CN104109719B - A kind of method differentiating spirit quality based on box traction substation - Google Patents
A kind of method differentiating spirit quality based on box traction substation Download PDFInfo
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Abstract
The present invention relates to spirit quality authentication technique field, especially a kind of method differentiating spirit quality based on box traction substation, is screened variable by the microorganism frequency of occurrences and relative content method, and builds the discriminating of spirit quality grade in conjunction with box traction substation; Directly can carry out grade discriminating to unknown graded samples, and this model has, and method and thought is simple, easy and simple to handle, feasibility is high, differentiate the feature that accuracy rate is high.
Description
Technical field
The present invention relates to spirit quality authentication technique field, especially a kind of method differentiating spirit quality based on box traction substation.
Background technology
Daqu is the required raw material of White wine brewing process, has the title of " bone of wine ", plays extremely important effect.Its function comprises microbe-derived, the microbial enzyme system source provided in brewing process, flavour substances and prerequisite material thereof and supplementary material, first three microflora's Fauna construction of planting in function and Daqu has great cognation, and therefore Daqu microorganism composition difference can react spirit quality to a certain extent.
454 high-flux sequence methods are the DNA sequencing technology of new generation developed in the last few years, famous with its digitized signal, high data throughput, high feature such as the order-checking degree of depth, high-accuracy etc., the biological bacteria colony assay of extensive multiple sample can be realized, be widely used in the comparative analysis of human body and microbial population of animal intestinal tract, marine microorganism flora composition.Further, its achievement in research part is published on the magazine such as " Nature ", " PNAS ", " GenomeRes ", " Gut ", " Gastroenterology ".
Box traction substation is also called box palpus figure, boxlike figure or box-shaped figure, is a kind of statistical graph being used as display one group of data scatter situation data.To arbitrary group of data, box traction substation can identify outlier fast, and does not need prior tentative data) to obey specific distribution form, not to the requirement of imposing any restrictions property of data.Therefore, various field, particularly qualitative control is widely used in.
For in the discrimination method of spirit quality, the large quantifier elimination that had investigator to make, as " a kind of method differentiating spirit quality " that the patent No. is 201310201060.5, described method is that spirit quality is differentiated in application solid-phase microextraction and discriminatory analysis, is evaluated spirit quality by Daqu flavour substances; And for example the patent No. is " quality controlling means that a kind of yeast of white spirit is produced " of 200910228701.X, described method is the DGGE Criterion finger printing of application Daqu microorganism, and mensuration gained collection of illustrative plates and standard diagram are compared obtain qualified Daqu product, in the program, DGGE technology can only detect tens kinds of dominant bacterias in sample; If learn concrete bacterial classification information, also need to carry out cloning, checking order, experimental implementation is loaded down with trivial details; And adopt the abundance situation that can not reflect microorganism in this way, in addition, the collection of illustrative plates comparison in the program is simple comparison, does not set up correlation model and compares.
As can be seen here, traditional spirit quality Detection & Controling can not adopt unknown classification sample information Criterion, differentiate the quality grade of unknown Daqu sample, meanwhile, utilize 454 high throughput sequencing technologies to differentiate that the method for spirit quality have not been reported in the prior art in conjunction with box traction substation.
Summary of the invention
In order to solve the above-mentioned technical problem existed in prior art, the invention provides a kind of method differentiating spirit quality based on box traction substation, being achieved particular by following technical scheme:
(1) Daqu sample obtains and divides into groups
Obtain finished product Daqu sample, and using the finished product Daqu sample of 120/149 as modeling, the finished product Daqu sample of 29/149 is used for test and/or the checking of model, and then be divided into modeling sample and sample test two groups, stand-by;
(2) Daqu sample message obtains
Adopt 454 high throughput sequencing technologies, carry out sequencing analysis to modeling sample, obtain 580 quasi-microorganism kinds altogether, and be numbered as Xn to 580 quasi-microorganisms, wherein n is the integer of 1-580; Meanwhile, then measure each quasi-microorganism relative content in the sample to which, and keep a record;
(3) screening of microorganism variable
On the basis of step 2, the frequency occurred in modeling sample according to microorganism variable and content screen, and obtain and contribute large microorganism variable index to sample quality;
(4) microorganism variable range is determined
On the basis of step 3, will screen and the analysis of the microorganism variable index obtained employing box traction substation, according to analytical results rejecting abnormalities value, and then determine microorganism variable content distribution in the sample to which; Wherein the determination of outlier is defined as: the 3rd quartile numerical value, IQR=Q3-Q1 that the first quartile value that Q3+1.5IQR<Y<Q1-1.5IQR, Q1 are variable, Q3 are variable;
(5) key microorganisms variable is determined
According to traditional result of study, determine to play the local flavor of Daqu and character in Daqu that conclusive to act on microorganism be key microbial variable;
(6) foundation of discrimination model and sample test
The foundation of a discrimination model: based on step 4 and step 5, sets up spirit quality discrimination model, and related microorganisms variable >=83.33% of middle distribution per sample, and when key microorganisms variable does not lack, be defined as top grade Daqu; Related microorganisms variable≤83.33% distributed in sample, and when lacking key microorganisms variable, be defined as one-level Daqu; Wherein, in top grade Daqu, related microorganisms variable be distributed as 100%, and key microorganisms range of variables is Q1-Q3, is defined as superfine Daqu;
B sample test: sample test group is carried out spirit quality differentiation according to above-mentioned model, whether research test sample represents identical trend in modeling sample.
