CN117256843A

CN117256843A - Gastrodia elata ferment and preparation method thereof

Info

Publication number: CN117256843A
Application number: CN202311349010.1A
Authority: CN
Inventors: 和劲松; 高晴; 毕保良; 吕霞
Original assignee: Yunnan Agricultural University
Current assignee: Yunnan Agricultural University
Priority date: 2023-10-18
Filing date: 2023-10-18
Publication date: 2023-12-22

Abstract

The invention relates to the technical field of gastrodia elata ferment preparation. The invention provides a gastrodia elata ferment and a preparation method thereof, comprising the following steps: (1) Pulverizing rhizoma Gastrodiae with water, and homogenizing to obtain rhizoma Gastrodiae homogenate; (2) Adding enzyme into rhizoma Gastrodiae homogenate for enzymolysis to obtain rhizoma Gastrodiae enzymolysis solution; (3) Sterilizing the enzymolysis liquid of gastrodia elata, inoculating lactobacillus plantarum, adding sucrose, and fermenting to obtain fermentation liquid; (4) And inoculating saccharomycetes into the fermentation liquor, and continuing fermenting after adding sucrose to obtain the gastrodia elata ferment. The invention develops an enzyme product taking gastrodia elata as a raw material through researching a saccharification process of gastrodia elata homogenate, a process of fermenting gastrodia elata by lactobacillus plantarum, a process of fermenting gastrodia elata by saccharomycetes, a process of sensory evaluation and nutritional ingredient content of gastrodia elata ferment prepared by single-strain fermentation and a process of fermenting gastrodia elata by lactobacillus plantarum and saccharomycetes, provides a new idea for new application of gastrodia elata in food, and provides a certain technical basis for industrial production of gastrodia elata ferment.

Description

Gastrodia elata ferment and preparation method thereof

Technical Field

The invention relates to the technical field of gastrodia elata ferment preparation, in particular to gastrodia elata ferment and a preparation method thereof.

Background

The main chemical components of the underground tuber of gastrodia tuber comprise gastrodin, phenols, various amino acids, vitamin A, polysaccharide, protein, organic acid, various microelements such as iron, manganese, tin, chromium, copper and the like, and the active components of the gastrodia tuber have very wide functions. For example, enhancing learning memory, protecting nerves, and protecting cardiovascular and cerebrovascular vessels. In clinical medicine, can be used for treating infantile convulsion, partial numbness of limbs and relieving headache. The gastrodia polysaccharide has wide application range, such as resisting radiation, slowing aging, enhancing immunity, treating hypertension and inflammation. The gastrodia elata is a medicinal and edible substance with higher phenolic substance content, for example, gastrodin, also called p-hydroxybenzyl alcohol or 4-hydroxybenzyl alcohol, can play a role in protecting nerves, and the probability of occurrence of cerebral infarction is reduced.

The application forms of the gastrodia elata and related products are greatly expanded since the gastrodia elata is used as a medicine and food homologous substance for trial production, but the gastrodia elata is difficult to store for a long time after being picked, and mildewing and rotting can occur after the storage time exceeds 3-5 days, so that the gastrodia elata is mainly used as a medicine after being dried at present, and the application of the gastrodia elata is limited. How to increase the processing mode and change the rough processing technology of the food, and developing a novel product suitable for food application is a current problem to be solved urgently.

Disclosure of Invention

The invention aims to provide a gastrodia elata ferment and a preparation method thereof, and develop a novel product which is suitable for food application.

In order to achieve the above object, the present invention provides the following technical solutions:

the invention provides a preparation method of gastrodia elata ferment, which comprises the following steps:

(1) Pulverizing rhizoma Gastrodiae with water, and homogenizing to obtain rhizoma Gastrodiae homogenate;

(2) Adding enzyme into rhizoma Gastrodiae homogenate for enzymolysis to obtain rhizoma Gastrodiae enzymolysis solution;

(3) Sterilizing the enzymolysis liquid of gastrodia elata, inoculating lactobacillus plantarum, adding sucrose, and fermenting to obtain fermentation liquid;

(4) And inoculating saccharomycetes into the fermentation liquor, and continuing fermenting after adding sucrose to obtain the gastrodia elata ferment.

