CN115812951B - Gastrodia elata fermentation product with antioxidant activity and preparation method and application thereof - Google Patents
Gastrodia elata fermentation product with antioxidant activity and preparation method and application thereof Download PDFInfo
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- CN115812951B CN115812951B CN202211484596.8A CN202211484596A CN115812951B CN 115812951 B CN115812951 B CN 115812951B CN 202211484596 A CN202211484596 A CN 202211484596A CN 115812951 B CN115812951 B CN 115812951B
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Abstract
The invention provides a gastrodia elata fermentation product and a preparation method and application thereof. The gastrodia elata fermentation product provided by the invention comprises, by mass, 0.62-0.68% of gastrodin, 4.5-5.5% of glutamic acid, 2.9-3.5% of alanine and 3.1-4% of histidine. The gastrodia elata fermentation product provided by the invention has high active ingredients such as gastrodin, obvious antioxidant activity and proper taste.
Description
Technical Field
The invention relates to the field of plant fermented products, in particular to a gastrodia elata fermentation product with antioxidant activity, and a preparation method and application thereof.
Background
The gastrodia elata is a dry tuber of gastrodia elata belonging to orchidaceae, is a health-care food raw material approved by the national ministry of health, and has the effects of calming endogenous wind, relieving spasm, stabilizing liver yang, dispelling wind and dredging collaterals. The main chemical components of the gastrodia elata comprise phenolic compounds such as gastrodin and p-hydroxybenzyl alcohol, gastrodia elata polysaccharide, carotene, citric acid and various trace elements, and the main active substances are gastrodin. Research shows that gastrodin has various pharmacological activities of resisting oxygen free radical, tranquilizing, relieving pain, resisting brain injury, protecting cardiovascular system, improving intestinal tract absorption, enhancing body humoral immunity, etc.
Gastrodine and p-hydroxybenzyl alcohol rich in rhizoma Gastrodiae can block oxidation chain reaction and activate ERK2/1-Nrf2 passage by hydrogen ion transfer, and up-regulate Catalase (CAT) and superoxide dismutase (SOD) to achieve antioxidant effect. The efficacy of the gastrodia elata can be shown by processing, and the traditional processing technology needs to steam, dry and reshape and sweat for 3-4 times. The process is complicated, a large amount of energy is consumed, and the gastrodia elata product prepared by the processing process has special flavor and obvious traditional Chinese medicine taste. In addition, wild gastrodia elata has urine odor, and artificially planted gastrodia elata has slightly sour taste and bitter taste, and is not good in direct eating taste, so that consumers are also difficult to accept, and other treatments are needed in the development of products taking gastrodia elata as a raw material.
The ferment is a general term for products containing specific bioactive components prepared by taking vegetables and fruits, brown rice, medicinal and edible traditional Chinese medicines and other plants as main raw materials and adding or not adding auxiliary materials and fermenting by microorganisms. The active substances include enzymes such as lipase, amylase, superoxide dismutase, etc., organic acids such as lactic acid and acetic acid, and bioactive components such as phenols, flavonoids, polypeptides, etc. Has physiological functions of scavenging free radicals, resisting oxidation, inhibiting bacteria, relieving alcoholic intoxication, protecting liver, loosening bowel to relieve constipation, regulating immunity, promoting metabolism, and purifying blood.
Disclosure of Invention
Aiming at the technical problems of poor palatability and poor oxidation resistance of the conventional gastrodia elata raw material in the comprehensive development process, the invention provides a gastrodia elata fermentation product and a preparation method thereof.
The gastrodia elata fermentation product is prepared from gastrodia elata serving as a raw material through enzymolysis, fermentation and filtration, and key active ingredients of the gastrodia elata fermentation product are quantitatively controlled. The method firstly improves the taste of the gastrodia elata product through enzymolysis treatment, improves the extraction rate of active components, and secondly directionally enriches active components such as gastrodin through regulation and control fermentation, thereby realizing stable and controllable product efficacy and quality.
The invention provides a gastrodia elata fermentation product, which comprises, by mass, 0.62-0.68% of gastrodin, 4.5-5.5% of glutamic acid, 2.9-3.5% of alanine and 3.1-4% of histidine.
Preferably, the fermentation product contains, by mass, 0.67-0.68% of gastrodin, 5.1-5.2% of glutamic acid, 3.4-3.5% of alanine and 3.9-4% of histidine.
Preferably, the ethanol content of the fermentation product is 0.25-0.5wt%.
Preferably, the total acid content in the fermentation product is 2.2-2.7% by mass, calculated as lactic acid;
preferably, the pH of the fermentation product is 3.6-4.0.
Preferably, the para-hydroxybenzene methanol content of the fermentation product is 1.12-1.21wt% in mass percent.
