CN108795783B - Tea fruit wine yeast and preparation method and application thereof - Google Patents
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- 240000004808 Saccharomyces cerevisiae Species 0.000 title claims abstract description 98
- 235000019990 fruit wine Nutrition 0.000 title claims abstract description 22
- 238000002360 preparation method Methods 0.000 title claims abstract description 17
- 235000014680 Saccharomyces cerevisiae Nutrition 0.000 claims abstract description 82
- 244000269722 Thea sinensis Species 0.000 claims abstract description 64
- 235000011389 fruit/vegetable juice Nutrition 0.000 claims abstract description 50
- 238000000034 method Methods 0.000 claims abstract description 24
- 235000015203 fruit juice Nutrition 0.000 claims abstract description 22
- 241000756042 Polygonatum Species 0.000 claims abstract description 12
- 239000000203 mixture Substances 0.000 claims abstract description 3
- 244000184734 Pyrus japonica Species 0.000 claims abstract 6
- 235000013616 tea Nutrition 0.000 claims description 50
- 235000006468 Thea sinensis Nutrition 0.000 claims description 11
- 239000002244 precipitate Substances 0.000 claims description 11
- 230000001954 sterilising effect Effects 0.000 claims description 7
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- 239000006228 supernatant Substances 0.000 claims description 5
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- 229930006000 Sucrose Natural products 0.000 claims description 4
- 235000015197 apple juice Nutrition 0.000 claims description 4
- 230000004048 modification Effects 0.000 claims description 4
- 238000012986 modification Methods 0.000 claims description 4
- 235000020333 oolong tea Nutrition 0.000 claims description 4
- 239000005720 sucrose Substances 0.000 claims description 4
- 244000241235 Citrullus lanatus Species 0.000 claims description 3
- 235000012828 Citrullus lanatus var citroides Nutrition 0.000 claims description 3
- 235000013399 edible fruits Nutrition 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 235000009569 green tea Nutrition 0.000 claims description 3
- 235000015205 orange juice Nutrition 0.000 claims description 3
- 235000015206 pear juice Nutrition 0.000 claims description 3
- 230000004913 activation Effects 0.000 claims description 2
- 239000000843 powder Substances 0.000 claims description 2
- 238000003756 stirring Methods 0.000 claims description 2
- 235000020334 white tea Nutrition 0.000 claims description 2
- 235000020338 yellow tea Nutrition 0.000 claims description 2
- 241000205407 Polygonum Species 0.000 claims 1
- 238000000605 extraction Methods 0.000 claims 1
- 238000009928 pasteurization Methods 0.000 claims 1
- 230000009466 transformation Effects 0.000 claims 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 abstract description 14
- 238000000855 fermentation Methods 0.000 abstract description 14
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- 230000000694 effects Effects 0.000 abstract description 6
- 230000036541 health Effects 0.000 abstract description 3
- 230000025938 carbohydrate utilization Effects 0.000 abstract description 2
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- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
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- 238000007254 oxidation reaction Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
- C12N1/16—Yeasts; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12G—WINE; PREPARATION THEREOF; ALCOHOLIC BEVERAGES; PREPARATION OF ALCOHOLIC BEVERAGES NOT PROVIDED FOR IN SUBCLASSES C12C OR C12H
- C12G3/00—Preparation of other alcoholic beverages
- C12G3/02—Preparation of other alcoholic beverages by fermentation
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/36—Adaptation or attenuation of cells
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Abstract
本发明提供了一种茶果酒酵母及其制备方法和应用，所述方法包括：采用茶汁、果汁和玉竹提取液的混合液驯化葡萄酒酵母，得到所述茶果酒酵母。本发明的制备方法采用驯化的方法制备茶果酒酵母，得到的茶果酒酵母的糖利用率高、发酵能力强、酒精度高，具有药物保健作用。The invention provides a tea fruit wine yeast and a preparation method and application thereof. The method comprises: using a mixture of tea juice, fruit juice and Polygonatum japonica extract to domesticate wine yeast to obtain the tea fruit wine yeast. The preparation method of the invention adopts the domestication method to prepare the tea fruit wine yeast, and the obtained tea fruit wine yeast has high sugar utilization rate, strong fermentation ability and high alcohol content, and has the effect of medicine and health care.
Description
Technical Field
The invention belongs to the technical field of fermentation, and relates to a tea fruit wine yeast and a preparation method and application thereof.
