CN113061500A - Fermentation brewing process of musalaisi with high caramel aroma yield - Google Patents
Fermentation brewing process of musalaisi with high caramel aroma yield Download PDFInfo
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Abstract
The invention provides a fermentation brewing process of musalaisi with high caramel aroma yield. The invention belongs to the technical field of fermentation industry, and relates to a fermentation brewing process of musalaisi with high caramel aroma yield. The method comprises the following steps: s1: raw material screening and cleaning, S2: raw material stem removal and crushing, S3: squeezing to obtain juice, S4: concentration, S5: primary inoculation, S6: fermentation of blood sugar and nitrogen reducing elements, S7: secondary inoculation, S8: producing caramel fragrance and fermenting. Compared with the prior art, firstly, the Musalace is fermented by compounding the saccharomyces cerevisiae F2-P85-3 and the F2-B163-2 strain with high yield of furanone, so that the content of caramel aroma substances in the Musalace can be effectively increased, and the taste of the Musalace is improved; secondly, the process adopts a fermentation mode to promote the caramel flavor substances in the musalaisi, does not generate byproducts, and is safer and more environment-friendly; finally, the production process is simple in flow, low in cost and convenient to popularize.
Description
Technical Field
The invention belongs to the technical field of fermentation industry, and relates to a fermentation brewing process of musalaisi with high caramel aroma yield.
Background
Musalaisi is a special local product in Xinjiang, has a main local product in Avati county, is a brewed delicious and mellow grape beverage, has high medicinal value, and is rich in amino acids, multiple vitamins, glucose, iron and other nutritional ingredients and trace elements required by a human body. The musalaisi belongs to warm property and is a pure natural green drink. The caramel fragrance endows the wine with unique and aromatic flavor and improves the wine body quality. At present, substances with caramel aroma are mainly furan substances, and the current caramel aroma synthesis method comprises the following steps: natural extraction and chemical synthesis, but the cost of the natural extraction is too high, and the chemical synthesis can generate byproducts, which are not suitable for increasing the content of caramel aroma substances in the musalaisi. The microbial fermentation has the most outstanding advantages, has the advantages of simple synthesis and no pollution, is very suitable for increasing the content of caramel aroma substances in the production of the musalaisi, but the prior art does not adopt the microbial fermentation technology for the production process of the musalaisi to increase the content of the caramel aroma substances in the musalaisi.
Disclosure of Invention
In order to solve the above problems, the primary object of the present invention is to provide a fermentation brewing process of musalaisi with high caramel aroma yield, which can increase the content of caramel aroma substances in the musalaisi.
The invention also aims to provide a fermentation brewing process of the musalaisi with high caramel aroma yield, which does not produce by-products and is environment-friendly and pollution-free.
The last purpose of the invention is to provide a fermentation brewing process of the musalaisi with high caramel aroma yield, which has simple production flow, low cost and convenient popularization.
In order to achieve the purpose, the technical scheme of the invention is as follows:
the invention provides a fermentation brewing process of musalaisi with high caramel aroma yield, which comprises the following steps:
s1: screening and cleaning raw materials: ripe Hetian red grapes are picked and washed.
S2: raw material stem removal and crushing: and (3) removing stems of the selected grapes, crushing the grapes, and conveniently squeezing the grapes to obtain juice.
S3: squeezing to obtain juice: the grapes are pressed to obtain grape juice.
S4: concentration: concentrating the grape juice to a sugar degree of 23-35 Brix, cooling to room temperature, and then pumping into a fermentation tank.
S5: primary inoculation: inoculating the activated fresh culture solution of the F2-P85-3 strain into concentrated grape juice of a fermentation tank for primary inoculation according to the inoculation amount of 2%.
S6: fermenting the blood sugar-reducing and nitrogen-reducing elements: and (3) performing temperature-controlled fermentation on the concentrated grape juice subjected to primary inoculation at 28 ℃ by using a static fermentation method, and monitoring the sugar content, the alcohol content and the nitrogen level every day, so that the sugar content is not more than 196g/L, the nitrogen content is not more than 230mg/L, and the alcohol content is not more than 8%.
S7: secondary inoculation: after the fermentation of the glucose-reducing and nitrogen-reducing elements is finished, inoculating the activated fresh culture solution of the F2-B163-2 strain into concentrated grape juice of a fermentation tank for secondary inoculation according to the inoculation amount of 2 percent.
S8: caramel aroma producing fermentation: and (3) fermenting the concentrated grape juice subjected to secondary inoculation to generate caramel aroma at the fermentation temperature of 28 ℃ until the foam is completely disappeared, returning the product temperature to the room temperature, and finishing the fermentation to obtain the finished product of the musalaisi.