Preferably, the fermentation product further comprises 0.27-0.3% of threonine, 0.5-0.65% of serine, 0.7-1.2% of glycine, 2.9-3.5% of alanine, 3.1-4% of histidine, 0.8-1.2% of aspartic acid, 4.5-5.5% of glutamic acid, 0.5-1.2% of lysine, 0.6-0.8% of tyrosine and 0.7-1.2% of phenylalanine by mass percent.
The invention provides a preparation method of a gastrodia elata fermentation product, which comprises the following steps:
Mixing rhizoma Gastrodiae powder with water to obtain mixed solution, adding enzyme preparation for enzymolysis, and fermenting for 20-25 days to obtain the fermentation product;
The enzyme preparation is cellulase, pectase and papain with the mass ratio of 1-2:1-2:1.
Preferably, the mass ratio of the gastrodia elata powder to the water is 1:5-10.
Preferably, the enzymolysis reaction temperature is 50-55 ℃,
Preferably, the pH value of the enzymolysis reaction is 5.5-6.5;
Preferably, the enzymolysis reaction time is 3-5h.
Preferably, the mass of the added enzyme preparation is 1% -1.5% of the mass of the mixed solution.
Preferably, the enzyme preparation is cellulase, pectase and papain in a mass ratio of 2:1:1.
Preferably, the solution after enzymolysis is sterilized and then the microbial inoculum is added.
Preferably, the addition of the carbon source is also included before the addition of the microbial inoculum,
Preferably, the mass of the added carbon source is 10% -15% of the mass of the mixed solution;
preferably, the carbon source is sucrose.
Preferably, the mass of the microbial inoculum is 1% -2% of the mass of the mixed liquid.
Preferably, the fermentation temperature is 35-40 ℃;
Preferably, the fermentation mode is anaerobic fermentation.
Preferably, the fermenting agent comprises saccharomycetes and lactobacillus, and preferably, the mass ratio of saccharomycetes to lactobacillus is 2:3;
preferably, the yeast is one or more than two of Saccharomyces cerevisiae and Kluyveromyces marxianus;
Preferably, the lactobacillus is one or more than two of lactobacillus acidophilus, lactobacillus plantarum and bifidobacterium.
Preferably, the preparation method further comprises a filtration process;
preferably, the fermented solution is circulated through a 150-300 mesh plate frame for filtration until the solution is clear.
The invention also provides application of the fermentation product or the fermentation product prepared by the preparation method in drinks, skin whitening products, cosmetics and daily chemical washing and caring.
The gastrodia elata is subjected to microbial fermentation treatment, so that functional substances of the gastrodia elata can be revealed, functional components can be further enriched in the fermentation process, the taste problem of the gastrodia elata is effectively improved, and the gastrodia elata is convenient for consumers to accept.
The gastrodia elata fermentation product provided by the invention has good taste. Ether substances in gastrodia elata and bitter amino acid and bitter peptide substances such as arginine (4.88%), valine (4.05%) and the like contained in gastrodia elata cause odor and poor palatability of the gastrodia elata. In the method, the amino acids and the bitter peptides are subjected to biotransformation in combination with proteolytic pretreatment and probiotic fermentation links, so that the delicate flavor and sweet taste components, such as glutamic acid (5.19%), alanine (3.40%), histidine (3.96%) and the like, are increased, and the taste of the gastrodia elata is obviously improved. In addition, the fermentation link can reduce ether substances, increase alcohols and esters substances, eliminate the odor of medicines and urine in the gastrodia elata raw material, improve the palatability and have pleasant aromatic odor.
The gastrodia elata fermentation product provided by the invention has high active ingredients such as gastrodin, obvious antioxidant activity and proper taste, is suitable for commercial comprehensive development value and is easy to realize industrial production.
The preparation method of the gastrodia elata fermentation product provided by the invention is used for carrying out micro-crushing and enzymolysis treatment on gastrodia elata, so that the cell wall of the gastrodia elata can be effectively destroyed, the cell tissues are softened, active substances in the gastrodia elata can be better dissolved out, and the active substances are extracted to the greatest extent; in addition, the enzymolysis can convert macromolecular cellulose, protein and other organic matters into fermentable nutrient sources, so as to provide directly absorbable nutrient sources for fermentation of probiotics, promote rapid growth and shorten the fermentation period.
Compared with the existing methods of direct fermentation of gastrodia elata, re-fermentation after leaching of gastrodia elata and the like, the method is simple in process route and short in ferment preparation period.
Detailed Description
The invention provides a gastrodia elata fermentation product which has high active ingredients such as gastrodin, obvious antioxidant activity and proper taste.
The gastrodia elata fermentation product provided by the invention comprises, by mass, 0.62-0.68% of gastrodin, 4.5-5.5% of glutamic acid, 2.9-3.5% of alanine and 3.1-4% of histidine.
