentum is Lactobacillus fermentum B153, whose preservation number is CGMCC No. 16454.

[0010] In one embodiment, when the lactic acid bacteria powder is any two of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum, the lactic acid bacteria powder comprises one of the following proportions: Lactobacillus paracasei: Lactobacillus plantarum: 1-3:1-5; Lactobacillus paracasei: Lactobacillus fermentum: 1-3:1-7; The ratio of plant lactobacillus: fermented lactobacillus is 1-5:1-7.

[0011] When the lactic acid bacteria powder is any three of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum, the formula includes the following ratios: The mass ratio of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum is 1-3:1-5:1-7.

[0012] Preferably, the ratio of Lactobacillus plantarum to Lactobacillus fermentum is 1:1.

[0013] Preferably, the mass ratio of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum is 3:2:1 or 3:1:1.

[0014] In one embodiment, in the fermentation step, the fermentation temperature is 35-39° C. and the fermentation time is 18-40 hours.

[0015] In one embodiment, in the fermentation step, the carbon source is any one of white sugar, fructooligosaccharide, xylooligosaccharide, polyglucose or isomaltooligosaccharide; and the amount of carbon source added is 5% to 10% of the mass of water.

[0016] Preferably, the carbon source is white sugar.

[0017] Preferably, the amount of white sugar added is 10%.

[0018] In one embodiment, in the hydrolysis step, the hydrolysis temperature is 50-60° C., the hydrolysis time is 1-3 hours, and the mass of the cellulase is 0.3%-0.8% of the mass of the fermentation broth.

[0019] In one embodiment, in the extraction step, the extraction temperature is 85-95° C., the extraction time is 30-90 min, and an extract is obtained; the extract is centrifuged to separate the solid and the liquid, and the supernatant is taken and microfiltered through a membrane with a pore size of 0.22-0.45 μm to obtain the emblica fermentation liquid.

[0020] The invention also provides a phyllanthus emblica fermentation liquid, which is prepared by adopting the preparation method of the phyllanthus emblica fermentation liquid as mentioned above.

[0021] The emblica emblica fermented liquid provided by the present invention has at least one of the following effects: (1) Possessing α-glucosidase inhibitory activity; (2) Possess xanthine oxidase inhibitory activity; (3) Possess pancreatic lipase inhibitory activity; (4) Possess tyrosinase inhibitory activity; (5) Possesses antioxidant properties.

[0022] Based on the above, compared with the prior art, the preparation method of the emblica fermentation liquid of the present invention has the following beneficial effects: The invention prepares the emblica fermentation liquid by microbial fermentation, enzymolysis and extraction of emblica. The method of the invention can prepare the emblica fermentation liquid with high gallic acid, polyphenol and flavonoid content. Gallic acid, polyphenol and flavonoid are the sources of the astringency and bitterness of emblica, and are also one of the sources of the sweetness and fruity flavor of emblica. Therefore, the emblica fermentation liquid prepared by the invention has obvious characteristic flavor.

[0023] In a preferred embodiment, the combined fermentation of three bacteria, namely, Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum, can significantly improve the gallic acid, polyphenol and flavonoid contents in the emblica phyllanthus fermented liquid. These three bacteria are matched together according to a certain ratio, and the synergistic fermentation can increase the quantity and diversity of the beneficial bacteria, inhibit the growth of harmful bacteria, and thus improve the gallic acid, polyphenol and flavonoid contents in the emblica phyllanthus fermented liquid.

[0024] The emblica emblica fermentation broth prepared by the method of the present invention has xanthine oxidase inhibitory activity, pancreatic lipase inhibitory activity, alpha-glucosidase inhibitory rate, tyrosinase inhibitory activity and antioxidant property, and can be used as a raw material component of functional food and applied in functional products.

[0025] In a preferred embodiment, the emblica emblica fermentation liquid produced by co-fermentation of Lactobacillus plantarum and Lactobacillus fermentum has a better α-glucosidase inhibition rate.