Further, the high performance liquid pre-column derivatization method is adopted to detect the concentration content of the total amino acids in the grape juice of the raw material in S1, and the N element level of the red grape selected in S1 is ensured to be not lower than 230 mg/L.
Further, the juice is squeezed in the S3 to ensure that the juice yield is over 75 percent.
Further, in the S5, a YPD culture medium or fresh concentrated grape juice is used for activating the F2-P85-3 strain, the activation temperature is 28 ℃, and the activation time is 24 hours. The F2-P85-3 strain is a saccharomyces cerevisiae strain with excellent caramel aroma and flavor.
Further, when the sugar concentration is 189g/L, the alcoholic strength is 6.9 percent and the nitrogen concentration is 230mg/L in the step S6, performing secondary fermentation until the foam falls back, and cooling the product temperature to room temperature to finish the fermentation.
Further, in the S7, the F2-B163-2 strain is activated by YPD medium at 28 ℃ for 24 h. The F2-B163-2 strain is a strain with high yield of furanone.
Furthermore, in the S8 fermentation process, coating observation is carried out every day, so that the normal growth of the F2-B163-2 fermentation strain inoculated twice is ensured, and the smooth completion of fermentation is ensured.
Further, the YPD medium comprises: 2% of glucose, 2% of peptone, 1% of yeast extract powder and the balance of distilled water, wherein the pH value of the YPD medium is a natural pH value, and the YPD medium is sterilized at high temperature and high pressure by using a high-temperature high-pressure vertical sterilizing pot. The sterilization temperature of the high-temperature and high-pressure sterilization is 121 ℃, the sterilization pressure is 0.12MPa, and the sterilization time is 15 min.
Further, the biological preservation information of the F2-P85-3 strain is as follows:
and (3) classification and naming: saccharomyces cerevisiae, latin name: saccharomyces cerevisiae;
the name of the depository: institute of microbiology, national academy of sciences;
the address of the depository: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North;
the preservation date is as follows: 03 month 10 in 2021;
the preservation number is: CGMCC No. 21886.
Further, the biological preservation information of the F2-B163-2 strain is:
and (3) classification and naming: kluyveromyces pichia, latin name: pichia pastoris;
the name of the depository: institute of microbiology, national academy of sciences;
the address of the depository: xilu No. 1 Hospital No. 3, Beijing, Chaoyang, North;
the preservation date is as follows: 03 month 10 in 2021;
the preservation number is: CGMCC No. 21887.
The invention has the beneficial effects that: compared with the prior art, firstly, the Musalace is fermented by compounding the saccharomyces cerevisiae F2-P85-3 and the F2-B163-2 strain with high yield of furanone, so that the content of caramel aroma substances in the Musalace can be effectively increased, and the taste of the Musalace is improved; secondly, the process adopts a fermentation mode to promote the caramel flavor substances in the musalaisi, does not generate byproducts, and is safer and more environment-friendly; finally, the production process is simple in flow, low in cost and convenient to popularize.
Drawings
FIG. 1 is a flow diagram of a fermentation process of the present invention;
FIG. 2 is a graph showing the number concentration of mixed bacteria (F2-P85-3 and F2-B163-2) in the fermentation process of the present invention 7 days before fermentation;
FIG. 3 is a graph showing the data of the number of fermentation bacteria of a control strain EC1118 7 days before fermentation in the fermentation brewing process of Musalace of the present invention;
FIG. 4 is a graph showing the data on the change in sugar concentration of mixed bacteria (F2-P85-3 and F2-B163-2) and a control group (EC1118 strain) during fermentation according to the present invention;
FIG. 5 is a graph showing the data on the alcohol concentrations produced by the mixed bacteria (F2-P85-3 and F2-B163-2) and the control group (EC1118 strain) in the fermentation process of the present invention;
FIG. 6 is a graph of sensory score data of the mixed bacteria (F2-P85-3 and F2-B163-2) fermented wine of the present invention versus the control (EC1118) fermented wine;
FIG. 7 is a graph showing the furan substance concentration of the mixed bacteria (F2-P85-3 and F2-B163-2) fermented liquors of the present invention with respect to the control (EC1118) fermented liquor;
FIG. 8 is a data graph showing the degradation amount and residual amount of 5-hydroxymethylfurfural in the mixed-bacteria (F2-P85-3 and F2-B163-2) fermented liquors of the present invention and the control (EC1118) fermented liquor;
FIG. 9 is a plate morphology of the high caramel aroma-producing strain of the present invention (F2-B163-2) WL;
FIG. 10 is a morphology of sensory evaluation strain (F2-P85-3) WL plate of the present invention;
FIG. 11 is a graph of the data of the tolerance analysis of the caramel highly productive strain (F2-B163-2) of the present invention to OD value variation at different sugar concentrations;
FIG. 12 is a graph of the tolerance analysis of the caramel highly-producing strain of the present invention (F2-B163-2) to the production of a furan species at different sugar concentrations;
FIG. 13 is a graph of the tolerance analysis of the caramel highly productive strain of the invention (F2-B163-2) with data on the change in OD values at different N levels;
FIG. 14 is a graph of tolerance analysis of a caramel highly-producing strain of the present invention (F2-B163-2) to the production of a furan species at different levels of N;
FIG. 15 is a graph of the tolerance analysis of the caramel highly-producing strain (F2-B163-2) of the present invention with OD value variation data at different alcohol concentrations;
FIG. 16 is a graph of tolerance analysis of a high caramel aroma-producing strain of the present invention (F2-B163-2) to the production of a furan species at different alcohol concentrations;
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail with reference to the following embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
To achieve the above object, the present invention is implemented as follows:
screening high-yield caramel aroma strains:
s21: a total of 65 strains of Saccharomyces cerevisiae and non-Saccharomyces cerevisiae were selected and numbered as in Table 1.