Background
In recent years, the tea fruit wine is favored by consumers due to the unique flavor and the health care effect, and the tea fruit wine is rich in various amino acids, vitamins and mineral substances and has the effects of resisting oxidation, radiation, cancer, blood fat and the like. Yeast varieties are key factors for brewing tea fruit wine, and yeast properties directly influence the taste and flavor of brewed tea fruit wine, so domestication and screening of tea fruit wine yeast are always regarded as important.
In the development of tea wine yeast, Zhang Shuai adopts a method of domestication and ultraviolet mutagenesis, develops tea wine yeast, separates two yeasts from distiller's yeast, utilizes sugar-containing tea wort and malt extract culture mediums with different proportions to domesticate, screens out the yeast which can better adapt to the tea juice environment, and then carries out ultraviolet mutagenesis treatment on the yeast, thereby preparing the tea wine yeast with stronger growth ability in sugar-containing tea juice (Zhang Shu, the development of tea wine yeast, food industry science and technology, 2008, 29(1):162 and 164.). However, the yeast adopted by the method needs to be separated and purified from the fermentation process firstly, the time and the cost are consumed, the sugar utilization degree of the obtained saccharomyces cerevisiae is not high, sugar needs to be added into the tea juice and the wort additionally, and the cost is increased.
The high sugar resistance, the alcohol resistance and other performances of the cider yeast Y02 are domesticated by Song Andong and the like, the domesticated strain can resist 35 percent of high sugar and 25 percent of alcohol, and the alcoholic strength of the fermented cider reaches 10.9 percent (domestication test of the cider yeast of Song Andong, conwybin, plum blossom plum and the like, brewing, 2003, 30(2): 44-45.). However, the fruit wine brewed by the apple wine yeast domesticated by the method has single taste and does not have the faint scent of tea wine.
Therefore, the saccharomyces cerevisiae with rich taste and high fermentation efficiency is prepared and has an important role in the field of brewing.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides the tea fruit wine yeast and the preparation method and the application thereof, the tea fruit wine yeast is prepared by the preparation method through an domestication method, and the obtained tea fruit wine yeast has high carbon source utilization rate, strong fermentation capacity and high alcoholic strength.
In order to achieve the purpose, the invention adopts the following technical scheme:
in a first aspect, the present invention provides a method for preparing a yeast for wine of camellia sinensis fruit, comprising:
and domesticating wine yeast by adopting a mixed solution of the tea juice, the fruit juice and the polygonatum odoratum extract to obtain the tea fruit wine yeast.
According to the invention, the mixed liquor of the tea juice, the fruit juice and the polygonatum odoratum extract is adopted to domesticate the wine yeast, so that the utilization rate of the wine yeast to a carbon source is obviously improved, additional sugar and wort do not need to be additionally added in the domestication process, the wine yeast can utilize limited energy sources in the tea juice, the fruit juice and the polygonatum odoratum extract to produce alcohol, and the wine yeast has the advantages of high utilization rate of the carbon source, strong fermentation capacity and high alcohol degree.
In the invention, the yeast product special for the commercial Angel grape wine fruit wine is directly used as the fermentation yeast, and compared with the yeast separated in the fermentation process, the yeast has high purity, strong fermentation capacity, simple and convenient process and lower cost.
Preferably, the ratio of the tea juice, the fruit juice and the polygonatum extract in parts by weight is (3-8): (1-3): (1-2), and may be, for example, 3:1:1, 5:1:1, 8:1:1, 3:2:1, 5:2:1, 8:2:1, 3:3:1, 5:3:1, 8:3:1, 3:1:2, 5:1:2, 8:1:2, 3:2:2, 5:2:2, 8:2:2, 3:3:2, 5:3:2 or 8:3:2, preferably (3-5): 2-3): 1-2.
Preferably, the tea juice comprises any one of oolong tea juice, black tea juice, green tea juice, black tea juice, white tea juice or yellow tea juice or a combination of at least two of them.
Preferably, the juice comprises any one or a combination of at least two of apple juice, pear juice, orange juice or watermelon juice.
Preferably, the acclimation time is 12-24h, for example, 12h, 13h, 14h, 15h, 16h, 17h, 18h, 19h, 20h, 21h, 22h, 23h or 24h, preferably 20-24 h.
Preferably, the step of modifying the wine yeast is further included after the domestication of the wine yeast.
