Screening device and method for ester-producing yeast in Maotai-flavor yeast for making hard liquor
By designing a sauce-flavored Daqu middle-yield ester yeast screening device integrating temperature control, stirring, pH detection and multi-stage screening, using specific culture medium and lyophilization protection, the problems of low screening efficiency and inaccurate control in traditional methods are solved, and the problem of improved performance and improvement of the problem of low screening efficiency and low control. The quality and production efficiency of sauce-flavored Daqu are screened out to select high yield and diversity of ester-producing yeasts.
Patent Information
- Application Number
- CN202411275958.1
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2024-09-12
- Publication Date
- 2024-12-10
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
The screening method of traditional sauce-flavored Daqu middle-produced ester yeast is inefficient and complex in operation, making it difficult to ensure that the selected yeast is optimal in yield and type, and the temperature and pH value control during the culture process is not accurate enough, which affects the improvement of the wine flavor. .
A screening device integrating temperature control, stirring, pH detection, multi-stage screening and efficient lyophilization protection was designed. A 10-12°Bx corn saccharified mash or saccharified liquid with wheat as raw material was used as culture medium. A multi-stage screening process screened out high yield and diversity of ester-producing yeasts, and strain identification was performed using molecular biological identification devices.
The quality of the sauce-flavored Daqu was improved, the production process was optimized, and the strains with high ester production types and yields were successfully screened, laying a solid foundation for industrial production and improving the esterification capacity and growth rate.
Smart Images
Figure CN119101585A_ABST
Abstract
Description
Technical Field
[0001] The invention relates to the technical field of microbial resource utilization of Maotai-flavor Daqu, and in particular to a screening device and method for ester-producing yeast in Maotai-flavor Daqu. Background Art
[0002] In the brewing process of Maotai-flavor Daqu, the screening and utilization of ester-producing yeast plays a vital role in improving the flavor of the wine and enriching the aroma components. However, traditional screening methods often have problems such as low screening efficiency, complex operation, and unstable results. Specifically, traditional methods usually rely on manual screening and cultivation, which is not only time-consuming and labor-intensive, but also difficult to ensure that the screened ester-producing yeast is optimal in terms of yield and type. At the same time, the control of key parameters such as temperature and pH value during the cultivation process is not precise enough, resulting in limited yeast growth rate and esterification ability, which further affects the improvement of wine flavor.
[0003] In addition, the traditional method is also relatively single in the selection of culture medium, which often cannot meet the specific needs of different ester-producing yeasts. Especially for the special environment of Maotai-flavor Daqu, how to simulate its unique microbial ecosystem and improve the adaptability and activity of ester-producing yeasts is a problem that traditional methods are difficult to overcome.
[0004] In summary, the urgency of technological innovation is obvious. Developing a new ester-producing yeast screening device and method that integrates temperature control, stirring, pH detection, multi-stage screening and efficient freeze-drying protection is of great significance for improving the quality of Maotai-flavor Daqu, optimizing the production process and realizing industrial production. Summary of the invention
[0005] The present invention provides a screening device and method for ester-producing yeast in sauce-flavored Daqu, innovatively introducing corn saccharification mash with a Bx of 10 to 12° and saccharification liquid with wheat as raw material as culture medium, which can not only better simulate the fermentation environment of sauce-flavored Daqu, but also provide more abundant nutrients for ester-producing yeast, thereby promoting its growth and improving its esterification ability.
[0006] In order to achieve the above-mentioned purpose, the present invention provides the following technical solutions: a screening device for ester-producing yeast in sauce-flavored Daqu, comprising: a culture device, the culture device is a cavity structure with a stirring device, a temperature control device, and a pH detection device inside, and a bacterial powder adding device is provided on the top of the culture device, and an output end is provided on the side; a first flat plate culture unit; a single bacteria primary screening culture unit, the input end of the single bacteria primary screening culture unit is connected to the output end of the culture device through the first flat plate culture unit, and the single bacteria primary screening culture unit is provided with multiple output ends, and is respectively connected to a second flat plate culture unit, an esterification power detection device, and a single bacteria re-screening culture unit; the single bacteria re-screening culture unit is provided with multiple output ends, and is respectively connected to a flavor detection device, a total ester detection device and a single bacteria re-screening culture unit, and the single bacteria re-screening culture unit is provided with multiple output ends; a molecular biology identification device and a freeze-drying protection device, and the molecular biology identification device and the freeze-drying protection device are respectively connected to the output end of the single bacteria re-screening culture unit.
