CN104931443A - Method for measuring yellow rice wine yeast growth curve by separation and purification - Google Patents
Method for measuring yellow rice wine yeast growth curve by separation and purification Download PDFInfo
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- CN104931443A CN104931443A CN201510353721.5A CN201510353721A CN104931443A CN 104931443 A CN104931443 A CN 104931443A CN 201510353721 A CN201510353721 A CN 201510353721A CN 104931443 A CN104931443 A CN 104931443A
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Abstract
The invention relates to a method for measuring a yellow rice wine yeast growth curve by separation and purification. The method includes the following steps: (1) performing separation and purification; (2) measuring absorbancy, adopting a dry cell weight method as a measurement method, numbering centrifuge tubes for dry weighing, taking 4mL of samples for centrifuging with 4000rpm for five minutes, removing supernate, washing cells with diluted hydrochloric acid, continuing washing and centrifuging with distilled water after centrifugation, and drying at the temperature of 50 DEG C for 10 hours prior to weighing; (3) adding all the components in a 25mL colorimetric tube, sizing the volume to 25mL, performing boiling water bath for 5 minutes, cooling to room temperature and measuring absorbancy under the condition of 520m, wherein a linear expression of the absorbancy and standard glucose concentration is y=1.2289X-0.4568, R<2>=0.9966, X is the standard glucose concentration (mg/mL), and y is the absorbancy. By the method, the growth curve of yeast can be measured accurately, production state of the yeast can be grasped, and a foundation is laid for making high-quality yellow rice wine.
Description
Technical field
The present invention relates to a kind of method being measured yellow wine yeast growth curve by separation and purification, belong to the technical field of brewing yellow rice wine.
Background technology
The saccharifying agent in brewing yellow rice wine is not only by yeast, fraction leaven, also gives the local flavor of yellow rice wine uniqueness simultaneously.Because yeast occupies extremely important status in brewing yellow rice wine, the quality of its quality, directly has influence on the Quality and yield of yellow rice wine, is therefore likened to visually " bone of wine ".Therefore, prepare to measure the growth curve of yeast, the production status of grasp yeast is very necessary.The present invention produces thus.
Summary of the invention
For the above-mentioned technical matters of prior art, the object of this invention is to provide a kind of method being measured yellow wine yeast growth curve by separation and purification, the growth curve of Accurate Determining yeast, the production status of grasp yeast, lay the foundation for brewageing high-quality yellow rice wine.
For achieving the above object, the present invention is achieved by the following technical solutions:
Measured a method for yellow wine yeast growth curve by separation and purification, comprise the following steps:
(1) step of separation and purification:
A, get preservation strain, add the flat lining out of 2% agar sterilizing at brewer's wort and be separated, cultivate two days for 30 DEG C, then repeat line and be separated once, be then transferred on the inclined-plane of same medium, preservation in refrigerator;
B, to get in inclined-plane in bacterial strain access 250mL sterilizing wheat juice, 100rpm, temperature 30 DEG C of shaking tables are cultivated every 2 hours sampling 5mL, and wherein 1mL is with measuring absorbance method determination growth curve, and 4mL is used for dry cell weight method determination growth curve in addition;
(2) mensuration of absorbance: sampling 1mL dilutes 20 times and measures absorbance to 620m place, using same dilution wheat juice as blank;
Described assay method is dry cell weight method: the drying of centrifuge tube numbering is weighed, sampling 4mL in 4000rpm centrifugal 5 minutes, supernatant discarded night, watery hydrochloric acid washing thalline, and centrifugal rear continuation distilled water washing is centrifugal, within 10 hours, weighs in 50 DEG C of oven dry;
(3) add each component in 25mL color comparison tube, be settled to 25mL, boiling water bath 5 minutes, cool to room temperature, measures absorbance under 520m;
The linear representation of absorbance and standard glucose concentration is y=1.2289X-0.4568,
R
2=0.9966, X is standard glucose concentration (mg/mL), y is absorbance;
(4) inoculation yeast is in 170ml malt extract medium, does a blank simultaneously, 30 DEG C of cultivations, as initial time, and the cell number of record start solution, and in cultivation, 2,4,6,8,10,12,14,16,20,22, within 24 hours, measure the light absorption value of saccharomycete at 660nm place; Then make ordinate with yeast cell extinction matter, growth time makes horizontal ordinate, draws growth curve.
Beneficial effect of the present invention is as follows:
A kind of method being measured yellow wine yeast growth curve by separation and purification of the present invention, can Accurate Determining yeast growth curve, grasp the production status of yeast, lay the foundation for brewageing high-quality yellow rice wine.
Accompanying drawing explanation
Fig. 1 is the growth curve chart of yeast of the present invention.
Embodiment
Below in conjunction with specific embodiment, the present invention is further illustrated, but protection scope of the present invention is not limited to this.
