CN117378719A - Microorganism screening method and fermentation process for improving bitter and astringent feeling of sophora flower water extract - Google Patents
Microorganism screening method and fermentation process for improving bitter and astringent feeling of sophora flower water extract Download PDFInfo
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- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/38—Other non-alcoholic beverages
- A23L2/382—Other non-alcoholic beverages fermented
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
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Abstract
The invention discloses a microorganism screening method for improving bitter taste of a sophora flower water extract, a fermentation process and application, wherein ten edible microorganisms are used as fermentation strains, inoculated into the sophora flower water extract, 3 strains with better effect of improving the sophora flower taste are screened out through single-strain fermentation, then the screened 3 strains are used as fermentation strains, different strain mixing ratios are designed, the optimal strain combination for improving the taste through fermentation is screened out, then the process condition for improving the taste through mixed strain fermentation is optimized by using a response surface analysis method, the optimal process condition for improving the taste through mixed strain fermentation is obtained, the bitter and astringent uncomfortable taste of the sophora flower water extract can be obviously improved, the content of flavone in the effective component of a fermentation product is increased, the product can be manufactured into food additive raw materials, the product can be better applied to the development of the sophora flower beverage, a new development path is provided for the traditional Chinese medicine for the conversion of the traditional Chinese medicine to the high-end food industry.
Description
Technical Field
The invention relates to a microorganism screening method and a fermentation process for improving bitter and astringent feeling of a sophora flower water extract, belonging to the technical field of fermentation of medicines and foods.
Background
Along with the rapid development of science and technology, fermentation engineering is also receiving more and more attention. At present, people have high definition on food and focus on color, smell and taste. The pagodatree flower is used as a traditional medicine and food dual-purpose traditional Chinese medicine, and the problems of bitter, astringent and the like are also commonly existed. The pagodatree flower fermentation is to change the traditional processing mode of pagodatree flower, and the problem of bitter taste of pagodatree flower eating is improved by applying modern microbiology and some existing fermentation technologies. In the past, the traditional Chinese medicine fermentation processing is mainly natural fermentation, and has the problem of mixed bacteria pollution.
At present, research on the fermentation and utilization of traditional Chinese medicine pagodatree flower mainly stays at the combined fermentation of pagodatree flower and various substances, such as other traditional Chinese medicine, tea and wine compositions, the combination modes are complex, the change of effective components of pagodatree flower is uncontrollable, and the influence factors on experiments are increased. The method for screening the microorganism for improving the taste of the pagodatree flower by fermentation and the application thereof are not available. Therefore, a new technical solution is needed to solve the above technical problems.
Disclosure of Invention
The invention aims to solve the technical problem of providing a microorganism screening method for improving bitter taste of a sophora flower water extract, a fermentation process and application thereof, and the microorganism screening method is used for screening out strains capable of effectively improving bitter taste of the sophora flower and improving application and resource utilization efficiency of the sophora flower.
In order to solve the technical problems, the invention adopts the following technical scheme:
a microorganism screening method and a fermentation process for improving bitter taste of a sophora flower water extract comprise the following steps:
(1) Weighing appropriate amount of flos Sophorae Immaturus, soaking in tap water for 30 min, decocting with 10 times of tap water for 1 hr, filtering with gauze, collecting filtrate, boiling the residue with 8 times of water for 1 hr, and filtering; mixing the two filtrates, concentrating to obtain flos Sophorae Immaturus water extract with a feed-liquid ratio of 1:10;
(2) Packaging the pagodatree flower water extract obtained in the step (1) into 11 triangular bottles, and sterilizing at 121 ℃ under high pressure;
(3) And (3) taking 11 bottles of sterilized sophora flower water extract in the step (2), wherein 1 bottle of triangular bottles are not inoculated with bacteria, and the rest 10 bottle of triangular bottles are respectively inoculated with the following ten edible fungi: aspergillus oryzae, aspergillus niger, saccharomyces cerevisiae, bacillus subtilis, lactobacillus pentosus, lactobacillus helveticus, lactobacillus plantarum, lactobacillus bulgaricus, lactobacillus acidophilus, streptococcus thermophilus, and the inoculation amount is 4%;
(4) Culturing 11 bottles of pagodatree flower water extract in the step (3) in a shaking incubator with the rotating speed of 200r/min and the temperature of 25 ℃ for 24 hours;