[0007] Preferably, the culture medium in the culture device is YPD culture medium.
[0008] Preferably, the single bacteria primary screening culture unit, the single bacteria secondary screening culture unit and the single bacteria re-screening culture unit all use corn saccharification mash with a Bx of 10 to 12°, or saccharification liquid using wheat as raw material.
[0009] Preferably, a screening method comprises the following steps:
[0010] Step 1: adding sauce-flavored Daqu powder and a selected culture medium to a culture device, and controlling the temperature at 25° C. by a temperature control device; stirring the culture solution by a stirring device, and controlling the pH value of the culture solution at 4.0 by a pH detection device;
[0011] Step 2: sampling the culture solution of the Maotai-flavor Daqu after culture to the first plate culture unit for spreading culture to obtain a single colony;
[0012] Step 3, picking a single bacterial colony from the first flat plate culture unit and placing it in the single bacterial primary screening culture unit for fermentation culture;
[0013] Step 4: Use an esterification power detection device to detect the esterification power of the primary screening fermentation broth, and select strains with higher esterification power to be placed in a single bacteria re-screening culture device;
[0014] Step 5: Ferment the strains in the rescreening culture device, and use a flavor detection device and a total ester detection device to detect the esterified liquid after fermentation, and select strains with high ester production types and yields to be placed in a single bacteria rescreening culture device;
[0015] Step 6: using a molecular biology identification device to identify the strain genus of the single-bacteria fermentation liquid in the re-screening culture device;
[0016] Step 7: Add freeze-drying protective agent to the freeze-drying protective device, freeze-dry the identified strains, and preserve the strains.
[0017] Preferably, the lyophilization protective agent is a 15% (w / v) trehalose solution, which is mixed with the ester-producing yeast suspension in a ratio of 2:1.
[0018] The beneficial effect of the present invention is that: corn saccharification mash with 10-12°Bx and saccharification liquid made from wheat are innovatively introduced as culture medium, which can not only better simulate the fermentation environment of sauce-flavored Daqu, but also provide more abundant nutrients for ester-producing yeast, promote its growth and esterification ability. At the same time, the preparation process of wheat saccharification liquid is optimized, and the quality and stability of saccharification liquid are ensured by accurately controlling parameters such as gelatinization time, saccharification temperature and time.
[0019] A high-precision temperature control device and heating wire system were introduced to achieve precise control of the culture temperature through a closed-loop control algorithm, ensuring that the ester-producing yeast grows efficiently within the optimal temperature range. At the same time, the addition of a stirring device and a pH detection device further improved the stability and controllability of the culture process. Ultimately, through a multi-stage screening process, strains with high ester production types and yields were successfully screened out, and the genus was identified through a molecular biology identification device, laying a solid foundation for subsequent industrial production and application. BRIEF DESCRIPTION OF THE DRAWINGS
[0020] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings required for use in the embodiments or the description of the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.
[0021] Figure 1 It is a schematic diagram of the overall structure of the present invention.
[0022] In the figure: 1. Cultivation device; 2. Stirring device; 3. Temperature control device; 4. pH detection device; 5. Bacterial powder adding device; 6. First flat plate cultivation unit; 7. Single bacteria primary screening cultivation unit; 8. Second flat plate cultivation unit; 9. Esterification power detection device; 10. Single bacteria re-screening cultivation unit; 11. Flavor detection device; 12. Total ester detection device; 13. Single bacteria re-screening cultivation unit; 14. Molecular biology identification device; 15. Freeze-drying protection device. DETAILED DESCRIPTION
[0023] The technical solution of the present invention will be described clearly and completely below in conjunction with the accompanying drawings of the present invention. Obviously, the described embodiments are only part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention.