The reagents and instrument source information used in the present invention are shown in table 1 below.
TABLE 1
Kluyveromyces KW-6 (Kluyveromyces marxious KW-6) is preserved in China Center for Type Culture Collection (CCTCC) on the 02 th year 2020, the preservation number is CCTCC NO: M2020173, and the preservation address is: chinese, wuhan, university of Wuhan, postal code: 430072; telephone: (027) -68754052. This strain is described in prior art CN114107082 a.
Lactobacillus plantarum S2 strain (Lactobacillus plantarum S2) was deposited at the China Center for Type Culture Collection (CCTCC) at 4-8 of 2021 with a deposit number of cctcccno: m2021340, deposit address: chinese, wuhan, university of Wuhan, postal code: 430072; telephone: 027-68754052. This strain is described in prior art CN113755360 a.
The following detailed description of the invention in conjunction with the specific examples is merely illustrative of the technical solution of the invention, and not limiting thereof.
Example 1
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 80 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking 1kg of gastrodia elata powder, adding 5 times of water, stirring and uniformly mixing to obtain a mixed solution, heating to 50 ℃, adjusting the pH to 5.5, adding mixed enzyme (the mass ratio, cellulase: pectase: papain=1:1:1) accounting for 1% of the total mass of the mixed solution, and carrying out enzymolysis for 3 hours.
Fermentation: sterilizing the solution after enzymolysis, cooling to 35 ℃, adding sucrose with the mass accounting for 10% of the total mass of the mixed solution, and adding a fermentation inoculant with the mass accounting for 2% of the total mass of the mixed solution (mass ratio, high-temperature resistant saccharomyces cerevisiae, lactobacillus acidophilus D24673, lactobacillus plantarum S2 strain, bifidobacterium D51579=2:1:1:1), and uniformly mixing. Sealing and fermenting at 40 ℃ for 20 days.
Filtering and canning: and (3) circulating the fermented solution through a 150-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
Example 2
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 100 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking 1kg of gastrodia elata powder, adding 10 times of water, stirring and uniformly mixing to obtain a mixed solution, heating to 50 ℃, adjusting pH to 6.0, adding mixed enzyme (the mass ratio, cellulase: pectase: papain=2:1:1) accounting for 1.2% of the total mass of the mixed solution, and carrying out enzymolysis for 3 hours.
Fermentation: sterilizing the solution after enzymolysis, cooling to 35 ℃, adding sucrose with the mass accounting for 10% of the total mass of the mixed solution, and adding a fermentation inoculant (Kluyveromyces KW-6: lactobacillus acidophilus D24673: lactobacillus plantarum S2 strain: bifidobacterium D51579=2:1:1:1) with the mass accounting for 1.5% of the total mass of the mixed solution, and uniformly mixing. Sealing and fermenting at 35 ℃ for 25 days.
Filtering and canning: and (3) circulating the fermented solution through a 200-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
Example 3
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 100 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking a certain amount of rhizoma Gastrodiae powder, adding 10 times of water, stirring, mixing to obtain mixed solution, heating to 55deg.C, adjusting pH to 6.5, adding mixed enzyme (mass ratio, cellulase: pectase: papain=2:1:1) with mass of 1.5% of the total mass of the mixed solution, and performing enzymolysis for 4 hr.
Fermentation: sterilizing the solution after enzymolysis, cooling to 35 ℃, adding sucrose with the mass being 15% of the total mass of the mixed solution, and adding a fermentation inoculant (mass ratio, kluyveromyces marxianus: lactobacillus acidophilus: lactobacillus plantarum = bifidobacterium = 2:1:1:1) with the mass being 1% of the total mass of the mixed solution, and uniformly mixing. Sealing and fermenting at 35 ℃ for 20 days.
Filtering and canning: and (3) circulating the fermented solution through a 300-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 1
Slicing fresh rhizoma Gastrodiae, placing into a tank, filling with one layer of rhizoma Gastrodiae tablet and one layer of sucrose, sealing the tank opening with eight layers of gauze, fermenting at room temperature and ventilation for 1 month, sealing the tank opening, fermenting at room temperature for 6 months, and filtering and clarifying to obtain rhizoma Gastrodiae ferment.
