CN112961746A - Brewing method for improving flavor characteristics of cider - Google Patents
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Abstract
The invention discloses a brewing method for improving the flavor characteristics of cider, which adopts Schizosaccharomyces pombe (Schizosaccharomyces pombe) and Saccharomyces cerevisiae (Saccharomyces cerevisiae) to sequentially inoculate and ferment, and belongs to the technical field of cider brewing. The invention provides a sequential inoculation method for inoculating the schizosaccharomyces pombe in concentrated apple juice for fermentation, and inoculating the saccharomyces cerevisiae for secondary fermentation after 4 days. Compared with the cider fermented by singly inoculating the saccharomyces cerevisiae, the cider prepared by the method has the advantages of improved flavor characteristics of wine body, reduced acidity and increased alcoholic strength.
Description
Technical Field
The invention relates to the technical field of brewing of cider, and particularly relates to a brewing method for improving flavor characteristics of cider.
Background
Cider is the second largest fruit wine in the world after wine, has the flavor of fruit juice and the aroma of good wine, and can also increase the value of a large number of non-commercial apples. The production of the cider has no seasonality, short period, low price, mellow taste, rich nutrients such as vitamins, amino acids, calcium, iron, potassium and the like, and organic acids such as malic acid and the like which are not contained in other wines can regulate the metabolism of human bodies and promote blood circulation.
The concentrated apple juice is a main product for deep processing of apples in China, but has the problem of insufficient development and utilization, thereby causing resource waste. Therefore, the apple wine brewed by the concentrated apple juice fully utilizes apple resources, effectively improves the technical level of fruit deep processing, creates considerable economic benefit and has wide prospect.
Saccharomyces cerevisiae is widely used for cider fermentation due to its characteristics of high alcohol conversion rate, thorough fermentation and the like. However, the problem of single aroma, homogenization of flavor and the like of the cider is often caused by the independent fermentation of saccharomyces cerevisiae, the existing cider in the market of China has poor flavor and mouthfeel, the characteristics of the fruit wine cannot be fully reflected, the wine body is generally peracid, light in aroma, thin in mouthfeel and lack of aftertaste, and the acceptance of consumers in China to the cider is directly reduced.
It is reported that non-saccharomyces cerevisiae can participate in the formation of volatile substances such as esters, alcohols, terpenes and the like through biochemical processes such as metabolism, autolysis and the like, and improve the aroma of cider.
Schizosaccharomyces pombe (Schizosaccharomyces pombe) is a rod-shaped yeast of the genus Schizosaccharomyces, and research shows that the main fermentation characteristic of the Schizosaccharomyces pombe is the capability of converting malic acid into ethanol and CO2The alcohol has strong fermentation capacity, can be metabolized to generate glycerol and pyranoid anthocyanin in the fermentation process, and increases the content of aroma substances such as esters, terpenes and the like, so the alcohol has application potential of reducing acidity, improving aroma complexity and the like in the aspect of cider fermentation.
Therefore, how to utilize schizosaccharomyces pombe and saccharomyces cerevisiae to ferment cider to improve the flavor characteristics of wine is a problem to be solved urgently.
Disclosure of Invention
The invention provides a brewing method for millet to improve the flavor characteristic of apple wine in order to solve the technical problems.
In order to achieve the purpose, the invention adopts the following technical scheme:
a brewing method of Schizosaccharomyces pombe for improving the flavor characteristics of cider, comprising the following steps:
(1) treating raw material, adding sodium pyrosulfite into concentrated apple juice to make effective SO of sodium pyrosulfite2Adding pectinase to obtain effective content of 60mg/L, adjusting sugar degree of apple juice to 20Brix and pH to 3.8, and standing for 24 hr.
(2) Inoculation: inoculating Schizosaccharomyces pombe bacterial liquid into the concentrated apple juice, wherein after inoculation, the final concentration of Schizosaccharomyces pombe in the concentrated apple juice is 1 x 106cfu/mL；
(3) Carrying out primary fermentation culture on the concentrated apple juice, wherein the culture conditions are as follows: culturing at 28 deg.C and 120rpm for 4 days;
(4) adding Saccharomyces cerevisiae bacterial solution into concentrated apple juice after the first fermentation culture, performing secondary fermentation, and inoculating to obtain concentrated apple juice with final concentration of Saccharomyces cerevisiae of 1 × 106cfu/mL, total fermentation period 12 days.
