A method for predicting root and bud loss based on sucrose metabolic enzyme activity during malting and its application

Abstract

The present invention proposes a method for predicting root and bud loss based on the activity of the sucrose metabolic enzyme system in the malting process and its application, which belongs to the field of malting loss prediction and can solve the technical problems that traditional root and bud loss calculation methods are time-consuming, have large hysteresis, and cannot achieve precise control of the malting process. The technical solution includes: the raw barley is subjected to a soaking treatment to obtain dew point barley, the enzyme activity of key enzymes for sucrose synthesis and decomposition in the dew point barley is detected, and the root and bud loss is obtained by using a root and bud loss calculation formula to predict the actual root and bud loss of the raw barley after the entire malting process. The present invention has the characteristics of easy operation, short time consumption, accurate results, and the ability to predict the root and bud loss in the malting process in advance. The present invention can be applied to the prediction of malting loss.

Description

Technical Field

[0001] The present invention belongs to the field of malting loss prediction, and in particular relates to a root and bud loss prediction method based on the activity of sucrose metabolic enzymes in the malting process and an application thereof. Background Art

[0002] Malting loss refers to the percentage of dry matter lost in barley after steeping, germination, drying, and de-rooting. Measuring malting loss can be used to assess the quality of malting methods. Malting loss primarily includes steeping loss, respiratory loss, and root and sprout loss.

[0003] Among them, soaking loss refers to the loss of dry matter content caused by the removal of dust, partial filtration of raw materials and metabolism of grains (releasing carbon dioxide and ethanol) during the soaking stage. The presence of microorganisms may promote the dissolution of grain husks. Respiratory loss mainly comes from the formation of carbon dioxide (a small amount of other volatile metabolites), germination and drying. Respiratory loss is the loss of dry matter caused by the conversion of glucose, the final product of the dissolution of macromolecules including starch, protein, cellulose, etc. through central carbon metabolism into carbon dioxide, ethanol, lactic acid, etc. Root bud loss refers to the loss of dry matter after the root buds are removed after the malting, dehumidification and kilning are completed. Soaking loss accounts for approximately 0.3-0.6%, respiratory loss accounts for 6.5-8.9%, and root bud loss accounts for 2.1-3.8%.

[0004] The maltster's primary goal is to maximize the utilization of starchy raw materials with minimal losses, labor, and expense. It has long been recognized that while producing quality malt, it is essential to understand the sources of malting losses and find ways to reduce them. However, predicting these losses is crucial before minimizing them. While respiratory losses contribute the most, predicting root and sprout losses is equally important.

[0005] Currently, the traditional method for calculating root and sprout loss can only be done by weighing the total root and sprout weight of the malted barley after root removal to calculate the proportion of root and sprout loss during the malting process. This method is time-consuming (generally, the complete malting process takes about 7 days) and has a large lag, making it impossible to predict malting losses in advance and thus guide subsequent malting processes. Therefore, providing a fast and accurate method for predicting root and sprout loss is the key to solving the above problems. Summary of the Invention

[0006] In response to the technical problems that traditional root and bud loss calculation methods are time-consuming, have large hysteresis, and cannot achieve precise control of the malting process, the present invention proposes a root and bud loss prediction method based on the activity of sucrose metabolic enzymes in the malting process and its application. In this method, the raw barley only needs to go through the soaking stage. By measuring the enzyme activity of key enzymes in the barley at the dew point after the soaking, the root and bud loss of the raw barley in the entire malting process can be predicted. The method is easy to operate, short in time (about 4 hours), accurate in results, and can predict the root and bud loss in the malting process in advance, providing data support for subsequent malting technology and process monitoring.

[0007] In order to achieve the above object, the technical solution adopted by the present invention is:

[0008] A method for predicting root and sprout loss based on the activity of sucrose metabolic enzymes during the malting process. Raw barley is subjected to a soaking treatment to obtain dew-point barley. The activity of key enzymes for sucrose synthesis and decomposition in the dew-point barley is detected, and the root and sprout loss is obtained using a root and sprout loss calculation formula to predict the actual root and sprout loss of the raw barley after the entire malting process.