S22: activation of the strain: the 65 types of the culture solution preserved with 25% glycerol were inoculated and activated in a sterilized medium containing 1ml of YPD. The inoculation amount is 2 percent, and the culture is carried out for 24 hours at the constant temperature of (28 +/-1) DEG C. The YPD medium comprises the following components: 2% of glucose, 2% of peptone, 1% of yeast extract powder and distilled water, wherein the pH value of the culture medium is a natural pH value, and autoclaving at 121 ℃ for 15 min.
S23: preparing concentrated grape juice:
s231: and selecting raw materials of the red grape: ripe and red grape raw materials, and selecting Chinese olive, foreign matters, rotten fruits and the like. And cleaning the selected grapes.
S232: removing stems: and (3) manually removing stems of the cleaned grapes.
S233: squeezing to obtain juice: and squeezing the grapes subjected to stem removal by using a screw press to obtain juice.
S234: concentrating grape juice: boiling and concentrating the Sucus Vitis viniferae until the sugar degree reaches 28 Brix.
S235: packaging and sterilizing: and (3) filling the concentrated grape juice into triangular bottles, sealing the triangular bottles by 300ml each, placing the triangular bottles into a vertical pressure steam sterilizer for sterilization at the sterilization temperature of 121 ℃ for 15min, and cooling to room temperature for later use.
S24: inoculating and fermenting: inoculating the activated 65 bacterial liquids to be tested into sterilized concentrated grape juice respectively according to the inoculation amount of 2%, putting the grape juice into an incubator at 28 ℃ for fermentation for 14 days, and measuring the generation amount of furanone by HLPC.
S25: detecting the generation amount of caramel fragrant substances: and (3) respectively measuring the concentrations of furanone, acetylfuran and 5-methylfurfural in the fermented concentrated grape juice by using a high performance liquid chromatography, wherein the substances of the furanone, the acetylfuran and the 5-methylfurfural have caramel aroma characteristics and can be used for judging the content of the caramel aroma substances in the product. The data for the concentrations of furanone, acetylfuran and 5 methylfurfural are shown in table 2.
S26: analysis of caramel aroma OVA value results: the OAV value is the aroma activity value of the yeast fermented wine and can be used for representing the caramel aroma content in the musalaisi. The ratio between the content of OAV value in a food product and its aroma threshold in a particular substrate (water, oil, etc.). The threshold value of furanone in water is 0.16mg/L, the threshold value of acetylfuran in water is 10mg/L, and the threshold value of 5-methylfurfural in water is 466.32 mg/L. The data indicating the sum of OAV of furanone, acetylfuran and 5 methylfurfural among the 65 strains are shown in Table 3, and the OAV sum is the highest among F2-B163-2 strains, and the OAV value is 996.21.