Preferably, in the step (1), the gastrodia elata is cleaned and peeled, and the dosage ratio of the gastrodia elata to the water is 1kg: 2.8-3.2L;

and (3) in the step (1), a colloid mill is adopted during homogenization, the number of times of homogenization is 2-4, and the time of each time of homogenization is 2-4 min.

Preferably, in the step (2), the mass concentration of the enzyme in the gastrodia elata homogenate is 0.5-0.7%, the enzyme is alpha-amylase and saccharifying enzyme, and the enzyme activity ratio of the alpha-amylase to the saccharifying enzyme is 1:13 to 15.

Preferably, in the step (2), the pH value of the enzymolysis is 5.3-5.7, the temperature of the enzymolysis is 65-75 ℃, and the time of the enzymolysis is 55-65 min.

Preferably, the sterilization method in the step (3) is as follows: heating the enzymolysis liquid of gastrodia elata to 83-87 ℃ and preserving heat for 28-32 s.

Preferably, in the step (3), lactobacillus plantarum bacterial liquid is adopted when lactobacillus plantarum is inoculated, and the preparation method of the lactobacillus plantarum bacterial liquid comprises the following steps: culturing lactobacillus plantarum in an MRS liquid culture medium for 46-50 h, wherein the inoculating amount of lactobacillus plantarum bacterial liquid is 0.8-1.2% of the volume of the gastrodia elata enzymolysis liquid.

Preferably, the sucrose is added in the amount of 3.5-4.5% in the step (3), the fermentation temperature is 35-39 ℃, the pH value of the fermentation is 6.2-6.6, and the fermentation time is 46-50 h.

Preferably, in the step (4), a saccharomycete liquid is adopted when the saccharomycete is accessed, and the preparation method of the saccharomycete liquid is as follows: activating saccharomycete in glucose water in 1.8-2.2% at 35-39 deg.c for 25-35 min, with saccharomycete liquid being connected in 0.5-0.7% of the volume of the fermented liquid.

Preferably, the sucrose is added in the amount of 3.5-4.5% in the step (4), the fermentation temperature is 30-34 ℃, the pH value of the fermentation is 3.8-4.2, and the fermentation time is 94-98 h.

The invention also provides the gastrodia elata ferment prepared by the preparation method.

The invention provides a gastrodia elata ferment and a preparation method thereof, comprising the following steps: (1) Pulverizing rhizoma Gastrodiae with water, and homogenizing to obtain rhizoma Gastrodiae homogenate; (2) Adding enzyme into rhizoma Gastrodiae homogenate for enzymolysis to obtain rhizoma Gastrodiae enzymolysis solution; (3) Sterilizing the enzymolysis liquid of gastrodia elata, inoculating lactobacillus plantarum, adding sucrose, and fermenting to obtain fermentation liquid; (4) And inoculating saccharomycetes into the fermentation liquor, and continuing fermenting after adding sucrose to obtain the gastrodia elata ferment. The invention develops an enzyme product taking gastrodia elata as a raw material through researching a saccharification process of gastrodia elata homogenate, a process of fermenting gastrodia elata by lactobacillus plantarum, a process of fermenting gastrodia elata by saccharomycetes, a process of sensory evaluation and nutritional ingredient content of gastrodia elata ferment prepared by single-strain fermentation and a process of fermenting gastrodia elata by lactobacillus plantarum and saccharomycetes, provides a new idea for new application of gastrodia elata in food, and provides a certain technical basis for industrial production of gastrodia elata ferment.

Drawings

FIG. 1 is a standard curve of total phenol content;

FIG. 2 is a polysaccharide concentration standard curve;

FIG. 3 shows a standard curve of gastrodin content;

FIG. 4 shows the concentration of Lactobacillus plantarum in each of the enzyme solutions;

FIG. 5 shows the variation of lactic acid content in each group of enzyme solutions;

FIG. 6 shows the change in concentration of yeast in each group of enzyme solutions;

FIG. 7 shows the variation of ethanol concentration in each group of enzyme solutions;

FIG. 8 is a graph of individual groups of enzyme fluid sensory scores;

FIG. 9 is a radar chart of sensory evaluation of each group of enzyme solutions;

FIG. 10 shows the total phenol content of each group of enzyme solutions;

FIG. 11 shows polysaccharide content of each group of enzyme solutions;

fig. 12 shows the gastrodin content of each group of enzyme solutions.