(5) Carrying out sensory scoring on the fermented sample in the step (4), and screening three strains with higher export sensory scoring and top three ranks, namely lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus respectively;
(6) Carrying out mixed fermentation on the lactobacillus helveticus, the lactobacillus bulgaricus and the streptococcus thermophilus screened in the step (5), wherein the inoculation amount is 4%, and culturing the lactobacillus helveticus, the lactobacillus bulgaricus and the streptococcus thermophilus in a shaking incubator with the rotation speed of 200r/min and the temperature of 25 ℃ for 24 hours;
(7) Screening lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus as mixed bacteria with good export feel and high content of main active ingredient total flavonoids according to the sensory score and the content change of main active ingredient total flavonoids, wherein the mixed bacteria are lactobacillus helveticus: lactobacillus bulgaricus: streptococcus thermophilus is 1:1:1;
(8) Inoculating the mixed bacteria combination of the best lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus screened in the step (7) into the aqueous extract of the pagodatree flower, and carrying out single factor investigation analysis on fermentation time, fermentation temperature and inoculation amount;
(9) Taking sensory scores and total flavone content as evaluation indexes, and determining the optimal fermentation time, the optimal fermentation temperature and the optimal inoculation amount when the single factor of the sophora flower water extract is inspected in the step (8);
(10) Designing a three-factor three-level experiment based on determining the optimal fermentation time, the optimal fermentation temperature and the optimal inoculation quantity by utilizing a Box-Benhnken test scheme of Design Expert V8.0.5 b software, and obtaining the optimal fermentation process condition of the sophora flower water extract for improving the bitter and astringent feeling by taking the sensory score and the total flavone content as response values and carrying out secondary regression equation analysis on a response surface and predicting by the Design Expert V8.0.5 b software;
(11) The optimal fermentation process conditions for improving the bitter taste of the sophora flower water extract obtained in the step (10) are verified for three times, sensory scores and total flavone content are used as evaluation indexes, the relative errors of the verified actual measurement values and the theoretical prediction values in the step (10) are compared, and the optimal fermentation process conditions for improving the bitter taste of the sophora flower water extract by multi-strain fermentation are combined with industrial production practice, wherein the optimal fermentation process conditions comprise: fermenting for 20h at 24 ℃ with 3% of inoculation amount.
The invention also relates to application of lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus in preparing pagodatree flower products.
The invention relates to application of lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus in improving bitter taste of pagodatree flowers.
The invention has the beneficial effects that: the unique flavor and the drug effect of the pagodatree flower are the Chinese medicines which are favored by consumers as both medicine and food, but the pagodatree flower water extract has certain bitter and uncomfortable taste and severely limits the development and utilization of resources. The pagodatree flower is widely used as a beverage additive raw material, but the bitter and astringent taste of the pagodatree flower is covered by sweet or delicious substances. The existing fermentation research of the pagodatree flower is that the pagodatree flower and other traditional Chinese medicinal materials are combined for fermentation, and the influence factors of the fermentation are many. At present, no research report for improving the taste of the sophora flower water extract by utilizing a microbial fermentation method exists. According to the invention, ten edible microorganisms are used as fermentation strains, inoculated into the aqueous extract of the pagodatree flower, 3 strains with better effect of improving the taste of the pagodatree flower are screened out through single-strain fermentation, then the screened 3 strains are used as fermentation strains, different strain mixing ratios (single-strain, double-strain and three-strain mixing) are designed, the optimal strain combination for improving the taste through fermentation is screened out, and then the process condition for improving the taste through mixed strain fermentation is optimized by using a response surface analysis method, so that the optimal process condition for improving the taste of the aqueous extract of the pagodatree flower through mixed strain fermentation is obtained. Can obviously improve bitter and astringent uncomfortable taste of the pagodatree flower extract, increase the content of flavone which is an active ingredient of a fermentation product, can be made into food additive raw materials, and is better applied to development of pagodatree flower drinks. According to the invention, only one medicinal material water extract of the pagodatree flower is subjected to fermentation to improve the taste, the taste of the pagodatree flower water extract can be obviously improved, the obtained pagodatree flower fermentation liquid can be used as a subsequent food additive, and a new development way is provided for the traditional Chinese medicine for both food and medicine. The invention optimizes the fermentation process by adopting the response surface analysis method, and the obtained optimal fermentation process can be applied to industrial production.