In the invention, the domesticated wine yeast is transformed by adopting an ultraviolet lamp irradiation method, wherein the power of the ultraviolet lamp is 30-50W, for example, 30W, 31W, 32W, 33W, 34W, 35W, 36W, 37W, 38W, 39W, 40W, 41W, 42W, 43W, 44W, 45W, 46W, 47W, 48W, 49W or 50W, preferably 35-40W.
Preferably, the modification time is 30-180s, for example 30s, 40s, 50s, 60s, 70s, 80s, 90s, 100s, 110s, 120s, 130s, 140s, 150s, 160s, 170s or 180s, preferably 120-.
In the invention, the time for modification can be adjusted by a person skilled in the art according to actual requirements.
As a preferred technical scheme, the invention provides a preparation method of tea fruit wine yeast, which comprises the following steps:
domesticating wine yeast by using a mixed solution of tea juice, fruit juice and polygonatum extract in a weight ratio of (3-8) to (1-3) to (1-2) for 12-24h, and modifying the domesticated wine yeast for 30-180s to obtain the tea fruit wine yeast.
In a second aspect, the present invention provides a wine yeast prepared by the method of the first aspect.
In a third aspect, the invention provides the use of the tea wine yeast of the second aspect in the preparation of tea wine.
In a fourth aspect, the present invention provides a method for producing a wine yeast budding rate of greater than 45% with a mortality rate of less than 5%.
In a fifth aspect, the present invention provides a method for producing a wine yeast having a residual sugar content of less than 8% and an alcoholic strength of greater than 9%.
Compared with the prior art, the invention has the following beneficial effects:
(1) in the invention, the wine yeast is domesticated under the condition that the weight ratio of the tea juice to the fruit juice to the polygonatum extract is (3-8) to (1-3) to (1-2), thereby promoting the vigorous growth of the wine yeast, and having high concentration of bacterial suspension, high budding rate and low mortality;
(2) the wine yeast prepared by the invention can fully utilize sugar in the tea juice, the fruit juice and the polygonatum odoratum extract to carry out alcohol fermentation, has strong fermentation capacity, low residual sugar content and high alcohol content, and has a health-care effect.
Detailed Description
To further illustrate the technical means and effects of the present invention, the present invention is further described with reference to the following examples. It is to be understood that the specific embodiments described herein are merely illustrative of the invention and are not limiting of the invention.
The examples do not show the specific techniques or conditions, according to the technical or conditions described in the literature in the field, or according to the product specifications. The reagents or apparatus used are conventional products commercially available from normal sources, not indicated by the manufacturer.
Example 1
(1) Extracting tea juice: weighing a certain amount of tea powder in a sterilized conical flask, adding purified water according to the material-liquid ratio of 1:60, leaching in a water bath kettle at 90 ℃ for 10min, and filtering to obtain tea juice;
(2) preparing juice: squeezing several cleaned fruits, filtering, and measuring sugar degree of the juice with a handheld sugar meter;
(3) preparing a polygonatum extract: refer to the 'Chinese pharmacopoeia' 2015 edition;
(4) activation of wine yeast: weighing 2g of sucrose, dissolving in 40mL of water, stirring for dissolving, performing high-pressure steam sterilization, weighing 4g of wine yeast, adding into the sucrose solution, incubating at 38 ℃ for 20min, and activating the wine yeast;
(5) and (3) sterilization: sterilizing with high pressure steam at 121 deg.C for 15min, and pasteurizing the tea juice and fruit juice at 65 deg.C for 30 min.
Example 2 preparation of wine Yeast
Taking 50mL of mixed solution of oolong tea juice, apple juice and radix polygonati officinalis extract with the weight ratio of 3:1:1 under aseptic condition, putting another 3mL of activated wine yeast into a 150mL conical flask, carrying out shake culture at room temperature for 24h, and then irradiating with a 35W ultraviolet lamp for 150 s. And (3) after the culture is finished, subpackaging the white precipitate at the bottom of the culture solution into centrifuge tubes, centrifuging at 4000rpm for 10min, discarding the supernatant, collecting the precipitate, and storing in a refrigerator at 4 ℃ for later use.