S27: sensory evaluation: seven testers are selected, the caramel aroma is distinguished according to the wine noses, the caramel aroma after 65 strains of bacteria are fermented is graded according to 1-5, and 1 represents that the caramel aroma is light and almost not available; 2-3 represents that the caramel has common fragrance and is not strong; 4-5 represents the strong fragrance of caramel. An alcohol aqueous solution of furanone. Preparing 5 furanone concentration gradients of 200mg/L, 40mg/L, 8mg/L, 1.6mg/L and 0.32mg/L respectively, and reference solution (concentration is obtained by detecting concentration in fermented wine), randomly numbering, presenting to an evaluator, identifying the furanone reference solution (1-5 points in scale), and asking the evaluator to score the wine sample according to the scoring rule and the scoring requirement, wherein the scoring results are shown in Table 4. As can be seen from the scoring Table 4, the fermentation broth of the strain No. F-B18-6 had the highest score, followed by the strain Nos. F2-C507-4 and F2-P85-3, (F-B18-6 and F2-C507-4 were non-Saccharomyces cerevisiae, and the strain F2-P85-3 was Saccharomyces cerevisiae).
In general, the F2-B163-2 strain was a non-Saccharomyces strain with excellent caramel aroma production, and the F2-P85-3 strain had the best sensory evaluation of caramel aroma and was a Saccharomyces cerevisiae strain. The production process of the musalaisi capable of producing the caramel incense with high yield is characterized in that saccharomyces cerevisiae F2-P85-3 and F2-B163-2 strain with high furanone yield are mixed and fermented in time sequence.
Table 1:
table 2:
table 3:
| name of bacterium | OAV value | Name of bacterium | OAV value |
| F2-B163-2 | 966.21±35.12 | F1-7d2-04 | 197.23±18.35 |
| F1-B256-8 | 559.81±24.14 | F2-P85-3 | 193.3±31.25 |
| C1-AR3-03 | 410.42±26.22 | T-344-1 | 192.63±27.21 |
| F2-B178-11 | 372.52±14.54 | A0-Y2-01 | 191.02±43.34 |
| F2-C507-4 | 332.46±34.24 | A3-18d3-01 | 189.88±23.74 |
| I0-15d3-03 | 327.11±39.41 | F0-T427-1 | 187.99±25.68 |
| I0-15d2-08 | 322.51±30.84 | GO-Y9 | 187.8±42.38 |
| A1-4d5 | 321.69±46.11 | G2-9d1-05 | 182.24±25.48 |
| A4-25d1-01 | 318.34±33.76 | G3-12d3-03 | 178.23±34.44 |
| EC1118 | 285.76±26.21 | F2-P88-6 | 176.69±31.24 |
| C1-7d3-02 | 282.82±24.33 | F1-X205-4 | 176.62±27.29 |
| HO-6d5-01 | 282.66±51.25 | G2-7d1-04 | 176.21±34.21 |
| B0-3d1-06 | 279.78±11.14 | G2-9d7-01 | 173.04±27.12 |
| H0-6d2-03 | 276.64±19.11 | A0-Y8-02 | 171.42±31.88 |
| F-B18-6 | 273.99±21.74 | G3-12d1-04 | 170.05±22.34 |
| A0-3d1-03 | 270.95±27.58 | F1-M127-3A | 164.04±23.65 |
| F2-P85-1 | 261.56±39.76 | G3-12d4-01 | 159.84±33.61 |
| B0-Y3-01 | 259.78±44.28 | G0-Y7 | 158.71±22.05 |
| E0-1Y2-004 | 258.72±50.74 | F0-Y2-02 | 154.27±11.08 |
| B1-8d1 | 258.36±16.34 | A3-18d1-02 | 152.17±27.17 |
| C0-1d1-05 | 256.18±31.34 | F2-A62-3 | 150.96±9.01 |
| B3-23d1-02 | 249.23±26.89 | F2-M93-20 | 142.78±11.35 |
| F2-16d3-01 | 249.21±16.21 | C2-12d2-03 | 130.3±11.44 |
| F0-Y3-02 | 246.52±36.08 | F0-B163-2 | 122.14±14.87 |
| I0-15d1-07 | 244.74±21.07 | F1-L193-6 | 118.98±21.48 |
| G2-9d2-02 | 238.41±52.14 | E1-12d2-01 | 118.55±11.38 |
| H0-6d3-03 | 234.07±33.19 | F1-S487-8 | 118.27±16.68 |
| H0-6d2-02 | 232.8±22.21 | C0-Y2-02 | 117.32±11.48 |
| F1-X208-4 | 229.99±11.31 | P-1L1-15 | 114.99±31.35 |
| G0-Y4 | 228.56±21.38 | B0-Y3-02 | 110.99±22.21 |
| F0-L368-1 | 227.07±31.58 | Blank space | 37.41±6.28 |
| E2-21d3-01 | 224.8±22.68 | Beijing | 219.46±31.84 |
| G1-3d1-06 | 221.47±21.56 | C2-12d1-03 | 216.24±20.77 |
Table 4:
fermentation and brewing of the musalaisi with high caramel aroma yield:
s1: screening and cleaning raw materials: ripe Hetian red grapes are picked and washed.
