阅读说明：本技术 一种丢糟酒曲的制备及其酿酒方法 (Preparation of distiller's yeast from waste lees and brewing method thereof ) 是由 任志强 黄科屹 黄治国 卫春会 邓杰 钟迟迪 于 2020-10-23 设计创作，主要内容包括：本发明涉及白酒酿造技术领域,具体涉及一种丢糟酒曲的制备及其酿酒方法。所述丢糟酒曲的制备方法,包括以下步骤：采用发酵用霉菌于麸皮中发酵培养,待出现大量菌丝后,将其烘干/干燥、粉碎,制备得到麸曲；将粮食粉经糊化后制成培养基,向培养基中接入小曲发酵培养得到酵母扩培菌液；将所述麸曲和扩培菌液混合,即得丢糟酒曲。本发明丢糟酒曲的制备方法简单,在获得高活力丢糟曲同时,大幅度节省了成本投入。利用本发明丢糟曲可实现对丢糟剩余淀粉的高效利用。本发明为白酒行业丢糟进一步绿色化回收利用产生经济效益提供一种新思路,对白酒行业发展具有极其深远的意义。(The invention relates to the technical field of white spirit brewing, in particular to a preparation method of distiller's yeast with waste lees and a brewing method thereof. The preparation method of the distiller's yeast with waste lees comprises the following steps: fermenting and culturing mould for fermentation in bran, and drying/drying and crushing after a large amount of hypha appears to prepare bran koji; gelatinizing grain powder to prepare a culture medium, and inoculating a Xiaoqu to the culture medium for fermentation culture to obtain a yeast expanded culture solution; and mixing the bran koji and the propagation culture liquid to obtain the waste distiller's yeast. The preparation method of the distiller's yeast with waste lees is simple, and greatly saves the cost investment while obtaining the distiller's yeast with high activity. The waste distiller's yeast can realize the high-efficiency utilization of the residual starch in the waste distiller's yeast. The invention provides a new idea for further green recycling of the waste lees in the white spirit industry to generate economic benefits, and has a profound significance for the development of the white spirit industry.)

1. A preparation method of distiller's grains distiller's yeast is characterized by comprising the following steps:

fermenting and culturing mould for fermentation in bran, and drying/drying and crushing after a large amount of hypha appears to prepare bran koji;

gelatinizing grain powder, preparing a yeast propagation culture medium, and inoculating a Xiaoqu into the culture medium for fermentation culture to obtain a yeast propagation culture solution;

and mixing the bran koji and the propagation culture liquid to obtain the waste distiller's yeast.

2. The method according to claim 1, further comprising activating the fermentation mold, inoculating the fermentation mold on PDA culture medium, culturing for 48-72h, observing the surface of hyphae to form obvious spores, eluting the spores with sterile water to obtain spore liquid, and inoculating the spore liquid in bran for fermentation culture.

3. The production method according to claim 1 or 2, wherein the mold for fermentation is Rhizopus oryzae or Aspergillus niger.

4. The method according to claim 1, wherein the preparation of the culture medium comprises: pulverizing one or more kinds of grain raw materials, sieving, gelatinizing, and mixing with water.

5. The method of claim 4, wherein the grain comprises sorghum, corn, rice, glutinous rice and wheat;

preferably, the sorghum is taken as a raw material to be crushed;

preferably, a mixture of five grains, namely sorghum, corn, rice, sticky rice and wheat, is used as a raw material to be crushed; wherein the mass ratio of the sorghum, the corn, the rice, the sticky rice and the wheat is 30-50:5-10:15-30:15-30: 10-20.

6. The preparation method of claim 4, wherein the grain raw material is crushed, sieved by a sieve of 80-100 meshes, gelatinized, mixed with a small amount of cold water into the grain, stirred uniformly, added into boiling water and cooked for a moment, and the gelatinized material-water ratio is controlled to be 1: 9, preparing a yeast expanding culture medium; and after cooling to normal temperature, inoculating the Xiaoqu to a culture medium for yeast amplification to obtain an expanded culture solution.

