Preparation method of high-concentration ethyl acetate high-ester flavoring wine

Abstract

The invention belongs to the technical field of biological fermentation, particularly relates to a preparation method of high-concentration ethyl acetate high-ester flavoring liquor, and provides the following scheme: the method comprises the following steps: step 1, expanding culture: expanding culture of saccharomyces cerevisiae by using a wort culture medium, and carrying out aerobic culture; step 2, concentration: carrying out reduced pressure distillation concentration on the acetic acid fermentation liquor obtained in the step 1; step three, esterification: mixing the concentrated acetic acid fermentation liquor obtained in the step two with fragrant tail liquor, adjusting the pH value to be below 4, adding immobilized esterifying enzyme, stirring at 40 DEG C, sealing and culturing, and carrying out esterification reaction; step 4, distillation: performing reduced pressure distillation on the esterified material in the step 3; and step 5, dehydration: carrying out dehydration treatment by adopting an inorganic vapor permeable membrane. The high-ester flavoring wine containing high-concentration ethyl acetate is produced through the steps of concentration, esterification, distillation, dehydration and the like, the whole process of the flavoring wine is completed through biological fermentation, the flavoring wine is green and safe, and diversified wine mixing requirements are met.

Description

Technical Field

[0001] This invention relates to the field of bio-fermentation technology, and in particular to a method for preparing a high-concentration ethyl acetate high-ester flavoring wine. Background Technology

[0002] In the brewing and blending system of baijiu, flavoring liquor plays a key role in compensating for the flavor defects of the base liquor, enhancing the main aroma, and improving the fullness and harmony of the liquor. Traditional high-ester flavoring liquor has a low ethyl acetate concentration, which generally does not exceed 20g / L. It also contains more substances and has a more complex aroma, which requires higher standards for blending.

[0003] In existing technologies, pure acetic acid and esterifying enzymes are used to produce ethyl acetate. The concentration of ethyl acetate in the esterification solution does not exceed 15 g / L, which is low. Furthermore, the acetic acid is not produced through pure fermentation, which does not meet the requirements for green and safe food safety. Summary of the Invention

[0004] Based on the technical problems in the background art, the present invention proposes a method for preparing high-concentration ethyl acetate high-ester flavoring wine.

[0005] This invention proposes a method for preparing a high-concentration ethyl acetate flavored liquor, comprising the following steps: Step 1, expansion culture: Brewing yeast is expanded using malt extract culture medium and subjected to aerobic culture to obtain acetic acid fermentation broth; Step 2, concentration: The acetic acid fermentation broth obtained in Step 1 is concentrated by vacuum distillation to obtain concentrated acetic acid fermentation broth; Step 3, esterification: The concentrated acetic acid fermentation broth obtained in Step 2 is mixed with light-aroma tail liquor, the pH is adjusted to below 4, immobilized esterifying enzyme is added, and the mixture is stirred and sealed at 40°C to carry out the esterification reaction; Step 4, distillation: The esterified material in Step 3 is subjected to vacuum distillation, and the distillate is collected; Step 5, dehydration: The distillate obtained in Step 4 is dehydrated using an inorganic vapor permeation membrane to obtain a high-concentration ethyl acetate flavored liquor.

[0006] Preferably, in step one, the brewing yeast is a brewing yeast that can produce high levels of acetic acid; the aerobic culture conditions are culture at 30℃ for 48 hours, and the concentration of acetic acid in the acetic acid fermentation broth reaches 8g / L-12g / L; the preparation method of the malt wort culture medium is as follows: add crushed barley malt to pure water, saccharify at 60℃ for 4 hours, filter, and then heat and sterilize the obtained malt wort and cool it.

[0007] Preferably, in step two, before vacuum distillation concentration, the pH of the acetic acid fermentation broth is adjusted to 8 using a food-grade sodium hydroxide solution; the vacuum distillation concentration conditions are 65°C, and the concentration target is to make the acetic acid content in the concentrated acetic acid fermentation broth reach more than 100g / L.

[0008] Preferably, in step three, the volume ratio of concentrated acetic acid fermentation broth to light-aroma tail liquor is 1:0.5, and the alcohol content of the light-aroma tail liquor is 30-35% vol; the amount of immobilized esterase added is 1% of the total mass of the mixture; the immobilized esterase is the Blue Ocean Bio IM-NE100 immobilized esterase, and it is recovered by filtration and reused after the reaction is completed; food-grade phosphoric acid is used to adjust the pH, and the esterification reaction time is 24 hours.