[0009] In one embodiment, the key enzymes for sucrose synthesis and decomposition include sucrose synthase in the SS-I decomposition direction and sucrose synthase in the SS-II synthesis direction.

[0010] In one embodiment, the root and bud loss calculation formula is:

[0011] Root and bud loss ratio (%) = 3.69-0.000362*A+0.00304*B;

[0012] In the above formula, A refers to the enzyme activity of the sucrose synthase in the decomposition direction of SS-I, and B refers to the enzyme activity of the sucrose synthase in the synthesis direction of SS-II.

[0013] In one embodiment, the root and bud loss calculation formula is obtained by the following method:

[0014] Select several enzymes from the enzyme systems related to sucrose synthesis and decomposition as candidate key enzymes;

[0015] Selecting different batches and varieties of raw barley for soaking, and obtaining barley with different dew points after soaking;

[0016] performing enzyme activity determination on the candidate key enzymes in barley of different batches and varieties with dew points after soaking;

[0017] A correlation analysis is performed between the enzyme activity data of the candidate key enzymes and the root bud loss, and the enzyme with a high correlation with the root bud loss is selected as the final key enzyme, and a regression equation is established to obtain the root bud loss calculation formula.

[0018] In one embodiment, the candidate key enzymes include sucrose phosphate synthase, sucrose synthase in the direction of SS-I decomposition, and sucrose synthase in the direction of SS-II synthesis.

[0019] In one embodiment, the enzyme activity of the candidate key enzyme is detected by any one of chemical method, enzyme-linked immunosorbent assay (ELISA), spectrophotometry or liquid chromatography.

[0020] In one embodiment, the accuracy verification step is further included:

[0021] Selecting some batches of raw barley to complete the entire malting process, the dry thousand-kernel weight of the raw barley before malting and the dry thousand-kernel weight of the malted barley after malting were weighed, and the actual root and sprout loss was calculated using the weighing method formula;

[0022] A correlation analysis was performed between the actual root sprout loss and the root sprout loss based on the activity of sucrose metabolic enzymes during the malting process to verify its accuracy.

[0023] In one embodiment, the gravimetric calculation formula is:

[0024]

[0025] In the above formula, M1 refers to the absolute dry thousand-grain weight of the malt that has not been derooted after malting, measured in grams, and M2 refers to the absolute dry thousand-grain weight of the malt that has been derooted after malting, measured in grams.

[0026] In one embodiment, the entire malting process includes a steeping stage, a germination stage, a drying stage, and a root removal stage.

[0027] The present invention also provides an application of the root bud loss prediction method based on the activity of the sucrose metabolic enzyme system in the malting process as described in any of the above embodiments in the regulation of the malting process, thereby reducing the root bud loss by reducing the amount of germination spray water and / or adjusting the germination stirring interval time and / or changing the temperature setting to high at the beginning and low at the end.

[0028] Compared with the prior art, the advantages and positive effects of the present invention are:

[0029] 1. The present invention provides a method for predicting root and bud loss based on the activity of sucrose metabolic enzymes during the malting process and its application. In this method, the raw barley only needs to go through the soaking stage, without going through the entire malting process. The enzyme activity of key enzymes for sucrose synthesis and decomposition in the barley at the dew point after soaking is first measured, and the enzyme activity data is then substituted into a root and bud loss calculation formula to obtain root and bud loss data. This method can predict the root and bud loss of the raw barley throughout the entire malting process.

[0030] 2. The present invention provides a method for predicting root and bud loss based on the activity of sucrose metabolic enzymes during the malting process and its application. This method screens key enzyme systems from multiple enzyme systems related to sucrose synthesis and decomposition, associates the key enzyme systems with the malting loss of the raw barley, and further establishes a formula for calculating root and bud loss to achieve accurate prediction of malting loss.