Detailed Description

In the present invention, the gastrodia elata in step (1) is preferably a washed peeled gastrodia elata.

In the invention, the dosage ratio of the gastrodia elata to the water is preferably 1kg:2.8 to 3.2L, more preferably 1kg:3L.

In the present invention, a colloid mill is preferably used for the homogenization in the step (1), and the number of homogenization is preferably 2 to 4, more preferably 3, and the time for each homogenization is preferably 2 to 4min, more preferably 3min.

In the present invention, the mass concentration of the enzyme in the gastrodia elata homogenate in the step (2) is preferably 0.5-0.7%, more preferably 0.6%, the enzyme is preferably an α -amylase and a saccharifying enzyme, and the enzyme activity ratio of the α -amylase to the saccharifying enzyme is preferably 1:13 to 15, more preferably 1:14.

in the present invention, the pH value of the enzymolysis in the step (2) is preferably 5.3 to 5.7, more preferably 5.5, the temperature of the enzymolysis is preferably 65 to 75 ℃, more preferably 70 ℃, and the time of the enzymolysis is preferably 55 to 65min, more preferably 60min.

In the present invention, the sterilization method in step (3) is preferably: heating the gastrodia elata enzymatic hydrolysate to 83-87 ℃ and preserving heat for 28-32 s, and further preferably heating the gastrodia elata enzymatic hydrolysate to 85 ℃ and preserving heat for 30s.

In the present invention, the lactobacillus plantarum bacterium solution is preferably used in the step (3) of inoculating lactobacillus plantarum, and the preparation method of the lactobacillus plantarum bacterium solution preferably includes: the lactobacillus plantarum is cultivated in an MRS liquid culture medium for 46-50 h, more preferably 48h, and the inoculating amount of the lactobacillus plantarum bacterial liquid is preferably 0.8-1.2% of the volume of the gastrodia elata enzymolysis liquid, more preferably 1%.

In the present invention, the sucrose is preferably added in an amount of 3.5 to 4.5%, more preferably 4%, the fermentation temperature is preferably 35 to 39 ℃, more preferably 37 ℃, the fermentation pH is preferably 6.2 to 6.6, more preferably 6.4, and the fermentation time is preferably 46 to 50 hours, more preferably 48 hours.

In the present invention, the step (4) is preferably performed by using a yeast liquid, and the preparation method of the yeast liquid is preferably: the saccharomycete is activated for 25-35 min under the conditions of 1.8-2.2% of glucose water and 35-39 ℃, more preferably the saccharomycete is activated for 30min under the conditions of 2% of glucose water and 37 ℃, and the access amount of the saccharomycete liquid is preferably 0.5-0.7% of the volume of the fermentation liquid, more preferably 0.6%.

In the present invention, the sucrose is preferably added in an amount of 3.5 to 4.5%, more preferably 4%, the fermentation temperature is preferably 30 to 34 ℃, more preferably 32 ℃, the fermentation pH is preferably 3.8 to 4.2, more preferably 4.0, and the fermentation time is preferably 94 to 98 hours, more preferably 96 hours.

The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.

Raw materials and instruments

Gastrodia elata Blume is purchased from Yi-Liang county small grass dam of Zhaotong, yunnan province and stored at 4deg.C for use;

yeast: the Wang Jiaomu fungus powder of the white wine is purchased from Angel Yeast company;

lactobacillus plantarum: lactobacillus plantarum (TR 22) powder was purchased from Sesamum indicum biological Co.

Example 1

A preparation method of gastrodia elata ferment comprises the following steps:

(1) Cleaning and peeling the tuber of Sichuan black-bone gastrodia of Yunnan Zhaotong Xiaoyouba according to 1kg:3.2L of dosage ratio is crushed by adding water, and then homogenized by a colloid mill for 2 times for 4min each time to obtain rhizoma gastrodiae homogenate;

(2) Adding alpha-amylase and saccharifying enzyme (the enzyme activity ratio of the alpha-amylase to the saccharifying enzyme is 1:15) into rhizoma Gastrodiae homogenate to obtain rhizoma Gastrodiae homogenate with the enzyme mass concentration of 0.5%, and performing enzymolysis at pH=5.7 at 75deg.C for 55min to obtain rhizoma Gastrodiae enzymolysis solution;