Compared with the prior art, the invention uses the modern fermentation technology, is different from the traditional natural fermentation, and ensures that the fermentation process is scientific and controllable. The sterilization treatment is strictly followed in the fermentation process, and the addition of edible fungi greatly avoids the pollution of mixed bacteria. The prepared pagodatree flower fermentation liquor can also be used for the research and development of the following food additive raw materials, and provides a new development approach for the traditional Chinese medicine for both food and medicine. Provides a new technical route for the conversion of traditional Chinese medicines to high-end food industry. The invention is different from other combined fermentation modes in the fermentation of the single medicinal material of the pagodatree flower, has less influence factors, and is beneficial to observing the taste of the pagodatree flower fermentation liquid and the change of main effective components.
Drawings
Fig. 1 is a process flow diagram of the present invention.
The invention is further described below with reference to the drawings and the detailed description.
Detailed Description
Example 1: the microbial screening and fermentation process optimizing method for improving bitter and astringent feeling of the sophora flower water extract comprises the following steps:
(1) Weighing appropriate amount of flos Sophorae Immaturus, soaking in tap water for 30 min, adding 10 times of tap water into flos Sophorae Immaturus, decocting for 1 hr, filtering with gauze, collecting filtrate, boiling residue with 8 times of water for 1 hr, filtering, mixing the two filtrates, and concentrating to obtain flos Sophorae Immaturus water extract with a ratio of 1:10;
(2) Packaging the pagodatree flower water extract in the step (1) into 11 triangular bottles (50 ml per bottle), and sterilizing at 121 ℃ under high pressure;
(3) The 11 bottles of sterilized water decoction in the step (2) are respectively inoculated with ten different edible strains (aspergillus oryzae, aspergillus niger, saccharomyces cerevisiae, bacillus subtilis, lactobacillus pentosus, lactobacillus helveticus, lactobacillus plantarum, lactobacillus bulgaricus, lactobacillus acidophilus and streptococcus thermophilus) in the other 10 bottles of triangular bottles, wherein 1 bottle of triangular bottles are not inoculated with bacteria (serving as blank control), and the inoculation amount is 4%; the ten edible fungi are purchased from China industry microbiological culture Collection center (CICC), and the preservation numbers are respectively: aspergillus oryzae (CICC 40786), aspergillus niger (CICC 41594), saccharomyces cerevisiae (CICC 1793), bacillus subtilis (CICC 20522), lactobacillus pentosus (CICC 21798), lactobacillus helveticus (CICC 22171), lactobacillus plantarum (CICC 21809), lactobacillus bulgaricus (CICC 20247), lactobacillus acidophilus (CICC 22162), streptococcus thermophilus (CICC 25079);
(4) Culturing 11 bottles of water decoction in the step (3) in a shaking incubator with the rotating speed of 200r/min and the temperature of 25 ℃ for 24 hours;
(5) Carrying out sensory scoring on the fermented sample in the step (4) according to a sensory evaluation table in table 1, screening strains with higher export sensory scores and 3 ranking the strains with the highest export sensory scores, wherein the results are shown in table 3, and the strains with the highest ranking 3 are lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus respectively;
(6) Carrying out mixed fermentation on 3 strains screened in the step (5), wherein the mixed bacteria proportion is shown in table 2, the inoculation amount is 4%, and culturing for 24 hours in a shaking incubator with the rotating speed of 200r/min and the temperature of 25 ℃;
table 1 sensory evaluation table
TABLE 2 mixing ratio of bacteria
TABLE 3 screening of Sophora japonica fermentation single bacteria
(7) Screening mixed bacteria combinations (see table 4) with good export feel and 3 before sensory scores according to the sensory scores and the content changes of main active ingredient total flavonoids, wherein the mixed bacteria combinations are respectively lactobacillus helveticus: streptococcus thermophilus: lactobacillus bulgaricus (2:1:1), lactobacillus helveticus: streptococcus thermophilus: lactobacillus bulgaricus (1:1:1), lactobacillus helveticus, and detecting total flavone content (see table 5) after fermentation of the 3 groups of mixed bacteria, wherein the 3 groups of sensory evaluation differences are not large, so that a mixed bacteria combination with high flavone content of active ingredients, namely lactobacillus helveticus, is selected: streptococcus thermophilus: lactobacillus bulgaricus (1:1:1);
TABLE 4 screening of mixed fermentation broths of Sophora japonica
TABLE 5 total flavone content of mixed fermentation broth