[0031] 3. The present invention provides a method for predicting root and bud loss based on the activity of sucrose metabolic enzymes in the malting process. The method is easy to operate, short in time (about 4 hours), accurate in results, and can predict the root and bud loss in the malting process in advance, providing data support for subsequent malting technology and process monitoring, and solving the technical problems of traditional malting loss calculation methods that are time-consuming, have large lags, and cannot achieve precise control of the malting process. BRIEF DESCRIPTION OF THE DRAWINGS

[0032] Figure 1 This is the correlation analysis result between the root bud loss calculated based on the key enzyme system and the actual root bud loss provided in the embodiment of the present invention. DETAILED DESCRIPTION

[0033] The following is a clear and complete description of the technical solutions in the embodiments of the present invention. Obviously, the embodiments described are only some embodiments of the present invention, not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative efforts are within the scope of protection of the present invention.

[0034] An embodiment of the present invention provides a method for predicting root and sprout loss based on the activity of sucrose metabolic enzymes during the malting process. Raw barley is subjected to a soaking treatment to obtain dew point barley. The activity of key enzymes for sucrose synthesis and decomposition in the dew point barley is detected, and the root and sprout loss is calculated using a root and sprout loss calculation formula to predict the actual root and sprout loss of the raw barley after the entire malting process.

[0035] The above embodiment provides a method for predicting root and sprout loss based on the activity of sucrose metabolism enzymes during the malting process. This method screens out key enzymes (only two) from numerous enzyme systems related to sucrose synthesis and decomposition (as many as 14-18 types), and associates the key enzymes with the root and sprout loss of the raw material barley during malting. A formula for calculating root and sprout loss is further established to achieve accurate prediction of malting loss. The advantages of the above prediction method are: (1) the raw barley only needs to go through the soaking stage (estimated to be 1-2 days), and does not need to go through the entire malting process like the traditional weighing method (the entire malting process generally takes about 7 days), so that the root bud loss of the raw barley in the entire malting process can be predicted, which shortens the detection time and is convenient and fast; (2) only the enzyme activity of two key enzyme systems needs to be measured, and there is no need to simultaneously detect the enzyme activity of dozens or even dozens of related enzyme systems, so that the root bud loss of the raw barley in the entire malting process can be predicted, which greatly saves workload and shortens detection time; (3) the key enzyme systems (two key enzyme systems) are successfully linked to the root bud loss in malting, and a root bud loss calculation formula is established, which shortens the calculation time and improves the convenience of the operation method; (5) the method provided by the present invention is used to predict the actual root bud loss of the raw barley in advance, providing data support for the malting process, so as to facilitate timely adjustment of the subsequent malting process to achieve the purpose of reducing malting loss.

[0036] In a specific embodiment, the key enzymes for sucrose synthesis and decomposition include sucrose synthase in the decomposition direction of SS-I and sucrose synthase in the synthesis direction of SS-II.

[0037] In the above embodiment, sucrose is one of the final small molecule sugars produced by starch decomposition during the malting process. Sucrose synthase (EC 2.4.1.13) is a bidirectional enzyme that catalyzes both sucrose synthesis and decomposition, making it a key enzyme in sucrose metabolism. Sucrose synthase that decomposes sucrose in the SS-I direction is also referred to as sucrose synthase (SS-I decomposition direction), while sucrose synthase that synthesizes sucrose in the SS-II direction is also referred to as sucrose synthase (SS-II synthesis direction). Specifically, sucrose synthase is one of the key enzymes that facilitates the entry of sucrose into various metabolic pathways and exists in two forms.

[0038] In one embodiment, the root and bud loss calculation formula is:

[0039] Root and bud loss ratio (%) = 3.69-0.000362*A+0.00304*B;

[0040] In the above formula, A refers to the enzyme activity of the sucrose synthase in the decomposition direction of SS-I, and B refers to the enzyme activity of the sucrose synthase in the synthesis direction of SS-II.

[0041] In one embodiment, the root and bud loss calculation formula is obtained by the following method:

[0042] S1. Select several candidate key enzymes from the enzyme systems related to sucrose synthesis and decomposition;

[0043] In step S1, the candidate key enzymes include sucrose phosphate synthase, sucrose synthase for SS-I degradation, and sucrose synthase for SS-II synthesis. The enzyme activity of the candidate key enzymes is detected by chemical method, enzyme-linked immunosorbent assay (ELISA), spectrophotometry, or liquid chromatography.

