Simple culture method of solid saccharifying enzyme

Abstract

The invention relates to the technical field of beer brewing, in particular to a simple culture method of solid saccharifying enzyme. The solid saccharifying enzyme comprises high-temperature-resistant alpha-amylase, barley amylolytic enzyme, wheat amylolytic enzyme, rice amylolytic enzyme and corn amylolytic enzyme, and the culture method comprises the following steps: step 1, primary culture of the solid saccharifying enzyme: preparing a mixture according to different saccharifying enzymes, and uniformly stirring; step 2, putting the mixture into a constant-temperature incubator, and culturing a primary saccharifying enzyme preparation for 20 days at the temperature of 30 DEG C in the constant-temperature incubator; step three, secondary culture of saccharifying enzyme: adding 1 part of the primary culture strain of the saccharifying enzyme prepared in the step one and the step two into 50 parts of crushed barley malt, 50 parts of crushed wheat malt, 1 part of crushed yeast paste and sterile water according to a material-water ratio of 1: 0.4, operating in a production workshop, and uniformly stirring; controlling the temperature of a constant temperature box to be 30 DEG C and culturing for 4-5 days; and 4, taking out the saccharifying enzyme cultured in the step 3 from the constant temperature box, and refrigerating and storing.

Description

Technical Field

[0001] The invention relates to the technical field of beer brewing, and in particular to a simple culture method of solid saccharifying enzyme. Background Art

[0002] Enzyme preparations are biological products with catalytic functions that are prepared, purified and processed from enzymes. Enzyme preparations are widely used in the fields of medicine, food, beer brewing and chemical industry.

[0003] The production of enzyme preparations includes several steps: raw material preparation, enzyme source screening and cultivation, enzyme fermentation, separation and purification, drying and packaging.

[0004] First of all, the production of enzyme preparations mainly includes extraction, fermentation and genetic engineering. The extraction method is the earliest method used to prepare enzyme preparations. This method extracts the required enzymes from the raw materials of animals, plants or microorganisms, and obtains enzyme preparations after separation and purification. The advantage of the extraction method is that the raw materials are widely available and a mixture of natural enzymes can be obtained, but the disadvantage is that the extraction process is complicated, the purity of the enzyme is not high, and it may contain impurities.

[0005] Secondly, the fermentation method is a method of preparing enzyme preparations by using the ability of microorganisms to produce enzymes under specific conditions. By optimizing the composition of the culture medium and controlling the fermentation conditions, specific enzyme preparations can be produced on a large scale. The advantage of the fermentation method is that it has high production efficiency and can be produced on an industrial scale. However, the disadvantage is that the fermentation process takes a certain amount of time and has high requirements for equipment and technology.

[0006] Finally, genetic engineering is a method of preparing enzyme preparations through genetic engineering technology. This method can introduce specific enzyme genes into host cells suitable for expression and produce enzyme preparations by culturing host cells. The advantage of genetic engineering is that it can produce enzymes that do not exist in nature, and the enzymes have high yields and high purity. The disadvantages are that the technology is complex, the cost is high, and it may involve ethical and safety issues.

[0007] In summary, the current methods for manufacturing enzyme preparations all have different defects. Summary of the invention

[0008] The main purpose of the present invention is to utilize the fermentation method, the ability of microorganisms to produce enzymes under specific conditions, to cultivate enzyme preparations by a simple method, to cultivate enzyme preparations by using beer production raw materials, and to control the fermentation temperature, so as to produce high temperature resistant α-amylase, barley starch decomposing enzyme, wheat starch decomposing enzyme, rice starch decomposing enzyme, and corn starch decomposing enzyme for specific saccharification of beer. A variety of cheap saccharifying enzymes are cultivated by a simple method, the saccharification process of brewing beer is simplified, and the proportion of auxiliary materials for brewing beer is increased.

[0009] Taking wheat white beer as an example, the raw material cost of beer can be reduced by about 40% by changing the ratio of barley malt and wheat malt to 60% unmalted barley and 40% wheat and adding 5% saccharifying enzyme to brew beer.

[0010] In beer brewing, different starch structures require different enzymes to break down. Cereals are the main raw materials for brewing beer, and the starch in them is a polysaccharide. For example, the starch in barley is mainly straight-chain starch and amylopectin. Strains of straight-chain starch are linear molecules formed by connecting glucose units with α-1,4-glycosidic bonds; amylopectin has a branched structure composed of α-1,6-glycosidic bonds in addition to α-1,4-glycosidic bonds.