[0026] In a more preferred embodiment, the inventors found that in the technical scheme of the present invention, by combining Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum for fermentation, the α-glucosidase inhibitory activity, xanthine oxidase inhibitory activity, pancreatic lipase inhibitory activity, tyrosinase inhibitory activity and antioxidant activity can be significantly improved.

[0027] The method of the invention can prepare the required emblica fermentation liquid through a simple combination of operations such as fermentation, enzymolysis, extraction and filtration, has low requirements on equipment, is simple in process and easy to operate, and is convenient for large-scale industrial production. BRIEF DESCRIPTION OF THE DRAWINGS

[0028] Figure 1 The amounts of gallic acid, polyphenols and flavonoids that can be extracted from 1 gram of Emblica officinalis pulp under different operations.

[0029] Figure 2 is the pancreatic lipase inhibition rate of different Emblica officinalis fermentation broths.

[0030] Figure 3 The xanthine oxidase inhibitory activity of different Emblica officinalis fermentation broths.

[0031] Figure 4 is the α-glucosidase inhibition rate of different embryonic fermentation broths.

[0032] Figure 5 is the tyrosinase activity inhibition rate of different Emblica officinalis fermentation broths. DETAILED DESCRIPTION

[0033] In order to make the purpose, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention are clearly and completely described below. Obviously, the described embodiments are part of the embodiments of the present invention, rather than all the embodiments; the technical features designed in different implementation modes of the present invention described below can be combined with each other as long as they do not conflict with each other; based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0034] In the description of the present invention, it should be noted that all terms (including technical terms and scientific terms) used in the present invention have the same meanings as those generally understood by ordinary technicians in the field to which the present invention belongs, and should not be understood as limitations on the present invention; it should be further understood that the terms used in the present invention should be understood to have the same meanings as these terms in the context of this specification and in the relevant fields, and should not be understood in an idealized or overly formal sense, unless explicitly defined in this invention.

[0035] The present invention provides an operational example of a method for preparing a fermented liquid of Phyllanthus emblica, and the specific steps are as follows: Step 1: Raw material pretreatment: The emblica pulp is crushed and sieved, and the sieved material is taken to obtain the emblica pulp powder, and the crushed emblica pulp powder is mixed with water and sterilized to obtain a slurry; Wherein, the mass ratio of the emblica emblica powder to water is 1:(7.5-30); Among them, the crushing and screening steps are specifically as follows: crushing the emblica pulp with a traditional Chinese medicine crusher, and passing the crushed powder through a stainless steel mesh sieve with a pore size of 0.85 mm to obtain the emblica pulp powder.

[0036] Step 2: Fermentation: adding lactic acid bacteria powder and a carbon source to the slurry for fermentation to obtain a fermentation liquid; In the fermentation process, the fermentation temperature is 35-39° C. and the fermentation time is 18-40 hours; In the fermentation process, the carbon source can be any one of white sugar, oligofructose, oligoxylose, polyglucose or oligomaltose; the added mass is 5% to 10% of the mass of water in step 1; In the fermentation process, the amount of lactic acid bacteria powder added is 0.003% to 0.03% of the mass of water in step 1; The lactic acid bacteria powder is one or more of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum; In the lactic acid bacteria powder, the mass ratio of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum is 1-3:1-5:1-7; Wherein, the Lactobacillus paracasei is Lactobacillus paracasei YYS-K1, Latin name: Lactobacillus paracasei , deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, deposit number: CGMCCNo.26405. Deposit address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, deposit date: January 6, 2023; The plant lactobacillus is plant lactobacillus BXM2, Latin name: Lactobacillus plantarum, deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, deposit number: CGMCC NO.16436. Deposit address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, deposit date: September 6, 2018; The fermented lactobacillus is fermented lactobacillus B153, Latin name: Lactobacillus fermentum , deposited in the General Microbiology Center of China Microbiological Culture Collection Administration, with the deposit number: CGMCC No.16454. Deposit address: No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, deposit date: September 10, 2018.