7. The preparation method according to claim 1, wherein the moldy bran and the expanded culture solution are mixed in a mass-to-volume ratio of 1-5:30-80 g/mL.

8. The method for brewing wine by using the distiller's yeast with lost lees prepared by any one of claims 1 to 7, wherein fermented grains are obtained by mixing the distiller's yeast with lost lees, the fermented grains are placed in a thermal insulation barrel for fermentation for 30 to 60 days, and distillation is performed after fermentation to produce wine.

9. The method of claim 8, wherein the fermentation temperature in the holding tank is 25-30 ℃.

Technical Field

The invention relates to the technical field of white spirit brewing, in particular to a preparation method of distiller's yeast with waste lees and a brewing method thereof.

Background

The utilization of vinasse, a byproduct of white spirit brewing, is also the key point of work while the yield of white spirit is improved. The content and proportion of the nutrient components of the vinasse have certain difference due to different raw material processes in wineries in different regions. The nutrient components of the vinasse mainly come from thalli, metabolic products thereof, autolysates, residual raw materials which are not completely formed by saccharification and fermentation and the like. Wherein the solid fermented grains of the Daqu liquor are rich in abundant nutrient substances such as protein, amino acid, fat, mineral substances and the like. In order to further utilize the abundant energy substances, some traditional breweries add distiller's yeast consisting of saccharifying enzyme and active dry yeast into solid fermented grains and then ferment the fermented grains again to obtain more economic benefit. However, the solid fermented grains have the characteristics of difficult utilization of nutrient substances and higher acidity, and cannot provide a good propagation environment for yeast, so that the cost and difficulty of brewing the wine by using the distiller's yeast are greatly improved.

The Chinese invention patent CN1970718B discloses a method for preparing white spirit by fermenting waste lees of the white spirit, and the preparation of the reinforced bran koji comprises the following steps: (1) and (3) expanded culture of TR12 engineering bacteria: selecting a proper amount of spores from an aspergillus candidus gene engineering bacterium TR12 slant test tube preserved at 0-4 ℃ in a Sichuan university laboratory, inoculating the spores on a starch culture medium containing 20g/L of corn starch, 20g/L of agar powder, 2g/L of potassium nitrate, 1.2g/L of magnesium sulfate heptahydrate and 2.7g/L, pH of monopotassium phosphate, culturing for 4-5 days at the temperature of 33-35 ℃ and the humidity of 75-95%, and preserving for later use at the temperature of 0-4 ℃; (2) preparing seed koji: respectively adding 40mL of sterile water and 1 drop of Tween 80 into an expanded TR12 engineering bacterium eggplant bottle, lightly scraping spores on the surface of a culture medium by using a sterile inoculating shovel to prepare spore bacterium suspension, adding 9.0g of bran, 1.0g of corn flour, 7.5mL of tap water, 0.2g of ammonium sulfate, 0.02g of monopotassium phosphate and 0.02g of magnesium sulfate into a 250mL triangular bottle, moistening for 2-3 hours, sterilizing at the temperature of 118-123 ℃ for 20-25 min, cooling to 38-42 ℃, inoculating 0.5-1.0 mL of the spore bacterium suspension into the seed bottle under sterile operation, culturing at the humidity of 75-95% and the temperature of a bending room of 30-32 ℃ for 3-4 days, and drying at the temperature of 40-45 ℃ for later use by hot air; (3) preparing reinforced bran koji: 50.0g of bran, 40.0mL of tap water, 0.1g of monopotassium phosphate, 0.1g of magnesium sulfate and 1.5g of sodium nitrate are added into a double-dish culture dish with the diameter of 150mm, materials are moistened for 2-3 h, the double-dish culture dish is sterilized at the temperature of 118-123 ℃ for 20-25 min, cooled to 38-42 ℃, 1-2% of dry seed koji is inoculated under the aseptic operation, the mixture is cultured at the humidity of 75-95% and the temperature of 37-39 ℃ in a koji chamber for 36-44 h, the temperature is reduced to 32-34 ℃ for continuous culture, the humidity is removed, the mixture is stirred once each time when the culture time is 36h, 40h and 44h, the total culture time is 46-50 h, and finally the mixture is dried at the temperature of 40-45 ℃ and stored in a shade place for later use. The method has complex steps, needs specific strains and is not beneficial to popularization and application.