The present invention measures the method for yellow wine yeast growth curve by separation and purification, comprises the following steps:
(1) step of separation and purification:
A, get preservation strain, add the flat lining out of 2% agar sterilizing at brewer's wort and be separated, cultivate two days for 30 DEG C, then repeat line and be separated once, be then transferred on the inclined-plane of same medium, preservation in refrigerator;
B, to get in inclined-plane in bacterial strain access 250mL sterilizing wheat juice, 100rpm, temperature 30 DEG C of shaking tables are cultivated every 2 hours sampling 5mL, and wherein 1mL is with measuring absorbance method determination growth curve, and 4mL is used for dry cell weight method determination growth curve in addition;
(2) mensuration of absorbance: sampling 1mL dilutes 20 times and measures absorbance to 620m place, using same dilution wheat juice as blank;
Described assay method is dry cell weight method: the drying of centrifuge tube numbering is weighed, sampling 4mL in 4000rpm centrifugal 5 minutes, supernatant discarded night, watery hydrochloric acid washing thalline, and centrifugal rear continuation distilled water washing is centrifugal, within 10 hours, weighs in 50 DEG C of oven dry;
(3) according to the form below 1 adds each component in 25mL color comparison tube, is settled to 25mL, boiling water bath 5 minutes, and cool to room temperature, measures absorbance under 520m;
The linear representation of absorbance and standard glucose concentration is y=1.2289X-0.4568,
R
2=0.9966, X is standard glucose concentration (mg/mL), y is absorbance;
(4) inoculation yeast is in 170ml malt extract medium, does a blank simultaneously, 30 DEG C of cultivations, as initial time, and the cell number of record start solution, and in cultivation, 2,4,6,8,10,12,14,16,20,22, within 24 hours, measure the light absorption value of saccharomycete at 660nm place; Then make ordinate with yeast cell extinction matter, growth time makes horizontal ordinate, obtained growth curve as shown in Figure 1.
A kind of method being measured yellow wine yeast growth curve by separation and purification of the present invention, can Accurate Determining yeast growth curve, grasp the production status of yeast, lay the foundation for brewageing high-quality yellow rice wine.
Above-described embodiment only illustrates inventive concept of the present invention for explaining, but not the restriction to rights protection of the present invention, all changes utilizing this design the present invention to be carried out to unsubstantiality, all should fall into protection scope of the present invention.
Claims (1)
1. measured a method for yellow wine yeast growth curve by separation and purification, it is characterized in that comprising the following steps:
(1) step of separation and purification:
A, get preservation strain, add the flat lining out of 2% agar sterilizing at brewer's wort and be separated, cultivate two days for 30 DEG C, then repeat line and be separated once, be then transferred on the inclined-plane of same medium, preservation in refrigerator;
B, to get in inclined-plane in bacterial strain access 250mL sterilizing wheat juice, 100rpm, temperature 30 DEG C of shaking tables are cultivated every 2 hours sampling 5mL, and wherein 1mL is with measuring absorbance method determination growth curve, and 4mL is used for dry cell weight method determination growth curve in addition;
(2) mensuration of absorbance: sampling 1mL dilutes 20 times and measures absorbance to 620m place, using same dilution wheat juice as blank;
Described assay method is dry cell weight method: the drying of centrifuge tube numbering is weighed, sampling 4mL in 4000rpm centrifugal 5 minutes, supernatant discarded night, watery hydrochloric acid washing thalline, and centrifugal rear continuation distilled water washing is centrifugal, within 10 hours, weighs in 50 DEG C of oven dry;
(3) add each component in 25mL color comparison tube, be settled to 25mL, boiling water bath 5 minutes, cool to room temperature, measures absorbance under 520m;
The linear representation of absorbance and standard glucose concentration is y=1.2289X-0.4568,
R2=0.9966, X are standard glucose concentration (mg/mL), y is absorbance;
(4) inoculation yeast is in 170ml malt extract medium, does a blank simultaneously, 30 DEG C of cultivations, as initial time, and the cell number of record start solution, and in cultivation, 2,4,6,8,10,12,14,16,20,22, within 24 hours, measure the light absorption value of saccharomycete at 660nm place; Then make ordinate with yeast cell extinction matter, growth time makes horizontal ordinate, draws growth curve.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106290162A (en) * | 2016-10-10 | 2017-01-04 | 承启医学（深圳）科技有限公司 | A kind of apparatus and method of continuous detecting growth curve of bacteria |
| CN107764754A (en) * | 2017-08-31 | 2018-03-06 | 厦门博露环保科技有限公司 | A kind of online test method of microbes biomass |
Citations (2)
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| CN102766580A (en) * | 2011-05-05 | 2012-11-07 | 中国科学院生态环境研究中心 | Yeast strain for producing biosurfactant and application thereof |
| CN103740807A (en) * | 2013-11-06 | 2014-04-23 | 中国热带农业科学院环境与植物保护研究所 | Yeast separating and screening method and biocontrol bacterial agent preparation method |
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2015
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102766580A (en) * | 2011-05-05 | 2012-11-07 | 中国科学院生态环境研究中心 | Yeast strain for producing biosurfactant and application thereof |
| CN103740807A (en) * | 2013-11-06 | 2014-04-23 | 中国热带农业科学院环境与植物保护研究所 | Yeast separating and screening method and biocontrol bacterial agent preparation method |
Non-Patent Citations (3)
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| 王冰: "青霉素菌渣制取饲料酵母与酵母膏的工艺研究", 《中国优秀硕士学位论文全文数据库农业科技辑》 * |
| 王金娜: "生物破乳剂生产菌生长特性及破乳效能研究", 《中国优秀硕士学位论文全文数据库工程科技I辑》 * |
| 邱佩: "魔芋飞粉蛋白酶解物对酿酒酵母生长影响的研究", 《中国优秀硕士学位论文全文数据库•工程科技I辑》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106290162A (en) * | 2016-10-10 | 2017-01-04 | 承启医学（深圳）科技有限公司 | A kind of apparatus and method of continuous detecting growth curve of bacteria |
| CN106290162B (en) * | 2016-10-10 | 2019-09-06 | 承启医学（深圳）科技有限公司 | A kind of device and method of continuous detection bacterium growth curve |
| CN107764754A (en) * | 2017-08-31 | 2018-03-06 | 厦门博露环保科技有限公司 | A kind of online test method of microbes biomass |
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