(3) Culturing lactobacillus plantarum in an MRS liquid culture medium for 46 hours to obtain lactobacillus plantarum bacterial liquid;

(4) Heating rhizoma Gastrodiae enzymolysis solution to 87deg.C, maintaining the temperature for 28s for sterilization, inoculating lactobacillus plantarum bacterial solution accounting for 0.8% of the volume of rhizoma Gastrodiae enzymolysis solution, adding sucrose accounting for 4.5%, and fermenting at 39deg.C for 50h to obtain fermentation liquor;

(5) Activating yeast in 2.2% glucose water at 35deg.C for 25min to obtain yeast liquid;

(6) And inoculating saccharomycete liquid accounting for 0.7% of the volume of the fermentation liquor into the fermentation liquor, adding 4.5% of sucrose, and continuing fermenting for 98 hours at the pH value of=4.2 and the temperature of 30 ℃ to obtain the gastrodia elata ferment.

Example 2

A preparation method of gastrodia elata ferment comprises the following steps:

(1) Cleaning and peeling the tuber of Sichuan black-bone gastrodia of Yunnan Zhaotong Xiaoyouba according to 1kg:2.8L of dosage ratio is crushed by adding water, and then homogenized by a colloid mill for 4 times for 2min each time to obtain rhizoma gastrodiae homogenate;

(2) Adding alpha-amylase and saccharifying enzyme (the enzyme activity ratio of the alpha-amylase to the saccharifying enzyme is 1:13) into the rhizoma gastrodiae homogenate to obtain rhizoma gastrodiae homogenate with the enzyme mass concentration of 0.7%, and carrying out enzymolysis for 65min at the pH value of=5.3 and the temperature of 65-75 ℃ to obtain rhizoma gastrodiae enzymolysis liquid;

(3) Culturing lactobacillus plantarum in an MRS liquid culture medium for 50 hours to obtain lactobacillus plantarum bacterial liquid;

(4) Heating rhizoma Gastrodiae enzymolysis solution to 83 deg.C, maintaining the temperature for 32s for sterilization, inoculating lactobacillus plantarum bacterial solution accounting for 1.2% of the volume of the rhizoma Gastrodiae enzymolysis solution, adding sucrose accounting for 3.5%, and fermenting at 35 deg.C for 46h to obtain fermentation liquor;

(5) Activating saccharomycete in 1.8% glucose water at 39 deg.c for 35min to obtain saccharomycete liquid;

(6) And inoculating saccharomycete liquid accounting for 0.5% of the volume of the fermentation liquor into the fermentation liquor, adding 3.5% of sucrose, and continuing fermenting for 94 hours at the pH value of = 3.8 and the temperature of 34 ℃ to obtain the gastrodia elata ferment.

Example 3

A preparation method of gastrodia elata ferment comprises the following steps:

(1) Cleaning and peeling the tuber of Sichuan black-bone gastrodia of Yunnan Zhaotong Xiaoyouba according to 1kg:3L of dosage ratio is crushed by adding water, and homogenized by a colloid mill for 3 times and 3min each time to obtain rhizoma Gastrodiae homogenate;

(2) Adding alpha-amylase and saccharifying enzyme (the enzyme activity ratio of the alpha-amylase to the saccharifying enzyme is 1:14) into rhizoma Gastrodiae homogenate to obtain rhizoma Gastrodiae homogenate with the enzyme mass concentration of 0.6%, and performing enzymolysis at 70 deg.C for 60min at pH value=5.5 to obtain rhizoma Gastrodiae enzymolysis solution;

(3) Culturing lactobacillus plantarum in an MRS liquid culture medium for 48 hours to obtain lactobacillus plantarum bacterial liquid;

(4) Heating rhizoma Gastrodiae enzymolysis solution to 85deg.C, maintaining the temperature for 30s for sterilization, inoculating lactobacillus plantarum bacterial solution accounting for 1% of the volume of rhizoma Gastrodiae enzymolysis solution, adding sucrose accounting for 4%, and fermenting at 37deg.C for 48 hr to obtain fermentation liquor;

(5) Activating saccharomycete in 2% glucose water at 37 deg.c for 30min to obtain saccharomycete liquid;

(6) And (3) inoculating saccharomycete liquid accounting for 0.6% of the volume of the fermentation liquid into the fermentation liquid, adding 4% of sucrose, and continuing fermenting for 96 hours at the pH value of = 4 and the temperature of 32 ℃ to obtain the gastrodia elata ferment.

