CN115474641A - Lactobacillus acidophilus fermented broadleaf holly leaf and fermentation process and application thereof - Google Patents
Lactobacillus acidophilus fermented broadleaf holly leaf and fermentation process and application thereof Download PDFInfo
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23F—COFFEE; TEA; THEIR SUBSTITUTES; MANUFACTURE, PREPARATION, OR INFUSION THEREOF
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- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/23—Lactobacillus acidophilus
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Abstract
The invention provides lactobacillus acidophilus fermented broadleaf holly tea, which is prepared by fermenting yuqing lobular broadleaf holly tea by lactobacillus acidophilus, and the adopted fermentation process comprises the following steps: 1) Preparing a culture medium; 2) Culturing the first-level strain of lactobacillus acidophilus to obtain a second-generation seed suspension; 3) Inoculating the second-generation seed suspension for fermentation, wherein the dosage ratio of the broadleaf holly leaf to the pure water to the lactobacillus acidophilus suspension is 45-55g. 4) And (5) drying. The invention belongs to the technical field of tea fermentation, and provides lactobacillus acidophilus fermented broadleaf holly leaf which has double physiological effects of lactobacillus acidophilus and lobular broadleaf, can effectively improve the content of active ingredients, improves the taste of the broadleaf holly leaf, and is easy to brew and absorb by a human body.
Description
Technical Field
The invention belongs to the technical field of tea fermentation, and particularly relates to lactobacillus acidophilus fermented broadleaf holly leaf, a fermentation process and application thereof.
Background
According to the record of Chinese medicine dictionary, kuding tea has the functions of clearing away heat and toxic material, resisting bacteria, relieving inflammation, relieving pain, stopping diarrhea, promoting the production of body fluid, quenching thirst, regulating body function, etc. The Yuqing lobular Kuding tea is a unique rare Kuding tea in Guizhou, belongs to perennial evergreen plants, mainly grows on the coasts of Wujiang and Miao Ling mountains with the altitude of 400-800 m, is completely different from Aquifoliaceae lobular Kuding tea in Guangdong, hainan, guangxi and other places, has tender and lasting fragrance, and has the characteristics of 'three greens' of dry tea green, green leaf bottom and green liquor color.
According to the reports of the literature, the Yuqing lobular ilex latifolia tea contains 17 amino acids necessary for human bodies, contains various substances such as selenium, zinc, magnesium, tea polyphenol, reducing sugar and the like, has higher flavonoid content and low caffeine content, finds more than 70 compounds beneficial to human health in the developed oil, and has the effects of reducing blood pressure, blood fat, aging, clearing heat and toxic materials, invigorating stomach and removing food stagnation, promoting urination and losing weight, preventing and inhibiting cancers and the like. Yuqing lobular Kuding tea is listed in Chinese pharmacopoeia in 2008, has dual purposes of medicine and food, has cool property, slight bitter taste and sweet taste, has refreshing taste after drinking, has lasting aftertaste, and integrates health care and medicinal effects.
Yuqing Kuding tea belongs to the plant of the family of wood titaniaceae, and because of containing more phenolic acid substances, tea soup has strong bitter taste and is not easy to be accepted by the public. Therefore, it is necessary to improve the taste of the lobular Kuding tea by fermentation or the like. CN 110742155A discloses a production method of fermented broadleaf holly leaf, which comprises the following steps: picking and cleaning, deactivating enzymes, performing primary fermentation, cleaning tea leaves, performing secondary fermentation, screening and drying; wherein, the primary fermentation is to fully mix the protective agent and the tea leaves and ferment for 5 hours at 16-22 ℃, the secondary fermentation is to spray fermentation liquor on the cleaned tea leaves and place the fermentation liquor at room temperature of 25-30 ℃ for continuous fermentation for 3 hours, although the secondary fermentation is used, the improvement effect on the taste of the broadleaf holly leaf is limited due to shorter fermentation time.
Therefore, the fermented broadleaf holly leaf and the fermentation process thereof, which can effectively improve the content of active ingredients and improve the taste of the broadleaf holly leaf, have important significance.
Disclosure of Invention
In order to solve the problems in the prior art, the invention carries out extensive research on the aspects of strain screening, ilex latifolia thunb fermentation, fermented product detection and the like, preferably selects suitable strains and fermentation conditions, and provides lactobacillus acidophilus fermented ilex latifolia thunb tea and a fermentation process and application thereof.
The objects of the present invention will be further explained by the following detailed description.