S2: raw material stem removal and crushing: and (3) removing stems of the selected grapes, crushing the grapes, and conveniently squeezing the grapes to obtain juice.
S3: squeezing to obtain juice: the grapes are pressed to obtain grape juice.
S4: concentration: concentrating the grape juice to a sugar degree of 23-35 Brix, cooling to room temperature, and then pumping into a fermentation tank.
S5: primary inoculation: inoculating the activated fresh culture solution of the F2-P85-3 strain into concentrated grape juice of a fermentation tank for primary inoculation according to the inoculation amount of 2%.
S6: fermenting the blood sugar-reducing and nitrogen-reducing elements: and (3) performing temperature-controlled fermentation on the concentrated grape juice subjected to primary inoculation at 28 ℃ by using a static fermentation method, and monitoring the sugar content, the alcohol content and the nitrogen level every day, so that the sugar content is not more than 196g/L, the nitrogen content is not more than 230mg/L, and the alcohol content is not more than 8%.
S7: secondary inoculation: after the fermentation of the glucose-reducing and nitrogen-reducing elements is finished, inoculating the activated fresh culture solution of the F2-B163-2 strain into concentrated grape juice of a fermentation tank for secondary inoculation according to the inoculation amount of 2 percent.
S8: caramel aroma producing fermentation: and (3) fermenting the concentrated grape juice subjected to secondary inoculation to generate caramel aroma at the fermentation temperature of 28 ℃ until the foam is completely disappeared, returning the product temperature to the room temperature, and finishing the fermentation to obtain the finished product of the musalaisi.
Detecting the concentration content of total amino acids in the grape juice of the raw material in S1 by using a high performance liquid pre-column derivatization method, and ensuring that the N element level of red grapes selected in S1 is not lower than 230 mg/L.
The juice is squeezed in the S3 to ensure that the juice yield is over 75 percent.
In the S5, a YPD culture medium or fresh concentrated grape juice is used for activating the F2-P85-3 strain, the activation temperature is 28 ℃, and the activation time is 24 hours.
And in the S6, when the sugar concentration reaches 189g/L, the alcoholic strength reaches 6.9 percent and the nitrogen concentration reaches 230mg/L, performing secondary fermentation until foam falls back, and cooling the product to room temperature to finish the fermentation.
In the S7, a YPD culture medium is used for activating the F2-B163-2 strain, the activation temperature is 28 ℃, and the activation time is 24 hours.
In the S8 fermentation process, coating observation is carried out every day, so that the normal growth of the F2-B163-2 fermentation strain inoculated for the second time is ensured, and the smooth completion of fermentation is ensured.
The YPD medium comprises the following components: 2% of glucose, 2% of peptone, 1% of yeast extract powder and the balance of distilled water, wherein the pH value of the YPD medium is a natural pH value, and the YPD medium is sterilized at high temperature and high pressure by using a high-temperature high-pressure vertical sterilizing pot. The sterilization temperature of the high-temperature and high-pressure sterilization is 121 ℃, the sterilization pressure is 0.12MPa, and the sterilization time is 15 min.
The biological preservation information of the F2-P85-3 strain is as follows:
the biological preservation information of the JFB2 strain is as follows:
evaluation of high-yield caramel aroma of fermented concentrated grape juice by mixing and compounding of an excellent strain F2-B163-2 and a Saccharomyces cerevisiae strain F2-P85-3:
the number of mixed bacteria (F2-P85-3 and F2-B163-2) before 7 days of fermentation, the number of fermentation bacteria, the growth data are shown in FIG. 2, and the number of fermentation bacteria of the strain EC1118 before 7 days of fermentation, the number of fermentation bacteria, the growth data are shown in FIG. 3. As is clear from FIGS. 2 and 3, the mixed strains (F2-P85-3) and (F2-B163-2) reached the maximum strain concentration on day 3, and the concentrations of the strains (F2-B163-2) were 3.02X108(CFU/ml) and 2.9X108(CFU/ml), respectively, and then the cell number concentration began to gradually decrease. The concentration of the number of bacteria in the control group (EC1118) reached a maximum of 3.1X108(CFU/ml) at 3d, and then the concentration of the number of bacteria began to decrease slowly.
The change in the sugar consumption concentrations of the mixed bacteria (F2-P85-3, F2-B163-2) and the control (EC1118) during fermentation is shown in FIG. 4. The sugar concentration in the mixed strain fermentation process is obviously higher than that in the control strain EC1118 fermentation process due to the change of the sugar concentration in 30 days, and after 30 days of fermentation, the sugar concentration in the fermented wine of the mixed strains (F2-P85-3) and (F2-B163-2) is 122g/L, and the sugar concentration in the fermented wine of the control strain EC1118 is 131 g/L.