[0024] according to Figure 1 As shown, a screening device for ester-producing yeast in sauce-flavored Daqu comprises: a culture device 1, wherein the culture device 1 is a cavity structure with a sauce-flavored Daqu powder adding device on the top, a stirring device 2, a temperature control device 3, and a pH detection device 4 inside, and a bacterial powder adding device 5 is provided on the top of the culture device 1, and an output end is provided on the side; a first flat culture unit 6; a single bacteria primary screening culture unit 7, wherein the input end of the single bacteria primary screening culture unit 7 is connected to the output end of the culture device 1 through the first flat culture unit 6, and the single bacteria primary screening culture unit 7 is connected to the output end of the culture device 1. Unit 7 is provided with multiple output ends, and is respectively connected to the second flat plate culture unit 8, the esterification power detection device 9, and the single bacteria re-screening culture unit 10; the single bacteria re-screening culture unit 10 is provided with multiple output ends, and is respectively connected to the flavor detection device 11, the total ester detection device 12 and the single bacteria re-screening culture unit 13, and the single bacteria re-screening culture unit 13 is provided with multiple output ends; a molecular biology identification device 14 and a freeze-drying protection device 15, and the molecular biology identification device 14 and the freeze-drying protection device 15 are respectively connected to the output ends of the single bacteria re-screening culture unit 13. Among them, the culture medium in the culture device 1 is selected from YPD culture medium. The single bacteria primary screening culture unit 7, the single bacteria re-screening culture unit 10 and the single bacteria re-screening culture unit 13 all use corn saccharification mash with 10-12°Bx, or saccharification liquid with wheat as raw material.
[0025] In the above-mentioned setting, the Maotai-flavor Daqu powder adding device is used to add Maotai-flavor Daqu powder to the culture device 1, and the inoculation amount is 10%; the culture device 1 is used to accommodate and cultivate the culture medium containing Maotai-flavor Daqu powder; the stirring device 2 is used to stir the culture solution to ensure that the Daqu powder and the culture medium are fully mixed and provide sufficient oxygen for the ester-producing yeast fermentation; the pH detection device 4 is used to detect the pH value of the culture solution and control it to maintain at about 4.0; the flat plate culture device 1 is used to spread the culture solution of Maotai-flavor Daqu to obtain a single colony; the single bacteria initial screening culture device 1 is used to pick up a single colony in the flat plate culture device 1 for fermentation culture; the esterification force The detection device 9 is used to detect the esterification power of the single-bacteria fermentation liquid fermented in the single-bacteria primary screening culture device 1; the single-bacteria re-screening culture device 1 is used to cultivate strains with higher esterification power; the flavor detection device 11 is used to detect various ester substances in the esterified liquid fermented in the re-screening culture device 1; the total ester detection device 12 is used to detect the total ester content of the esterified liquid fermented in the re-screening culture device 1; the single-bacteria re-screening culture device 1 is used to ferment strains with high ester types and yields; the molecular biology identification device 14 is used to identify the strain genus of the single-bacteria fermentation liquid in the re-screening culture device 1; the freeze-drying protection device 15 is used to perform freeze-drying protection on the strain after adding a freeze-drying protective agent.
[0026] The screening method includes the following steps:
[0027] Step 1: add sauce-flavored Daqu powder and a selected culture medium to the culture device 1, and control the temperature at 25° C. by the temperature control device 3; stir the culture solution by the stirring device 2, and control the pH value of the culture solution at 4.0 by the pH detection device 4;
[0028] Step 2: sampling the culture solution of the Maotai-flavor Daqu after culture to the first flat plate culture unit 6 for coating culture to obtain a single bacterial colony;
[0029] Step 3, picking up a single bacterial colony in the first flat plate culture unit 6 and placing it in the single bacteria primary screening culture unit 7 for fermentation culture;
[0030] Step 4: Use the esterification power detection device 9 to detect the esterification power of the primary screening fermentation liquid, and select strains with higher esterification power to be added to the single bacteria re-screening culture device 1;
[0031] Step 5: Ferment the strains in the re-screening culture device 1, and use the flavor detection device 11 and the total ester detection device 12 to detect the esterified liquid after fermentation, and select the strains with high ester production type and yield to be added to the single bacteria re-screening culture device 1;
[0032] Step 6: using the molecular biology identification device 14 to identify the strain genus of the single-bacteria fermentation liquid in the re-screening culture device 1;
[0033] Step 7: Add a freeze-drying protective agent to the freeze-drying protective device 15 to freeze-dry the identified strains and preserve the strains. The freeze-drying protective agent is a 15% w / v trehalose solution mixed with the ester-producing yeast suspension at a ratio of 2:1.
