CN104062293B - A kind of detection in barley or Fructus Hordei Germinatus causes the method that yeast shifts to an earlier date the coherency factor - Google Patents
A kind of detection in barley or Fructus Hordei Germinatus causes the method that yeast shifts to an earlier date the coherency factor Download PDFInfo
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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 41
- 238000000034 method Methods 0.000 title claims abstract description 33
- 235000007340 Hordeum vulgare Nutrition 0.000 title claims abstract description 30
- 238000001514 detection method Methods 0.000 title claims abstract description 18
- 240000005979 Hordeum vulgare Species 0.000 title 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 43
- 241000209219 Hordeum Species 0.000 claims abstract description 29
- SRBFZHDQGSBBOR-IOVATXLUSA-N Xylose Natural products O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 claims description 24
- 238000006243 chemical reaction Methods 0.000 claims description 22
- 229960003487 xylose Drugs 0.000 claims description 22
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 claims description 14
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 claims description 14
- 239000006228 supernatant Substances 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 11
- 238000002156 mixing Methods 0.000 claims description 11
- 238000001556 precipitation Methods 0.000 claims description 11
- SRBFZHDQGSBBOR-LECHCGJUSA-N alpha-D-xylose Chemical compound O[C@@H]1CO[C@H](O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-LECHCGJUSA-N 0.000 claims description 8
- 239000000376 reactant Substances 0.000 claims description 8
- 239000012086 standard solution Substances 0.000 claims description 8
- 229960000583 acetic acid Drugs 0.000 claims description 7
- 238000009835 boiling Methods 0.000 claims description 7
- 239000008367 deionised water Substances 0.000 claims description 7
- 229910021641 deionized water Inorganic materials 0.000 claims description 7
- 235000019441 ethanol Nutrition 0.000 claims description 7
- 239000012362 glacial acetic acid Substances 0.000 claims description 7
- 150000004823 xylans Chemical class 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 5
- 238000002835 absorbance Methods 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 229920001221 xylan Polymers 0.000 claims description 4
- 125000000969 xylosyl group Chemical group C1([C@H](O)[C@@H](O)[C@H](O)CO1)* 0.000 claims description 4
- 230000003750 conditioning effect Effects 0.000 claims description 3
- 239000012153 distilled water Substances 0.000 claims description 3
- 230000031700 light absorption Effects 0.000 claims description 3
- QCDYQQDYXPDABM-UHFFFAOYSA-N phloroglucinol Chemical compound OC1=CC(O)=CC(O)=C1 QCDYQQDYXPDABM-UHFFFAOYSA-N 0.000 claims description 3
- 229960001553 phloroglucinol Drugs 0.000 claims description 3
- 238000002360 preparation method Methods 0.000 claims description 3
- 238000005204 segregation Methods 0.000 claims description 3
- 238000012360 testing method Methods 0.000 claims description 3
- 239000002253 acid Substances 0.000 claims description 2
- 238000010790 dilution Methods 0.000 claims description 2
- 239000012895 dilution Substances 0.000 claims description 2
- 238000001914 filtration Methods 0.000 claims description 2
- 230000001681 protective effect Effects 0.000 claims description 2
- 238000000855 fermentation Methods 0.000 abstract description 12
- 230000004151 fermentation Effects 0.000 abstract description 12
- 230000002349 favourable effect Effects 0.000 abstract description 5
- 230000000694 effects Effects 0.000 abstract description 4
- 238000004737 colorimetric analysis Methods 0.000 abstract description 3
- 230000003287 optical effect Effects 0.000 abstract description 3
- 238000011179 visual inspection Methods 0.000 abstract description 3
- 241000209140 Triticum Species 0.000 description 23
- 235000021307 Triticum Nutrition 0.000 description 23
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 12
- 238000005189 flocculation Methods 0.000 description 4
- 230000016615 flocculation Effects 0.000 description 4
- 238000004519 manufacturing process Methods 0.000 description 3
- 238000006213 oxygenation reaction Methods 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 238000005457 optimization Methods 0.000 description 2
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- 230000005070 ripening Effects 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- GXCLVBGFBYZDAG-UHFFFAOYSA-N N-[2-(1H-indol-3-yl)ethyl]-N-methylprop-2-en-1-amine Chemical compound CN(CCC1=CNC2=C1C=CC=C2)CC=C GXCLVBGFBYZDAG-UHFFFAOYSA-N 0.000 description 1
- 238000013124 brewing process Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 210000003981 ectoderm Anatomy 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 239000011888 foil Substances 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 208000015181 infectious disease Diseases 0.000 description 1
- 238000007689 inspection Methods 0.000 description 1
- 238000013507 mapping Methods 0.000 description 1
- 230000000873 masking effect Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
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- 230000006920 protein precipitation Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
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- 239000000126 substance Substances 0.000 description 1
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- 238000005406 washing Methods 0.000 description 1
- 210000005253 yeast cell Anatomy 0.000 description 1
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- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The present invention relates to brewing field, it causes in particular to a kind of detection in barley or Fructus Hordei Germinatus the method that yeast shifts to an earlier date the coherency factor.The present invention does not need to use yeast, the PYF vigor of barley, Fructus Hordei Germinatus is directly reflected by measuring Water soluble pentosan content, can visual inspection barley and Fructus Hordei Germinatus PYF vigor, relative to existing fermentation method and optical density colorimetric method, do not use yeast, not by the impact of the objective condition such as yeast activity, there is the data precision high, the advantage of favorable reproducibility.