EXAMPLE 3 preparation of wine Yeast
Taking 50mL of mixed solution of black tea juice, pear juice and polygonatum odoratum extract with the weight ratio of 3:3:1 in a 150mL conical flask under the aseptic condition, taking 3mL of activated wine yeast, transferring the activated wine yeast into the conical flask, performing shake culture at room temperature for 24h, and then irradiating by using a 40W ultraviolet lamp for 120 s. After the culture is finished, the white precipitate at the bottom of the culture solution is subpackaged into centrifuge tubes, centrifuged at 4000rpm for 10min, the supernatant is discarded, the precipitate is collected and stored in a refrigerator at 4 ℃ for later use.
EXAMPLE 4 preparation of wine Yeast
Aseptically placing 50mL of a mixture of green tea juice, orange juice and rhizoma Polygonati Odorati extract at a weight ratio of 8:1:2 in a 150mL Erlenmeyer flask, adding 3mL of activated wine yeast into the Erlenmeyer flask, shake culturing at room temperature for 20h, and irradiating with 30W ultraviolet lamp for 180 s. After the culture is finished, the white precipitate at the bottom of the culture solution is subpackaged into centrifuge tubes, centrifuged at 4000rpm for 10min, the supernatant is discarded, the precipitate is collected and stored in a refrigerator at 4 ℃ for later use.
EXAMPLE 5 preparation of wine Yeast
Taking 50mL of mixed solution of black tea juice, watermelon juice and radix polygonati officinalis extract with the weight ratio of 8:3:2 in a 150mL conical flask under the aseptic condition, taking 3mL of activated wine yeast, transferring into the conical flask, performing shake culture at room temperature for 12h, and then irradiating by a 50W ultraviolet lamp for 30 s. After the culture is finished, the white precipitate at the bottom of the culture solution is subpackaged into centrifuge tubes, centrifuged at 4000rpm for 10min, the supernatant is discarded, the precipitate is collected and stored in a refrigerator at 4 ℃ for later use.
Comparative example 1
The temperature was raised to 40 ℃ during the culture as compared with example 2, and other conditions were the same as in example 2.
Comparative example 2
Compared with example 2, apple juice is not added in the culture process, and other conditions are the same as example 2.
Comparative example 3
In comparison with example 2, no oolong tea juice was added during the cultivation, and the other conditions were the same as in example 2.
Comparative example 4
In comparison with example 2, no extract of Yuzhu was added during the culture, and the other conditions were the same as in example 2.
Identification method
The culture solution in the conical flasks of the examples and comparative examples was observed and microscopic, and the number of dead and viable cells of yeast was identified by the meyer's blue staining solution water immersion method, and counted by a hemocytometer, and the suspension concentration, budding rate and death rate were calculated.
Taking a counting plate with 25 middle squares as an example, the total number of bacteria in 5 middle squares is set as A0The total number of budding bacteria is A1The total number of dead bacteria is A2And if the bacterial liquid dilution multiple is B, then:
growth status of wine yeast
Table 1 shows the growth of wine yeast, the culture liquid of the wine yeast of the example is darker in color, the bottom of the culture liquid generates thicker white precipitate, a large number of small bubbles are generated, the growth is vigorous, and a large number of buds grow.
Compared with example 2, the culture temperature of comparative example 1 is too high, and comparative example 2 contains no fruit juice and has insufficient sugar content, thus severely inhibiting the growth of wine yeast; the culture conditions of comparative examples 3 and 4 were not good, reducing the growth rate of wine yeast.
TABLE 1 growth conditions of Angel wine yeasts
Bacterial suspension concentration, germination and mortality
Table 2 shows the bacterial suspension concentration, the budding rate and the death rate of the wine yeast, and example 2 provides the best conditions for the growth of the wine yeast, the tea juice, the fruit juice and the polygonatum extract are all suitable for the growth of the yeast, the bacterial suspension concentration of the wine yeast is high, the budding rate reaches the highest value, and the death rate is low.
In comparison with example 2, comparative example 2 contains no fruit juice, the sugar degree does not satisfy the growth of yeast, and the concentration of the bacterial suspension is at least (0.12 + -0.03). times.106cfu/mL, the germination rate is 0, and the mortality rate is high; comparative example 3 the highest concentration of fruit juice and the highest concentration of bacterial suspension was (12.18 + -0.94) × 106cfu/mL, but too large a sugar degree inhibits the budding of yeast, and the death rate is low; comparative example 4 juice concentration was higher and yeast mortality was low.