The invention provides lactobacillus acidophilus fermented broadleaf holly leaf, which is prepared by fermenting yuqing lobular broadleaf holly leaf by lactobacillus acidophilus, and the adopted fermentation process comprises the following steps:
1) Preparing a culture medium;
2) Culturing the first-level strain of lactobacillus acidophilus to obtain a second-generation seed suspension;
3) Inoculating the second-generation seed suspension for fermentation, wherein the dosage ratio of the broadleaf holly leaf to the pure water to the lactobacillus acidophilus suspension is 45-55g, 2-5mL, the fermentation temperature is 18-30 ℃, and the fermentation time is 12-18d.
4) And (5) drying.
Correspondingly, the invention also provides a fermentation process for fermenting the broadleaf holly leaf by the lactobacillus acidophilus, which comprises the following steps:
1) Preparing a culture medium;
2) Culturing the first-class strain of lactobacillus acidophilus to obtain a second-generation seed suspension;
3) Inoculating the second-generation seed suspension, and fermenting at 21-30 ℃ for 12-18 days according to the dosage ratio of the broadleaf holly leaf to the pure water to the lactobacillus acidophilus suspension of 45-55g.
4) And (5) drying.
Preferably, the dosage ratio of the broadleaf holly leaf to the pure water to the lactobacillus acidophilus suspension is 48-52g.
More preferably, the ratio of the amount of broadleaf holly leaf, purified water and lactobacillus acidophilus suspension is 50g. The amount of the lactobacillus acidophilus suspension cannot be too low, and if the amount is too low (such as less than 2 mL), the lactobacillus acidophilus suspension is not beneficial to become a dominant flora, so that the fermentation effect is not good.
Preferably, the temperature of the fermentation is 22-27 ℃, and the time of the fermentation is 13-16d.
More preferably, the temperature of the fermentation is 25 ℃ and the time of fermentation is 15d.
Preferably, the air humidity of the fermentation is 50% -60%.
Preferably, the culture medium is MRS broth culture medium or is prepared from MRS broth.
Preferably, the culturing of the first grade strain of lactobacillus acidophilus comprises the steps of activating, rejuvenating and expanding culturing of lactobacillus acidophilus.
In addition, the invention also provides application of the lactobacillus acidophilus fermented broadleaf holly leaf in preparing health-care food.
Preferably, the health food has effects including: has antioxidant, antiinflammatory, intestinal flora improving, and odor eliminating effects.
Compared with the prior art, the invention has the beneficial effects that:
(1) The invention preferably selects suitable strains and fermentation conditions, and provides the lactobacillus acidophilus fermented broadleaf holly leaf which has double physiological effects of lactobacillus acidophilus and lobular broadleaf holly leaf, can effectively improve the content of active ingredients, improves the taste of the broadleaf holly leaf, and is easy to brew and absorb by a human body.
(2) The fermentation process for fermenting the broadleaf holly leaves by the lactobacillus acidophilus adopts a microbial fermentation method, has simple process and low energy consumption, avoids the use of complex flora, does not need secondary fermentation, is easier to control the fermentation environment and effect, has longer fermentation time, ensures the fermentation effect, and achieves the aims of deeply processing and improving the quality of the Yuqing lobular broadleaf holly leaves.
(3) The invention also provides the application of the lactobacillus acidophilus fermented broadleaf holly leaf in preparing health-care food, and the health-care food has the following effects: antioxidant, anti-inflammatory, improving intestinal flora, and eliminating odor.
Drawings
FIG. 1 is a schematic view of a fermentation process of the present invention.
Detailed Description
The present invention will be described in further detail with reference to the accompanying drawings and examples.
In the invention, the bacterial strains, materials and reagents are all conventional commercial products or can be obtained by means of conventional technical means in the field.
In this example, the preparation method of the culture medium includes the following steps:
(1) First-class seed liquid culture medium
Weighing 48.3g of MRS broth, adding 1000mL of pure water, and heating and boiling until the MRS broth is fully dissolved; subpackaging the test tubes, wherein each test tube should not exceed 5mL, sterilizing, and cooling to room temperature for use.
(2) Second-stage seed liquid culture medium
Weighing 48.3g of MRS broth, adding 100mL of pure water, and heating and boiling until the MRS broth is fully dissolved; sterilizing, and cooling to room temperature.
(3) Solid culture medium
Weighing 48.3g of commercial product MRS broth culture medium and 15g of agar, adding 1000mL of pure water, stirring, heating and boiling until the MRS broth culture medium is completely dissolved; sterilizing at 121 deg.C under high pressure for 15min, cooling to 45 deg.C, and pouring into plate.