The Chinese invention patent CN101921689B discloses a method for brewing liquor by secondary fermentation of distiller's grains of Luzhou-flavor liquor and the liquor brewed by the method, which comprises the following steps: (1) spreading and air-drying the waste lees to 10-40 deg.C, adding yeast, stirring, and fermenting the waste lees with yeast for 4-12 days at 10-45 deg.C; (2) adding wine tail, esterifying enzyme and yellow water into the waste lees after the stacking fermentation in the step (1), and uniformly stirring the caproic acid fermentation liquor; (3) sealing the waste lees mixture obtained in the step (2) by using a plastic film, and carrying out stacking fermentation for 15d-50d, wherein the temperature of the stacking fermentation is controlled at 10-50 ℃; white spirit distilled under normal pressure after fermentation is finished; the weight percentage of the starter added in the step (1) is 0.5 to 8 percent of the weight of the waste lees; the weight percentage contents of the wine-tailed wine, the yellow water, the esterifying enzyme and the caproic acid fermentation liquid added in the step (2) are respectively 3-12 percent, 2-8 percent, 0.8-4 percent and 4-7.5 percent of the weight of the waste lees. In the method, after the waste lees are fermented, the wine tail, the esterifying enzyme, the yellow water, the caproic acid fermentation liquor, the esterifying enzyme and other components are added, so that the raw material investment cost is greatly increased.

Based on this, there is still a need to find a method for making full use of the waste lees and reducing the production cost.

Disclosure of Invention

The invention mainly aims to provide a preparation method of distiller's yeast from waste lees and a brewing method thereof. The wine can be mixed with the waste lees for brewing wine without adding other materials, and the higher wine yield can be achieved, thereby overcoming the defects of the prior art.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides a preparation method of distiller's yeast of spent grains, comprising the following steps:

fermenting and culturing mould for fermentation in bran, and drying/drying and crushing the bran after a large amount of hypha appears to prepare bran koji;

gelatinizing grain powder to prepare a culture medium, inoculating xiaoqu into the culture medium, wherein saccharifying enzyme in the xiaoqu can decompose starch in grains and is used as an energy source for yeast fermentation culture to obtain yeast expanded culture solution;

and mixing the bran koji and the yeast propagation culture solution to obtain the waste distiller's yeast.

Preferably, before the fermentation culture, the method also comprises activation culture of the fermentation mould, the fermentation mould is inoculated on a PDA culture medium for culture for 48-72h, obvious spores are observed on the surface of hyphae, the spores are eluted by sterile water to prepare spore liquid, and the spore liquid is inoculated in the bran for fermentation culture.

Preferably, the fermentation mold used should ensure high saccharification ability to utilize the more difficult-to-decompose starch in the spent grain (most of the starch in the spent grain is amylopectin), and includes, but is not limited to, Rhizopus oryzae or Aspergillus niger.

Preferably, the preparation of the culture medium: pulverizing one or more kinds of grain raw materials, sieving, gelatinizing, and mixing with water.

Preferably, the foodstuffs include sorghum, corn, rice, glutinous rice and wheat;

preferably, the sorghum is taken as a raw material to be crushed;

preferably, a mixture of five grains, namely sorghum, corn, rice, sticky rice and wheat, is used as a raw material to be crushed; wherein the mass ratio of the sorghum, the corn, the rice, the sticky rice and the wheat is 30-50:5-10:15-30:15-30: 10-20.

Preferably, the grain can be one or more grains, and the expanding culture yeast cultured by the sorghum culture medium is used for distiller's grains of sorghum for brewing raw materials. The propagation yeast cultured by the five-grain culture medium is used for making wine and using the waste lees of the five grains as raw materials.