[0037] Step 3: Hydrolysis: adding cellulase to the fermentation broth for hydrolysis to obtain a hydrolyzate; Specifically, cellulase is added to the fermentation broth, hydrolyzed at 50-60° C. for 1-3 hours, and then the enzyme is inactivated at 85-90° C. for 20-30 minutes to obtain a hydrolyzate; The mass of the cellulase is 0.3% to 0.8% of the mass of the fermentation liquid.

[0038] Step 4: Extraction: leach the hydrolyzate at a certain temperature for a certain time to obtain an extract; Specifically, the hydrolyzate is heated to 85-95° C. and leached for 30-90 minutes to obtain an extract.

[0039] Step 5: Separation and filtration: The extract is subjected to solid-liquid separation, and the supernatant is taken for suction filtration to obtain the emblica fermentation liquid.

[0040] Specifically, the extract is centrifuged, and the supernatant is filtered through a membrane with a pore size of 0.22 to 0.45 μm to obtain the emblica fermentation liquid.

[0041] The present invention provides the following embodiments and comparative examples Example 1 (1) Raw material pretreatment: The emblica pulp was crushed with a traditional Chinese medicine crusher, and 10 g of the emblica pulp powder passed through a stainless steel mesh sieve with a pore size of 0.85 mm was taken, and 300 g of water was added and stirred for 5 min until completely mixed to obtain a mixed slurry.

[0042] (2) Fermentation: The mixed slurry was sterilized at 95° C. for 30 min, 24 g of white sugar and 0.03 g of Lactobacillus paracasei YYS-K1 powder were added, and the mixture was fermented at 37° C. for 38 h to obtain a fermentation liquid.

[0043] (3) Hydrolysis: Add 1.5 g of cellulase to the fermentation broth and hydrolyze at 55° C. for 2 h to obtain a hydrolyzate; (4) Extraction: The hydrolyzate is extracted at 90° C. for 2 h to obtain an extract.

[0044] (5) Separation and filtration: The extract is centrifuged to separate the solid and liquid, and the supernatant is filtered through a membrane with a pore size of 0.22 to 0.45 μm to obtain the emblica fermentation liquid.

[0045] Example 2 The powder of Lactobacillus paracasei YYS-K1 was replaced with an equal amount of Lactobacillus plantarum BXM2 powder, and the other operations and steps were the same as those in Example 1.

[0046] Example 3 The powder of Lactobacillus paracasei YYS-K1 was replaced with an equal amount of Lactobacillus fermentum B153 powder, and the other operations and steps were the same as those in Example 1.

[0047] Example 4 The powder of Lactobacillus paracasei YYS-K1 was replaced with a mixed powder of equal amounts of Lactobacillus plantarum BXM2 and Lactobacillus fermentum B153, with a mass ratio of 1:1. Other operations and steps were the same as those in Example 1.

[0048] Example 5 The powder of Lactobacillus paracasei YYS-K1 was replaced with a mixed powder of equal amounts of Lactobacillus paracasei YYS-K1, Lactobacillus plantarum BXM2 and Lactobacillus fermentum B153 in a mass ratio of 3:2:1. Other operations and steps were the same as those in Example 1.

[0049] Example 6 The powder of Lactobacillus paracasei YYS-K1 was replaced with a mixed powder of equal amounts of Lactobacillus paracasei YYS-K1, Lactobacillus plantarum BXM2 and Lactobacillus fermentum B153 in a mass ratio of 3:1:1. Other operations and steps were the same as those in Example 1.

[0050] Example 7 The mass of the white sugar in Example 1 was increased to 30 g, and the other operations and steps were the same as in Example 1.

[0051] Example 8 No white sugar was added during the fermentation process in Example 1, and other operations and steps were the same as in Example 1.

[0052] Example 9 The white sugar in Example 1 was replaced with an equal amount of oligofructose, and the other operations and steps were the same as those in Example 1.