Description
Technical field
The present invention relates to brewing field, it causes in particular to a kind of detection in barley or Fructus Hordei Germinatus the method that yeast shifts to an earlier date the coherency factor.
Background technology
Yeast flocculate in advance (prematureyeastflocculation, PYF) refer to yeast ferment not yet during the fermentation at the end of occur Yeast Flocculation phenomenon.The PYF factor is a negative barley, malt quality index, derives from barley and Fructus Hordei Germinatus, and its stable chemical nature is heat-resisting, and being difficult to dispel its negative effect once enter brewing process, is the factor affecting latter stage of ripening yeast number most critical.
At present, in Fructus Hordei Germinatus, yeast shifts to an earlier date the detection major part of AgF is be based upon in yeast-leavened mechanism, and these methods are totally divided into two classes from principle, and a class is rapid fermentation method, and ultimate principle is: Fructus Hordei Germinatus PYF factor pair yeast flocculence has an impact; Using active good production yeast to be added in the agreement act wheat juice of improvement, by detecting the suspended yeast cell number after fermentation, determining the impact of Fructus Hordei Germinatus on yeast flocculence; Another kind of is optical density colorimetric method, by extracting the PYF factor in agreement wheat juice, itself and yeast is reacted in specific reaction system, by the turbidity value of supernatant after assaying reaction, and the PYF factor of reflection Fructus Hordei Germinatus.The common point of these two kinds of methods all needs to use yeast, reflection be Fructus Hordei Germinatus PYF vigor, inoculation yeast state combined influence that Yeast Flocculation is showed, be applicable to, for the flocculation performance of prediction inoculation yeast large production latter stage of ripening under formula Fructus Hordei Germinatus, being not suitable for for objective evaluation Fructus Hordei Germinatus PYF quality and instructing malthouse to carry out malt-making technique optimization.
This detection method, provide one can visual inspection barley and Fructus Hordei Germinatus PYF vigor (not needing to utilize yeast), accurately, the PYF factors check method of favorable reproducibility, the method is not by the impact of the objective condition such as yeast activity, oxygenation, compared with fermentation method, data accurately, favorable reproducibility, the time short (shortening to 9 hours by 40 hours).
Summary of the invention
The present invention is directed to the deficiencies in the prior art, the object of the invention is to develop a kind of do not need to use yeast can visual inspection barley and Fructus Hordei Germinatus PYF vigor, accurately, the detection method of favorable reproducibility, cause with its prediction barley and Fructus Hordei Germinatus the ability that yeast flocculates in advance.
The PYF factor is because barley ectoderm is subject to microbial infection, a kind of water-soluble character produced, the xylan fragments with certain molecular weight, therefore can extract, be purified into the xylan fragments that this part can characterize barley, Fructus Hordei Germinatus PYF vigor, directly be reflected the PYF vigor of barley, Fructus Hordei Germinatus by xylan content.