Preferably, the grain raw materials are crushed and then sieved by a sieve of 80-100 meshes, the grain raw materials are gelatinized, a small amount of cold water is mixed into the grains and stirred uniformly, then the grains are added into boiling water for cooking for a moment, and the gelatinized material-water ratio is controlled to be 1: 9, preparing the yeast expanding culture medium. And after cooling to normal temperature, inoculating the Xiaoqu to a culture medium for yeast amplification to obtain an expanded culture solution.

Preferably, the moldy bran and the expanded culture solution are mixed according to the mass-volume ratio of 1-5:30-80 g/mL.

Preferably, the fermentation temperature in the heat-preserving barrel is 25-30 ℃.

The invention also provides a method for brewing wine by using the distiller's yeast with waste lees prepared by the method, which comprises the following steps: mixing the distiller's yeast and the distiller's grains to obtain fermented grains, fermenting in a heat insulating barrel for 30-60 days, and distilling to obtain wine.

Compared with the prior art, the invention has the following advantages:

the preparation method of the distiller's yeast with waste lees is simple, and greatly saves the cost investment while obtaining the distiller's yeast with high activity. The waste distiller's yeast can realize the high-efficiency utilization of the residual starch in the waste distiller's yeast. The invention provides a new idea for further green recycling of the waste lees in the white spirit industry to generate economic benefits, and has a profound significance for the development of the white spirit industry.

Detailed Description

It is to be understood that the following detailed description is exemplary and is intended to provide further explanation of the invention as claimed. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.

It is noted that the terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of exemplary embodiments according to the invention. As used herein, the singular forms "a", "an" and "the" are intended to include the plural forms as well, and it should be understood that when the terms "comprises" and/or "comprising" are used in this specification, they specify the presence of the stated features, steps, operations, and/or combinations thereof, unless the context clearly indicates otherwise.

In order to make the technical solutions of the present invention more clearly understood by those skilled in the art, the technical solutions of the present invention will be described in detail below with reference to specific embodiments.

The rhizopus oryzae and aspergillus niger AS3.4309 used in the embodiment of the invention are all commercially available strains.

Example 1

A preparation method of distiller's yeast with waste lees comprises the following steps:

after rhizopus oryzae is selected and inoculated on a PDA culture medium for culturing for 48-72h, obvious cyan black spores are observed on the surface of hyphae. The spores were eluted with sterile water to obtain a spore liquid, inoculated into sterilized bran, and cultured at 30 ℃. Drying/drying and pulverizing to obtain bran koji after a large amount of mycelia appear. A small part of the crushed sample is taken and the glucoamylase activity of the small part of the crushed sample is measured by a Fehling reagent method to be 1091.2U/g.

Five grains of sorghum, corn, rice, glutinous rice and wheat are taken as raw materials, and the five grains are prepared from the following raw materials in percentage by mass: 36% of sorghum, 8% of corn, 22% of rice, 18% of glutinous rice and 16% of wheat are mixed to be used as raw materials. Crushing the raw materials, sieving the crushed raw materials through a 100-mesh sieve, and pasting, wherein the ratio of the pasted materials to water is controlled to be 1: 9, preparing the yeast expanding culture medium. Weighing 1% of commercially available Xiaoqu, inoculating, stirring uniformly, sealing, and culturing at 30 deg.C for 48h to obtain the culture expanding solution.The statistics shows that the number of the yeast in the expanding culture liquid is about 2.4 multiplied by 10⁸cfu/ml。

Taking 50ml of the prepared expanded culture solution per kg, adding 2g of bran koji, and uniformly mixing to prepare the distiller's yeast with waste lees.

