CN115372545A - Method for analyzing saccharifying power of wine brewing yeast - Google Patents
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Abstract
The invention provides a method for analyzing saccharifying power of wine making Daqu, and relates to the technical field of saccharifying power analysis. The method for analyzing the saccharifying power of the brewing yeast comprises the following experimental analysis steps of S1: preparing a yeast sample solution, placing a yeast sample in a beaker, adding a sodium acetate buffer solution, adding water into the beaker, stirring the solution in the beaker, placing the solution in a constant-temperature water bath, filtering the solution in the beaker, and collecting filtrate to obtain the yeast sample solution for later use; s2: and (5) measuring the Daqu sample solution. According to the method for analyzing the glycation force, through two different experiments of a blank experiment and a formal experiment, the glucose consumption volume under different experimental states is recorded and counted, and then the data of the glycation force is calculated through a specific formula, so that relatively accurate glycation force data is obtained, the measurement is not influenced by a plurality of factors in the traditional manual bottom dropping method, and the data accuracy is high.
Description
Technical Field
The invention relates to the technical field of saccharifying power analysis, in particular to a saccharifying power analysis method of a brewing yeast.
Background
Saccharification, which refers to the process of hydrolyzing starch into sweet products, is the main process of starch sugar manufacturing process and also the main process of many intermediates in the fermentation process of distiller's yeast, and is basically divided into three types according to the sweetness and the corresponding physicochemical properties of the required products: acid method, enzyme method, and enzyme-binding method.
When the saccharifying power of the brewing yeast is analyzed and detected, the traditional analysis and detection mode is a manual titration method, but the measurement of the manual titration method is influenced by various factors such as titration speed, evaporation capacity, stirring speed, heating power, end point judgment and the like, so that the problems that certain deviation occurs in measured data of the brewing yeast, the saccharifying power value is inaccurate and the like easily exist, and therefore, the technical personnel in the field provide the saccharifying power analysis method for the brewing yeast.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a method for analyzing the saccharifying power of a brewing yeast, which solves the problems of inaccurate saccharifying power value and easy deviation caused by more influenced factors in the traditional analysis mode.
In order to achieve the purpose, the invention is realized by the following technical scheme: a method for analyzing saccharifying power of brewing yeast comprises the following experimental analysis steps:
s1: preparing a yeast sample solution, placing a yeast sample in a beaker, adding a sodium acetate buffer solution, adding water into the beaker, stirring the solution in the beaker, placing the solution in a constant-temperature water bath, filtering the solution in the beaker, and collecting filtrate to obtain the yeast sample solution for later use;
s2: measuring Daqu sample liquid, and respectively obtaining the volume V of glucose consumed in each test through two test operations of blank test and formal test 1 And V 2 ；
S3: analyzing and calculating the result, namely obtaining the result in the step S2Glucose data V of consumption 1 And V 2 Carrying out calculation in a calculation formula;
s4: and obtaining the saccharification capacity value.
Preferably, the specific step of preparing the Daqu sample solution in S1 is to weigh 10g of an absolutely dry sample amount according to the measured moisture of the Daqu sample, the absolute dry sample amount is accurate to 0.001g, the weighed sample amount is put into a 250mL beaker, 20mL of sodium acetate buffer solution is added, water is added, a glass rod is used for stirring uniformly, the volume is constant to 200mL, then the beaker is placed into a 35 ℃ constant temperature water bath for heat preservation and immersion for 1h, and filtration and filtrate collection are carried out to obtain the Daqu sample solution for later use.
Preferably, the specific operation steps of the S2 step hollow white test are adding 25.0mL soluble starch solution into a test tube, adding 5.0mL Daqu sample solution, shaking up, adding 20% NaOH solution lmL, absorbing 5.0mL blank solution into 150mL conical flasks containing 5.0mL each of the first solution and the second solution of the feilin, adding 50mL water, shaking up, heating to boiling, keeping the solution in the flasks slightly boiling for 2min, adding 2 drops of methylene blue indicator, continuously titrating with glucose standard solution in the lmin under boiling state until blue just disappears as an end point, and recording the volume V of consumed glucose 1 。。
Preferably, the specific operation of the fixing experiment in the step S2 is that 25.0mL of soluble starch solution is added into another test tube, 5.0mL of Daqu sample solution is added, the solution is shaken up and placed in a warm water bath at 35 ℃, the solution is saccharified for 1h, after 1mL of 20-percent NaOH solution is added, 5.0mL of saccharified solution is absorbed into 150mL conical flasks containing 5.0mL of each of the first solution and the second solution of the fiolin, 50mL of water is added, the mixture is shaken up and then heated to boiling, the solution in the flasks is kept slightly boiled for 2min, 2 drops of methylene blue indicator are added, the titration is continued in lmin in a boiling state until blue just disappears, and the volume V of consumed glucose is recorded 2 。
Preferably, the glycation ability calculation formula in the step S3 is
In the formula, the saccharifying power of X samples is U; v1 one-to-one titrationThe volume of the glucose standard solution is consumed in milliliters (mL) when the sample is titrated by V2; 2.5 one-to-one milligram (mg) of glucose per milliliter of glucose standard solution; 30, the total volume of the saccharified mixed solution (soluble starch solution and yeast sample solution) is 0.25 to 5 milliliters (mL) of yeast sample solution which is equivalent to the mass of yeast, and the unit is gram (g); 5 titration the volume of the mash withdrawn in milliliters (mL).