Comparative example 2
Crushing gastrodia elata, sieving with a 200-mesh sieve to obtain gastrodia elata powder, weighing 1kg of gastrodia elata powder, adding 10 times of pure water, leaching for 2 hours at 60 ℃ and pH6.0, centrifuging to obtain leaching liquor, and repeating the leaching step once. And mixing the two leaching solutions, adding 15% sucrose, stirring uniformly, sterilizing, cooling to room temperature, inoculating 1% composite microbial inoculum (mass ratio, saccharomycetes: lactobacillus=1:1), sealing and fermenting at 35 ℃ for 5 days, filtering with a plate frame, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 3
Fresh gastrodia elata and 5 times of pure water are mixed and homogenized, and after the temperature is raised to 50 ℃, 1% of compound enzyme (the mass ratio is that of cellulase and pectase=1:1) is added for enzymolysis, and the enzymolysis time is 3 hours. Then adding 15% sucrose into the enzymolysis liquid, stirring uniformly, sterilizing, cooling to room temperature, inoculating 1% composite microbial inoculum (mass ratio, saccharomycetes: lactobacillus=1:1), fermenting at 35 ℃ for 60 days, filtering with a plate frame, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 4
Fresh gastrodia elata and 5 times of pure water are mixed and homogenized, and after the temperature is raised to 50 ℃, 1% of compound enzyme (the mass ratio is that of cellulase and pectase and amylase=1:1:1) is added for enzymolysis, and the enzymolysis time is 3 hours. Then adding 15% sucrose into the enzymolysis liquid, stirring uniformly, sterilizing, cooling to room temperature, inoculating 1% composite microbial inoculum (mass ratio, saccharomycetes: lactobacillus=1:1), fermenting at 35 ℃ for 60 days, filtering with a plate frame, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 5
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 100 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking 1kg of gastrodia elata powder, adding 10 times of water, stirring and mixing uniformly, heating to 50 ℃, adjusting pH to 6.0, adding 0.5% mixed enzyme (the mass ratio of cellulase to pectase to papain=2:1:1), and carrying out enzymolysis for 3h.
Fermentation: and (3) sterilizing the solution after enzymolysis, cooling to 35 ℃, adding 10% of sucrose, and adding a fermentation inoculant (mass ratio, saccharomycetes: lactobacillus=1:1) with a mass ratio of 1.5%, and uniformly mixing. Sealing and fermenting at 35 ℃ for 25 days.
Filtering and canning: and (3) circulating the fermented solution through a 200-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 6
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 100 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking 1kg of gastrodia elata powder, adding 10 times of water, stirring and mixing uniformly, heating to 50 ℃, adjusting pH to 6.0, adding 1.2% of mixed enzyme (the mass ratio of cellulase to pectase to papain=2:1:1), and carrying out enzymolysis for 3h.
Fermentation: sterilizing the solution after enzymolysis, cooling to 35 ℃, adding 10% of sucrose, and adding lactobacillus plantarum with the mass ratio of 1.5% for uniform mixing. Sealing and fermenting at 35 ℃ for 25 days.
Filtering and canning: and (3) circulating the fermented solution through a 200-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 7
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 100 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking 1kg of gastrodia elata powder, adding 10 times of water, stirring and mixing uniformly, heating to 50 ℃, adjusting pH to 6.0, adding 1.2% of mixed enzyme (the mass ratio of cellulase to pectase to papain=2:1:1), and carrying out enzymolysis for 3h.
Fermentation: and (3) sterilizing the solution after enzymolysis, cooling to 35 ℃, adding 10% of sucrose, and adding a mixed microbial inoculum (the mass ratio of lactobacillus plantarum: acetic acid bacteria=1:1) with the mass ratio of 1.5%, and uniformly mixing. Sealing and fermenting at 35 ℃ for 25 days.
Filtering and canning: and (3) circulating the fermented solution through a 200-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 8
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 100 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking 1kg of gastrodia elata powder, adding 10 times of water, stirring and mixing uniformly, heating to 50 ℃, adjusting pH to 6.0, adding 1.2% mixed enzyme (the mass ratio of cellulase to pectase to papain=2:1:1), and carrying out enzymolysis for 3h.
Fermentation: and (3) sterilizing the solution after enzymolysis, cooling to 35 ℃, adding 10% of sucrose, and adding a mixed microbial inoculum (the mass ratio, lactobacillus plantarum and saccharomycetes: acetic acid bacteria=1:1:1) with the mass ratio of 1.5%, and uniformly mixing. Sealing and fermenting at 35 ℃ for 25 days.
Filtering and canning: and (3) circulating the fermented solution through a 200-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
Comparative example 9
Pretreatment of raw materials: slicing fresh rhizoma Gastrodiae, oven drying, pulverizing with pulverizer, and sieving with 100 mesh sieve to obtain rhizoma Gastrodiae powder.
Enzymolysis: taking 1kg of gastrodia elata powder, adding 10 times of water, stirring and mixing uniformly, heating to 50 ℃, adjusting pH to 6.0, adding 1.2% mixed enzyme (the mass ratio of cellulase to pectase to papain=2:1:1), and carrying out enzymolysis for 3h.