Example A Lactobacillus acidophilus fermented broadleaf holly leaf
Lactobacillus acidophilus fermented ilex is prepared by fermenting petqing lobular ilex with Lactobacillus acidophilus, the preparation process is schematically shown in figure 1, and the adopted fermentation process comprises the following steps:
1) Preparing a culture medium, namely preparing a primary seed liquid culture medium, a secondary seed liquid culture medium and a solid culture medium as described above;
2) Culturing the first-level strain of lactobacillus acidophilus to obtain a second-generation seed suspension; the method specifically comprises the following steps: (1) primary seed activation: inoculating lactobacillus acidophilus into the first-stage seed liquid culture medium according to 1.5% of the seed culture medium, and culturing in a shaking incubator at 37 deg.C for 36h to obtain first-generation seed liquid; (2) rejuvenation of the first-level strain: dipping the activated primary strain liquid by using the fired inoculating loop, carrying out zone-division scribing on the surface of a sterile flat plate, and then putting the sterile flat plate into a 37 ℃ incubator for culturing for 36h to obtain a single colony; picking a single colony to a liquid culture medium for culturing for 36 hours; (3) the amplification culture of the primary seeds: inoculating the rejuvenated first-stage lactobacillus acidophilus seeds into a second-stage seed liquid culture medium according to 1.5 percent of the seed culture medium, and culturing for 36 hours in a shaking incubator at the temperature of 37 ℃ to obtain second-stage seed suspension;
3) Inoculating second-generation seed suspension, and fermenting according to the proportion of Yuqing lobular ilex tea to pure water to Lactobacillus acidophilus suspension =50g to 3mL, wherein the fermentation temperature is (27 +/-2) DEG C, and the fermentation time is 15d; the humidity is kept between 50 and 60 percent, and the tea is turned over once a day to ferment the tea uniformly;
4) And (3) drying: pouring the fermented tea leaves into a tray, and putting the tray into a drying oven at 60 ℃ for drying for 3 hours to obtain lactobacillus acidophilus fermented broadleaf holly leaves.
Example Lactobacillus acidophilus fermented ilex
Lactobacillus acidophilus fermented ilex is prepared by fermenting Yuqing lobular ilex with Lactobacillus acidophilus, and the adopted fermentation process comprises the following steps:
1) Preparing a culture medium, namely preparing a primary seed liquid culture medium, a secondary seed liquid culture medium and a solid culture medium as described above;
2) Culturing the first-class strain of lactobacillus acidophilus to obtain a second-generation seed suspension; the method specifically comprises the following steps: (1) primary seed activation: inoculating lactobacillus acidophilus into a first-stage seed liquid culture medium according to 1.2% of the seed culture medium, and culturing in a shaking incubator at 37 deg.C for 36h to obtain a first-generation seed liquid; (2) rejuvenation of the first-level strain: dipping the activated primary strain liquid by using the fired inoculating loop, carrying out zone-division scribing on the surface of a sterile flat plate, and then putting the sterile flat plate into a 37 ℃ incubator for culturing for 36h to obtain a single colony; picking a single colony to a liquid culture medium for culturing for 36 hours; (3) expanding and culturing the primary seeds: inoculating the rejuvenated first-level lactobacillus acidophilus seeds into a second-level seed liquid culture medium according to the proportion of 1.2 percent of the seed culture medium, and culturing for 36 hours in a shaking incubator at 37 ℃ to obtain second-generation seed suspension;
3) Inoculating second-generation seed suspension, and fermenting according to the proportion of Yuqing lobular ilex tea to pure water to Lactobacillus acidophilus suspension =50g in volume of 3mL, wherein the fermentation temperature is (23 +/-2) DEG C, and the fermentation time is 17d; the humidity is kept between 50 and 60 percent, and the tea is turned over once a day to ferment the tea uniformly;
4) And (3) drying: pouring the fermented tea leaves into a tray, and putting the tray into a drying oven at 58 ℃ for drying for 3h to obtain the lactobacillus acidophilus fermented broadleaf holly leaf tea.