Technical scheme of the present invention is:
A kind of detection in barley or Fructus Hordei Germinatus causes the method that yeast shifts to an earlier date the coherency factor (PYF), and the step of described method is as follows:
Step 1: get the sample that moisture is Wc, pulverizes, and weighs sample W
2for 25g is in the triangular flask of 500mL, add 300mL deionized water, mixing; 150rpm vibrates 1-2h, and the constant weight that adds water is to W
1for 350g, Filter paper filtering, filtered fluid detects proportion S
g, extract Me; Get filtered fluid 10mL and add 30mL absolute ethyl alcohol, mixing; Ice bath alcohol precipitation reaction 1h; The centrifugal 10min of sample 10000rpm after reaction, removes supernatant, and precipitation adds the mixing of 10mL deionized water, and 85 DEG C of water-bath 2h dissolve; After dissolving, 10000rpm carries out centrifugal segregation precipitation, supernatant freezen protective, for subsequent use;
Step 2: drawing standard curve, utilizes wood sugar and phloroglucin chromogenic reaction in acid condition, forms red material, and measure content, computing formula is as follows:
Y＝aX
552+b
Wherein: X
552for sample absorbance during 552nm; A is the slope of typical curve; B is the intercept of typical curve; Y is xylose concentration in supernatant, mg/L;
Wherein: G is Water soluble pentosan content in sample, mg/100g over dry Fructus Hordei Germinatus; N is extension rate; Y is xylose concentration in supernatant, mg/L; Wc is the moisture of sample, %; Me is the extract content of brewer's wort, g; S
gfor the proportion of brewer's wort; W
1for sample wheat juice weight, g; W
2for example weight, g; 0.65 is wood sugar and xylan conversion coefficient.
Preferably, the sample described in step 1 is barley or Fructus Hordei Germinatus.
Preferably, in step 1 in ice bath alcohol precipitation course of reaction, within every 20 minutes, shake up once.
Preferably, in step 1, sample comminution adopts EBC comminutor.
Preferably, in step 2, xylose standard curve method for drafting is: the configuration of 500mg/L xylose standard solution: 50mg wood sugar deionized water is settled to 100mL; Take out 1.0 respectively, 2.0,3.0,4.0,6.0, xylose standard solution described in 8.0mL, be diluted to 10mL, the xylose standard solution of preparation variable concentrations, get the standard solution 1mL after dilution respectively, after adding phloroglucin-glacial acetic acid reactant liquor 5mL, mixing, boiling water bath reaction 25min, ice bath cessation reaction; Add reactant liquor for contrast with water, measure each concentration wood sugar at 552nm absorbance, according to known xylose concentration value, Criterion curve.
Preferably, the step of step 2 chromogenic reaction is: get 3 color-comparison tubes and also identify respectively, number; Blank: in No. 1 color comparison tube, to add 1mL distilled water; Sample: add 1mL and prepare sample in 2, No. 3 color comparison tubes; Phloroglucin-glacial acetic acid reactant liquor 5mL is respectively added in color comparison tube; Immediately 3 color comparison tubes are put into boiling water bath and react 25 minutes, need cap at stage of reaction color comparison tube; Ice bath cessation reaction; With blank conditioning instrumentation zero point, under 552nm wavelength, detect the light absorption value of testing sample.
A lot of to the research of the Fructus Hordei Germinatus PYF factor at present, also do not come to a conclusion about its composition and molecular structure, the result that different research institutions provides is all not identical.At this, we are not that specifically to study the PYF factor be what on earth, and its molecular structure how, just hopes the material extracting and can cause Yeast Flocculation in barley or Fructus Hordei Germinatus, represents the number of the PYF factor with the xylan content in this material.
Compared with prior art, the present invention has following features:
One: the present invention is relative to current Rapid Fermentation detection method, and detection time shortens, and shortens to 9 hours by original 40 hours;
Its two: the present invention, relative to existing fermentation method and optical density colorimetric method, does not use yeast, not by the impact of the objective condition such as yeast activity, has the data precision high, the advantage of favorable reproducibility;
Its three: the present invention establishes the relation of Water soluble pentosan and PYF, specify its can substitute PYF detect.
Therefore, the present invention can detect the PYF factor in barley, Fructus Hordei Germinatus, is the incoming inspection of Brewery Fructus Hordei Germinatus, and the daily quality management of Fructus Hordei Germinatus factory provides a kind of effective method.
Accompanying drawing explanation
Accompanying drawing 1 is Water soluble pentosan and PYF related coefficient.
Embodiment
The specific embodiment of the present invention is as follows:
Embodiment 1:
Get 12 Fructus Hordei Germinatus samples of different manufacturer, Fructus Hordei Germinatus kind comprises and adds wheat, Australia wheat, state wheat, with 20 DEG C, 40 hours fermentation (rapid fermentation method) measure the PYF of Fructus Hordei Germinatus respectively, record the PYF of sample from 20% ~ 159%.