Fermentation: and (3) sterilizing the solution after enzymolysis, cooling to 35 ℃, adding 10% of sucrose, and adding 3% of mixed microbial inoculum (mass ratio, lactobacillus plantarum: saccharomycetes: acetic acid bacteria=1:1:1) for uniformly mixing. Sealing and fermenting at 35 ℃ for 25 days.
Filtering and canning: and (3) circulating the fermented solution through a 200-mesh plate frame, filtering until the solution is clear, sterilizing and canning to obtain the gastrodia elata ferment.
The free amino acid content of the gastrodia elata ferment obtained in the example 1-3 and the comparative example 1-9 and the pH value, ethanol content, gastrodin content and p-hydroxybenzyl alcohol content of the gastrodia elata ferment obtained in the example 1-3 and the comparative example 1-9 are detected. The detection method of gastrodin and p-hydroxybenzyl alcohol is according to RP-HPLC method of Chinese pharmacopoeia. The results of the detection are shown in tables 3 and 4.
Free amino acid detection:
0.5 g-1 g (accurate to 0.001 g) of the sample is weighed and placed in a 50mL volumetric flask. Adding 20mL of sulfosalicylic acid, carrying out ultrasonic treatment until the sulfosalicylic acid is fully dissolved, carrying out constant volume to a 50mL scale, fully and uniformly mixing, standing for 1h, accurately sucking 1mL of supernatant into a 25mL volumetric flask, adding sodium citrate buffer solution, carrying out constant volume to the scale, uniformly mixing, filtering to a sample injection bottle through a 0.45 mu m microporous filter membrane, and detecting by adopting an amino acid automatic analyzer.
Sensory evaluation:
randomly selecting 40 people to score according to the sensory evaluation criteria shown in table 2.
TABLE 2
Table 3 shows the free amino acid content of the gastrodia elata enzymes obtained in examples 1 to 3 and comparative examples 1 to 9.
TABLE 3 Table 3
Table 4 below shows the pH, ethanol content, total acid content, gastrodin content, p-hydroxybenzyl alcohol content and sensory evaluation scores of the gastrodia elata ferments obtained in examples 1-3 and comparative examples 1-9.
TABLE 4 detection results
The data in Table 4 shows that the pH and ethanol content of the gastrodia elata ferment product prepared by the method meet the industry standard (QB/T5323-2018). While the ethanol content in both comparative example 2 and comparative example 3 did not meet the standards. The total acid content of each group of products is compared, and the total acid content in the comparative example is higher than that in the examples, so that the taste is sour and astringent, and subsequent taste preparation is needed. The total content of gastrodine and p-hydroxybenzyl alcohol is not less than 0.25%, and the total content of the two active components is higher than that of the comparative example. And the total amount of gastrodin and p-hydroxybenzyl alcohol in the example 2 with optimal condition is highest and reaches 18.82mg/g.
Application evaluation:
The related index detection of antioxidant efficacy was performed for examples 1-3 and comparative examples 1-9 as follows:
1. DPPH radical scavenging Capacity determination
The gastrodia elata ferment samples prepared in examples 1-3 and comparative examples 1-9 are diluted to 1mg/mL by deionized water respectively to obtain a diluted solution, 1mL of the diluted solution is taken, 1mL of a DPPH-absolute ethanol solution is added into the diluted solution respectively, the diluted solution is uniformly mixed, the mixture is reacted for 30min at room temperature under the dark condition to obtain a sample solution, the light absorption value (marked as A1) of the sample solution at the wavelength of 517nm is measured, the light absorption value (marked as A0) of the sample solution is replaced by absolute ethanol in a blank group, the DPPH-absolute ethanol solution is replaced by absolute ethanol in a control group, and the light absorption value (marked as A2) of the solution is obtained after the measurement reaction. The DPPH radical scavenging rate was calculated according to the following formula, and the detection results are shown in table 5.
DPPH radical clearance (%) = [1- (A1-A2)/A0 ] ×100
2. Superoxide anion radical scavenging capability assay
The gastrodia elata ferment samples prepared in examples 1-3 and comparative examples 1-9 are diluted to 1mg/mL by deionized water respectively to obtain a diluent, 1mL of the diluent is taken, 5mL of 50mmol/L Tris-HCl buffer (pH 8.2) is added into the diluent respectively, the mixture is uniformly mixed, 0.4mL of 25mmol/L pyrogallol solution which is also preheated to 25 ℃ is added after the mixture is subjected to water bath in a constant temperature water bath kettle at 25 ℃ for 20min, the mixture is uniformly mixed and reacted for 5min, and finally 0.5mL of concentrated hydrochloric acid is added to terminate the reaction to obtain a sample liquid, and the absorbance value (recorded as A1) of the sample liquid is measured at a wavelength of 325 nm. The absorbance of the solution obtained after the reaction was measured by using distilled water instead of the sample solution (designated as A0) in the blank group and by using distilled water instead of the pyrogallol solution in the control group (designated as A2). The superoxide anion radical scavenging rate was calculated according to the following formula, and the detection results are shown in table 5.