(5) And carrying out suction filtration on the concentrated apple juice obtained by secondary fermentation to remove precipitated substances and fungi, thus obtaining the apple wine.
Further, the suction filtration adopts an organic microporous filtering membrane with the diameter of 50mm and the pore diameter of 0.45 μm.
Further, the culture method of the schizosaccharomyces pombe bacterial liquid comprises (1) strain activation, namely inoculating the schizosaccharomyces pombe and saccharomyces cerevisiae which are stored at ultralow temperature to a YPD liquid culture medium, and culturing for 24 hours at the temperature of 28 ℃ and the rotating speed of a shaking table of 120rpm for later use;
(2) preparing a seed solution: respectively transferring the activated schizosaccharomyces pombe and saccharomyces cerevisiae into YPD liquid culture medium with the culture amount of 8 percent to prepare seed liquid, wherein the seed culture conditions are as follows: culturing at 28 deg.C and 120rpm for 24h to logarithmic phase;
(3) preparing a bacterial liquid: and centrifuging the schizosaccharomyces pombe and saccharomyces cerevisiae seed liquid to remove supernatant liquid to obtain bacterial liquid.
Further, the parameters of a refrigerated centrifuge used for centrifugation are 4 ℃, and the refrigerated centrifuge is centrifuged at 6000rpm/min for 15 min;
further, the YPD liquid medium was prepared as follows: uniformly mixing 10g of peptone, 5g of yeast extract powder, 20g of glucose and 1L of water, and sterilizing at 121 ℃ for 30 min;
the schizosaccharomyces pombe has the strain preservation number as follows: 1760, the fission yeast schizosaccharomyces pombe is purchased from China Center of Industrial Culture Collection (CICC).
The saccharomyces cerevisiae has the strain preservation number as follows: 31084, a Saccharomyces cerevisiae available from China Center of Industrial Culture Collection (CICC).
The invention provides an application of schizosaccharomyces pombe and saccharomyces cerevisiae in cider fermentation by taking concentrated cider juice as a raw material.
The invention provides a processing and fermenting method of cider, which comprises the following steps: during the fermentation process, the bacteria liquid containing the fission yeast schizosaccharomyces pombe is added firstly for fermentation, and the bacteria liquid of the saccharomyces cerevisiae is added at intervals of 4 days for fermentation. The total fermentation period was 12 days.
The brewing method for improving the flavor characteristic of the cider by using the schizosaccharomyces pombe in the embodiment of the invention has the beneficial effects that:
the invention takes the concentrated apple juice as the raw material, is not limited by seasons and regions, and the concentrated apple juice is more convenient for long-time storage, so as to improve the quality of the apple wine, improve the flavor characteristic of the wine body, reduce the acidity and improve the alcoholic strength.
In addition, compared with the single inoculated saccharomyces cerevisiae, the finished wine has good flavor, reduced acidity and increased alcoholic strength.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings in the embodiments of the present invention will be briefly introduced below, the drawings in the following description are merely examples of the present invention, and it is obvious to those skilled in the art that other drawings can be obtained according to the drawings without making any inventive changes:
FIG. 1 is a process flow diagram of an embodiment of the invention;
FIG. 2 shows the change of alcoholic strength of cider wine in 12 days per day after fermentation;
FIG. 3 shows the change of soluble granules (Brix) per day for 12 days of fermentation of cider according to the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. The examples, in which specific conditions are not specified, were conducted under conventional conditions or conditions recommended by the manufacturer. The reagents or instruments used are not indicated by the manufacturer, and are all conventional products available commercially.