Example Lactobacillus acidophilus fermented Folum Ilicis
Lactobacillus acidophilus fermented broadleaf holly leaf is prepared by fermenting petqing lobular broadleaf holly leaf with Lactobacillus acidophilus, and the adopted fermentation process comprises the following steps:
1) Preparing a culture medium, namely preparing a primary seed liquid culture medium, a secondary seed liquid culture medium and a solid culture medium as described above;
2) Culturing the first-level strain of lactobacillus acidophilus to obtain a second-generation seed suspension; the method specifically comprises the following steps: (1) primary seed activation: inoculating lactobacillus acidophilus into a first-stage seed liquid culture medium according to 1.8% of the seed culture medium, and culturing in a shaking incubator at 37 deg.C for 36h to obtain a first-generation seed liquid; (2) rejuvenation of the first-level strain: dipping the activated primary strain liquid by using the fired inoculating loop, carrying out zone-division scribing on the surface of a sterile flat plate, and then putting the sterile flat plate into a 37 ℃ incubator for culturing for 36h to obtain a single colony; picking a single colony to be cultured in a liquid culture medium for 36 hours; (3) the amplification culture of the primary seeds: inoculating the rejuvenated first-stage lactobacillus acidophilus seeds into a second-stage seed liquid culture medium according to 1.8 percent of the seed culture medium, and culturing for 36 hours in a shaking incubator at the temperature of 37 ℃ to obtain second-stage seed suspension;
3) Inoculating a second-generation seed suspension, and fermenting according to the proportion of the Yuqing lobular Kuding tea to pure water to Lactobacillus acidophilus suspension =50g 3mL, wherein the fermentation temperature is (25 +/-2) DEG C, and the fermentation time is 15d; the humidity is kept between 50 and 60 percent, and the tea is turned over once a day to ferment the tea uniformly;
4) And (3) drying: pouring the fermented tea leaves into a tray, and putting the tray into a drying oven at 60 ℃ for drying for 3 hours to obtain lactobacillus acidophilus fermented broadleaf holly leaves.
Example detection of Lactobacillus acidophilus fermented broadleaf holly leaf
The comparison of the content of the components such as tea polyphenol and the like in the Lactobacillus acidophilus fermented lobular Kuding tea of the third embodiment and the microbially unfermented lobular Kuding tea are respectively shown in the following tables 1 and 2. The detection of tea polyphenol and catechin refers to GB/T8313-2018 detection method for tea polyphenol and catechin content in tea, the detection of total flavone refers to SN/T4592-2016 determination of total flavone in export food, and the detection of amino acid refers to GB 5009.124-2016 determination of amino acid in food.
TABLE 1 Table of the change of tea polyphenols etc. of ilex latifolia thunb fermented by lactobacillus acidophilus
Note: "-" indicates that the Lactobacillus acidophilus group was decreased in content compared to the control group; "+" indicates increased Lactobacillus acidophilus content compared to the control group.
As can be seen from Table 1, the tea polyphenol content of the Yuqing lobular Kuding tea in the Lactobacillus acidophilus group is reduced by 7.4% compared with the control group, which shows that the tea polyphenol content of the tea can be reduced through the fermentation of the Lactobacillus acidophilus, and the bitter taste is improved. Meanwhile, the flavone is one of the main bioactive components which are beneficial to the human body and contained in the tea leaves, is the main substance component forming the yellow green color of the green tea soup, and has the functions of oxidation resistance and the like. From the detection result, the total flavone content of the Lactobacilli acidophilus lobular Kuding tea is reduced by 13.6 percent. Catechin is the main polyphenol component in tea, and research reports indicate that the catechins have multiple effects of resisting oxidation, resisting inflammation, reducing the probability of heart diseases, preventing cancer, reducing blood fat, reducing body fat formation, resisting bacteria, changing intestinal bacteria, eliminating odor and the like. The catechin content of the Yuqing lobular Kuding tea in the lactobacillus acidophilus fermentation group is increased by 34.1 percent, which indicates that beneficial catechin components are increased by the lactobacillus acidophilus fermentation.
TABLE 2 amino acid content change table of Lactobacillus acidophilus fermented broadleaf holly leaf
As can be seen from Table 2, the L.acidophilus group tea leaves showed a 3.0% increase in glutamic acid content and a 20.0% increase in methionine content, but the contents were low. The content of alanine, glycine and proline is kept unchanged after fermentation by lactobacillus acidophilus. The content of other amino acids such as aspartic acid, arginine, lysine, histidine, phenylalanine, etc. is reduced after fermentation, wherein lysine is reduced by 9.8%. Research shows that amino acid is an important component for forming aroma and taste of tea and is also a key substance for presenting fresh and sweet taste of tea soup. The content of amino acid in the tea leaves accounts for 1-4% of the dry weight of the tea leaves, wherein phenylalanine, leucine, isoleucine, methionine, valine and threonine are essential amino acid for human bodies. Glutamic acid is an umami characteristic amino acid, alanine and glycine are sweet characteristic amino acids, and tyrosine and phenylalanine are fragrant characteristic amino acids.
The foregoing is a more detailed description of the invention in connection with specific preferred embodiments and it is not intended that the invention be limited to these specific details. For those skilled in the art to which the invention pertains, numerous simple deductions or substitutions may be made without departing from the spirit of the invention, which shall be deemed to belong to the scope of the invention.