Superoxide anion radical clearance (%) = [1- (A1-A2)/A0 ] ×100
3. Determination of the scavenging ability of hydroxyl radicals
The gastrodia elata ferment samples prepared in examples 1-3 and comparative examples 1-9 are diluted to 1mg/mL by deionized water to obtain a diluent, 1mL of the diluent is taken, 1mL of a 6mmol/L FeSO 4 solution and 1mL of a 2.4mmol/L H 2O2 solution are added into the diluent respectively, the mixture is uniformly mixed, after reaction for 10min, 1mL of a 6mmol/L salicylic acid solution is added, reaction is carried out for 30min at 37 ℃, and the absorbance of the sample liquid at the wavelength of 510nm is measured (marked as A1). The absorbance of the sample solution was measured by using distilled water instead of the sample solution in the blank group (denoted as A0), and the absorbance of the solution obtained after the reaction was measured by using distilled water instead of the H 2O2 solution in the control group (denoted as A2). The hydroxyl radical scavenging rate was calculated according to the following formula, and the detection results are shown in table 5.
Hydroxyl radical clearance (%) = [1- (A1-A2)/A0 ] ×100
4. Superoxide dismutase (SOD) activity determination
The gastrodia elata ferment samples prepared in examples 1 to 3 and comparative examples 1 to 9 were diluted to 1mg/mL with deionized water to obtain diluted samples, 1mL of the diluted samples (V) were taken, 15mL of a 0.05mol/L phosphate buffer (pH 7.8), 3mL of 130mmol/L methionine solution, 3mL of 750. Mu. Mol/L NBT solution, 3mL of 20. Mu. Mol/L riboflavin solution, 3mL of 100. Mu. Mol/L EDTA-Na 2 solution and 2mL of purified water were added thereto, and the mixture was uniformly mixed, reacted at 25℃under 4500lx sunlight for 20 minutes, and after the reaction was completed, covered with black cloth to terminate the reaction. The absorbance of the reaction solution (designated A1) was measured at 560 nm. The blank was replaced with phosphate buffer and left in the dark for detection zeroing. The control group was replaced with phosphate buffer, and the same sample was left under sunlight for 20min and covered with black cloth, and absorbance was measured (designated as A0). Superoxide dismutase activity was calculated according to the following formula, and the detection results are shown in table 5.
Superoxide dismutase enzyme Activity (U/mL) = (A0-A1)/(0.5XA0 XV)
TABLE 5
The data in table 5 above demonstrates that the overall level of the characteristic index of the 4 oxidation resistance properties is superior to the comparative example, and that the oxidation resistance effect of example 2 is most pronounced. Proved that the gastrodia elata ferment prepared by the invention has an antioxidation function.
Claims (67)
1. The gastrodia elata fermentation product is characterized by comprising, by mass, 0.62-0.68% of gastrodin, 4.5-5.5% of glutamic acid, 2.9-3.5% of alanine and 3.1-4% of histidine;
wherein, the fermentation product is prepared by the following preparation method: mixing rhizoma Gastrodiae powder with water to obtain mixed solution, adding enzyme preparation for enzymolysis, and fermenting for 20-25 days to obtain the fermentation product; the enzyme preparation is cellulase, pectase and papain with the mass ratio of 1-2:1-2:1;
Wherein the microbial inoculum comprises saccharomycetes and lactobacillus, and the saccharomycetes is one or two of high-temperature-resistant saccharomyces cerevisiae and kluyveromyces KW-6; the lactobacillus is one or more of Lactobacillus acidophilus, lactobacillus plantarum and Bifidobacterium.
2. The gastrodia elata fermentation product according to claim 1, wherein the fermentation product comprises, by mass, 0.67-0.68% of gastrodin, 5.1-5.2% of glutamic acid, 3.4-3.5% of alanine and 3.9-4% of histidine.
3. The gastrodia elata fermentation product according to claim 1, wherein the ethanol content of the fermentation product is 0.25-0.5wt%.
4. The gastrodia elata fermentation product according to claim 2, characterized in that the ethanol content of the fermentation product is 0.25-0.5wt%.
5. The gastrodia elata fermentation product according to claim 1, wherein the total acid content in the fermentation product is 2.2-2.7% in terms of lactic acid in terms of mass percent.
6. The gastrodia elata fermentation product according to claim 2, wherein the total acid content in the fermentation product is 2.2-2.7% in terms of lactic acid in terms of mass percent.