The change in the concentration of alcohol produced by the mixed bacteria (F2-P85-3, F2-B163-2) and the control (EC1118) during fermentation is shown in FIG. 5. The alcohol content in the mixed strain fermentation process is slightly higher than that in the control strain EC1118 fermentation process due to different alcohol content changes within 30 days, the alcohol content in the mixed strain (F2-P85-3) and (F2-B163-2) fermented wine is 16.52%, and the alcohol content in the control strain EC1118 fermented wine is 15.82%. As can be seen from the sugar concentration fermentation graph and the alcohol fermentation graph in the fermentation process of the mixed inoculated strains (F2-P85-3) and (F2-B163-2) and the control group EC1118, the sugar fermentation capacity of the mixed inoculated strains (F2-P85-3) and (F2-B163-2) is higher than that of the control group EC1118, so that the alcohol fermentation capacity of the mixed inoculated strains (F2-P85-3) and (F2-B163-2) is naturally higher than that of the control group EC 1118.
Sensory scores of the mixed bacteria (F2-P85-3, F2-B163-2) fermented wine and the control (EC1118) fermented wine are shown in FIG. 6. The furan content of the mixed bacteria (F2-P85-3, F2-B163-2) fermented wine and the control (EC1118) fermented wine are shown in FIG. 7. Therefore, the following steps are carried out: 168.31mg/L of furan, 2.25mg/L of acetylfuran and 21.43mg/L of 5-methylfurfural, which are generated by fermenting two strains of F2-P85-3 and F2-B163-2. 56.83mg/L of furanone, 1.83mg/L of acetylfuran and 9.93mg/L of 5-methylfurfural are generated by the fermentation of the control group EC 1118. The furan substance produced by the mixed fermentation of the two strains F2-P85-3 and F2-B163-2 is higher than that produced by the fermentation of the control group EC 1118. As can be seen from the sensory evaluation chart in FIG. 6, the aroma of the caramel fermented mixture of the two strains F2-P85-3 and F2-B163-2 is stronger than that of the control EC 1118.
The degradation amount and residual amount of 5-hydroxymethylfurfural in the mixed-bacteria (F2-P85-3, F2-B163-2) fermented wine and the control (EC1118) fermented wine are shown in FIG. 8. When the strains F2-P85-3 and F2-B163-2 are fermented for 30d in a mixed mode, the residual 5HMF of the 5-hydroxymethylfurfural in the fermented wine is 0.15g/L, the concentration of the degraded 5-hydroxymethylfurfural is 0.66g/L, and the degradation rate is 81.4%. The residual 5HMF of the 5-hydroxymethylfurfural in the control EC1118 fermented wine is 0.16g/L, the concentration of the degraded 5-hydroxymethylfurfural is 0.65g/L, and the degradation rate is 80.2%. Compared with a control group EC1118, the F2-P85-3 and JFB2 mixed fermentation has a small difference in the amplitude of degrading 5-hydroxymethylfurfural.
The high caramel aroma-producing strain (F2-B163-2) WL plate morphology is shown in 9: large volume, rough surface, white color with light green color.
Sensory evaluation of the strain (F2-P85-3) WL plate morphology is shown in FIG. 10: large volume, smooth surface, white creamy.