[0009] Preferably, in step four, the conditions for vacuum distillation are 60°C, and distillation is stopped when the volume of the distillate accounts for one-quarter of the total volume of the material to be distilled.

[0010] Preferably, in step five, the inorganic vapor permeation membrane module operates under vacuum negative pressure; the goal of the dehydration treatment is to reduce the water content of the final flavored wine to below 2%.

[0011] The beneficial effects of this invention are as follows: In this invention, a high-ethyl acetate flavoring liquor is produced through steps such as concentration, esterification, distillation, and dehydration. The entire process of this flavoring liquor is completed using biological fermentation, which is green and safe. The ethyl acetate concentration in the high-ethyl acetate flavoring liquor can reach more than 100.0 g / L. The high-ethyl acetate flavoring liquor has no off-odor and a pure aroma, which meets the requirements for blending and production of baijiu (Chinese liquor).

[0012] In this invention, after the acetic acid fermentation broth is treated in step two, the concentration of acetic acid can reach more than 100.0 g / L.

[0013] In this invention, after the acetic acid fermentation broth is treated in steps two and three, the ethyl acetate content in the esterification liquid can reach more than 25.0 g / L, and the esterification rate of acetic acid can reach more than 25.0%. Attached Figure Description

[0014] Figure 1 This is a schematic diagram of the overall structure of the production apparatus for a method of preparing high-concentration ethyl acetate high-ester flavoring wine proposed in this invention; Figure 2 This is a schematic diagram of the distillate dehydration device for a method of preparing high-concentration ethyl acetate high-ester flavoring wine proposed in this invention. Detailed Implementation

[0015] Reference Figures 1-2 A method for preparing high-concentration ethyl acetate and high-ester flavoring wine. This invention solves the problems of low ethyl acetate concentration, low esterification rate, and impure aroma when using pure biological fermentation to produce high-ester flavoring wine rich in ethyl acetate. This invention is achieved by the following technologies: Step 1: Expand the acetic acid fermentation broth. Use malt extract medium to expand the brewing yeast until the acetic acid concentration reaches about 10g / L. Step 2: Concentrate the acetic acid fermentation broth by vacuum distillation until the acetic acid content reaches 100g / L or higher. Step 3: Esterification of acetic acid fermentation broth. Mix 1 part concentrated acetic acid fermentation broth with 0.5 parts 30-35% vol light aroma tail wine. Adjust the pH to below 4 using food-grade phosphoric acid. Add 1% of Blue Ocean Bio IM-NE100 immobilized esterification enzyme. Stir and seal at 40℃ for 24 hours. Step 4: Distillation of the acetic acid esterification solution. Ethyl acetate is extracted from the acetic acid esterification solution by vacuum distillation. Step 5: Dehydration of the distillate. The distillate is dehydrated using an inorganic vapor permeation membrane until the water content is below 2%, resulting in a high-ester flavoring wine rich in ethyl acetate.

[0016] The method for step one of the acetic acid fermentation broth cultivation is as follows: The brewing yeast used is the brewing yeast purchased by our company from Zhongke Quality Inspection Company, with the product number ZKCC36025, which can produce high levels of acetic acid. The culture medium consists of 250 kg of crushed barley malt added to 1000 L of purified water, saccharified at 60℃ for 4 hours, filtered to obtain wort, heated to 100℃ and held for 15 minutes for sterilization, then cooled to about 30℃, and 30 L of the cultured yeast fermentation broth was added. The mixture was then cultured aerobically at 30℃ for 48 hours to obtain the acetic acid fermentation broth.

[0017] The method for the second step of acetic acid fermentation broth concentration is as follows: First, adjust the pH of the cultured acetic acid fermentation broth to 8 using a 20% food-grade sodium hydroxide solution, then concentrate it by vacuum distillation at 65℃ with cooling water at 4℃, and control the concentration ratio by using a level gauge in the concentration tank.

[0018] The method for step three of esterification of acetic acid fermentation broth is as follows: The IM-NE100 esterifying enzyme from Blue Ocean Biotechnology can be reused approximately 100 times. After each esterification is completed, the esterifying enzyme is filtered out using a filter screen, and then the next batch of esterification is continued according to step three of the method.