[0053] Comparative Example 1 The emblica flesh powder in Example 1 was subjected to enzymolysis and extraction without microbial fermentation, and other operations and steps were the same as those in Example 1.

[0054] In the embodiments and comparative examples: Lactobacillus paracasei ( Lactobacillus paracasei )YYS-K1, deposited on January 6, 2023 in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO.26405.

[0055] Lactobacillus plantarum Lactobacillus plantarum )BXM2 was deposited in the General Microbiology Center of China Microorganism Culture Collection Administration on September 6, 2018. The deposit address is No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is CGMCC NO.16436.

[0056] Lactobacillus fermentum ( Lactobacillus fermentum )B153, deposited on September 10, 2018 in the General Microbiology Center of China Microorganism Culture Collection Administration, with the deposit address at No. 3, Yard No. 1, Beichen West Road, Chaoyang District, Beijing, and the deposit number is: CGMCC No.16454.

[0057] These strains are all preserved and maintained at the General Microbiology Center of the China Microbiological Culture Collection Administration. Technicians can obtain them from the collection center based on the collection number.

[0058] The cellulase is specifically Pombo cellulase, and the enzyme activity is 20,000 Unit / g.

[0059] The performance test of the emblica fermentation broth products obtained in the above examples and comparative examples was performed: 1. The amount of gallic acid, polyphenols and flavonoids extracted from 1 gram of Phyllanthus emblica pulp under different operations In the detection examples 1-9 and comparative example 1, the amounts of gallic acid, polyphenols and flavonoids that can be extracted from 1 gram of emblica pulp under different operations were determined.

[0060] Among them, the determination of gallic acid content refers to the LC-DAD of Phyllanthus emblica in Part I of the 2020 edition of the Chinese Pharmacopoeia; the determination of flavonoids content refers to the second method of determination of total flavonoids in health foods in the Technical Guidelines for Testing and Evaluation of Physical, Chemical and Hygiene Indicators of Health Foods (2020 Edition); the determination of polyphenols content refers to the spectrophotometric method for determination of total polyphenols in plant extracts and their products in T / AHFIA005-2018. The specific results are shown in Figure 1 .

[0061] Depend on Figure 1 It can be seen that: The contents of gallic acid, polyphenols and flavonoids in the emblica fermentation broth obtained in Examples 5 and 6 are higher than those in other Examples and Comparative Examples, indicating that specific strains and the proportion of specific strains have a great influence on the extraction of effective ingredients in emblica, among which the content of polyphenols has the greatest influence.

[0062] Among them, in Example 7, when the proportion of white sugar was increased to 10% (30 g), compared with Example 1, the polyphenol content was significantly increased, and the gallic acid and flavonoid contents did not change significantly.

[0063] Compared with Examples 1-3 and Examples 8-9, the contents of gallic acid, polyphenols and flavonoids in the emblica fermentation broth prepared in Examples 5 and 6 were significantly increased. This result shows that although the effect of adding a single bacterium or changing the carbon source is improved to a certain extent compared with Comparative Example 1, it cannot reach the effect of the emblica fermentation broth prepared by fermenting three bacteria in a certain ratio. Only when the three bacteria are implemented in a specific ratio can the effect of significantly increasing the contents of gallic acid, polyphenols and flavonoids in the emblica fermentation broth be achieved.

[0064] The difference between Example 9 and Example 1 is that the carbon source is different, the carbon source of Example 9 is oligofructose, and the carbon source of Example 1 is white sugar. Compared with Example 9, the polyphenol content in the emblica fermentation broth after adding white sugar in Example 1 is higher, indicating that using white sugar as a carbon source is more preferred.

[0065] It is explained that the embodiment of the present invention adopts a process combining lactic acid bacteria fermentation, enzymatic hydrolysis and extraction, as well as fermentation of specific strains and specific strains in proportion, which can improve the extraction rate of gallic acid, polyphenols and flavonoids in emblica pulp, and when the white sugar content is 10%, the polyphenol content is significantly improved.