Preferably, the calculation result in the step S4 is retained to one decimal place, and the absolute difference between the two independent measurement results obtained in the step S4 under the repeated condition should not exceed 10% of the arithmetic average value.
Preferably, the specific preparation steps of the feilin solution are that 69.28g of copper sulfate (CuS04.5H20) is weighed firstly and then dissolved and diluted to 1000mL by water; finally obtaining the solution of the fibrin.
Preferably, the fibulin solution b is prepared by firstly weighing 346g of potassium sodium tartrate and 100g of sodium hydroxide, then dissolving and diluting with water to 1000mL, then shaking up the aqueous solution, and finally filtering the aqueous solution after shaking up to obtain the fibulin solution b.
Preferably, the heating device used after shaking the mixed liquid prepared in the blank experiment and the main experiment in the step S2 is an electric furnace, and the mixed liquid prepared in the blank experiment and the main experiment is heated to boiling by the electric furnace.
Preferably, the PH of the sodium acetate monobasic buffer solution added to the yeast sample solution prepared in the step S1 is 4.6.
The invention provides a saccharifying power analysis method of a brewing yeast. The method has the following beneficial effects:
according to the method, through two different experiments of a blank experiment and a formal experiment, the glucose consumption volumes under different experimental states are recorded and counted, then the data of the glycation force is calculated through a specific formula, then the calculated data of the glycation force is reserved to one position behind a decimal point, and finally the absolute value of the measured data does not exceed 10% of the arithmetic mean value by comparing the data measured independently twice, so that relatively accurate glycation force data is obtained, the measurement is not influenced by a plurality of factors in the traditional manual bottom dropping method, and the data accuracy is high.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The embodiment is as follows:
the embodiment of the invention provides a saccharifying force analysis method of brewing yeast for making wine, which comprises the following experimental analysis steps of:
s1: preparing a yeast sample solution, placing a yeast sample in a beaker, adding a sodium acetate buffer solution, adding water into the beaker, stirring the solution in the beaker, placing the solution in a constant-temperature water bath, filtering the solution in the beaker, and collecting filtrate to obtain the yeast sample solution for later use;
s2: measuring Daqu sample liquid, and respectively obtaining the volume V of glucose consumed in each test through two test operations of blank test and formal test 1 And V 2 ；
S3: analyzing and calculating the result, and analyzing and calculating the consumed glucose data V obtained in the step S2 1 And V 2 Carrying out calculation in a calculation formula;
s4: and obtaining the value of the saccharifying power.
Sampling mode of Daqu sample: respectively extracting 1 bent block at four corners and central positions of the top layer and the lower layer of the stack of the bent blocks of the Qukuqu, extracting 10 bent blocks in total, and respectively checking the sensory quality condition of the 10 bent blocks. After sensory test, 10 pieces of yeast samples are crushed, the yeast powder is evenly mixed and then is subjected to sample separation and sieving, and small samples are reserved for detecting full quality indexes.
The sensory quality standard of the Daqu is shown in Table 1
Table 1: daqu sensory quality standard
The sensory quality of the third Daqu is shown in Table 2:
table 2:2021 year third round Daqu sensory quality report
| Serial number | Sample number | Tracing number | First stage | Second stage | Three-stage |
| 1 | 21090730808 | 80％ | 20％ | ||
| 2 | 21090530909 | 80％ | 10％ | 10％ | |
| 3 | 21090331010 | 70％ | 30％ | ||
| 4 | 21083031313 | 80％ | 10％ | 10％ | |
| 5 | 21082631717 | 60％ | 30％ | 10％ | |
| 6 | 21082831554 | 80％ | 10％ | 10％ | |
| 7 | 21082320255 | 80％ | 20％ | ||
| 8 | 21082120356 | 90％ | 10％ | ||
| 9 | 21081920457 | 80％ | 20％ | ||
| 10 | 21090131458 | 70％ | 30％ |
The third round of Daqu full quality index condition is shown in Table 3.