7. A gastrodia elata fermentation product according to claim 3, characterized in that the total acid content in the fermentation product is 2.2-2.7% in terms of lactic acid in mass percent.
8. The gastrodia elata fermentation product according to claim 4, wherein the total acid content in the fermentation product is 2.2-2.7% in terms of lactic acid in terms of mass percent.
9. The gastrodia elata fermentation product according to any one of claims 1-8, wherein the pH of the fermentation product is 3.6-4.0.
10. The gastrodia elata fermentation product according to any one of claims 1-8, wherein the p-hydroxybenzyl alcohol content of the fermentation product is 1.12-1.21wt% in terms of mass percent.
11. The gastrodia elata fermentation product according to claim 9, wherein the p-hydroxybenzene methanol content of the fermentation product is 1.12-1.21wt% in terms of mass percent.
12. The gastrodia elata fermentation product according to any one of claims 1-8, wherein the fermentation product further comprises, in mass percent, 0.27-0.3% threonine, 0.5-0.65% serine, 0.7-1.2% glycine, 2.9-3.5% alanine, 3.1-4% histidine, 0.8-1.2% aspartic acid, 4.5-5.5% glutamic acid, 0.5-1.2% lysine, 0.6-0.8% tyrosine and 0.7-1.2% phenylalanine.
13. The gastrodia elata fermentation product according to claim 9, wherein the fermentation product further comprises, in mass percent, 0.27-0.3% of threonine, 0.5-0.65% of serine, 0.7-1.2% of glycine, 2.9-3.5% of alanine, 3.1-4% of histidine, 0.8-1.2% of aspartic acid, 4.5-5.5% of glutamic acid, 0.5-1.2% of lysine, 0.6-0.8% of tyrosine and 0.7-1.2% of phenylalanine.
14. The gastrodia elata fermentation product according to claim 10, wherein the fermentation product further comprises, in mass percent, 0.27-0.3% of threonine, 0.5-0.65% of serine, 0.7-1.2% of glycine, 2.9-3.5% of alanine, 3.1-4% of histidine, 0.8-1.2% of aspartic acid, 4.5-5.5% of glutamic acid, 0.5-1.2% of lysine, 0.6-0.8% of tyrosine and 0.7-1.2% of phenylalanine.
15. The gastrodia elata fermentation product according to claim 11, wherein the fermentation product further comprises, in mass percent, 0.27-0.3% of threonine, 0.5-0.65% of serine, 0.7-1.2% of glycine, 2.9-3.5% of alanine, 3.1-4% of histidine, 0.8-1.2% of aspartic acid, 4.5-5.5% of glutamic acid, 0.5-1.2% of lysine, 0.6-0.8% of tyrosine and 0.7-1.2% of phenylalanine.
16. The method for preparing a gastrodia elata fermentation product according to any one of claims 1-15, characterized in that the method comprises the steps of:
Mixing rhizoma Gastrodiae powder with water to obtain mixed solution, adding enzyme preparation for enzymolysis, and fermenting for 20-25 days to obtain the fermentation product;
The enzyme preparation is cellulase, pectase and papain with the mass ratio of 1-2:1-2:1;
Wherein the microbial inoculum comprises saccharomycetes and lactobacillus, and the saccharomycetes is one or two of high-temperature-resistant saccharomyces cerevisiae and kluyveromyces KW-6; the lactobacillus is one or more of Lactobacillus acidophilus, lactobacillus plantarum and Bifidobacterium.
17. The method according to claim 16, wherein the mass ratio of the gastrodia elata powder to the water is 1:5-10.
18. The method of claim 16, wherein the enzymatic hydrolysis reaction temperature is 50 ℃ to 55 ℃.
19. The method of claim 17, wherein the enzymatic hydrolysis reaction temperature is 50 ℃ to 55 ℃.
20. The method of claim 16, wherein the pH of the enzymatic reaction is between 5.5 and 6.5.
21. The method of claim 17, wherein the pH of the enzymatic hydrolysis reaction is between 5.5 and 6.5.
22. The method of claim 18, wherein the enzymatic hydrolysis reaction has a pH of 5.5 to 6.5.
23. The process of claim 19, wherein the enzymatic hydrolysis reaction has a pH of 5.5 to 6.5.
24. The method of claim 20, wherein the enzymatic hydrolysis reaction time is 3-5 hours.
25. The method of claim 21, wherein the enzymatic hydrolysis reaction time is 3-5 hours.
26. The method of claim 22, wherein the enzymatic hydrolysis reaction time is 3-5 hours.
27. The method of claim 23, wherein the enzymatic hydrolysis reaction time is 3-5 hours.
28. The preparation method according to claim 16, wherein the mass of the added enzyme preparation is 1% -1.5% of the mass of the mixed liquor.