The method for brewing wine by adopting the distiller's yeast of waste lees prepared in the embodiment comprises the following steps:

taking 6kg of distiller's grains, adding distiller's yeast (prepared by uniformly mixing 300ml of yeast liquid and 12g of bran yeast), and fermenting the fermented grains in a stainless steel heat-preserving container (30 ℃) for 30 days. And after the fermentation is finished, putting fermented grains into a steamer for steaming, detecting the starch consumption before and after the fermentation, and detecting the liquor yield by distilling the liquor.

TABLE 1 method of example 1 starch consumption and liquor yield results

Example 2

A preparation method of distiller's yeast with waste lees comprises the following steps:

aspergillus niger AS3.4309 is selected to be inoculated on a PDA culture medium for culturing for 48 to 72 hours, and obvious black spores are observed on the surface of hyphae. The spores were eluted with sterile water to obtain a spore liquid, inoculated into sterilized bran, and cultured at 30 ℃. Drying/drying and pulverizing to obtain bran koji after a large amount of mycelia appear. A small part of the crushed sample is taken and the glucoamylase activity of the small part of the crushed sample is measured by a Fehling reagent method to be 1285.7U/g.

Five grains of sorghum, corn, rice, glutinous rice and wheat are taken as raw materials, and the five grains are prepared from the following raw materials in percentage by mass: 36% of sorghum, 8% of corn, 22% of rice, 18% of glutinous rice and 16% of wheat are mixed to be used as raw materials. Crushing the raw materials, sieving the crushed raw materials through a 100-mesh sieve, and pasting, wherein the ratio of the pasted materials to water is controlled to be 1: 9, preparing the yeast expanding culture medium. Weighing 1% of commercially available Xiaoqu, inoculating, stirring uniformly, sealing, and culturing at 30 deg.C for 48h to obtain the culture expanding solution. The statistics shows that the number of the yeast in the expanding culture liquid is about 2.4 multiplied by 10⁸cfu/ml。

Taking 50ml of the prepared expanded culture solution per kg, adding 2g of bran koji, and uniformly mixing to prepare the distiller's yeast with waste lees.

The method for brewing wine by adopting the distiller's yeast of waste lees prepared in the embodiment comprises the following steps:

taking 6kg of waste lees, adding distiller's yeast (prepared by uniformly mixing 300ml of yeast liquid and 12g of bran yeast), and then putting the fermented grains into a stainless steel heat-preserving container for 30 days for fermentation. And after the fermentation is finished, putting fermented grains into a steamer for steaming, detecting the starch consumption before and after the fermentation, and detecting the liquor yield by distilling the liquor.

Table 2 example 2 method starch consumption and liquor yield results

Example 3

A preparation method of distiller's yeast with waste lees comprises the following steps:

aspergillus niger AS3.4309 is selected to be inoculated on a PDA culture medium for culturing for 48 to 72 hours, and obvious black spores are observed on the surface of hyphae. The spores were eluted with sterile water to obtain a spore liquid, inoculated into sterilized bran, and cultured at 30 ℃. Drying/drying and pulverizing to obtain bran koji after a large amount of mycelia appear. A small part of the crushed sample is taken and the glucoamylase activity of the small part of the crushed sample is measured by a Fehling reagent method to be 1285.7U/g.

The sorghum is taken as a raw material, crushed, sieved by a sieve with 100 meshes, and gelatinized, wherein the material-water ratio of the gelatinization is controlled to be 1: 9, preparing the yeast expanding culture medium. Weighing 1% of commercially available Xiaoqu, inoculating, stirring uniformly, sealing, and culturing at 30 deg.C for 48h to obtain the culture expanding solution. The statistics shows that the number of yeast in the expanding culture liquid is about 1.8 multiplied by 10⁸cfu/ml。

Taking 50ml of the prepared expanded culture solution per kg, adding 2g of bran koji, and uniformly mixing to prepare the distiller's yeast with waste lees.

The method for brewing wine by adopting the distiller's yeast of waste lees prepared in the embodiment comprises the following steps:

taking 6kg of waste lees, adding distiller's yeast (prepared by uniformly mixing 300ml of yeast liquid and 12g of bran yeast), and then putting the fermented grains into a stainless steel heat-preserving container for 30 days for fermentation. And after the fermentation is finished, putting fermented grains into a steamer for steaming, detecting the starch consumption before and after the fermentation, and detecting the liquor yield by distilling the liquor.