[0034] The above is only a specific embodiment of the present invention, but the protection scope of the present invention is not limited thereto. Any person skilled in the art can easily think of changes or substitutions within the technical scope disclosed by the present invention, which should be included in the protection scope of the present invention. Therefore, the protection scope of the present invention should be based on the protection scope of the claims.
Claims
1. A screening device for ester-producing yeast in Maotai-flavor Daqu, characterized in that: include: A culture device (1), wherein the culture device (1) is a cavity structure having a stirring device (2), a temperature control device (3), and a pH detection device (4) disposed therein, and a bacterial powder adding device (5) is disposed at the top of the culture device (1) and an output end is disposed at the side; A first plate culture unit (6); A single bacteria primary screening culture unit (7), wherein the input end of the single bacteria primary screening culture unit (7) is connected to the output end of the culture device (1) via the first flat plate culture unit (6), and the single bacteria primary screening culture unit (7) is provided with a plurality of output ends, which are respectively connected to the second flat plate culture unit (8), the esterification force detection device (9), and the single bacteria secondary screening culture unit (10); The single bacteria re-screening and culturing unit (10) is provided with a plurality of output ends, and is respectively connected to a flavor detection device (11), a total ester detection device (12) and a single bacteria re-screening and culturing unit (13); the single bacteria re-screening and culturing unit (13) is provided with a plurality of output ends; A molecular biology identification device (14) and a freeze-drying protection device (15), wherein the molecular biology identification device (14) and the freeze-drying protection device (15) are respectively connected to the output end of the single bacteria re-screening culture unit (13).
2. The screening device for ester-producing yeast in Maotai-flavor Daqu according to claim 1, characterized in that: The culture medium in the culture device (1) is YPD culture medium.
3. The screening device for ester-producing yeast in Maotai-flavor Daqu according to claim 2, characterized in that: The single bacteria primary screening culture unit (7), the single bacteria secondary screening culture unit (10) and the single bacteria re-screening culture unit (13) all use corn saccharification mash with a Bx of 10 to 12°, or saccharification liquid using wheat as raw material.
4. A screening method, used for the screening device of ester-producing yeast in the Maotai-flavor Daqu according to claim 1, characterized in that: The following steps are involved: Step 1: adding sauce-flavored Daqu powder and a selected culture medium to a culture device (1), and controlling the temperature at 25° C. by a temperature control device (3); stirring the culture solution by a stirring device (2), and controlling the pH value of the culture solution at 4.0 by a pH detection device (4); Step 2: sampling the culture solution of the Maotai-flavor Daqu after culturing and placing it in the first plate culture unit (6) for spreading and culturing to obtain a single colony; Step 3, picking a single bacterial colony from the first flat plate culture unit (6) and placing it in the single bacterial primary screening culture unit (7) for fermentation culture; Step 4: Use the esterification power detection device (9) to detect the esterification power of the primary screening fermentation broth, and select strains with higher esterification power to be placed in the single bacteria re-screening culture device (1); Step 5, fermenting and culturing the strains in the rescreening culture device (1), and using the flavor detection device (11) and the total ester detection device (12) to detect the esterified liquid after fermentation, and selecting strains with high ester production types and yields to be placed in the single bacteria rescreening culture device (1); Step 6: using a molecular biology identification device (14) to identify the strain genus of the single bacterial fermentation liquid in the rescreening culture device (1); Step 7: Add a freeze-drying protective agent to the freeze-drying protective device (15), freeze-dry the identified strains, and preserve the strains.
5. A screening method according to claim 4, characterized in that: The freeze-drying protective agent is a 15% (w / v) trehalose solution, which is mixed with the ester-producing yeast suspension in a ratio of 2:1.