29. The preparation method according to claim 17, wherein the mass of the added enzyme preparation is 1% -1.5% of the mass of the mixed liquor.
30. The preparation method according to claim 18, wherein the mass of the added enzyme preparation is 1% -1.5% of the mass of the mixed liquor.
31. The preparation method according to claim 20, wherein the mass of the added enzyme preparation is 1% -1.5% of the mass of the mixed liquor.
32. The method according to claim 16, wherein the enzyme preparation comprises cellulase, pectase and papain in a mass ratio of 2:1:1.
33. The method according to claim 17, wherein the enzyme preparation comprises cellulase, pectase and papain in a mass ratio of 2:1:1.
34. The method of claim 18, wherein the enzyme preparation comprises cellulase, pectase and papain in a mass ratio of 2:1:1.
35. The method according to claim 20, wherein the enzyme preparation comprises cellulase, pectase and papain in a mass ratio of 2:1:1.
36. The method of claim 28, wherein the enzyme preparation comprises cellulase, pectase and papain in a mass ratio of 2:1:1.
37. The method of claim 16, wherein the solution after enzymatic hydrolysis is sterilized and then added with a microbial inoculum.
38. The method of claim 17, wherein the solution after enzymatic hydrolysis is sterilized and then added with a microbial inoculum.
39. The method of claim 18, wherein the solution after enzymatic hydrolysis is sterilized and then added with a microbial inoculum.
40. The method of claim 20, wherein the solution after enzymatic hydrolysis is sterilized and then added with a microbial inoculum.
41. The method of claim 28, wherein the solution after enzymatic hydrolysis is sterilized and then added with a microbial inoculum.
42. The method of claim 32, wherein the solution after enzymatic hydrolysis is sterilized and then added with a microbial inoculum.
43. The method of claim 16, further comprising adding a carbon source prior to adding the microbial inoculum.
44. The method of claim 17, further comprising adding a carbon source prior to adding the microbial inoculum.
45. The method of claim 18, further comprising adding a carbon source prior to adding the microbial inoculum.
46. The method of claim 20, further comprising adding a carbon source prior to adding the microbial inoculum.
47. The method of claim 28, further comprising adding a carbon source prior to adding the microbial inoculum.
48. The method of claim 32, further comprising adding a carbon source prior to adding the microbial inoculum.
49. The method of claim 37, further comprising adding a carbon source prior to adding the microbial inoculum.
50. The process according to claim 43, wherein the mass of the carbon source is 10 to 15% of the mass of the mixed solution.
51. The method of claim 44, wherein the carbon source is added in an amount of 10-15% by mass of the mixture.
52. The process of claim 45, wherein the carbon source is added in an amount of 10 to 15% by mass of the mixture.
53. The process of claim 46 wherein the carbon source is added in an amount of 10% to 15% by mass of the mixture.
54. The method of claim 47, wherein the carbon source is added in an amount of 10-15% by mass of the mixture.
55. The process of claim 48 wherein the carbon source is added in an amount of 10% to 15% by mass of the mixture.
56. The process of claim 49, wherein the carbon source is added in an amount of 10 to 15% by mass of the mixture.
57. The method of claim 43, wherein the carbon source is sucrose.
58. The method of any one of claims 16 to 57, wherein the microbial inoculum is added in an amount of 1% -2% of the mass of the mixed liquor.
59. The process of any one of claims 16 to 57 wherein the fermentation temperature is from 35 ℃ to 40 ℃.
60. The process of any one of claims 16 to 57, wherein the fermentation is anaerobic fermentation.
61. The method according to any one of claims 16 to 57, wherein the mass ratio of the fermentation tubes to the lactobacillus is 2:3.
62. The method of claim 58, wherein the mass ratio of yeast to lactobacillus is 2:3.
63. The method of claim 59, wherein the mass ratio of yeast to Lactobacillus is 2:3.
64. The process of claim 60 wherein the mass ratio of yeast to lactobacillus is 2:3.
65. The method of any one of claims 16-57, wherein the method further comprises a filtration process.
66. The method of claim 65, wherein the fermented solution is circulated through a 150-300 mesh frame and filtered until the solution is clear.
67. Use of the fermentation product of claims 1-15 or the fermentation product of claims 16-66 in beverages, skin whitening products, cosmetics and daily chemical care.
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| CN113057320A (en) * | 2021-04-06 | 2021-07-02 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Gastrodia elata enzyme with effects of aiding sleep and improving intelligence and preparation process and application thereof |
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| CN113057320A (en) * | 2021-04-06 | 2021-07-02 | 贵州省中国科学院天然产物化学重点实验室(贵州医科大学天然产物化学重点实验室) | Gastrodia elata enzyme with effects of aiding sleep and improving intelligence and preparation process and application thereof |
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