Test examples

1. Sensory evaluation and nutritional ingredient analysis in gastrodia elata fermentation process

(1) Gastrodia elata ferment sensory evaluation

Sensory evaluation can be used for expressing the quality of ferment products to a great extent, is an important index of ferment type products, and establishes a proper sensory scoring standard of gastrodia elata ferment for the experiment, and is shown in table 1.

TABLE 1 Gastrodia elata ferment sensory evaluation criteria

(2) Determination of total phenol content

And measuring the total phenol content in the gastrodia elata ferment by adopting Fu Lin Fenfa. First, a gallic acid standard solution of 0.01mg/mL was prepared and diluted as shown in Table 2.

TABLE 2 gallic acid standard solution

Test tube number	1	2	3	4	5	6	7
								Gallic acid standard solution (mL)	1	2	3	4	5	6	7
Ultrapure water (mL)	6	5	4	3	2	1	0

To each tube was added 1mL of Fu Lin Fen reagent (1N) and 2mL of 15% sodium carbonate solution, respectively. The reaction was conducted in the dark for 2 hours, and the absorbance was measured at 760nm with 7mL of pure water as a reference solution instead of the standard solution, to prepare a total phenol content standard curve (FIG. 1).

And (3) centrifuging the gastrodia elata ferment liquid at 4000rmp for 5min, taking supernatant to dilute according to experimental requirements, repeating the steps, and calculating the total phenol content according to a standard curve.

(3) Determination of polysaccharide content

The content of crude polysaccharide in the sample was determined using the phenol sulfuric acid method. Glucose standard solutions of 0.1mg/mL were accurately prepared and diluted as in Table 3.

TABLE 3 glucose standard solution

The glucose standard solution is diluted according to the above table, 5% phenol solution and 5mL concentrated sulfuric acid are added in sequence, the mixture is stood for 10min, the boiling water bath is carried out for 15min, and the mixture is quickly placed into ice water prepared in advance for cooling after the boiling water bath is finished. The absorbance of the treated sample was measured at 490nm and a standard curve was drawn (FIG. 2).

Accurately transferring 4mL of ferment sample into a centrifuge tube, and using 1:7 (V: V) the feed liquid ratio is added with ultrapure water, and ultrasonic extraction is carried out for 30min after uniform mixing. Taking 5mL of supernatant after centrifugation, adding 20mL of ethanol solution with concentration of 80% (V/V), standing for 12h, centrifuging at 8000rpm for 10min, discarding supernatant, re-dissolving the precipitate with 4mL of pure water, properly diluting the obtained crude polysaccharide extract to replace the standard solution, repeating the steps, and calculating the crude polysaccharide content in the sample according to the standard curve.

(4) Determination of Gastrodin content

And (3) referring to the Chinese pharmacopoeia method, and measuring the gastrodin content in the sample by adopting a High Performance Liquid Chromatography (HPLC) method.

(1) Chromatographic column conditions: syncronis C18 (250X 46mm X5. Mu.g); column temperature is 30 ℃; mobile phase: acetonitrile: 0.05% water = 3:97 (V: V); sample injection amount: 10. Mu.L; sample injection mode: constant current; flow rate: 1.000mL/min; wavelength: 220nm.

(2) Establishment of a standard curve: preparing 0.2mg/mL gastrodin standard solution, respectively taking 1, 2, 3, 4 and 5mL, fixing volume into 5mL volumetric flask with acetonitrile water of 3:97 (acetonitrile: water), and filtering with 0.22 μm filter membrane to be detected. And taking the gastrodin mass as an abscissa and the peak area as an ordinate to make a standard curve. The standard curve is shown in fig. 3.

(3) Determination of gastrodin content in sample

Accurately transferring 5mL of fermentation liquor into a 50mL centrifuge tube, adding 3.5mL of absolute ethyl alcohol, sealing, performing ultrasonic extraction at 60 ℃ for 40min, centrifuging the extract for 5min at 3000r/min, concentrating the supernatant at 70 ℃ to dryness, washing the supernatant into a 5mL volumetric flask with acetonitrile water (acetonitrile: water=3:97), filtering the liquid with a fixed volume by a 0.22 μm filter membrane, injecting the liquid into a sample bottle for testing, and calculating the gastrodin content in the sample according to a standard curve after testing.