(8) And (3) inoculating the mixed bacteria to the water extract of the pagodatree flower by using the best mixed bacteria combination screened in the step (7), and performing single factor investigation analysis on fermentation time (12 h, 24h, 36h, 48h, 60h and blank control), fermentation temperature (15 ℃, 20 ℃, 25 ℃, 30 ℃, 35 ℃, 40 ℃ and blank control) and inoculation amount (1%, 2%, 3%, 4%, 5% and blank control). When the fermentation time is examined, the fermentation temperature is 25 ℃, and the inoculation amount is 4%. When the fermentation temperature is examined, the fermentation time is 24 hours, and the inoculation amount is 4%. When the inoculation amount is inspected, the fermentation temperature is 25 ℃, and the fermentation time is 24 hours;
(9) The sensory evaluation is taken as the main part, and the total flavone detection result (see table 6-table 8) is combined, so that the fermentation temperature of mixed bacteria fermentation of the water extract of the sophora japonica is 20 ℃, the bacterial inoculation amount is 3%, and the fermentation time is 24 hours;
TABLE 6 temperature investigation of mixed fermentation broths of Sophora japonica
TABLE 7 investigation of the inoculation quantity of Sophora japonica mixed fermentation broth
TABLE 8 investigation of fermentation time of mixed fermentation broth of Sophora japonica
(10) Taking the optimal fermentation temperature, the inoculation amount and the fermentation time determined in the step (9) as central test points, designing a three-factor three-level experiment by utilizing a Box-Benhnken test scheme of Design Expert V8.0.5 b software, and taking the sensory scores and the total flavone content as response values, wherein the test results are shown in Table 9;
TABLE 9Box-Benhnken experiment design and results
(11) Obtaining by regression fit analysis of the test results in step (10)Predictive regression equation for sensory scores: mouth feel score = 78.87+0.21 a+0.13 b+0.000 c+0.46 b c-0.032 a c+0.55 b c-1.21 a 2 -0.95*B 2 -0.95*C 2 The analysis of variance of the regression equation was also obtained (see Table 10). By analyzing the analysis result of variance analysis, the P value of the regression equation model is 0.0018 #<0.01 The model has good significance, and the mismatch value P is 0.3758%>0.05 Less pronounced loss, i.e., indicating that other factors have little impact on sensory scores. From the F value, the influence degree of the pagodatree flower fermentation influence factors on taste grading is known: temperature (A)>Inoculation quantity (B)>Time (C); influence of two factor interactions on sensory scores of pagodatree flower fermentation broths: BC (BC) type>AB>AC。
TABLE 10 response surface sensory score regression equation analysis of variance
(12) And (3) obtaining a predictive regression equation for the flavone content through regression fit analysis of the test results in the step (10):
total flavone concentration = 0.32-4.750E-005 a +5.363e-004 b-6.288EE-004 c +5.725e-004 a b-1.298E-003 a c +1.735e-003 b c-0.010 a 2 -7.394E-003*B 2 -7.773E-003*C 2 The analysis of variance of the regression equation was also obtained (see Table 11).
The analysis of variance results show that the model P value is 0.0073 (< 0.01), which shows that the model is good in significance, the mismatch value P value is 0.9076 (> 0.05), the mismatch is not significant, and other factors have little influence on the sensory scores. From the F value, the influence degree of the pagodatree flower fermentation influence factors on the total flavone content can be known: fermentation time (C) > inoculation amount (B) > fermentation temperature (A); influence of interaction of two factors on total flavone content of pagodatree flower fermentation broth: BC > AC > AB.
TABLE 11 regression equation analysis of variance of total flavone content of response surface
(13) The sensory scores and the total flavone concentrations in the regression equations of the steps (11) and (12) are the highest, the optimal fermentation time, the optimal fermentation temperature and the optimal fermentation inoculation amount for improving the taste of the sophora flower water extract are respectively 20.23h, 23.84 ℃ and 3.06%, and under the three optimal factors, the theoretical prediction sensory scores and the total flavone concentrations are respectively 78.8781 mg/ml and 0.322324mg/ml. The practical consideration is integrated, the optimal fermentation time predicted by theory is 20 hours, the fermentation temperature is 24 ℃, and the inoculation amount is 3 percent.
(14) 3 verification tests were performed according to the theoretical prediction of the optimal fermentation time, fermentation temperature, and inoculation amount in step (13), and the results are shown in table 12. The average value of the sensory scores of the three groups is 79.01, the relative error with the theoretical sensory score is 0.17%, the average concentration of the total flavone content of the three groups is 0.31920mg/ml, and the relative error with the theoretical total flavone content is 0.97%. In conclusion, the model established by the Design Expert V8.0.5 b response surface analysis software is accurate and reliable.