[0011] The enzymes that decompose starch are mainly α-amylase and β-amylase. α-amylase can randomly hydrolyze α-1,4-glycosidic bonds inside starch molecules, which can quickly reduce the viscosity of starch and produce dextrin and a small amount of maltose and other sugars. β-amylase hydrolyzes maltose units from the non-reducing end of starch in sequence and acts on α-1,4-glycosidic bonds, but it cannot continue to act beyond the α-1,6-glycosidic bond branch point. This is because the difference in starch structure in the raw materials will lead to different modes of action and effects of the enzymes required for decomposition. Therefore, when using different auxiliary materials to brew beer in the saccharification process, it is necessary to use amylase cultivated with the corresponding raw materials.

[0012] When cultivating amylase, adding enzymes to induce enzyme production is a common method. The synthesis of enzymes is affected by both genetic and environmental factors. In some microorganisms, amylase is an induced enzyme. When specific inducers exist in the environment, the expression of related genes will be induced, so that the microorganisms can synthesize amylase.

[0013] Commonly used inducers are mostly starch-containing substances, such as starch, dextrin, etc. If bacteria that can synthesize amylase are cultured in a culture medium containing these substances, they will produce amylase with higher activity. If they are cultured in a glucose solution without starch, they will not produce amylase.

[0014] The advantage of induced enzyme production is that it can significantly increase the production of amylase, thereby improving production efficiency, reducing production costs, and meeting the needs of large-scale industrial production.

[0015] Practical Application Cases In beer brewing, amylase is needed to decompose the starch in malt into fermentable sugars. By adding inducers to cultivate amylase-producing microorganisms, a large amount of amylase can be obtained, thereby better completing the starch hydrolysis process and improving the yield and quality of beer.

[0016] Different raw materials require different amylases to break down the starch they contain due to differences in structure and properties.

[0017] To achieve the above object, the present invention adopts the following technical scheme: a simple culture method of solid saccharifying enzyme, wherein the solid saccharifying enzyme comprises: thermostable α-amylase, barley starch decomposing enzyme, wheat starch decomposing enzyme, rice starch decomposing enzyme, corn starch decomposing enzyme, and the culture method comprises the following steps:

[0018] Step 1: Solid saccharifying enzyme primary culture

[0019] 1) Thermoresistant α-amylase medium: 25 parts of barley malt powder, 25 parts of wheat malt, 1 part of brewer's yeast mud, 1 part of thermoresistant α-amylase, 20 parts of sterile water, stir evenly under aseptic operation;

[0020] 2) Barley starch decomposing enzyme medium: 50 parts of barley flour, 1 part of brewer's yeast paste, 20 parts of sterile water, stir evenly under aseptic operation;

[0021] 3) Wheat starch decomposition enzyme medium: 50 parts of wheat flour, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under aseptic operation;

[0022] 4) Rice starch decomposing enzyme medium: 50 parts of rice flour, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under aseptic operation;

[0023] 5) Corn starch decomposing enzyme culture medium: 50 parts of corn starch, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under sterile operation;

[0024] Step 2: Place the stirred mixture in a constant temperature incubator to culture the first-level saccharifying enzyme preparation. The temperature of the incubator is controlled at 30° C. for 20 days.

[0025] Step 3: Secondary culture of saccharifying enzyme

[0026] 50 parts of crushed barley malt, 50 parts of wheat malt, 1 part of yeast paste, sterile water, and a material-water ratio of 1:0.4 are used as raw materials for culturing enzyme preparations, and five parts are prepared. One part of one of the five strains of primary culture of saccharifying enzymes prepared in step 1 and step 2 is added to each part, and the mixture is operated in a production workshop and stirred evenly; the temperature is controlled at 30° C. in a constant temperature box and the culturing time is 4-5 days;

[0027] Step 4: Take out the saccharifying enzyme cultured in step 3 from the incubator and store it in a refrigerator.

[0028] The invention has the beneficial effects of using raw materials such as barley malt, wheat malt, yeast mud and the like for beer production, and the cultured crude saccharifying enzyme emits a light fragrance, has no other peculiar smell, is safe, simple, reliable, and low in cost, and can be directly used in production without purification. The addition amount of the enzyme preparation is controlled to be 0.5%-5% of the dry material amount, the addition amount of ungerminated wheat and barley is controlled to be 40%-100%, and the homemade saccharifying enzyme is added to brew beer, and the wort is filtered by a rotary vibrating screen or a filter press, and the raw material leaching rate can reach about 64%. DETAILED DESCRIPTION

[0029] The technical solutions in the embodiments of the present invention will be clearly and completely described below in conjunction with the embodiments of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, rather than all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0030] A simple method for culturing solid saccharifying enzymes, the solid saccharifying enzymes comprising: thermostable α-amylase, barley starch decomposing enzyme, wheat starch decomposing enzyme, rice starch decomposing enzyme, corn starch decomposing enzyme, and the culturing method comprising the following steps:

[0031] Step 1: Solid saccharifying enzyme primary culture

[0032] 1) Thermoresistant α-amylase medium: 25 parts of barley malt powder, 25 parts of wheat malt, 1 part of brewer's yeast mud, 1 part of thermoresistant α-amylase, 20 parts of sterile water, stir evenly under aseptic operation;

[0033] 2) Barley starch decomposing enzyme medium: 50 parts of barley flour, 1 part of brewer's yeast paste, 20 parts of sterile water, stir evenly under aseptic operation;

[0034] 3) Wheat starch decomposition enzyme medium: 50 parts of wheat flour, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under aseptic operation;

[0035] 4) Rice starch decomposing enzyme medium: 50 parts of rice flour, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under aseptic operation;

[0036] 5) Corn starch decomposition enzyme culture medium: 50 parts of corn starch, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under sterile operation;

[0037] Step 2: Place the stirred mixture in a constant temperature incubator to culture the first-level saccharifying enzyme preparation. The temperature of the incubator is controlled at 30° C. for 20 days.

[0038] Step 3: Secondary culture of saccharifying enzyme

[0039] 50 parts of crushed barley malt, 50 parts of wheat malt, 1 part of yeast paste, sterile water, and a material-water ratio of 1:0.4 are used as raw materials for culturing enzyme preparations, and five parts are prepared. One part of one of the five strains of primary culture of saccharifying enzymes prepared in step 1 and step 2 is added to each part, and the mixture is operated in a production workshop and stirred evenly; the temperature is controlled at 30° C. in a constant temperature box and the culturing time is 4 days;

[0040] Step 4: Take out the saccharifying enzyme cultured in step 3 from the incubator and store it in a refrigerator.

[0041] Experimental analysis and comparison

[0042] Comparison of the raw material formula of traditional wheat white beer and the improved wheat white beer formula:

[0043] 1. Traditional wheat white beer raw material formula: 120kg barley malt, 80kg wheat malt (saccharification process omitted), output 1000L, original wort concentration 12.6°P, leaching rate 63%.

[0044] 2. The raw material formula of improved wheat white beer is: 120kg barley malt, 80kg wheat, 0.4kg wheat amylolytic enzyme (saccharification process omitted), output 1000L, original wort concentration 12.8°P, and leaching rate 64%.

[0045] Saccharifying enzyme is used in the saccharification process. The cultured enzyme preparation is refrigerated at low temperature and has a shelf life of 180 days.

[0046] The wort extraction rate of the improved wheat white beer formula is slightly higher than that of the traditional wheat white beer raw material formula. There is no obvious difference in the taste of the finished beers of the two. However, the cost is reduced by changing from 80kg of wheat malt to 80kg of wheat.

[0047] Although the present invention has been described in detail with reference to the aforementioned embodiments, it is still possible for those skilled in the art to modify the technical solutions described in the aforementioned embodiments, or to make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. A simple method for culturing solid saccharifying enzyme, characterized in that: The solid saccharifying enzyme includes: thermostable α-amylase, barley starch decomposing enzyme, wheat starch decomposing enzyme, rice starch decomposing enzyme, corn starch decomposing enzyme, and the culturing method includes the following steps: Step 1: Solid saccharifying enzyme primary culture 1) Thermoresistant α-amylase medium: 25 parts of barley malt powder, 25 parts of wheat malt, 1 part of brewer's yeast mud, 1 part of thermoresistant α-amylase, 20 parts of sterile water, stir evenly under aseptic operation; 2) Barley starch decomposing enzyme medium: 50 parts of barley flour, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under aseptic operation; 3) Wheat starch decomposition enzyme medium: 50 parts of wheat flour, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under aseptic operation; 4) Rice starch decomposing enzyme medium: 50 parts of rice flour, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under aseptic operation; 5) Corn starch decomposing enzyme culture medium: 50 parts of corn starch, 1 part of brewer's yeast mud, 20 parts of sterile water, stir evenly under sterile operation; Step 2: Place the stirred mixture in a constant temperature incubator to culture the first-level saccharifying enzyme preparation. The temperature of the incubator is controlled at 30° C. for 20 days. Step 3: Secondary culture of saccharifying enzyme 50 parts of crushed barley malt, 50 parts of wheat malt, 1 part of yeast paste, sterile water, and a material-water ratio of 1:0.4 are used as raw materials for culturing enzyme preparations, and five parts are prepared. One part of one of the five strains of primary culture of saccharifying enzymes prepared in step 1 and step 2 is added to each part, and the mixture is operated in a production workshop and stirred evenly; the temperature is controlled at 30° C. in a constant temperature box and the culturing time is 4-5 days; Step 4: Take out the saccharifying enzyme cultured in step 3 from the incubator and store it in a refrigerator.