[0044] S2. Selecting different batches and varieties of raw barley for soaking, and obtaining barley with different dew points after the soaking;

[0045] In the above step S2, the selection criteria for raw barley of different batches and varieties are: barley of different varieties from different origins is selected in order to make the obtained data applicable to different varieties.

[0046] S3. Determining the enzyme activity of the candidate key enzyme in barley of different batches and varieties at the dew point after soaking;

[0047] S4. Perform correlation analysis on the enzyme activity data of the candidate key enzymes and root bud loss, select the enzyme with high correlation with root bud loss as the final key enzyme, and establish a regression equation to obtain the root bud loss calculation formula.

[0048] In one embodiment, the accuracy verification step is further included:

[0049] (1) Selecting some batches of raw barley to complete the entire malting process, weighing the absolute dry thousand-kernel weight of the raw barley before malting and the absolute dry thousand-kernel weight of the malt after malting without root removal, and calculating the actual root and bud loss using the weighing method formula;

[0050] (2) A correlation analysis was conducted between the actual root bud loss and the root bud loss based on the activity of sucrose metabolism enzymes during the malting process to verify its accuracy.

[0051] In a specific embodiment, the weighing method calculation formula is:

[0052]

[0053] In the above formula, M1 refers to the absolute dry thousand-grain weight of the malt that has not been derooted after malting, measured in grams, and M2 refers to the absolute dry thousand-grain weight of the malt that has been derooted after malting, measured in grams.

[0054] In one embodiment, the entire malting process includes a steeping stage, a germination stage, a drying stage, and a root removal stage.

[0055] The present invention also provides an application of the root bud loss prediction method based on the activity of the sucrose metabolic enzyme system in the malting process as described in any of the above embodiments in the regulation of the malting process, thereby reducing the root bud loss by reducing the amount of germination spray water and / or adjusting the germination stirring interval time and / or changing the temperature setting to high at the beginning and low at the end.

[0056] In order to more clearly and in detail introduce the root and bud loss prediction method based on the activity of sucrose metabolic enzymes in the malting process and its application provided by the embodiments of the present invention, the following description will be made in conjunction with specific examples.

[0057] Example 1

[0058] This embodiment provides a method for establishing a formula for calculating root and bud loss, specifically:

[0059] (1) Sucrose is the most abundant starch in barley during germination and is a key substance for synthesis and decomposition during the malting process. The distribution of sucrose synthesis and decomposition capacity indicates the direction of the malting process. The stronger the sucrose decomposition, the more vigorous the malting dissolution; the stronger the sucrose synthesis, the weaker the malting dissolution. Sucrose is transported to the roots for the synthesis of root buds. Therefore, the enzyme system related to sucrose synthesis and decomposition is crucial for predicting the ability to generate root buds. There are many enzyme systems related to sucrose synthesis and decomposition in plants. The key precursors for sucrose synthesis and decomposition mainly come from two pathways, namely the starch hydrolysis pathway and the phosphorylation pathway. Therefore, the intermediate metabolites and key enzyme systems in these two pathways affect sucrose synthesis through positive and negative metabolic regulation. For example, in the Calvin cycle, there are phosphofructoisomerase (PGI), phosphoglucomutase (PGM), pyrophosphorylase (AGPase), sucrose phosphate synthase (SPS), as well as phosphosucrose phosphatase (SPP), α-galactosidase, galactosidase, sucrose invertase, fructokinase, hexokinase, adenosine diphosphate glucose pyrophosphorylase, starch phosphorylase, sucrose synthase, etc.

[0060] In this example, several enzymes related to sucrose synthesis and decomposition are selected as candidate key enzymes. The enzymes related to sucrose synthesis and decomposition are shown in the following table:

[0061] Table 1 Enzymes related to sucrose synthesis and decomposition

[0062]

[0063]

[0064] Based on the importance of enzyme systems in different reaction stages and the difficulty of detection, three enzymes were selected as candidate key enzymes, namely sucrose phosphate synthase, sucrose synthase in the direction of SS-Ⅰ decomposition, and sucrose synthase in the direction of SS-Ⅱ synthesis.