Table 12 validates the test results
From the above, the best fermentation strain combination for improving the taste of the aqueous extract of the sophorae flower by screening out the microbial mixed fermentation by the method is lactobacillus helveticus: streptococcus thermophilus: lactobacillus bulgaricus (1:1:1), wherein the optimal mixed bacteria fermentation condition is that the fermentation time is 20 hours, the fermentation temperature is 24 ℃, and the inoculation amount is 3%.
The above description is only of the preferred embodiments of the present invention and is not intended to limit the present invention, but various modifications and variations can be made to the present invention by those skilled in the art. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (3)
1. The microorganism screening method for improving the bitter and astringent feeling of the sophora flower water extract and the fermentation process are characterized by comprising the following steps:
(1) Activating and culturing strains of ten edible microorganisms such as Aspergillus oryzae, aspergillus niger, saccharomyces cerevisiae, bacillus subtilis, lactobacillus pentosus, lactobacillus helveticus, lactobacillus plantarum, lactobacillus bulgaricus, lactobacillus acidophilus and Streptococcus thermophilus;
(2) Weighing appropriate amount of flos Sophorae Immaturus, soaking in tap water for 30 min, decocting with 10 times of tap water for 1 hr, filtering with gauze, collecting filtrate, boiling the residue with 8 times of water for 1 hr, and filtering; mixing the two filtrates, concentrating to obtain flos Sophorae Immaturus water extract at a ratio of 1:10, packaging into 11 triangular flasks, and sterilizing at 121deg.C under high pressure;
(3) The 11 bottles of sterilized sophora flower water extract in the step (2) are respectively inoculated with ten edible fungi which are subjected to activation culture in the step (1) by using 1 bottle of triangular flask without inoculating bacteria as a blank control and the other 10 bottles of triangular flasks with 4% of inoculated bacteria;
(4) Culturing 11 bottles of pagodatree flower water extract in the step (3) in a shaking incubator with the rotating speed of 200r/min and the temperature of 25 ℃ for 24 hours;
(5) Carrying out sensory scoring on the fermented sample in the step (4), and screening strains with higher export sensory scoring and top three ranks, namely lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus respectively;
(6) Carrying out mixed fermentation on the lactobacillus helveticus, the lactobacillus bulgaricus and the streptococcus thermophilus screened in the step (5), wherein the inoculation amount is 4%, and culturing the lactobacillus helveticus, the lactobacillus bulgaricus and the streptococcus thermophilus in a shaking incubator with the rotation speed of 200r/min and the temperature of 25 ℃ for 24 hours;
(7) According to the sensory scores and the content changes of main active ingredient total flavonoids, screening the mixed bacterium combination with good export feel and high content of main active ingredient total flavonoids from Lactobacillus helveticus, lactobacillus bulgaricus and Streptococcus thermophilus, wherein the mixed bacterium proportion is that of the Lactobacillus: lactobacillus bulgaricus: streptococcus thermophilus is 1:1:1;
(8) Inoculating the mixed bacteria combination of the best lactobacillus helveticus, lactobacillus bulgaricus and streptococcus thermophilus screened in the step (7) into the aqueous extract of the pagodatree flower, and carrying out single factor investigation analysis on fermentation time, fermentation temperature and inoculation amount;
(9) Taking sensory scores and total flavone content as evaluation indexes, and determining the optimal fermentation time, the optimal fermentation temperature and the optimal inoculation amount when the single factor of the sophora flower water extract is inspected in the step (8);
(10) Designing a three-factor three-level experiment based on determining the optimal fermentation time, the optimal fermentation temperature and the optimal inoculation quantity by using a Box-Benhnken test scheme of Design Expert V8.0.5 b software, and obtaining the optimal fermentation process condition of the sophora flower water extract for improving the taste by using the sensory score and the total flavone content as response values and performing secondary regression equation analysis of a response surface and prediction by the Design Expert V8.0.5 b software;
(11) The optimal fermentation process conditions for improving the taste of the sophora flower water extract obtained in the step (10) are verified for three times, sensory scores and total flavone content are used as evaluation indexes, the relative errors of the verified actual measurement values and the theoretical prediction values in the step (10) are compared, and the optimal fermentation process conditions for improving the taste of the sophora flower water extract by multi-strain fermentation are combined with industrial production practice, wherein the optimal fermentation process conditions comprise: fermenting for 20h at 24 ℃ with 3% of inoculation amount.
2. The application of Lactobacillus helveticus, lactobacillus bulgaricus and Streptococcus thermophilus in preparing flos Sophorae Immaturus products.
3. The application of Lactobacillus helveticus, lactobacillus bulgaricus and Streptococcus thermophilus in improving bitter taste of flos Sophorae Immaturus is provided.
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