One, Water soluble pentosan detects
By each accurate weighing 25.0g of these 12 samples, add 300mL water, the shaking table of 150rpm shook after 1 hour, and balance constant weight is to 350.0g, filter with Medium speed filter paper, get 10mL filtrate and add 30mL absolute ethyl alcohol, alcohol precipitation 1 hour in ice bath, centre shakes up twice (every 20 minutes once), the centrifugal 10min of sample 10000rpm after reaction, remove supernatant, precipitation adds the mixing of 10mL deionized water, and 85 DEG C of water-bath 2h dissolve.After dissolving, 10000rpm carries out centrifugal segregation precipitation, gets supernatant for subsequent use.
Phloroglucin-glacial acetic acid reactant liquor preparation (developer): take 1.28g phloroglucin and be dissolved in 5mL absolute ethyl alcohol, use 55mL glacial acetic acid, 1mL concentrated hydrochloric acid, 5mL phloroglucin-ethanolic solution, the D/W mixing of 0.5mL1.75%.
Get 3 color comparison tubes and carry out respectively identifying, numbering, 1mL distilled water is added in 1# color comparison tube, 2#, 3# color comparison tube adds 1mL respectively and prepares sample, 5mL phloroglucin-glacial acetic acid reactant liquor is respectively added in 3 color comparison tubes, react 25 minutes in boiling water bath, ice bath cessation reaction, with blank conditioning instrumentation zero point, under 552nm wavelength, detect the light absorption value of testing sample, obtain the Water soluble pentosan content (mg/100g over dry Fructus Hordei Germinatus) of sample as table 1 according to formulae discovery:
Table 1:
Two, traditional fermentation process detects Fructus Hordei Germinatus PYF
Use saccharifying instrument to prepare agreement act wheat juice in 12 samples, get filtrate 280mL in SCOTT bottle, build bottle cap, put into pot, after boiling, Steam Heating 40min, makes wheat juice occur protein precipitation, is then cooled to 20 ~ 30 DEG C, filters by double-layer filter paper.
Measuring 220mL wheat juice with graduated cylinder pours in SCOTT bottle, adds appropriate syrup according to wort concentration, be made into pol be 12% wheat juice carry out dissolving mixing, screw bottle cap, then put into pot, wet sterilization 20min after boiling, cools for subsequent use.
Yeast prepares: the yeast paste (2 ~ 4 generation) of production and application is got in sterile working, fetches in rear placement refrigerator, uses in 8hr.Get a 40mL centrifuge tube drawing alcohol swab wiping, move into yeast paste, in the centrifugal 10min of 4000rpm, incline supernatant; Get a 40mL centrifuge tube drawing alcohol swab wiping, take centrifugal after yeast paste 1.10g.
The wheat juice prepared is added about 10-20mL in the centrifuge tube weighing yeast, and after using vortex oscillator fully to shake, rejoin in wheat juice, and utilize wheat juice to rinse 3 times to centrifuge tube, washing fluid all returns in SCOTT bottle; Screw oxygenation.
Fermentation: oxygenation is complete, pours into wheat juice after wheat juice shakes gently in drying, clean EBC color comparison tube, uses masking foil to be sealed by mouth, the incubator being uprightly placed in 9.5 DEG C carries out cultivation 5 days, carry out yeast counts, and calculate PYF value (%), as table 2.
Table 2:
| Numbering | 1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 |
| PYF(％) | 159 | 120 | 105 | 102 | 98 | 82 | 64 | 53 | 29 | 28 | 16 | 13 |
Three, mapping calculates
Water soluble pentosan and PYF related coefficient, as described in Figure 1, as can be seen from above data and Fig. 1, use the prediction Fructus Hordei Germinatus developed of the present invention to cause method that yeast flocculates in advance and traditional fermentation process have higher correlativity, can answer the size of PYF value by Fructus Hordei Germinatus Water soluble pentosan content back.
Embodiment 2:
Utilize detection method of the present invention, to adding wheat Metcalfe1212, Australia wheat Vlamingh1302, Australia wheat Gairdner1306 tri-kind wheat process Water soluble pentosan Changing Patterns are analyzed, and experimental data is as table 3:
Table 3: wheat process Water soluble pentosan Changing Pattern processed
Can find out by analyzing data:
Although barley variety is different, Water soluble pentosan content no significant difference in raw material barley, namely the PYF of barley and barley variety have nothing to do;
Water soluble pentosan content constantly increases along with wheat process processed, reaches peak value germination second or the 3rd day, starts afterwards to reduce, and namely in wheat process processed, the PYF of Fructus Hordei Germinatus first raises, then reduces.
As can be seen from example 2, utilize detection method of the present invention can carry out Fructus Hordei Germinatus PYF detection to barley, wheat process processed and finished product Fructus Hordei Germinatus, for the improvement of malt-making technique and optimization provide Data support.