[0066] 2. Inhibition of pancreatic lipase activity by Phyllanthus emblica fermentation broth The inhibition rate of the Emblica officinalis fermentation broth on pancreatic lipase in Examples 1-9 and Comparative Example 1 was detected.

[0067] Among them, the determination of pancreatic lipase inhibitory activity of Phyllanthus emblica fermentation broth refers to GB / T 23535-2009 National Standard: Lipase Preparations. The specific results are shown in Figure 2 .

[0068] Depend on Figure 2 It can be seen that: Compared with other embodiments and comparative examples, the inhibition rate of pancreatic lipase activity of the emblica fermented liquid prepared in Embodiment 5 and Embodiment 6 is significantly improved, which is several times that of other embodiments and comparative examples, indicating that specific strains and specific ratios have a greater influence on the inhibition rate of pancreatic lipase activity of the emblica fermented liquid; Comparative Example 1 is the emblica fermented liquid prepared without fermentation treatment, and its inhibition rate of pancreatic lipase activity is lower than that of Embodiments 1-9.

[0069] Pancreatic lipase plays a vital role in the digestive process. It is responsible for breaking down the fat in food into glycerol and fatty acids for absorption by the small intestine. Substances that inhibit the activity of pancreatic lipase can reduce the absorption of fat by inhibiting the activity of pancreatic lipase. The index of pancreatic lipase inhibition activity is an important index for evaluating the anti-obesity effect. It is explained that the embodiment of the present invention adopts a process combining lactic acid bacteria fermentation, enzymolysis and extraction, as well as specific strains and specific strains of fermentation. The obtained emblica fermentation liquid has a more excellent inhibition rate on lipase activity, and has potential application value in the treatment of obesity.

[0070] 3. Xanthine oxidase inhibitory activity of Phyllanthus emblica fermentation broth In Examples 1-9 and Comparative Example 1, the xanthine oxidase inhibitory activity of the Emblica officinalis fermentation broth was detected.

[0071] The determination of xanthine oxidase inhibitory activity of Phyllanthus emblica fermentation broth was based on the results of the screening, structural identification and preliminary study of xanthine oxidase inhibitors in Smilax china. Figure 3 .

[0072] Depend on Figure 3 It can be seen that: Compared with other embodiments and comparative examples, the inhibition rate of xanthine oxidase activity of the emblica fermentation broth prepared in Embodiment 5 and Embodiment 6 is higher, indicating that specific strains and specific ratios have a greater influence on the inhibition rate of xanthine oxidase activity of the emblica fermentation broth; Comparative Example 1 is the emblica fermentation broth prepared without fermentation treatment, and its inhibition rate of xanthine oxidase activity is lower than that of Embodiments 1-9.

[0073] Xanthine oxidase participates in purine metabolism and catalyzes the oxidation of hypoxanthine and xanthine to produce uric acid. It is a key enzyme in the production of uric acid. Therefore, the inhibitory activity of xanthine oxidase is an important indicator for evaluating the potential effect of drugs or natural products in treating diseases such as hyperuricemia and gout. It is explained that the embodiment of the present invention adopts a process combining lactic acid bacteria fermentation, enzymolysis and extraction, as well as specific strains and specific strains of fermentation ratios. The obtained Phyllanthus emblica fermentation liquid has a more excellent inhibition rate on xanthine oxidase activity, and has potential application value in the treatment of hyperuricemia.

[0074] 4. α-Glucosidase inhibitory activity of different embryonic fermentation broths The inhibition rate of emblica emblica fermentation broth on α-glucosidase in Examples 1-9 and Comparative Example 1 was detected.

[0075] Among them, the determination of α-glucosidase inhibitory activity of Phyllanthus emblica fermentation broth was based on the Characterization of antidiabetic and antihypertensive properties of canary seed ( Phalaris canariensis L. ) peptides”, and the specific results can be found in Figure 4 .