2. Lactobacillus plantarum and saccharomycetes compound fermentation gastrodia elata ferment

The test selects to insert saccharomycetes for segmented fermentation respectively at 24 hours, 48 hours, 72 hours, 96 hours and 120 hours of lactobacillus plantarum fermentation. Under the optimal technological conditions determined by technological research of fermenting gastrodia elata with single lactobacillus plantarum and saccharomycete, namely under the conditions that the pH is 6.5, the temperature is 37 ℃, and 6 percent of sucrose is added, 1 percent (V/V) lactobacillus plantarum is inoculated. After lactobacillus plantarum fermentation reaches a set time, the fermentation temperature is adjusted to 32 ℃, the pH value is 4.0, the sucrose addition amount is 4%, the inoculation amount is 0.6%, the fermentation substrate is ensured to be fully dissolved with oxygen, the saccharomycetes are inoculated for fermentation, and the total fermentation time is set to 168 hours. And determining the optimal time for inoculating the lactobacillus plantarum through sensory evaluation and determination analysis of the total phenol content, the polysaccharide content and the gastrodin content in the fermentation liquor.

Results:

(1) The influence of the yeast on the lactobacillus plantarum fermentation is shown in fig. 4 and 5 when lactobacillus plantarum fermentation is carried out, and as can be seen from fig. 4, the influence on the lactobacillus plantarum concentration is small no matter how long the lactobacillus plantarum fermentation is carried out after the yeast is inoculated for two-stage fermentation in all the compound fermentation groups. At the end of fermentation, the concentration of Lactobacillus plantarum in the experimental group of Lactobacillus plantarum fermentation 24h for inoculating yeast is the lowest, and is 8.87lg (CFU/mL), and the concentration of Lactobacillus plantarum in the experimental group of Lactobacillus plantarum fermentation 96h for inoculating yeast is the highest, and is 8.99lg (CFU/mL). As can be seen from fig. 5, the lactic acid content in the gastrodia elata ferment is obviously reduced when the gastrodia elata ferment is inoculated with saccharomycetes, which is caused by the following steps: the pH value at the end of the lactobacillus plantarum fermentation is lower than 3.0, and in order to make the saccharomycetes function better, sodium bicarbonate is used for adjusting the pH value of the fermentation substrate to 4.0 in the experiment. Thus, sodium bicarbonate reacts with some of the lactic acid present in the enzyme solution. And as can be seen from the figure, the lactic acid content in the ferment liquid is hardly changed after the yeast is inoculated for fermentation. At the end of fermentation, the group with the highest lactic acid content is the experimental group inoculated with saccharomycetes by lactobacillus plantarum fermentation for 72 hours, and the maximum value is 2.84mg/mL. From the above results, it can be found that: after the yeast is inoculated, metabolic products such as ethanol produced by the yeast do not kill lactobacillus plantarum in the ferment liquid, but inhibit the metabolic activity of the lactobacillus plantarum.

(2) The influence of the indirect saccharomycetes on the saccharomycetes fermentation at different times of the lactobacillus plantarum fermentation is shown in fig. 6 and 7, and as can be seen from fig. 6, the saccharomycetes concentration in the ferment liquid shows a trend of rising and then falling after the saccharomycetes are inoculated for fermentation in all experimental groups, and when the fermentation is finished, the experimental group with the highest saccharomycetes concentration is the experimental group with 48h of inoculated saccharomycetes for fermentation, and the maximum value is 6.30lg (CFU/mL); as can be seen from fig. 7, the ethanol content of all experimental groups showed a tendency of fast rise followed by slow rise, and when the fermentation was completed, the ethanol content of the experimental group, in which lactobacillus plantarum was fermented for 48 hours and inoculated with yeast, was the highest, and was 0.962%. According to the results, the yeast can grow and propagate more vigorously in a certain time during the composite fermentation, and then metabolism is inhibited.