Table 3 method of example 3 starch consumption and liquor yield results

Example 4

A preparation method of distiller's yeast with waste lees comprises the following steps:

after rhizopus oryzae is selected and inoculated on a PDA culture medium for culturing for 48-72h, obvious cyan black spores are observed on the surface of hyphae. The spores were eluted with sterile water to obtain a spore liquid, inoculated into sterilized bran, and cultured at 30 ℃. Drying/drying and pulverizing to obtain bran koji after a large amount of mycelia appear. A small part of the crushed sample is taken and the glucoamylase activity of the small part of the crushed sample is measured by a Fehling reagent method to be 1091.2U/g.

The sorghum is taken as a raw material, crushed, sieved by a sieve with 100 meshes, and gelatinized, wherein the material-water ratio of the gelatinization is controlled to be 1: 9, preparing the yeast expanding culture medium. Weighing 1% of commercially available Xiaoqu, inoculating, stirring uniformly, sealing, and culturing at 30 deg.C for 48h to obtain the culture expanding solution. The statistics shows that the number of yeast in the expanding culture liquid is about 1.8 multiplied by 10⁸cfu/ml。

Taking 100ml of the prepared expanded culture solution per kg, adding 4g of bran koji, and uniformly mixing to prepare the distiller's yeast with distilled grains.

The method for brewing wine by adopting the distiller's yeast of waste lees prepared in the embodiment comprises the following steps:

taking 6kg of waste lees, adding distiller's yeast (prepared by mixing 600ml of yeast liquid and 24g of bran yeast uniformly), and then putting the fermented grains into a stainless steel heat-preserving container for 30 days for fermentation. And after the fermentation is finished, putting fermented grains into a steamer for steaming, detecting the starch consumption before and after the fermentation, and detecting the liquor yield by distilling the liquor.

Table 4 example 4 starch consumption and liquor yield results

Example 5

A preparation method of distiller's yeast with waste lees comprises the following steps:

aspergillus niger AS3.4309 is selected to be inoculated on a PDA culture medium for culturing for 48 to 72 hours, and obvious black spores are observed on the surface of hyphae. And (3) eluting the spores with sterile water to prepare a spore solution, inoculating the spore solution into sterilized bran, and culturing at the temperature of 25-30 ℃. Drying/drying and pulverizing to obtain bran koji after a large amount of mycelia appear. A small part of the crushed sample is taken and the glucoamylase activity of the small part of the crushed sample is measured by a Fehling reagent method to be 1285.7U/g.

The sorghum is taken as a raw material, crushed, sieved by a sieve with 100 meshes, and gelatinized, wherein the material-water ratio of the gelatinization is controlled to be 1: 9, preparing the yeast expanding culture medium. Weighing 1% of commercially available Xiaoqu, inoculating, stirring uniformly, sealing, and culturing at 30 deg.C for 48h to obtain the culture expanding solution. The statistics shows that the number of yeast in the expanding culture liquid is about 1.8 multiplied by 10⁸cfu/ml。

Taking 100ml of the prepared expanded culture solution per kg, adding 4g of bran koji, and uniformly mixing to prepare the distiller's yeast with distilled grains.

The method for brewing wine by adopting the distiller's yeast of waste lees prepared in the embodiment comprises the following steps:

taking 6kg of waste lees, adding the waste lees distiller's yeast (prepared by uniformly mixing 600ml of yeast liquid and 24g of bran yeast), and then putting the fermented grains into a stainless steel heat-preserving barrel for fermenting for 30 days. And after the fermentation is finished, putting fermented grains into a steamer for steaming, detecting the starch consumption before and after the fermentation, and detecting the liquor yield by distilling the liquor.

TABLE 5 example 5 starch consumption and liquor yield results

The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.