High caramel incense-producing strains (F2-B163-2) tolerance analysis:
1. tolerance analysis for different sugar degrees: the simulated grape juice is used as a fermentation substrate, the sugar concentrations are respectively 4g/L, 68g/L, 132g/L, 196g/L and 260g/L, and the other components are unchanged, and the simulated grape juice is set into 5 treatment groups of 4S, 68S, 132S, 196S and 260S. Each treatment volume is 32mL, the treatment volume is filtered and sterilized through a filter membrane with the aperture of 0.22um, sterilized simulated grape juice is respectively filled into 410 mL sterilized centrifuge tubes, 8mL of each centrifuge tube is respectively connected with F2-B163-2 fresh culture solution, the inoculation amount is 2%, the constant temperature culture is carried out at 28 ℃, the start of 7 days is carried out, the OD value (absorbance OD600nm) is measured every 24 hours, the growth condition of the strain is observed, the fermentation is carried out for 30d, the concentration of furan substances is detected by using HPLC, and the generation amount is calculated. The OD value change data under different sugar concentrations in the fermentation process is shown in figure 13, simulated grape juice is used as a fermentation substrate, the sugar concentrations in the prepared fermentation substrate are respectively 4g/L, 68g/L, 132g/L, 196g/L and 260g/L, the growth OD value of the strain F2-B163-2 is detected by a microplate reader every day 7 days before the constant-temperature fermentation at 28 ℃ by controlling the different changes of the sugar concentrations in the simulated grape juice, and figure 11 shows that the strain F2-B163-2 has different growth capacities under different sugar concentrations and is influenced by the culture time. It is clear that low sugar (4g/L) is the least favorable for bacterial growth, 68g/L is inferior in sugar degree, and the low sugar degree obviously delays a specific growth peak value. The OD values of other sugar degree treatments are not obviously different in the fermentation period, and the OD values of all treatment groups in the 4d fermentation period reach the maximum; among them, the OD value was highest in the 196g/L treatment group and reached approximately 1. The change of the production concentration of the furan substance under different sugar concentrations in the fermentation process is shown in figure 12, after the fermentation liquor is placed at a standstill of 28 ℃ for 30 days, the production amount of the furan substance is detected by HPLC, as shown in figure 12, it is obvious that when the sugar degree is less than or equal to 196g/L, the production amount of the furan substance is increased along with the increase of the sugar degree until the sugar concentration is 196g/L, the production amount of the furan substance reaches the maximum (the concentration of furanone is 30.48mg/L, the concentration of acetylfuran is 1.088mg/L, and the concentration of 5 methylfurfural is 4.78mg/L), and the production amount of the furan substance is slightly reduced in the following 260S, but has no significant difference with 196S.
2. Tolerance analysis for different nitrogen levels: the simulated grape juice is used as a fermentation substrate, the nitrogen concentrations are respectively 140mg/L, 230mg/L, 330mg/L, 410mg/L and 500mg/L, and other components are unchanged, and the simulated grape juice is set into 140N, 230N, 330N, 410N and 500N 5 treatment groups. Each treatment group is 32mL in volume, the treatment groups are filtered and sterilized through a filter membrane with the aperture of 0.22um, sterilized simulated grape juice is respectively put into 410 mL sterilized centrifuge tubes, each centrifuge tube is 8mL, fresh culture solution of F2-B163-2 yeast is respectively added, the inoculation amount is 2%, the constant temperature culture is carried out at 28 ℃, the OD value is measured every 24 hours after the start for 7 days, the growth condition of the strain is observed, the strain is fermented for 30d, the concentration of furans is detected by HPLC, the OD value changes under different N concentrations are shown in figure 13, the simulated grape juice is used as a fermentation substrate, different nitrogen concentrations in the fermentation substrate are 140mg/L, 230mg/L, 330mg/L, 410mg/L and 500mg/L, F2-B163-2 bacteria are added, the constant temperature fermentation is carried out at 28 ℃, the previous 7 days, the OD values of the strain are detected by an enzyme labeling instrument every day, the OD value changes of the low nitrogen level N140 and the strain OD value changes in the treatment, a growth peak value; the OD values for the medium nitrogen level N330 and N410 treatment groups varied approximately, and the value for the high nitrogen level treatment group N500OD was generally slightly lower than the other treatment groups, indicating that the high nitrogen level was particularly slightly consistent. All treatment groups, 3d, reached a growth peak with N140 higher than the other treatment groups. The variation of the production concentration of the furanic substances at different concentrations of the N element during the fermentation is shown in FIG. 14.
3. Tolerance analysis of different alcohol amounts: the simulated grape juice was used as a fermentation substrate, and the alcohol concentrations of 8%, 10%, 12%, 14% and 16% were prepared, and the other components were unchanged, and the treatment groups were set to 8A, 10A, 12A, 14A and 16A 5. Each treatment volume is 32mL, the treatment volume is filtered and sterilized through a filter membrane with the aperture of 0.22um, sterilized simulated grape juice is respectively filled into 410 mL sterilized centrifuge tubes, 8mL of each centrifuge tube is respectively connected with fresh culture solution of F2-B163-2 yeast, the inoculation amount is 2%, the constant temperature culture is carried out at 28 ℃, the OD value is measured every 24 hours after the beginning of 7 days, the growth condition of the strain is observed, the fermentation is carried out for 30d, the concentration of furan substances is detected by using HPLC, and the generation amount is calculated. The change in OD values at different alcohol concentrations during fermentation is shown in FIG. 15. The variation of the production concentration of furanic substances at different alcohol concentrations during fermentation is shown in FIG. 16.
The above embodiments are only for illustrating the present invention, and the scope of the present invention is not limited to the above embodiments. The objectives of the present invention can be achieved by the ordinary skilled person in the art according to the disclosure of the present invention and the ranges of the parameters.