[0065] (2) 10 batches of different varieties of raw barley were selected for soaking. After the soaking, the dew point barley was obtained. The enzyme activity of three candidate key enzymes in the dew point barley was detected. The correlation analysis between the three enzymes and root bud loss was conducted and it was found that the correlation analysis results between different enzyme systems and root bud loss were as follows: Figure 1 As shown in Figure 2 ): The correlation between sucrose synthase in the decomposition direction of SS-I and sucrose synthase in the synthesis direction of SS-II and root bud loss is higher. Therefore, in this example, these two enzymes are selected to construct the root bud loss calculation formula. Specifically, based on the enzyme activity test results, the sucrose synthase in the decomposition direction of SS-I and the sucrose synthase in the synthesis direction of SS-II were subjected to regression analysis, and the regression equation was established as follows:

[0066] Root and bud loss ratio (%) = 3.69-0.000362*A+0.00304*B;

[0067] In the above formula, A refers to the enzyme activity of the sucrose synthase in the decomposition direction of SS-I, and B refers to the enzyme activity of the sucrose synthase in the synthesis direction of SS-II.

[0068] Based on the above-mentioned establishment method and regression equation, it can be seen that in this example, several candidate key enzymes are first selected from a variety of sucrose synthesis and decomposition-related enzyme systems, and then two key enzymes are screened from the candidate key enzymes. By simply measuring the enzyme activities of the two key enzymes, including the sucrose synthase in the SS-I decomposition direction and the sucrose synthase in the SS-II synthesis direction, and substituting them into the above-mentioned calculation formula, the root sprout loss of the raw barley can be calculated, thereby predicting its root sprout loss throughout the entire malting process.

[0069] Example 2

[0070] This example provides a method for detecting the enzyme activity of sucrose synthase (decomposition direction of SS-I), using sucrose synthase (decomposition direction; SS-I) from Gres, catalog number: G0513F. The specific operation is as follows:

[0071] (1) Take approximately 0.1 g of tissue (0.5 g for well-hydrated samples), add 1 mL of pre-cooled 95% ethanol, homogenize on ice, and incubate at 4°C for 10 min.

[0072] (2) Centrifuge at 12000 rpm, 4°C for 5 min; discard the supernatant and retain the precipitate;

[0073] (3) Add pre-cooled 80% ethanol to the precipitate, mix well, and let stand at 4°C for 5 min;

[0074] (4) Centrifuge at 12000 rpm, 4°C for 5 min; discard the supernatant and retain the precipitate;

[0075] (5) Add 1 mL of pre-cooled extract to the precipitate, vortex mix, and incubate at 4°C for 10 min.

[0076] (6) Centrifuge at 12000 rpm, 4°C for 10 min; retain the supernatant and discard the precipitate. Place the supernatant on ice for testing.

[0077] (7) Preheat the visible spectrophotometer for more than 30 minutes, adjust the wavelength to 540 nm, and zero with distilled water;

[0078] (8) All reagents were thawed to room temperature;

[0079] (9) Add the following reagents to the EP tube in sequence:

[0080] Table 2 Reagents

[0081]

[0082] (10) Calculated based on the fresh weight of the sample:

[0083] Unit definition: One unit of enzyme activity is the amount of fructose produced per gram of tissue per minute.

[0084] SS-Ⅰ activity (μg / min / g fresh weight) = [(ΔA+0.0076)÷0.0111]÷(W×V1÷V)÷T×D=150.2×(ΔA+0.0076)÷W×D.

[0085] Example 3

[0086] This example provides a method for detecting the enzyme activity of sucrose synthase (SS-II synthesis direction), using sucrose synthase (synthesis direction; SS-II) from Gres, catalog number: G0512F. The specific operation is as follows:

[0087] (1) Take approximately 0.1 g of tissue (0.5 g for well-hydrated samples), add 1 mL of pre-cooled 95% ethanol, homogenize on ice, and incubate at 4°C for 10 min.