[0076] Depend on Figure 4 It can be seen that: α-glucosidase is an enzyme that catalyzes the decomposition of carbohydrates into glucose in the small intestine, and the activity of α-glucosidase plays an important role in regulating blood sugar levels. Compared with other embodiments and comparative examples, the α-glucosidase activity inhibition rate of the emblica fermented liquid obtained in Examples 5 and 6 is higher, indicating that specific strains and specific ratios have a greater influence on the α-glucosidase activity inhibition rate of the emblica fermented liquid; Comparative Example 1 is an emblica fermented liquid obtained without fermentation treatment, and its α-glucosidase activity inhibition rate is lower than that of Examples 1-9.

[0077] It is explained that the embodiment of the present invention adopts a process combining lactic acid bacteria fermentation, enzymatic hydrolysis and extraction, as well as fermentation of specific bacteria species and specific bacteria species in proportion, and the obtained emblica fermentation liquid has a more excellent inhibition rate on α-glucosidase activity, and has potential application value in lowering blood sugar.

[0078] 5. Inhibition of tyrosinase activity by different emblica fermentation broths The inhibition rate of tyrosinase by the emblica fermentation broth in Examples 1-9 and Comparative Example 1 was detected.

[0079] Among them, the determination of tyrosinase inhibition activity of Phyllanthus emblica fermentation broth adopts T / SHRH015~2018 cosmetics tyrosinase activity inhibition test, and the specific results are shown in Figure 5 .

[0080] Depend on Figure 5 It can be seen that: Compared with other embodiments and comparative examples, the tyrosinase activity inhibition rate of the emblica fermentation broth prepared in Embodiment 5 and Embodiment 6 is higher, indicating that specific strains and specific ratios have a greater influence on the tyrosinase activity inhibition rate of the emblica fermentation broth; Comparative Example 1 is the emblica fermentation broth prepared without fermentation treatment, and its tyrosinase activity inhibition rate is lower than that of Embodiments 1-9.

[0081] It is explained that the embodiment of the present invention adopts a process combining lactic acid bacteria fermentation, enzymolysis and extraction, as well as fermentation of specific bacteria species and specific bacteria species in a certain proportion, so that the obtained Emblica emblica fermentation liquid has a more excellent inhibition rate on tyrosinase activity.

[0082] 6. Antioxidant activity of different emblica fermentation broths The hydroxyl radical scavenging rate, DPPH radical scavenging rate and ABTS radical scavenging rate of the Emblica officinalis fermentation broth of Examples 1-9 and Comparative Example 1 were detected.

[0083] Among them, the DPPH free radical and ABTS free radical scavenging rate detection method of the fermentation broth of Phyllanthus emblica refers to "GB / T39100-2020", and the hydroxyl free radical scavenging rate detection method refers to "Research on the Antioxidant Activity of Panax Notoginseng Polysaccharides".

[0084] See Table 1 for specific data: Table 1 Antioxidant capacity of different emblica fermentation broths project Hydroxyl free radical scavenging rate / % DPPH free radical scavenging rate / % ABTS free radical scavenging rate / % Example 1 90.10±0.65 99.87±0.50 95.63±1.25 Example 2 88.67±1.07 99.52±0.96 98.74±0.31 Example 3 88.97±1.09 100.11±0.48 99.08±0.12 Example 4 87.18±1.02 99.90±0.49 100.35±0.81 Example 5 100.30±1.41 99.80±0.55 98.78±0.87 Example 6 87.44±3.97 99.98±0.57 99.72±0.24 Example 7 88.60±1.09 99.27±0.61 99.46±0.31 Example 8 86.57±0.51 99.99±0.46 98.78±1.60 Example 9 87.61±0.49 99.55±0.57 98.74±1.44 Comparative Example 1 76.66±1.63 100.60±0.81 99.24±0.34 From Table 1, we can see that: Different emblica fermented liquids all have good antioxidant capacity, especially in DPPH free radical and ABTS free radical scavenging capacity. However, it is worth mentioning that the hydroxyl radical scavenging rate of the emblica fermented liquid in the embodiment is higher than that in the comparative example, indicating that from the comprehensive evaluation index of antioxidant activity, the embodiment of the present invention adopts the process of fermentation of mixed lactic acid bacteria powder, enzymolysis and extraction, and fermentation of specific strains and specific strains, and the obtained emblica fermented liquid has more excellent antioxidant activity.