Claims (9)
1. A fermentation brewing process of musalaisi with high caramel aroma yield is characterized by comprising the following steps:
s1: screening and cleaning raw materials: selecting and cleaning mature Hetian red grapes;
s3: squeezing to obtain juice: squeezing grapes to obtain grape juice;
s4: concentration: concentrating the grape juice to a sugar degree of 23-35 Brix, cooling to room temperature, and then pumping into a fermentation tank;
s5: primary inoculation: inoculating the activated fresh culture solution of the F2-P85-3 strain into concentrated grape juice of a fermentation tank for primary inoculation according to the inoculation amount of 2 percent;
s6: fermenting the blood sugar-reducing and nitrogen-reducing elements: performing temperature-controlled fermentation at 28 deg.C on the concentrated grape juice after primary inoculation by using static fermentation method, and monitoring sugar degree, alcohol content and nitrogen level every day to make sugar degree not more than 196g/L, nitrogen not more than 230mg/L, and alcohol content not more than 8%;
s7: secondary inoculation: after the fermentation of the glucose-reducing and nitrogen-reducing elements is finished, inoculating the activated fresh culture solution of the F2-B163-2 strain into concentrated grape juice of a fermentation tank for secondary inoculation according to the inoculation amount of 2 percent;
s8: caramel aroma producing fermentation: and (3) fermenting the concentrated grape juice subjected to secondary inoculation to generate caramel aroma at the fermentation temperature of 28 ℃ until the foam is completely disappeared, returning the product temperature to the room temperature, and finishing the fermentation to obtain the finished product of the musalaisi.
2. The process of brewing by fermentation mousse with high caramel aroma yield according to claim 1, further comprising the step of S2: raw material stem removal and crushing: and (3) removing stems of the selected grapes, crushing the grapes, and conveniently squeezing the grapes to obtain juice.
3. The fermentation brewing process of musalaisi with high caramel aroma yield of claim 1, wherein the juice pressing in S3 ensures that the juice yield of grapes is over 75%.
4. The fermentation and brewing process of musalaisi with high caramel aroma yield of claim 1, wherein the total amino acid concentration content in the grape juice of the raw material is detected by high performance liquid pre-column derivatization in S1, ensuring that the N element level of the red grapes selected in S1 is not lower than 230 mg/L.
5. The fermentation brewing process of Musalace of high caramel aroma yield according to claim 1, wherein F2-P85-3 strain is activated by YPD medium or fresh concentrated grape juice at 28 ℃ for 24h in S5.
6. The fermentation and brewing process of musalaisi with high caramel aroma yield of claim 1, wherein in S6, when the sugar concentration is 189g/L, the alcoholic strength is 6.9%, and the nitrogen concentration is 230mg/L, the secondary fermentation is performed until the foam falls back, and the product temperature is reduced to room temperature to finish the fermentation.
7. The fermentation brewing process of Musalace of high caramel aroma yield according to claim 1, wherein the F2-B163-2 strain is activated by YPD medium in S7 at 28 ℃ for 24 h.
8. The fermentation and brewing process of musalaisi with high caramel aroma yield of claim 1, wherein during the fermentation of S8, the F2-B163-2 fermentation strain inoculated twice is observed every day to ensure the normal growth of the fermentation strain, thereby ensuring the smooth completion of the fermentation.
9. The fermentation brewing process of musalaisi with high caramel aroma yield of claim 1, wherein the YPD medium comprises the following components: 2% of glucose, 2% of peptone, 1% of yeast extract powder and the balance of distilled water, wherein the pH value of the YPD culture medium is a natural pH value, and the YPD culture medium is subjected to high-temperature high-pressure sterilization by using a high-temperature high-pressure vertical sterilization pot, wherein the sterilization temperature of the high-temperature high-pressure sterilization is 121 ℃, the sterilization pressure is 0.12MPa, and the sterilization time is 15 min.
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| CN103243037A (en) * | 2013-05-24 | 2013-08-14 | 塔里木大学 | High temperature yeast for brewing msalais, preparation method thereof and brewed msalais |
| CN103343093A (en) * | 2013-05-24 | 2013-10-09 | 塔里木大学 | Low-temperature yeast for brewing musalaisi, preparation method and prepared musalaisi |
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| CN103243037A (en) * | 2013-05-24 | 2013-08-14 | 塔里木大学 | High temperature yeast for brewing msalais, preparation method thereof and brewed msalais |
| CN103343093A (en) * | 2013-05-24 | 2013-10-09 | 塔里木大学 | Low-temperature yeast for brewing musalaisi, preparation method and prepared musalaisi |
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