Table 3: full quality index report of third round of Daqu in 2021 years
The experimental principle of the analysis method is as follows: the saccharifying amylase in the yeast can hydrolyze starch to generate glucose. The glycation ability was expressed by sequentially hydrolyzing a-1, 4-glucosidic bonds of starch from the non-reducing end of the starch under a predetermined condition to produce glucose and measuring the amount of glucose produced by the Fisher method.
The specific steps for preparing the Daqu sample solution in the S1 are that according to the measured moisture of the Daqu sample, 10g of the absolute dry sample is weighed and is accurate to 0.001g, the absolute dry sample is placed into a 250mL beaker, 20mL of acetic acid-sodium acetate buffer solution is added, water is added, a glass rod is used for stirring uniformly, the volume is determined to be 200mL, then the beaker is placed into a 35 ℃ constant temperature water bath for heat preservation and immersion for 1h, filtration is carried out, and filtrate is collected to obtain the Daqu sample solution for later use.
Before blank experiments and formal experiments are started, firstly, preparing a used fiyline solution, wherein the specific preparation steps of a first solution and a second solution in the fiyline solution are as follows, and the first solution and the second solution are reserved after the preparation of the fiyline solution is finished;
the method comprises the following steps of calibrating by using prepared standby Fehling solution, wherein the calibration comprises a preliminary experiment and a formal experiment, and the preliminary experiment comprises the following specific steps: sucking 5.0mL of each of the Fehling's A and B solutions into a 150mL conical flask, adding 50mL of water, shaking up, heating on an electric furnace to boil, titrating with a prepared glucose standard solution in a boiling state, adding two drops of a methylene blue indicator when the blue color of the solution disappears, continuing to drip until the blue color just disappears as an end point, and recording the volume of the consumed glucose standard solution;
first-level and second-level curve index report (physicochemical)
First and second grade yeast sample index report (microorganism)
The specific operation steps of the formal experiment are as follows: sucking 5.0mL of each of the solutions of the Fehling A and the Fehling B into a 150mL conical flask, adding 50mL of water and a glucose standard solution which is less than lmL of a preliminary test, shaking up, heating on an electric furnace to boil, keeping slightly boiling for 2min, adding two drops of a methylene blue indicator, titrating the mixture to an end point in lmin with the glucose standard solution in a boiling state, and recording the total volume V of the consumed glucose standard solution;
acetic acid sodium acetate buffer solution (pH 4.6), 164g of anhydrous sodium acetate (CH 3 COONa) was weighed out, dissolved in water, 114mL of glacial acetic acid was added, and diluted with water to 1000mL. The pH of the buffer solution should be corrected with an acidimeter;
taking 2g of soluble starch solution (20 g/L), drying 2h of the soluble starch solution at 100-105 ℃ until the weight is accurate to 0.001g, mixing the soluble starch solution with water into paste, continuously stirring and injecting 70mL of boiling water, stirring and boiling for 2min until the solution is completely transparent, cooling the solution to room temperature, completely transferring the solution to a 100mL volumetric flask, and fixing the volume, wherein the solution is ready for use;
weighing 2.5g of anhydrous glucose which is dried to constant weight at 103-105 ℃ and is weighed to be accurate to 0.0001g by a glucose standard solution (2.5 g/L), dissolving the anhydrous glucose with water and fixing the volume to 1000mL. The solution needs to be prepared on the same day;
weighing 20g of sodium hydroxide solution (the mass fraction is 20%) into a beaker, adding 80mL of water for dissolving, and preparing a constant-temperature water bath for later use;
after the experimental preparation is completed, the saccharifying power analysis experiment of the brewing yeast for making wine can be carried out.