[0085] In summary, the method for preparing the emblica fermentation liquid provided by the present invention has at least the following innovative features, action mechanisms and technical effects: Its innovations: The present invention is different from the prior art in that: (1) The present invention is the first to use a specific microbial combination to ferment the emblica slurry, and further hydrolyze and extract it. This preparation method not only increases the content of gallic acid, polyphenols and flavonoids in the fermentation liquid, but also the obtained emblica slurry has a distinct characteristic flavor and no peculiar smell, and has a good taste and flavor.

[0086] (2) The phyllanthus emblica fermentation broth obtained by the synergistic action of fermentation, enzymolysis and extraction has α-glucosidase inhibitory activity, xanthine oxidase inhibitory activity, pancreatic lipase inhibitory activity, tyrosinase inhibitory activity and antioxidant activity.

[0087] Technical effects: (1) The method of the present invention uses mixed lactic acid bacteria powder to treat the emblica slurry, and then uses enzymatic hydrolysis and leaching treatment to obtain a fermented emblica slurry with high content of gallic acid, polyphenols and flavonoids; (2) The present invention processes the emblica pulp to obtain the fermented emblica liquid, which has multiple biological activities such as anti-cancer, hypoglycemic, lipid-lowering, uric acid-lowering, antibacterial, anti-inflammatory, antiviral, cardiovascular and cerebrovascular protection, liver protection, antioxidant, and whitening effects due to the polyphenols and flavonoids. Therefore, the emblica fermented liquid has the biological activities of hypoglycemic, uric acid-lowering, lipid-lowering, whitening, and antioxidant effects, and can be used as a functional factor in functional products (such as functional foods, cosmetics, medicines, and health products).

[0088] (3) The emblica emblica fermented liquid prepared by the present invention has obvious characteristic flavor, no foreign smell, good taste and good flavor.

[0089] (4) The method of the present invention adopts a simple combination of operations such as fermentation, enzymolysis, and extraction to prepare the desired embryonic phlox fermentation liquid, has low requirements on equipment, is simple and easy to operate, and is convenient for large-scale industrial production.

[0090] It should be noted that: (1) Definition: In this article, “~” is used to indicate a numerical range, and the range indicated by this expression includes two endpoint values.

[0091] The term "food" as used herein is used in a broad sense and includes food and drink for human beings.

[0092] The "DPPH" in this article is 1,1-diphenyl-2-trinitrophenylhydrazine, also known as 1,1-diphenyl-2-picrylhydrazyl (free radical).

[0093] Herein, “ABTS” refers to the free radical 2,2'-azino-bis-3-ethylbenzothiazoline-6-sulfonic acid.

[0094] (2) Raw materials used in implementation: The cellulase used is an existing commercially available enzyme, which can be purchased by those skilled in the art.

[0095] (3) Application of Phyllanthus emblica fermentation liquid: Emblica fermentation liquid has the following characteristics: 1) α-glucosidase inhibitory activity; 2) xanthine oxidase inhibitory activity; 3) pancreatic lipase inhibitory activity; 4) tyrosinase inhibitory activity; 5) antioxidant activity; based on this, according to the functional characteristics of 1)-5) above, Emblica fermentation liquid can also be used in functional products with effects such as skin care, weight loss, lowering blood sugar, anti-oxidation, and lowering uric acid (functional products can be food, health products, cosmetics, skin care products, etc., including any substances that provide preventive and / or other beneficial effects), including but not limited to the above effects.

[0096] In summary, the specific parameters or some commonly used reagents or raw materials in the above embodiments are specific embodiments or preferred embodiments under the concept of the present invention, but are not intended to limit the present invention; those skilled in the art can make adaptive adjustments within the concept and protection scope of the present invention.