Claims (10)
1. A Lactobacillus acidophilus fermented broadleaf holly leaf is characterized in that: the lactobacillus acidophilus is used for fermenting the Yuqing lobular Kuding tea, and the adopted fermentation process comprises the following steps:
1) Preparing a culture medium;
2) Culturing the first-class strain of lactobacillus acidophilus to obtain a second-generation seed suspension;
3) Inoculating the second-generation seed suspension for fermentation, wherein the dosage ratio of the broadleaf holly leaf to the pure water to the lactobacillus acidophilus suspension is 45-55g.
4) And (5) drying.
2. A fermentation process for fermenting broadleaf holly leaves by lactobacillus acidophilus is characterized by comprising the following steps: the method comprises the following steps:
1) Preparing a culture medium;
2) Culturing the first-class strain of lactobacillus acidophilus to obtain a second-generation seed suspension;
3) Inoculating the second-generation seed suspension, and fermenting at 21-30 ℃ for 12-18 days according to the dosage ratio of the broadleaf holly leaf to the pure water to the lactobacillus acidophilus suspension of 45-55g.
4) And (5) drying.
3. The fermentation process of lactobacillus acidophilus fermented broadleaf holly according to claim 2, characterized in that: the using amount ratio of the broadleaf holly leaf to the pure water to the lactobacillus acidophilus suspension is 48-52g.
4. The fermentation process of lactobacillus acidophilus fermented broadleaf holly according to claim 3, characterized in that: the dosage ratio of the broadleaf holly leaf, the pure water and the lactobacillus acidophilus suspension is 50g.
5. The fermentation process of lactobacillus acidophilus fermented ilex according to any of the claims from 2 to 4, characterized in that: the fermentation temperature is 22-27 ℃, and the fermentation time is 13-16d.
6. The fermentation process of Lactobacillus acidophilus fermented Folum Ilicis of claim 5, wherein: the fermentation temperature is 25 ℃, and the fermentation time is 15d.
7. The fermentation process of lactobacillus acidophilus fermented ilex according to any of the claims from 2 to 4, characterized in that: the air humidity of the fermentation is 50-60%.
8. The fermentation process of lactobacillus acidophilus fermented ilex according to any of the claims from 2 to 4, characterized in that: the culture medium is MRS broth culture medium or is prepared from MRS broth; the culture of the lactobacillus acidophilus first-level strain comprises the steps of activating, rejuvenating and expanding culture of the lactobacillus acidophilus.
9. Use of lactobacillus acidophilus fermented broadleaf holly according to any of claims 1 to 8 for the preparation of health food.
10. Use of lactobacillus acidophilus fermented broadleaf holly leaves according to claim 9 for the preparation of health foods having the effects comprising: antioxidant, anti-inflammatory, improving intestinal flora, and eliminating odor.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108813042A (en) * | 2018-07-04 | 2018-11-16 | 贵州省林艺波生态农业科技开发有限公司 | The processing method of Small-leaf Kuding tea fermented glossy ganoderma tea |
| CN109892440A (en) * | 2019-03-04 | 2019-06-18 | 宜宾学院 | A kind of coronoid process dissipate capsule bacterium fermentation Ilex Latifolia Thunb and preparation method thereof |
| CN111838369A (en) * | 2020-07-21 | 2020-10-30 | 余庆乌江泉小叶苦丁茶业有限公司 | Formula process of Yuqing lobular broadleaf holly leaf substitutional tea |
| CN114651888A (en) * | 2022-04-26 | 2022-06-24 | 贵州泰和现代生态农业科技有限公司 | Lactobacillus lobular kudingii fermented tea and preparation method thereof |
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108813042A (en) * | 2018-07-04 | 2018-11-16 | 贵州省林艺波生态农业科技开发有限公司 | The processing method of Small-leaf Kuding tea fermented glossy ganoderma tea |
| CN109892440A (en) * | 2019-03-04 | 2019-06-18 | 宜宾学院 | A kind of coronoid process dissipate capsule bacterium fermentation Ilex Latifolia Thunb and preparation method thereof |
| CN111838369A (en) * | 2020-07-21 | 2020-10-30 | 余庆乌江泉小叶苦丁茶业有限公司 | Formula process of Yuqing lobular broadleaf holly leaf substitutional tea |
| CN114651888A (en) * | 2022-04-26 | 2022-06-24 | 贵州泰和现代生态农业科技有限公司 | Lactobacillus lobular kudingii fermented tea and preparation method thereof |
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