[0088] (2) Centrifuge at 12000 rpm, 4°C for 5 min; discard the supernatant and retain the precipitate;

[0089] (3) Add pre-cooled 80% ethanol to the precipitate, mix well, and let stand at 4°C for 5 min;

[0090] (4) Centrifuge at 12000 rpm, 4°C for 5 min; discard the supernatant and retain the precipitate;

[0091] (5) Add 1 mL of pre-cooled extract to the precipitate, vortex mix, and incubate at 4°C for 10 min.

[0092] (6) Centrifuge at 12000 rpm, 4°C for 10 min; retain the supernatant and discard the precipitate. Place the supernatant on ice until assayed.

[0093] (7) Preheat the spectrophotometer for more than 30 minutes, adjust the wavelength to 480 nm, and zero with distilled water;

[0094] (8) Add the following reagents to the EP tube in sequence:

[0095] Table 3 Reaction reagents

[0096]

[0097] (9) Calculated based on the fresh weight of the sample:

[0098] Unit definition: One unit of enzyme activity is defined as the production of 1 μg of sucrose per gram of tissue per minute.

[0099] SS-Ⅱ activity (μg / min / g fresh weight) = [(ΔA+0.0016)÷0.0042]÷(W×V1÷V)÷T=297.6×(ΔA+0.0016)÷W.

[0100] Example 4

[0101] This example provides a verification test of a method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes during the malting process, specifically:

[0102] After 10 batches of raw barley were soaked, the enzyme activity of the two key enzymes was tested according to the method described in Examples 1-3. The enzyme activity data (results are shown in Table 4) were substituted into the root bud loss calculation formula to obtain the root bud loss data of the 10 batches of raw barley, as shown in Table 5. The root bud loss calculation formula is as follows:

[0103] Root and bud loss ratio (%) = 3.69-0.000362*A+0.00304*B;

[0104] In the above formula, A refers to the enzyme activity of the sucrose synthase in the decomposition direction of SS-I, and B refers to the enzyme activity of the sucrose synthase in the synthesis direction of SS-II.

[0105] The absolute dry thousand-kernel weight of the 10 batches of raw barley before malting and the absolute dry thousand-kernel weight of the malt without root removal after malting were weighed respectively, and the actual root and bud loss was calculated using the weighing method (the calculation results are shown in Table 5). The weighing method calculation formula is as follows:

[0106]

[0107] In the above formula, M1 refers to the absolute dry thousand-grain weight of the malt that has not been derooted after malting, measured in grams, and M2 refers to the absolute dry thousand-grain weight of the malt that has been derooted after malting, measured in grams.

[0108] Table 4 Enzyme activity data of two key enzymes in 10 batches of raw barley

[0109]

[0110]

[0111] Table 5 Detection results of root and sprout loss based on enzyme system and actual root and sprout loss of 10 batches of raw barley

[0112] Raw barley batch number Root and bud loss based on enzyme system / % Actual root and bud loss / % 1 2.43 2.43 2 3.34 3.36 3 3.60 3.75 4 2.60 2.64 5 3.43 3.42 6 3.86 3.85 7 4.33 4.13 8 3.23 3.25 9 3.67 3.82 10 2.44 2.27

[0113] Combined with the data calculated in Table 4-5, this example also conducted a correlation analysis based on the root bud loss obtained by the two key enzyme systems and the actual root bud loss measurement results (see the results in Figure 1 ) It can be seen that the correlation coefficient between the two is R 2 =0.9694, indicating that the root sprout loss obtained based on the two key enzyme systems of the present invention has a strong correlation with the actual root sprout loss in malting. The prediction method provided by the present invention is highly reliable and can reflect the actual root sprout loss in malting.