Described Daqu sample is all that high temperature finished product is bent.
The screening of described microorganism variable, its principle is the frequency of occurrences >=50% ~ 100%, average relative content >=0.6% ~ 5%.
Described key microorganisms variable is Bacillus (bacillus) and Lactobacillus (lactobacillus).
Compared with prior art, technique effect of the present invention is embodied in:
1. by utilizing 454 high throughput sequencing technologies, the composition information that first-generation sequencing technologies accurately can not reflect Daqu microorganism is comprehensively solved.
2. by the microorganism frequency of occurrences and relative content method, variable is screened, and build the discriminating of spirit quality grade in conjunction with box traction substation; Directly can carry out grade discriminating to unknown graded samples, and this model has, and method and thought is simple, easy and simple to handle, feasibility is high, differentiate the feature that accuracy rate is high.
Accompanying drawing explanation
Fig. 1 the present invention is based on box traction substation to differentiate the content distribution of content of microorganisms in Daqu after the method screening of spirit quality and outlier.
Fig. 2 the present invention is based on the schema that box traction substation differentiates the method for spirit quality.
Embodiment
Below in conjunction with accompanying drawing and concrete embodiment, further restriction is done to technical scheme of the present invention, but claimed scope is not only confined to done description.
Embodiment
As depicted in figs. 1 and 2, a kind of method differentiating spirit quality based on box traction substation, comprises the following steps:
(1) Daqu sample obtains and divides into groups
Sample is all high-temperature daqu, derives from Kweichow Moutai Co., Ltd., totally 149 samples, and wherein 120 samples are used for modeling, and all the other 29 samples are used for the test and validation of model.
(2) Daqu sample message obtains
Utilize 454 high throughput sequencing technologies, carry out sequencing analysis to sample, (microorganism is numbered x1, x2, x3, x4, x5, x6 to obtain 580 quasi-microorganism kinds altogether, x7, x8, x9, x10, x11, x12, x13, x14, x15, x16, x17, x18, x19, x20, x21, x22, x23, x24, x25, x26, x27, x28, x568, x569, x570, x571, x572, x573, x574, x575, x576, x577, x578, x579, x580) with the relative content of often kind of microorganism, and to keep a record.
(3) screening of microorganism variable
Based on the microorganism composition information of step 2 gained, the frequency occurred in modeling sample (120 samples) according to microorganism variable and content screen, and obtain the variable index large to sample quality classification contribution.The frequency of occurrences more than 50%, relatively average content more than 0.6% are analyzed, the results are shown in Table 1.
Table 1 different frequency and relative average content variations per hour (microorganism) quantity
As can be seen from Table 1, there are the 30 quasi-microorganism frequencies of occurrences more than 50%, and only have 8 quasi-microorganisms all to occur in all Daqu.When frequency is 80%, 19 quasi-microorganisms are had to occur.Find from microbial average content, different microorganisms content difference is remarkable, only have 3 quasi-microorganisms in Daqu relative average content more than 5%.
The frequency of occurrences of microorganism characterizes the distribution situation of this microorganism in Daqu sample, and the frequency of occurrences of 80% represents in the sample of 80% and all contains this microorganism, thus can ensure that most of sample all contains this microorganism.On content of microorganisms, if it is higher to screen the content of microorganisms obtained, then variable quantity can be caused few, Daqu sample message can not be reflected comprehensively, if but average content is too low, be unfavorable for the detection in later stage.At present level, general operation level can be identified and isolate the microorganism of content more than 1%.Consider, the screening frequency of occurrences is more than 80% with the microorganism variable of average relative content more than 1%.Have the 12 quasi-microorganism frequency of occurrences >=80% and in all Daqu average relative content >=1%, the information of this 12 quasi-microorganism is in table 2.