(3) As shown in FIG. 8 and FIG. 9, the effect of yeast on the sensory evaluation of the gastrodia elata ferment is shown in FIG. 8, and it is clear from FIG. 8 that the sensory evaluation of the gastrodia elata ferment by using the lactobacillus plantarum and the yeast together as a starter is 63 minutes at the maximum value of the sample of the lactobacillus plantarum fermented for 48 hours and is larger than that of the gastrodia elata ferment prepared by using all lactobacillus plantarum or a single strain of the yeast as a starter. As can be seen from fig. 9, the gastrodia elata ferment prepared by inoculating the saccharomycetes at 24 hours has low flavor score although the color, smell and taste reach the maximum value; the taste and flavor score of the sample fermented by the access saccharomycetes is lower at 72 hours; the taste score of the sample fermented by the access saccharomycetes is lower at 96 hours; the sensory scores of the samples accessed into the saccharomycetes fermentation for 120 hours are lower in all the sensory evaluation indexes; after lactobacillus plantarum fermentation for 48 hours, inoculating a yeast fermentation sample, and obtaining a higher score in each index of sensory evaluation.

(4) The influence of the indirect saccharomycetes on the nutritional ingredients in the gastrodia elata ferment liquid at different times of lactobacillus plantarum fermentation is shown in fig. 10-12, and fig. 10 shows that the total phenol content in all the compound bacterial ferment liquid is higher than that before fermentation, and when lactobacillus plantarum is inoculated after 48h of fermentation, the total phenol content in the fermentation liquid is up to 1.525mg/mL and higher than 118.481% before fermentation. As shown in FIG. 11, the polysaccharide concentration in the enzyme solution was reduced to a different degree after fermentation than before fermentation due to the metabolism of microorganisms, and was continuously reduced with the delay of the yeast access time. Wherein the polysaccharide content in the gastrodia elata ferment fermented by the access saccharomycetes is the lowest when lactobacillus plantarum is fermented for 120 hours, and the value is 21.123mg/100mL. In the fermentation sample, the content of the gastrodia elata ferment polysaccharide fermented by inoculating saccharomycetes is highest when lactobacillus plantarum is fermented for 24 hours, and the maximum value is 78.408mg/100mL. As can be seen from FIG. 12, after the composite fermentation of Lactobacillus plantarum and yeast, the gastrodin content of all experimental groups was higher than that in the pre-fermentation Gastrodia elata homogenate, wherein the maximum gastrodin content of the sample inoculated with the yeast fermentation at 96h of Lactobacillus plantarum inoculation was 6.5902 μg/mL. The phenomenon reflects that the gastrodia elata ferment obtained by sectionally fermenting lactobacillus plantarum and saccharomycetes can effectively retain gastrodin leached from gastrodia elata into fermentation liquid.

Conclusion(s)

The method is characterized in that the lactobacillus plantarum and saccharomycetes are subjected to composite fermentation, the optimal inoculation amount, fermentation temperature, initial pH and sucrose addition amount of the lactobacillus plantarum and saccharomycetes are selected, and the lactobacillus plantarum concentration, lactic acid content, saccharomycetes concentration and ethanol content in the gastrodia elata ferment liquid are measured and analyzed for different saccharomycetes inoculation time, so that when the gastrodia elata is subjected to composite fermentation by the lactobacillus plantarum and saccharomycetes, the two bacteria have the phenomenon of inhibiting growth metabolism mutually, and further nutrient substances such as phenolic substances, polysaccharide and gastrodin which are leached from the gastrodia elata to the ferment liquid are better reserved. The optimal inoculation time of the saccharomycetes is determined to be 48 hours of lactobacillus plantarum fermentation by analyzing sensory scores in lactobacillus plantarum and saccharomycetes fermentation processes and measuring and comparing total phenol, gastrodin and polysaccharide contents. The total sensory evaluation score and each nutrition index of the gastrodia elata ferment fermentation liquor prepared under the conditions are higher than the highest value in the fermentation process of two single strains, wherein the total sensory evaluation score is 63 minutes, the total phenol content is 1.525mg/mL, the polysaccharide content is 65.387mg/100mL, and the gastrodin content is 6.1031 mug/mL.