[0019] The method for step four of the acetic acid esterification distillation is as follows: distillation is carried out at 60°C under reduced pressure, with cooling water at 4°C, until the distillate accounts for one-quarter of the total volume, and the level is controlled by the level gauges in the concentration tank and the receiving tank.

[0020] The fifth step of the distillate dehydration process is as follows: The inorganic vapor permeation membrane module is a device that can control the size of the membrane pores, allowing substances with small molecular diameters, such as water molecules, to pass through, while preventing substances with large diameters, such as esters, from passing through. The dehydration and concentration of distillate rich in ethyl acetate is achieved by drawing a vacuum negative pressure.

[0021] The present invention will be further described below with reference to embodiments: Example 1: Refer to Figures 1-2A method for preparing a high-concentration ethyl acetate high-ester flavoring wine, the process is as follows: Take 1000L of pure water and 250kg of crushed barley malt, keep warm at 60℃ for 4h for saccharification, filter to obtain malt juice, heat to boiling and keep for 15min for sterilization, cool to about 30℃ with circulating water, add 30L of yeast fermentation liquid seed, and culture at 30℃ aerobic for 48h.

[0022] The fermented acetic acid broth was pumped into... Figure 1 In the apparatus, the pH was adjusted to 8 using a 20% sodium hydroxide solution, and the mixture was concentrated under reduced pressure at 65℃, approximately tenfold. The volume of the concentrated acetic acid fermentation broth was measured to be approximately 108L. 54L of a light-aroma tail liquor with an alcohol content of 30-35% vol was added, and the pH was adjusted to below 4 using food-grade phosphoric acid. 1.08kg of immobilized esterase from the Weilan Bio IM-NE100 reactor was added, and the mixture was incubated at 40℃ with stirring and sealed for 24 hours. After incubation, the esterase was filtered out and reserved for the next batch. Figure 1 The filtered esterification solution was subjected to vacuum distillation at 60°C to extract ethyl acetate, yielding 85 L of distillate, which was then used... Figure 2 The apparatus dehydrates the distillate, yielding approximately 56 L of high-ester flavored liquor. The above process is then repeated, using the same esterifying enzyme repeatedly. The following are the average physicochemical data from 10 repetitions.

[0023] Example 2: Refer to Figures 1-2 A method for preparing a high-concentration ethyl acetate high-ester flavoring wine, the process is as follows: Take 2000L of pure water and 500kg of crushed barley malt, keep warm at 60℃ for 4h for saccharification, filter to obtain malt juice, heat to boiling and keep for 15min for sterilization, cool to about 30℃ with circulating water, add 60L of yeast fermentation liquid seed, and culture at 30℃ aerobic for 48h.

[0024] The fermented acetic acid broth was pumped into... Figure 1 In the apparatus, the pH was adjusted to 8 using a 20% sodium hydroxide solution, and the mixture was concentrated under reduced pressure at 65℃, increasing in concentration by approximately 10 times. The volume of the concentrated acetic acid fermentation broth was measured to be approximately 220L. 110L of a light-aroma tail liquor with an alcohol content of 30-35% vol was added, and the pH was adjusted to below 4 using food-grade phosphoric acid. Then, 2.2kg of immobilized esterase from the Blue Ocean Biotechnology IM-NE100 reactor was added, and the mixture was incubated at 40℃ with stirring and sealed for 24 hours. After incubation, the esterase was filtered out and reserved for the next batch. Figure 1 The filtered esterification solution was subjected to vacuum distillation at 60°C to extract ethyl acetate, yielding 180 L of distillate, which was then used... Figure 2 The apparatus dehydrates the distillate, yielding approximately 120 L of high-ester flavored liquor. The above process is then repeated, using the same esterifying enzyme repeatedly. The following are the average physicochemical data from 10 repetitions.

[0025] Example 3: Reference Figures 1-2 A method for preparing a high-concentration ethyl acetate high-ester flavoring wine, the process is as follows: Take 3000L of pure water and 750kg of crushed barley malt, saccharify at 60℃ for 4h, filter to obtain malt juice, heat to boiling and maintain for 15min, sterilize, cool to about 30℃ with circulating water, add 90L of yeast fermentation liquid seed, and culture at 30℃ aerobic for 48h.