Table 2 frequency >=80% and the microorganism variable of average relative content >=1%
Know from table 2, the frequency of this 12 quasi-microorganism all >=80%, relative average content all >=1%; And this 12 quasi-microorganism total content accounts for 89.63% of the total content of microorganisms of Daqu, show that this 12 quasi-microorganism can represent the information of microorganism in Daqu comparatively comprehensively, therefore as the variable of discrimination standard.
(4) microorganism variable range is determined
In order to determine these ranges of variables, need rejecting abnormalities value, but due to unintelligible to the classification grade of sample, the multivariate statistical methods such as discriminatory analysis therefore can not be utilized to study.And box traction substation is we provide the standard identifying outlier: outlier is defined as the value being less than Q1-1.5IQR or being greater than Q3+1.5IQR.Therefore, utilize box traction substation, 12 variablees selected are analyzed, determines these 12 kinds of variable distributions in Daqu, outlier, the results are shown in Figure 1.
Known by Fig. 1, the relative content of these microorganisms (variable) differs greatly, and distribution situation is different.Except x11, its dependent variable has outlier to occur.Therefore next step needs to reject these outliers, calculates the first quartile (Q1) and the 3rd quartile (Q3), thus determines the upper and lower bound of Daqu microorganism variable.By box traction substation, determine this 12 kinds of ranges of variables, this results are shown in Table 3.
The determination of table 312 quasi-microorganism (variable) content range
(5) key microorganisms variable is determined
Certain micro-organisms plays conclusive effect to aromatic type Daqu local flavor and character, therefore needs to keep certain content ratio, if any one key microorganisms disappearance maybe can not arrive certain content ratio, all can affect the quality of Daqu.Therefore, when Modling model, using the content of functional microorganism as an important indicator.
In this method of discrimination, Bacillus (bacillus) and Lactobacillus (lactobacillus) is defined as the function stem important to aromatic type high-temperature daqu, and main reason is:
1. lot of documents and related experiment show, the sauce perfume (or spice) of aromatic type Daqu produced by genus bacillus, some important flavour substancess (as Pyrazine material) are also the main metabolites of genus bacillus, if therefore lack genus bacillus in Daqu, can cause the disappearance of high-temperature daqu style.
2. lactobacillus is the dominant microflora of aromatic type Daqu, and Bacterium lacticum can produce a large amount of acid in daqu fermentation process, on the one hand for flavour substances provides prerequisite, can suppress the growth of other miscellaneous bacterias on the other hand, thus play critical function.In addition, milk-acid bacteria also can produce other active substance and flavour substances, and these all show the importance of milk-acid bacteria, therefore also as the key variables in differentiating.
(6) foundation of discrimination model and sample test
By analysis and research above, establish a kind of Daqu microorganism discrimination standard: (1) is if the content range of 12 quasi-microorganisms all meets the differentiation scope obtained in step 4 in Daqu, and key microorganisms reaches certain proportion (namely content is between the first quartile Q1 and the 3rd quartile Q3), be then determined as superfine Daqu; (2) if the differentiation scope having at least the content range of 10 quasi-microorganisms to meet in Daqu to obtain in step 4, and key microorganisms do not lack be determined as top grade Daqu, otherwise be determined as one-level Daqu.
Utilize the discrimination standard of upper figure, test 120 samples, test-results is in table 4.
Table 4 differentiates the list of Daqu level results based on microbial information
Note: 1. modeling Daqu total number of samples amount is 120; 2. >=12 represent that at least 12 kinds of content of microorganisms are within scope, by that analogy.
Known by table 4, when having more than 10 classes (comprising 10 classes) content of microorganisms within scope in test sample, and during containing Bacillus (bacillus) and Lactobacillus (lactobacillus), this sample is top grade sample.Detect 129 Daqu samples, result shows have top grade rate to be 91.67%, and wherein superfine rate is 13.33%.
In order to verify Daqu microorganism method of discrimination, the method for discrimination set up above first utilizing carries out identification and classification to residue 29 Daqu sample, and whether research test sample represents identical trend in modeling sample, differentiates and the results are shown in Table 5.