[0114] Example 5

[0115] This embodiment provides an application example of a method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes during the malting process, specifically:

[0116] In this example, three batches of raw barley were selected to perform enzyme activity tests on two key enzymes according to the methods described in Examples 1-4. The enzyme activity data were substituted into the root and bud loss calculation formula to obtain root and bud loss data for the three batches of raw barley. The calculation results are shown in Table 6:

[0117] Table 6 Test results of enzyme activity, root and bud loss based on enzyme system and actual root and bud loss of three batches of raw barley

[0118]

[0119] Based on the data shown in Table 6, it can be seen that the root bud loss ratio is predicted after the soaking of wheat. After determining the root bud loss ratio, it is found that the calculated root bud loss is high, which may be related to the quality of barley in this season or the seasonal climate. It is necessary to make appropriate adjustments to the malting process. For example, by reducing the amount of germination spray water, adjusting the germination stirring interval, and changing the temperature setting to a high-first-low-last mode, the root bud loss ratio is finally obtained by the traditional weighing method (Example 4). The actual root bud loss ratio is compared with the calculated root bud loss ratio. The results show that the root bud loss ratio is reduced by about 0.38-0.44% after the germination process is adjusted. It can be seen that the root bud loss prediction method based on the activity of the sucrose metabolic enzyme system in the malting process provided by the present invention is easy to operate, short in time, accurate in results, and can predict the root bud loss in the malting process in advance, providing data support for subsequent malting process and process monitoring, and providing guidance for subsequent malting process.

Claims

1. A method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes in the malting process, characterized in that: The raw barley is subjected to a soaking treatment to obtain dew point barley, and the enzyme activities of key enzymes for sucrose synthesis and decomposition in the dew point barley are detected. The root sprout loss is calculated using a root sprout loss calculation formula to predict the actual root sprout loss of the raw barley after the entire malting process. The root and bud loss calculation formula is: Root and bud loss ratio (%) = 3.69-0.000362*A+0.00304*B; In the above formula, A refers to the enzyme activity of sucrose synthase in the direction of SS-Ⅰ decomposition, and B refers to the enzyme activity of sucrose synthase in the direction of SS-Ⅱ synthesis; The root and bud loss calculation formula is obtained by the following method: Select several enzymes from the enzyme systems related to sucrose synthesis and decomposition as candidate key enzymes; Selecting different batches and varieties of raw barley for soaking, and obtaining barley with different dew points after soaking; performing enzyme activity determination on the candidate key enzymes in barley of different batches and varieties with dew points after soaking; A correlation analysis is performed between the enzyme activity data of the candidate key enzymes and the root bud loss, and the enzyme with a high correlation with the root bud loss is selected as the final key enzyme, and a regression equation is established to obtain the root bud loss calculation formula.

2. The method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes in the malting process according to claim 1, characterized in that: The candidate key enzymes include sucrose phosphate synthase, sucrose synthase in the direction of SS-Ⅰ decomposition and sucrose synthase in the direction of SS-Ⅱ synthesis.

3. The method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes in the malting process according to claim 1, characterized in that: The enzyme activity of the candidate key enzyme is detected by any one of chemical method, enzyme-linked immunosorbent assay (ELISA), spectrophotometry or liquid chromatography.

4. The method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes in the malting process according to claim 1, characterized in that: An accuracy verification step is also included: Selecting some batches of raw barley to complete the entire malting process, the dry thousand-kernel weight of the raw barley before malting and the dry thousand-kernel weight of the malted barley after malting were weighed, and the actual root and sprout loss was calculated using the weighing method formula; A correlation analysis was performed between the actual root sprout loss and the root sprout loss based on the activity of sucrose metabolic enzymes during the malting process to verify its accuracy.

5. The method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes in the malting process according to claim 4, characterized in that: The weighing method calculation formula is: ； In the above formula, M1 refers to the absolute dry thousand-grain weight of the malt that has not been derooted after malting, measured in grams, and M2 refers to the absolute dry thousand-grain weight of the malt that has been derooted after malting, measured in grams.

6. The method for predicting root and bud loss based on the activity of sucrose metabolizing enzymes in the malting process according to claim 1, characterized in that: The entire malting process includes a malting treatment stage, a germination stage, a drying stage, and a root removal stage.

7. An application of the root and bud loss prediction method based on the activity of sucrose metabolizing enzymes in the malting process according to any one of claims 1 to 6 in malting process control, characterized in that: Reduce root and bud loss by reducing the amount of water sprayed during germination and / or adjusting the germination stirring interval and / or changing the temperature setting to high at the beginning and low at the end.

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