S2 step the specific procedure of the blank experiment is to add 25.0mL of soluble starch solution into a test tube, add 5.0mL of Daqu sample solution, shake it evenly, add 20% NaOH solution lmL, absorb 5.0mL as blank solutionAdding 50mL of water into a 150mL conical flask containing 5.0mL of each of the solutions A and B, shaking up, heating to boil, keeping the solution in the flask slightly boiling for 2min, adding 2 drops of a methine blue indicator, continuously titrating in lmin with a glucose standard solution in the boiling state until blue just disappears, and recording the volume V of glucose consumed by the bottles 1 。
The specific operation steps of the sample fixing experiment in the step S2 are that 25.0mL of soluble starch solution is added into another test tube, 5.0mL of Daqu sample solution is added, the mixture is shaken up and placed in a warm water bath at 35 ℃, the time is accurately counted, saccharification is carried out for 1h, after 1mL of 20-percent NaOH solution is added, 5.0mL of saccharification solution is absorbed into 150mL conical flasks containing 5.0mL of the first and second fibulin solutions respectively, 50mL of water is added, the mixture is shaken up and then heated to boil, the solution in the flasks is kept to be slightly boiled for 2min, 2 drops of methylene blue indicator are added, the glucose standard solution is continuously titrated in lmin in the boiling state until the blue color just disappears as the end point, and the volume V of the consumed glucose is recorded 2 。
The saccharifying power calculation formula in the step S3 is
In the formula, the saccharifying power of X samples is U; v1 is titrated one by one, the volume of the glucose standard solution is consumed in milliliters (mL); 2.5 one-to-one milligram (mg) of glucose per milliliter of glucose standard solution; 30, the total volume of the saccharified mixed solution (soluble starch solution and yeast sample solution) is 0.25 to 5 milliliters (mL) of yeast sample solution which is equivalent to the mass of yeast, and the unit is gram (g); 5 titration the volume of the mash withdrawn in milliliters (mL).
And (4) retaining the calculation result one digit after the decimal point in the step S4, and under the repeated condition in the step S4, obtaining the absolute difference value of the two independent measurement results which is not more than 10 percent of the arithmetic mean value.
The analysis method comprises the steps of recording and counting glucose consumption volumes under different experimental states through two different experiments of a blank experiment and a formal experiment, calculating the data of the glycation force through a specific formula, reserving the calculated data of the glycation force to one position behind a decimal point, and comparing the data of two independent determinations to obtain relatively accurate glycation force data, wherein the absolute value of the determined data does not exceed 10% of the arithmetic mean value, and the determination is not influenced by a plurality of factors in the traditional manual bottom dropping method, so that the data accuracy is high.
The specific preparation steps of the feilin solution are that 69.28g of copper sulfate (CuS04.5H20) is weighed firstly, and then dissolved and diluted to 1000mL by water; finally obtaining the Fehling solution A.
The specific preparation steps of the solution B of the Fehling solution are that firstly 346g of sodium potassium tartrate and 100g of sodium hydroxide are weighed, then dissolved by water and diluted to 1000mL, then the aqueous solution is shaken up, and finally the shaken up aqueous solution is filtered to obtain the solution B of the Fehling solution.
And (3) a heating device adopted after shaking up the mixed liquid prepared in the blank experiment and the formal experiment in the step (S2) is an electric furnace, the mixed liquid prepared in the blank experiment and the formal experiment is heated to boiling by the electric furnace, the mixed liquid can be replaced by other heating devices when being heated and prepared, and the specific device can be replaced by a specific device on the experiment site.
The pH value of the sodium acetate buffer solution added in the yeast solution prepared in the step S1 is 4.6, the subsequent experiment is convenient to carry out by accurately controlling the pH value of the sodium acetate buffer solution, and the accuracy of the data of the subsequent glycation ability experiment can be improved.
Finishing the tracking sampling of the Daqu and the preparation of a Daqu crushed sample, extracting 8 Daqu samples from 4 Daqu pools, and finishing the detection of 8 samples, wherein the detection results are detailed in tables 8 and 9.
Table 8: daqu detection data report (physicochemical)
Table 9: daqu detection data report (microorganism)
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (10)
1. The method for analyzing the saccharifying power of the brewing yeast is characterized by comprising the following experimental analysis steps of:
s1: preparing a yeast sample solution, placing a yeast sample in a beaker, adding a sodium acetate buffer solution, adding water into the beaker, stirring the solution in the beaker, placing the solution in a constant-temperature water bath, filtering the solution in the beaker, and collecting filtrate to obtain the yeast sample solution for later use;
s2: measuring Daqu sample liquid, and respectively obtaining the volume V of glucose consumed in each test through two test operations of blank test and formal test 1 And V 2 ；
S3: analyzing and calculating the result, and analyzing and calculating the consumed glucose data V obtained in the step S2 1 And V 2 Carrying out calculation in a calculation formula;
s4: and obtaining the value of the saccharifying power.