As shown in figure 1, the invention provides a brewing method for improving the flavor characteristics of cider by using schizosaccharomyces pombe, which comprises the following steps:
examples
A brewing method for improving the flavor characteristics of apple wine by using Schizosaccharomyces pombe adopts Schizosaccharomyces pombe mother and saccharomyces cerevisiae as fermentation strains to carry out sequential inoculation fermentation, wherein the preservation number of the Schizosaccharomyces pombe strain is as follows: 1760, and the Saccharomyces cerevisiae strain has the preservation number of: 31084, the method specifically comprises the following operations:
(1) treating raw materials by adding sodium pyrosulfite into concentrated apple juice to make available SO2Adding pectase to obtain effective content of 60mg/L, adjusting sugar degree of apple juice to 20Brix and pH to 3.8, standing for 24 hr, and packaging into 200mL sterile conical bottles. It is worth to be noted that the sodium metabisulfite and the pectinase are added to respectively play roles of bacteriostasis and clarification, so that the subsequent fermented wine has longer storage time and bright and beautiful color;
(2) and (3) activating strains, namely inoculating the ultralow-temperature-stored fission yeast schizosaccharomyces pombe to a YPD liquid culture medium, inoculating the ultralow-temperature-stored saccharomyces cerevisiae to another YPD liquid culture medium, and culturing for 24 hours at the temperature of 28 ℃ and the rotating speed of a shaking table of 120rpm for later use. The YPD liquid culture medium is prepared by the following method: uniformly mixing 10g of peptone, 5g of yeast extract powder, 20g of glucose and 1L of water, and sterilizing at 121 ℃ for 30 min;
(3) preparing a seed solution: transferring the activated fission yeast schizosaccharomyces pombe and saccharomyces cerevisiae in the step (2) into YPD liquid culture medium according to the culture amount of 8 percent (16mL) respectively to prepare seed liquid, wherein the seed culture conditions are as follows: the temperature is 28 ℃, the rotation speed of the shaking table is 120rpm, and the culture is carried out for 24 h.
(4) Preparing a bacterial liquid: and (4) centrifuging the fission yeast schizosaccharomyces pombe and saccharomyces cerevisiae seed liquid in the step (3) to remove supernatant liquid to obtain bacterial liquid. Wherein the parameters of a refrigerated centrifuge used for centrifugation are 4 ℃, and the refrigerated centrifuge is centrifuged at 6000rpm/min for 15 min;
(5) inoculation: inoculating the schizosaccharomyces pombe bacterial liquid prepared in the step (4) into the concentrated apple juice obtained in the step (1), and after inoculation, the final concentration of the schizosaccharomyces pombe in the concentrated apple juice is 1 multiplied by 106cfu/mL；
(6) And (3) first fermentation: culturing the concentrated apple juice obtained in the step (5), monitoring the change of sugar degree and alcoholic strength every day (as shown in figures 2 and 3), and culturing under the conditions as follows: culturing at 28 deg.C and 120rpm for 4 days;
(7) and (3) secondary fermentation: adding the saccharomyces cerevisiae liquid obtained in the step (4) into the apple juice after the primary fermentation in the step (6) for secondary fermentation, and after inoculation, concentrating the final concentration of saccharomyces cerevisiae in the apple juice to be 1 × 106cfu/mL, the change of the sugar degree and the alcoholic strength is monitored every day, and the fermentation is stopped when the sugar degree is not changed any more and the fermentation is stable (as shown in figures 2 and 3), and the total fermentation period is 12 days. The culture conditions were: the temperature is 28 ℃, and the rotating speed of the shaking table is 120 rpm;
(8) and (3) filtering: and (3) carrying out suction filtration on the fermented cider to remove precipitated substances and fungi, thus obtaining the cider, wherein the suction filtration uses an organic microporous filtering membrane with the diameter of 50mm and the pore diameter of 0.45 mu m.