In summary, the saccharified gastrodia elata homogenate is pasteurized, 4% of sucrose is added at pH 6.4, 1% of lactobacillus plantarum is inoculated at 37 ℃, and fermentation is carried out for 48 hours. The pH of the fermentation substrate is regulated to 4.0, 4 percent of sucrose is added, and under the condition of ensuring sufficient dissolved oxygen, 0.6 percent of saccharomyces cerevisiae is inoculated and fermented for 120 hours at 32 ℃. The gastrodia elata ferment prepared under the condition has higher sensory scores compared with the gastrodia elata ferment prepared by fermenting a single strain of saccharomycetes or lactobacillus plantarum; the total phenol, polysaccharide and gastrodin content in the ferment liquid is more balanced, and the total phenol and gastrodin content is increased compared with that before fermentation.

As can be seen from the above embodiments, the present invention provides a gastrodia elata ferment and a preparation method thereof, comprising the following steps: (1) Pulverizing rhizoma Gastrodiae with water, and homogenizing to obtain rhizoma Gastrodiae homogenate; (2) Adding enzyme into rhizoma Gastrodiae homogenate for enzymolysis to obtain rhizoma Gastrodiae enzymolysis solution; (3) Sterilizing the enzymolysis liquid of gastrodia elata, inoculating lactobacillus plantarum, adding sucrose, and fermenting to obtain fermentation liquid; (4) And inoculating saccharomycetes into the fermentation liquor, and continuing fermenting after adding sucrose to obtain the gastrodia elata ferment. The invention develops an enzyme product taking gastrodia elata as a raw material through researching a saccharification process of gastrodia elata homogenate, a process of fermenting gastrodia elata by lactobacillus plantarum, a process of fermenting gastrodia elata by saccharomycetes, a process of sensory evaluation and nutritional ingredient content of gastrodia elata ferment prepared by single-strain fermentation and a process of fermenting gastrodia elata by lactobacillus plantarum and saccharomycetes, provides a new idea for new application of gastrodia elata in food, and provides a certain technical basis for industrial production of gastrodia elata ferment.

The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims

1. The preparation method of the gastrodia elata ferment is characterized by comprising the following steps of:

2. The method according to claim 1, wherein the gastrodia elata is a cleaned and peeled gastrodia elata in the step (1), and the dosage ratio of the gastrodia elata to the water is 1kg: 2.8-3.2L;

3. The preparation method according to claim 2, wherein in the step (2), the mass concentration of the enzyme in the gastrodia elata homogenate is 0.5-0.7%, the enzyme is alpha-amylase and saccharifying enzyme, and the enzyme activity ratio of the alpha-amylase to the saccharifying enzyme is 1:13 to 15.

4. The method according to claim 3, wherein the pH value of the enzymolysis in the step (2) is 5.3-5.7, the temperature of the enzymolysis is 65-75 ℃, and the time of the enzymolysis is 55-65 min.

5. The method of claim 4, wherein the sterilization in step (3) is performed by: heating the enzymolysis liquid of gastrodia elata to 83-87 ℃ and preserving heat for 28-32 s.

6. The preparation method of claim 5, wherein the lactobacillus plantarum bacterial solution is adopted in the step (3) when the lactobacillus plantarum is inoculated, and the preparation method of the lactobacillus plantarum bacterial solution comprises the following steps: culturing lactobacillus plantarum in an MRS liquid culture medium for 46-50 h, wherein the inoculating amount of lactobacillus plantarum bacterial liquid is 0.8-1.2% of the volume of the gastrodia elata enzymolysis liquid.

7. The method according to claim 6, wherein the sucrose is added in the amount of 3.5 to 4.5% in the step (3), the fermentation temperature is 35 to 39 ℃, the pH of the fermentation is 6.2 to 6.6, and the fermentation time is 46 to 50 hours.

8. The method according to claim 7, wherein the step (4) is performed by using a yeast liquid, and the method comprises the steps of: activating saccharomycete in glucose water in 1.8-2.2% at 35-39 deg.c for 25-35 min, with saccharomycete liquid being connected in 0.5-0.7% of the volume of the fermented liquid.

9. The method according to any one of claims 1 to 8, wherein the sucrose is added in the amount of 3.5 to 4.5% in the step (4), the fermentation temperature is 30 to 34 ℃, the fermentation pH is 3.8 to 4.2, and the fermentation time is 94 to 98 hours.

10. The gastrodia elata ferment prepared by the preparation method according to any one of claims 1 to 9.