Table 5 is tested sample and is differentiated interpretation of result
Note: 1. testing Daqu total number of samples amount is 29; 2. >=12 represent that at least 12 quasi-microorganism content are within scope, by that analogy.
Can find out in table 5, test sample top grade rate is 89.66%, and superfine rate is 10.34%, with modeling sample closely, this shows that institute's construction method differentiates comparatively rationally effectively.
It is important to point out at this; above embodiment is only limitted to be further elaborated technical scheme of the present invention and understand; it is not limitation of the invention further; the essential characteristics of non-protruding made on this basis of those skilled in the art and the improvement of non-significant progress and amendment, still belong to protection category of the present invention for this reason.
Claims (4)
1. differentiate a method for spirit quality based on box traction substation, it is characterized in that, comprise the following steps:
(1) Daqu sample obtains and divides into groups
Obtain finished product Daqu sample, and using the finished product Daqu sample of 120/149 as modeling, the finished product Daqu sample of 29/149 is used for test and/or the checking of model, and then be divided into modeling sample and sample test two groups, stand-by;
(2) Daqu sample message obtains
Adopt 454 high throughput sequencing technologies, carry out sequencing analysis to modeling sample, obtain 580 quasi-microorganism kinds altogether, and be numbered as Xn to 580 quasi-microorganisms, wherein n is the integer of 1-580; Meanwhile, then measure each quasi-microorganism relative content in the sample to which, and keep a record;
(3) screening of microorganism variable
On the basis of step 2, the frequency occurred in modeling sample according to microorganism variable and content screen, and obtain and contribute large microorganism variable index to sample quality;
(4) microorganism variable range is determined
On the basis of step 3, will screen and the analysis of the microorganism variable index obtained employing box traction substation, according to analytical results rejecting abnormalities value, and then determine microorganism variable content distribution in the sample to which; Wherein outlier is defined as the value that is less than Q1-1.5IQR or is greater than Q3+1.5IQR, the 3rd quartile numerical value, IQR=Q3-Q1 that the first quartile value that Q1 is variable, Q3 are variable;
(5) key microorganisms variable is determined
According to traditional result of study, determine to play the local flavor of Daqu and character in Daqu that conclusive to act on microorganism be key microbial variable;
(6) foundation of discrimination model and sample test
The foundation of a discrimination model: based on step 4 and step 5, sets up spirit quality discrimination model, and related microorganisms variable >=83.33% of middle distribution per sample, and when key microorganisms variable does not lack, be defined as top grade Daqu; Related microorganisms variable≤83.33% distributed in sample, and when lacking key microorganisms variable, be defined as one-level Daqu; Wherein, in top grade Daqu, related microorganisms variable be distributed as 100%, and key microorganisms range of variables is Q1-Q3, is defined as superfine Daqu;
B sample test: sample test group is carried out spirit quality differentiation according to above-mentioned model, whether research test sample represents identical trend in modeling sample.
2. differentiate the method for spirit quality as claimed in claim 1 based on box traction substation, it is characterized in that, described Daqu sample is all that high temperature finished product is bent.
3. differentiate the method for spirit quality as claimed in claim 1 based on box traction substation, it is characterized in that, the screening of described microorganism variable, its principle is the frequency of occurrences >=50% ~ 100%, average relative content >=0.6% ~ 5%.
4. differentiate the method for spirit quality as claimed in claim 1 based on box traction substation, it is characterized in that, described key microorganisms variable is Bacillus (bacillus) and Lactobacillus (lactobacillus).
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| CN106897941A (en) * | 2017-01-03 | 2017-06-27 | 北京国能日新系统控制技术有限公司 | A kind of blower fan method for processing abnormal data and device based on quartile box traction substation |
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| CN102071124A (en) * | 2009-11-25 | 2011-05-25 | 贵州仁怀茅台镇金士酒业有限公司 | Quality control method for use in production of Maotai-flavor Daqu liquor |
| CN102382877A (en) * | 2010-08-30 | 2012-03-21 | 中国食品发酵工业研究院 | Method for studying structural diversity of daqu bacterial community |
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| CN102071124A (en) * | 2009-11-25 | 2011-05-25 | 贵州仁怀茅台镇金士酒业有限公司 | Quality control method for use in production of Maotai-flavor Daqu liquor |
| CN102382877A (en) * | 2010-08-30 | 2012-03-21 | 中国食品发酵工业研究院 | Method for studying structural diversity of daqu bacterial community |
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| 徐晓宇等.454测序法在环境微生物生态研究中的应用.《生物技术通报》.2010, * |
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