Comparative example
The difference from the embodiment is that: only adopting a saccharomyces cerevisiae strain as a fermentation strain for fermentation, and specifically comprising the following steps:
(1) and (3) treating the raw materials, namely adding sodium pyrosulfite into the concentrated apple juice to ensure that the effective sulfur dioxide content is 70mg/L, adding pectinase to ensure that the effective content is 60mg/L, adjusting the sugar degree of the apple juice to be 20Brix and the pH value to be 3.8, standing for 24 hours, and subpackaging 200mL of the apple juice in a sterile conical flask. It is worth to be noted that the sodium metabisulfite and the pectinase are added to respectively play roles of bacteriostasis and clarification, so that the subsequent fermented wine has longer storage time and bright and beautiful color;
(2) and (3) strain activation, namely inoculating the saccharomyces cerevisiae preserved at ultralow temperature to a YPD liquid culture medium, and culturing for 24 hours at the temperature of 28 ℃ and the rotating speed of a shaking table of 120rpm for later use. The YPD liquid culture medium is prepared by the following method: 10g of peptone, 5g of yeast extract powder, 20g of glucose and 1L of water are uniformly mixed and sterilized at 121 ℃ for 30 min. (ii) a
(3) Preparing a seed solution: transferring the saccharomyces cerevisiae activated in the step (2) into a YPD liquid culture medium by a culture amount of 8% (16mL) to prepare a seed liquid, wherein the seed culture conditions are as follows: culturing at 28 deg.C and 120rpm for 24 h;
(4) preparing a bacterial liquid: and (4) centrifuging the saccharomyces cerevisiae seed liquid obtained in the step (3) to remove the supernatant to obtain a bacterial liquid. Wherein the parameters of a refrigerated centrifuge used for centrifugation are 4 ℃, and the refrigerated centrifuge is centrifuged for 15min at 5000 rpm/min;
(5) inoculation: inoculating the saccharomyces cerevisiae bacterial liquid prepared in the step (4) into the concentrated apple juice obtained in the step (1), and after inoculation, the final concentration of saccharomyces cerevisiae in the concentrated apple juice is 1 multiplied by 106cfu/mL；
(6) Fermentation: culturing the concentrated apple juice of step (5), monitoring sugar degree and alcoholic strength change every day (as shown in figures 2 and 3), and terminating fermentation when sugar degree is stable, wherein total fermentation period is 12 days. The culture conditions were: the temperature is 28 ℃, and the rotating speed of the shaking table is 120 rpm;
(7) and (3) filtering: and (3) carrying out suction filtration on the fermented cider to remove precipitated substances and fungi, thus obtaining the cider. Wherein the above-mentioned suction filtration uses an organic microporous filtration membrane with a diameter of 50mm and a pore diameter of 0.45 μm.
The physical and chemical indexes of the ciders prepared by the method of the embodiment and the comparative example are detected as follows:
the total acid, reducing sugar and alcoholic strength analysis are measured according to GB/T15038-.
The experimental results are shown in the following table 1, and the analysis results show that the alcoholic strength of the cider is improved, and the contents of reducing sugar and total acid are reduced in the following table 1:
TABLE 1 physical and chemical indexes of cider
(Note: in the table, "+/-" indicates the standard deviation, P < 0.05)
The measurement of the organic acid in cider was carried out by the following method. Specifically, the conditions of sample pretreatment and high performance liquid chromatography analysis in the determination method are as follows:
(1) sample pretreatment: 1mL of cider sample was aspirated by syringe, filtered through 0.22 μm nylon filter, injected into HPLC sample vial, and then placed in an autosampler for sample analysis.
(2) High performance liquid chromatography conditions: HPLC chromatographic condition chromatographic column is Amethyl C18-H (4.6X250mm, 5 μm); the column temperature is 30 ℃; mobile phase A: b ═ water (containing 0.1% formic acid): methanol 97.5: 2.5 (volume ratio); the elution procedure is shown in the table; the flow rate is 0.6 mL/min; the sample injection amount is 20 mu L; the detection wavelength was 210 nm.
The HPLC elution procedure is shown in table 2.
TABLE 2 HPLC elution procedure
| Time (min) | A(％) | B(％) |
| 0 | 97.5 | 2.5 |
| 20 | 97.5 | 2.5 |
| 25 | 0 | 100 |
| 35 | 0 | 100 |
| 40 | 97.5 | 2.5 |
| 55 | 97.5 | 2.5 |
The measurement results are shown in Table 3. The malic acid content in the examples is significantly reduced (P < 0.05). This result may be related to the conversion of L-malic acid to ethanol by Schizosaccharomyces pombe.