TABLE 2 microscopic examination of Angel wine yeasts (n ═ 3)
| Numbering | Bacterial suspension concentration (× 10)6cfu/mL) | Germination rate/% | Mortality rate/%) |
| Example 2 | 2.28±0.18 | 46.18±4.99 | 1.47±0.81 |
| Example 3 | 2.89±0.19 | 45.32±3.56 | 1.91±0.78 |
| Example 4 | 1.78±0.18 | 41.22±2.99 | 4.54±0.81 |
| Example 5 | 3.45±0.28 | 44.34±3.15 | 1.99±0.67 |
| Comparative example 1 | 3.43±0.15 | 0.00±0.00 | 71.66±4.78 |
| Comparative example 2 | 0.12±0.03 | 0.00±0.00 | 61.11±4.21 |
| Comparative example 3 | 12.18±0.94 | 11.44±1.28 | 1.09±0.20 |
| Comparative example 4 | 10.85±0.20 | 12.29±1.69 | 1.45±0.59 |
Sugar and alcohol content of wine yeast
Table 3 shows the sugar content and alcohol content of wine yeast, and the wine yeast obtained in the examples had low residual sugar content and high alcohol content.
Compared with the example 2, the wine yeast of the comparative example 1 has overhigh culture temperature and has obvious influence on residual sugar content and alcoholic strength; the acclimatization conditions of comparative examples 2 to 4 were not good, affecting the alcoholic fermentation ability of wine yeast.
TABLE 3 sugar and alcohol content of Angel wine Yeast (n ═ 3)
| Numbering | Residual sugar degree/%) | Alcohol degree/% vol |
| Example 2 | 7.00±0.46 | 9.13±0.55 |
| Example 3 | 7.13±0.67 | 9.01±0.57 |
| Example 4 | 7.97±0.49 | 8.49±0.55 |
| Example 5 | 7.47±0.51 | 8.67±0.40 |
| Comparative example 1 | 15.97±0.67 | 4.27±0.31 |
| Comparative example 2 | 11.25±0.34 | 5.46±0.51 |
| Comparative example 3 | 15.47±0.55 | 6.11±0.26 |
| Comparative example 4 | 12.51±0.46 | 5.76±0.24 |
In conclusion, the wine yeast is domesticated under the condition that the weight ratio of the tea juice to the fruit juice to the polygonatum extract is (3-8) to (1-3) to (1-2), the vigorous growth of the wine yeast is promoted, the bacterial suspension concentration is high, the budding rate is high, the death rate is low, the obtained wine yeast can perform alcohol fermentation by fully utilizing the tea juice to the fruit juice and the polygonatum extract, the fermentation capacity is strong, the residual sugar content is low, the alcohol content is high, and the health care effect is achieved.
The applicant states that the present invention is illustrated in detail by the above examples, but the present invention is not limited to the above detailed methods, i.e. it is not meant that the present invention must rely on the above detailed methods for its implementation. It should be understood by those skilled in the art that any modification of the present invention, equivalent substitutions of the raw materials of the product of the present invention, addition of auxiliary components, selection of specific modes, etc., are within the scope and disclosure of the present invention.
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| CN101624560A (en) * | 2009-07-27 | 2010-01-13 | 山东俚岛海洋科技股份有限公司 | Preparation method of kelp fruit wine |
| CN203700336U (en) * | 2014-01-14 | 2014-07-09 | 陕西云智知识产权运营管理有限公司 | Yeast culture acclimatization system used for production of fruit wine |
| CN106244387A (en) * | 2016-10-26 | 2016-12-21 | 长江师范学院 | A kind of Composite fermentation type fruit wine and preparation method thereof |
| CN107312685A (en) * | 2017-08-30 | 2017-11-03 | 靖西市金峰贸易有限公司 | A kind of brewing method of emblic fruit wine |
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2018
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101624560A (en) * | 2009-07-27 | 2010-01-13 | 山东俚岛海洋科技股份有限公司 | Preparation method of kelp fruit wine |
| CN203700336U (en) * | 2014-01-14 | 2014-07-09 | 陕西云智知识产权运营管理有限公司 | Yeast culture acclimatization system used for production of fruit wine |
| CN106244387A (en) * | 2016-10-26 | 2016-12-21 | 长江师范学院 | A kind of Composite fermentation type fruit wine and preparation method thereof |
| CN107312685A (en) * | 2017-08-30 | 2017-11-03 | 靖西市金峰贸易有限公司 | A kind of brewing method of emblic fruit wine |
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