2. The method for analyzing saccharifying power of brewing yeast according to claim 1, characterized in that: the preparation method of the Daqu sample solution in the S1 comprises the specific steps of weighing 10g of absolutely dry sample according to the measured moisture of a Daqu sample, accurately measuring the sample to 0.001g, putting the sample into a 250mL beaker, adding 20mL of sodium acetate buffer solution, adding water, uniformly stirring by using a glass rod, metering the volume to 200mL, putting the beaker into a 35 ℃ constant-temperature water bath, preserving heat, immersing for 1h, filtering, and collecting filtrate to obtain the Daqu sample solution for later use.
3. The method for analyzing saccharifying power of brewing yeast according to claim 1, characterized in that: the specific operation steps of the hollow white test in the step S2 are that 25.0mL of soluble starch solution is added into a test tube, 5.0mL of Daqu sample solution is added, the test tube is shaken evenly, after 20% of NaOH solution lmL is added, 5.0mL of the Daqu sample solution is absorbed to be used as blank solution to be added into 150mL conical flasks containing 5.0mL of the first solution and the second solution respectively, 50mL of the blank solution is added, the mixture is shaken evenly and then heated to boiling, the solution in the flasks is kept to be slightly boiled for 2min, 2 drops of methylene blue indicator are added, the titration is continued in the lmin under the boiling state until the blue color just disappears to the end point, and the volume V of consumed glucose is recorded 1 。
4. The method for analyzing saccharifying power of brewing yeast according to claim 1, characterized in that: the specific operation steps of the sample fixing experiment in the step S2 are that 25.0mL of soluble starch solution is added into another test tube, 5.0mL of Daqu sample solution is added, the mixture is shaken up and placed in a warm water bath at 35 ℃, the time is accurately counted, saccharification is carried out for 1h, after 1mL of 20-percent NaOH solution is added, 5.0mL of saccharification solution is absorbed into 150mL conical flasks containing 5.0mL of the first and second fibaline solutions respectively, 50mL of water is added, the mixture is shaken up and then heated to boil, the solution in the flasks is kept to be slightly boiled for 2min, 2 drops of methylene blue indicator are added, the glucose standard solution is continuously titrated in lmin in a boiling state until blue disappears just as an end point, and the volume V of consumed glucose is recorded 2 。
5. The method for analyzing saccharifying power of brewing yeast according to claim 1, characterized in that: the saccharifying force calculation formula in the step S3 is
In the formula, the saccharifying power of X samples is U; v1 is titrated one by one, the volume of the glucose standard solution is consumed in milliliters (mL); 2.5 glucose mass in milligrams (mg) per milliliter of the glucose standard solution; 30, the total volume of saccharification mixed liquor (soluble starch solution and yeast sample liquid) is 0.25 milliliter (mL), 5mL of yeast sample liquid is equivalent to the mass of yeast, and the unit is gram (g); 5 titration the volume of the mash withdrawn in milliliters (mL).
6. The method for analyzing saccharifying power of brewing yeast according to claim 1, characterized in that: and the calculation result in the step S4 is reserved to one digit after the decimal point, and the absolute difference value of the two independent measurement results obtained in the step S4 is not more than 10% of the arithmetic mean value under the repeated condition.
7. The method for analyzing saccharifying power of brewing yeast according to claim 3, characterized in that: the specific preparation steps of the feilin solution are that 69.28g of copper sulfate (CuS04.5H20) is weighed firstly, and then dissolved and diluted to 1000mL by water; finally obtaining the solution of the fibrin.
8. The method for analyzing saccharifying power of brewing yeast according to claim 3, characterized in that: the method comprises the specific steps of firstly weighing 346g of potassium sodium tartrate and 100g of sodium hydroxide, then dissolving and diluting the potassium sodium tartrate and the sodium hydroxide to 1000mL with water, then shaking up the water solution, and finally filtering the water solution after shaking up to obtain the fibulin solution B.
9. The method for analyzing saccharifying power of brewing koji as claimed in any of claims 3 or 4, wherein: and (3) shaking the mixed liquid prepared in the blank experiment and the formal experiment in the step (S2) uniformly, and heating the mixed liquid prepared in the blank experiment and the formal experiment to boiling by using an electric furnace.
10. The method for analyzing saccharifying power of brewing yeast according to claim 1, characterized in that: and the pH value of the acetic acid-sodium acetate buffer solution added in the yeast sample liquid prepared in the step S1 is 4.6.
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