TABLE 3 cider organic acid concentration
(Note: in the table, "+/-" indicates the standard deviation, P < 0.05)
Sensory evaluation is carried out on the cider finished product:
the sensory evaluation average score of the product of the example was 95 points (full score of 100 points), and the flavor had a special beer flavor. This result may be related to changes in volatile alcohols, esters, aldehydes, terpenes during fermentation. Referring to the detailed sensory scoring criteria for cider in table 4, 10 panelists were invited to score, and 10 panelists received professional sensory scoring training for each half of the population.
TABLE 4 apple wine sensory evaluation table
In conclusion, the application of the fission yeast schizosaccharomyces pombe and the saccharomyces cerevisiae to sequential inoculation fermentation processing by taking concentrated apple juice as a raw material can improve the quality of apple wine, improve the flavor characteristic of wine body, reduce acidity and improve alcoholic strength. Therefore, the brewing method for improving the flavor characteristic of the cider by using the schizosaccharomyces pombe has good research and application prospects.
Claims (4)
1. A brewing method for improving the flavor characteristics of cider is characterized in that: the method comprises the following steps:
(1) treating raw material, adding sodium pyrosulfite into concentrated apple juice to make effective SO of sodium pyrosulfite2Adding pectinase to obtain effective content of 60mg/L, adjusting sugar degree of apple juice to 20Brix and pH to 3.8, and standing for 24 hr.
(2) First inoculation: inoculating Schizosaccharomyces pombe bacterial liquid into the concentrated apple juice, and adjusting the final concentration of Schizosaccharomyces pombe in the concentrated apple juice to be 1 x 10 after inoculation6cfu/mL；
(3) Carrying out primary fermentation culture on the concentrated apple juice, wherein the culture conditions are as follows: culturing at 28 deg.C and 120rpm for 4 days;
(4) and (3) second inoculation: adding the saccharomyces cerevisiae bacterial liquid into the concentrated apple juice after the primary fermentation culture is finished, performing secondary fermentation, and adjusting the final concentration of saccharomyces cerevisiae in the concentrated apple juice to be 1 × 10 after inoculation6cfu/mL, total fermentation period 12 days.
(5) And carrying out suction filtration on the concentrated apple juice obtained by secondary fermentation to remove precipitated substances and fungi, thus obtaining the apple wine.
2. The brewing method for improving the flavor characteristics of cider according to claim 1, wherein: the suction filtration adopts an organic microporous filtering membrane with the diameter of 50mm and the aperture of 0.45 mu m.
3. The brewing method for improving the flavor characteristics of cider according to claim 1, wherein the method for culturing the schizosaccharomyces pombe solution comprises:
(1) and (3) activating strains, namely inoculating the fission yeast schizosaccharomyces pombe and saccharomyces cerevisiae which are preserved at ultralow temperature to a YPD liquid culture medium, and culturing for 24 hours at the temperature of 28 ℃ and the rotating speed of a shaking table of 120rpm for later use.
(2) Preparing a seed solution: respectively transferring the activated schizosaccharomyces pombe and saccharomyces cerevisiae into YPD liquid culture medium with the culture amount of 8 percent to prepare seed liquid, wherein the seed culture conditions are as follows: culturing at 28 deg.C and 120rpm for 24 hr respectively;
(3) preparing a bacterial liquid: and centrifuging the schizosaccharomyces pombe and saccharomyces cerevisiae seed liquid to remove supernatant liquid to obtain bacterial liquid.
4. The brewing method for improving the flavor characteristics of cider according to claim 3, wherein: the parameters of the refrigerated centrifuge used for centrifugation are 4 ℃, and the refrigerated centrifuge is centrifuged at 6000rpm/min for 15 min.
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| CN115368994A (en) * | 2022-09-14 | 2022-11-22 | 山西农业大学 | Apple fortified wine and preparation process thereof |
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