[0026] The fermented acetic acid broth was pumped into... Figure 1 In the apparatus, the pH was adjusted to 8 using a 20% sodium hydroxide solution, and the mixture was concentrated under reduced pressure at 65℃, approximately tenfold. The volume of the concentrated acetic acid fermentation broth was measured to be approximately 330L. 165L of a light-aroma tail liquor with an alcohol content of 30-35% vol was added, and the pH was adjusted to below 4 using food-grade phosphoric acid. Then, 3.3kg of immobilized esterase from the Weilan Bio IM-NE100 reactor was added, and the mixture was incubated at 40℃ with stirring and sealed for 24 hours. After incubation, the esterase was filtered out and reserved for the next batch. Figure 1 The filtered esterification solution was subjected to vacuum distillation at 60°C to extract ethyl acetate, yielding approximately 265 L of distillate, which was then used... Figure 2 The apparatus dehydrates the distillate, yielding approximately 175 L of high-ester flavored liquor. The above process is then repeated, using the same esterifying enzyme repeatedly. The following are the average physicochemical data from 10 repetitions.

[0027] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A method for preparing a high-concentration ethyl acetate high-ester flavoring wine, characterized in that, Includes the following steps: Step 1, Expansion Cultivation: Expand the brewing yeast using malt extract medium and carry out aerobic culture to obtain acetic acid fermentation broth; Step 2, Concentration: The acetic acid fermentation broth obtained in Step 1 is concentrated by vacuum distillation to obtain concentrated acetic acid fermentation broth; Step 3, Esterification: Mix the concentrated acetic acid fermentation broth obtained in Step 2 with the light aroma tail wine, adjust the pH to below 4, add immobilized esterification enzyme, and carry out the esterification reaction by stirring and sealing at 40℃. Step 4: Distillation: The esterified material from Step 3 is subjected to vacuum distillation, and the distillate is collected. Step 5, Dehydration: The distillate obtained in Step 4 is dehydrated using an inorganic vapor permeation membrane to obtain a high-concentration ethyl acetate flavoring wine.

2. The method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to claim 1, characterized in that, In step one, the brewing yeast is a brewing yeast that can produce high levels of acetic acid; The aerobic culture conditions were 30℃ for 48 hours, and the concentration of acetic acid in the acetic acid fermentation broth reached 8g / L-12g / L.

3. The method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to claim 2, characterized in that, In step one, the preparation method of the malt extract culture medium is as follows: add crushed barley malt to pure water, saccharify at 60℃ for 4 hours, filter, and then heat the resulting malt extract to sterilize and cool.

4. The method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to claim 1, characterized in that, In step two, before vacuum distillation and concentration, the pH of the acetic acid fermentation broth is adjusted to 8 using food-grade sodium hydroxide solution.

5. The method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to claim 4, characterized in that, In step two, the vacuum distillation concentration is carried out at 65°C, and the goal of concentration is to make the acetic acid content in the concentrated acetic acid fermentation broth reach more than 100g / L.

6. A method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to any one of claims 1 to 5, characterized in that, In step three, the volume ratio of concentrated acetic acid fermentation liquid to light aroma tail liquor is 1:0.5, and the alcohol content of light aroma tail liquor is 30-35% vol. The amount of immobilized esterase added was 1% of the total mass of the mixture.

7. The method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to claim 6, characterized in that, In step three, the immobilized esterase is the IM-NE100 immobilized esterase from Blue Ocean Biotechnology, and it is recovered by filtration and reused after the reaction is completed.

8. A method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to any one of claims 1 to 5, characterized in that, In step three, food-grade phosphoric acid is used to adjust the pH, and the esterification reaction takes 24 hours.

9. A method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to any one of claims 1 to 5, characterized in that, In step four, the conditions for vacuum distillation are 60°C, and distillation is stopped when the volume of the distillate accounts for one-quarter of the total volume of the material to be distilled.

10. The method for preparing a high-concentration ethyl acetate high-ester flavoring wine according to claim 9, characterized in that, In step five, the inorganic vapor permeation membrane module operates under vacuum and negative pressure. The goal of the dehydration process is to reduce the water content of the final flavored liquor to below 2%.

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