Claims (6)
1. detect in barley or Fructus Hordei Germinatus and cause the method that yeast shifts to an earlier date the coherency factor (PYF), it is characterized in that the step of described method is as follows:
Step 1: get the sample that moisture is Wc, pulverizes, and weighs sample W
2for 25g is in the triangular flask of 500mL, add 300mL deionized water, mixing; 150rpm vibrates 1-2h, and the constant weight that adds water is to W
1for 350g, Filter paper filtering, filtered fluid detects proportion S
g, extract Me; Get filtered fluid 10mL and add 30mL absolute ethyl alcohol, mixing; Ice bath alcohol precipitation reaction 1h; The centrifugal 10min of sample 10000rpm after reaction, removes supernatant, and precipitation adds the mixing of 10mL deionized water, and 85 DEG C of water-bath 2h dissolve; After dissolving, 10000rpm carries out centrifugal segregation precipitation, supernatant freezen protective, for subsequent use;
Step 2: drawing standard curve, utilizes wood sugar and phloroglucin chromogenic reaction in acid condition, forms red material, and measure content, computing formula is as follows:
Y＝aX
552+b
Wherein: X
552for sample absorbance during 552nm; A is the slope of typical curve; B is the intercept of typical curve; Y is xylose concentration in supernatant, mg/L;
Wherein: G is Water soluble pentosan content in sample, mg/100g over dry Fructus Hordei Germinatus; N is extension rate; Y is xylose concentration in supernatant, mg/L; Wc is the moisture of sample, %; Me is the extract content of brewer's wort, g; S
gfor the proportion of brewer's wort; W
1for sample brewer's wort weight, g; W
2for example weight, g; 0.65 is wood sugar and xylan conversion coefficient.
2. cause the method that yeast shifts to an earlier date the coherency factor (PYF) in detection barley according to claim 1 or Fructus Hordei Germinatus, it is characterized in that the sample described in step 1 is barley or Fructus Hordei Germinatus.
3. cause the method that yeast shifts to an earlier date the coherency factor (PYF) in detection barley according to claim 1 or Fructus Hordei Germinatus, it is characterized in that in step 1 in ice bath alcohol precipitation course of reaction, within every 20 minutes, shake up once.
4. cause the method that yeast shifts to an earlier date the coherency factor (PYF) in detection barley according to claim 1 or Fructus Hordei Germinatus, it is characterized in that in step 1, sample comminution adopts EBC comminutor.
5. cause the method that yeast shifts to an earlier date the coherency factor (PYF) in detection barley according to claim 1 or Fructus Hordei Germinatus, it is characterized in that in step 2, xylose standard curve method for drafting is: the configuration of 500mg/L xylose standard solution: 50mg wood sugar deionized water is settled to 100mL; Take out 1.0 respectively, 2.0,3.0,4.0,6.0, xylose standard solution described in 8.0mL, be diluted to 10mL, the xylose standard solution of preparation variable concentrations, get the standard solution 1mL after dilution respectively, after adding phloroglucin-glacial acetic acid reactant liquor 5mL, mixing, boiling water bath reacts 25 minutes, ice bath cessation reaction; Add reactant liquor for contrast with water, measure each concentration wood sugar at 552nm absorbance, according to known xylose concentration value, Criterion curve.
6. cause the method that yeast shifts to an earlier date the coherency factor (PYF) in detection barley according to claim 1 or Fructus Hordei Germinatus, it is characterized in that the step of step 2 chromogenic reaction is: get 3 color-comparison tubes and also identify respectively, number; Blank: in No. 1 color comparison tube, to add 1mL distilled water; Sample: add 1mL step 1 and prepare sample in 2, No. 3 color comparison tubes; Phloroglucin-glacial acetic acid reactant liquor 5mL is respectively added in color comparison tube; Immediately 3 color comparison tubes are put into boiling water bath and react 25 minutes, ice bath cessation reaction; With blank conditioning instrumentation zero point, under 552nm wavelength, detect the light absorption value of testing sample.
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| JP3943122B1 (en) * | 2006-11-02 | 2007-07-11 | 麒麟麦酒株式会社 | Method for improving fermentability of fast-setting malt |
| CN101349636B (en) * | 2007-07-16 | 2010-09-29 | 青岛啤酒股份有限公司 | Method for testing germinated barley flocculating degree |
| CN101451958B (en) * | 2008-12-29 | 2010-11-10 | 东莞泛亚太生物科技有限公司 | Detecting method for Xylanase efficiency fordecomposing wheat |
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