[0097] In addition, unless otherwise specified, the raw materials used may also be conventional commercial products in the art, or prepared by conventional methods in the art; that is, the reagents and instruments used in this embodiment are not indicated with information such as the manufacturer, and are all conventional products that can be purchased on the market.

[0098] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for preparing a fermented liquid of Phyllanthus emblica, characterized in that , including the following steps: Raw material pretreatment: crushing the emblica pulp, sieving, taking the sieved material to obtain the emblica pulp powder, mixing the crushed emblica pulp powder with water, sterilizing, and obtaining a slurry; Fermentation: adding lactic acid bacteria powder and a carbon source to the slurry for fermentation to obtain a fermentation liquid; the lactic acid bacteria powder is one or more of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum; Hydrolysis: adding cellulase to the fermentation broth for hydrolysis to obtain a hydrolyzate; Extraction: extracting the hydrolyzate at a certain temperature for a certain time to obtain an extract; The extract is subjected to solid-liquid separation, and the supernatant is taken for suction filtration to obtain the emblica fermentation liquid.

2. The method for preparing the emblica emblica fermented liquid according to claim 1, wherein: In the raw material pretreatment step, the aperture of the sieve is 0.05-0.85 mm when the emblica flesh is crushed and sieved; the mass ratio of the emblica flesh powder to water is 1:(7.5-30); during the sterilization process, the sterilization temperature is 85-95° C. and the sterilization time is 20-30 min.

3. The method for preparing the emblica emblica fermented liquid according to claim 1, wherein: In the fermentation step, the lactic acid bacteria powder is added in an amount of 0.005% to 0.03% of the mass of water.

4. The method for preparing the emblica emblica fermented liquid according to claim 1, wherein: The Lactobacillus paracasei is Lactobacillus paracasei YYS-K1, and its deposit number is CGMCC No.26405; The plant lactobacillus is plant lactobacillus BXM2, and its deposit number is CGMCC NO.16436; The fermented lactobacillus is fermented lactobacillus B153, and its preservation number is CGMCC No.16454.

5. The method for preparing the emblica fermented liquid according to claim 1, wherein: When the lactic acid bacteria powder is any two of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum, the lactic acid bacteria powder is one of the following proportions: Lactobacillus paracasei: Lactobacillus plantarum: 1-3:1-5; Lactobacillus paracasei: Lactobacillus fermentum: 1-3:1-7; The ratio of plant lactobacillus: fermented lactobacillus is 1-5:1-7. When the lactic acid bacteria powder is any three of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum, the proportion is as follows: The mass ratio of Lactobacillus paracasei, Lactobacillus plantarum and Lactobacillus fermentum is 1-3:1-5:1-7.

6. The method for preparing the emblica emblica fermented liquid according to claim 1, wherein: In the fermentation step, the fermentation temperature is 35-39° C. and the fermentation time is 18-40 hours.

7. The method for preparing the emblica emblica fermented liquid according to claim 1, wherein: In the fermentation step, the carbon source is any one of white sugar, oligofructose, oligoxylose, polyglucose or oligomaltose; and the amount of carbon source added is 5% to 10% of the mass of water.

8. The method for preparing the emblica emblica fermented liquid according to claim 1, wherein: In the hydrolysis step, the hydrolysis temperature is 50-60° C., the hydrolysis time is 1-3 hours, and the mass of the cellulase is 0.3%-0.8% of the mass of the fermentation liquid.

9. The method for preparing the emblica emblica fermented liquid according to claim 1, wherein: In the extraction step, the extraction temperature is 85-95°C, the extraction time is 30-90 minutes, and an extract is obtained; the extract is centrifuged to separate the solid and the liquid, and the supernatant is taken and microfiltered through a membrane with a pore size of 0.22-0.45 μm to obtain the emblica fermentation liquid.

10. A fermented liquid of Phyllanthus emblica, characterized in that: The method is adopted to prepare the emblica fermentation liquid according to any one of claims 1 to 9.

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