Application of saussurea involucrata bacterial agent in wolfberry fermentation and product thereof

Abstract

The invention discloses application of a saussurea involucrata bacterial agent in fermentation of Chinese wolfberry fruits and a product thereof, and relates to the technical field of microbial fermentation. Research finds that when the saussurea involucrata bacterium agent is applied to fermentation of the Chinese wolfberry fruits, a novel Chinese wolfberry fruit deep processing product can be developed. Experiments prove that compared with the fructus lycii primary pulp, the fructus lycii deep processing product can remarkably improve the symptoms of the xerophthalmia, specifically, the tear secretion amount is increased, the tear film rupture time is prolonged, and corneal epithelium injury is reduced. According to the method, the saussurea involucrata is adopted as a fermentation strain to ferment the Chinese wolfberry primary pulp, a new Chinese wolfberry deep processing method is developed, and technical support is provided for further improving the commercial application value of the Chinese wolfberry.

Description

Technical Field

[0001] The invention relates to the technical field of microbial fermentation, in particular to application of a snow lotus fungus agent in wolfberry fermentation and products thereof. Background Art

[0002] Snow lotus mushroom, also known as Tibetan mushroom, is a milky white or light yellow gelatinous mass with irregular growth shape, most of which are spherical rice grains. Its surface curls up and can grow rapidly in milk, forming a cauliflower-like mass, which is elastic and sticky when squeezed by hand, similar to the appearance of snow lotus.

[0003] Wolfberry (Lycium barbarum L.) is a deciduous shrub of the genus Lycium in the family Solanaceae. It is rich in 32 trace elements and 18 amino acids needed by the human body, as well as various active ingredients such as wolfberry polysaccharides, betaine, carotenoids, etc. It has antioxidant, anti-fatigue, and immunity-enhancing effects, and has high medicinal health functions and nutritional value.

[0004] Dry eye syndrome, also known as keratoconjunctivitis sicca, is a common and complex ophthalmic disease caused by insufficient tear secretion, excessive evaporation or abnormal tear composition, which can cause eye discomfort and visual dysfunction.

[0005] The present invention intends to use snow lotus as a fermentation strain to ferment wolfberry pulp to develop a new wolfberry deep-processed product to play the function of improving eyesight and improving the symptoms of dry eyes. Summary of the invention

[0006] The purpose of the present invention is to provide an application of a snow lotus fungus agent in wolfberry fermentation and a product thereof to solve the problems existing in the above-mentioned prior art. The present invention has found that by applying the snow lotus fungus agent to wolfberry fermentation, a new wolfberry deep-processed product can be developed, and the wolfberry deep-processed product can significantly improve the symptoms of dry eye.

[0007] To achieve the above object, the present invention provides the following solutions:

[0008] The present invention provides a method for preparing a wolfberry fermented product, comprising the following steps:

[0009] Inoculating snow lotus fungus into wolfberry puree, and subjecting it to fermentation treatment to obtain fermentation liquid;

[0010] The fermentation liquid is centrifuged to obtain a supernatant, which is then concentrated to obtain a wolfberry extract, namely the wolfberry fermentation product.

[0011] Furthermore, the inoculation amount of the snow lotus fungus is 3-5%.

[0012] Furthermore, the fermentation treatment is carried out at a temperature of 25-28°C, a rotation speed of 120-140 rpm, and a time of 36-48 hours.

[0013] Furthermore, the solid content of the wolfberry extract is 40-60wt%.

[0014] The present invention also provides a wolfberry fermented product prepared according to the above preparation method.

[0015] The invention also provides application of the snow lotus fungus agent in wolfberry fermentation, and the wolfberry fermentation is used for preparing a product for treating dry eye.

[0016] The present invention also provides application of the wolfberry fermented product in preparing a product for treating dry eye.

[0017] Furthermore, the product for treating dry eyes is a medicine or a health product.

[0018] The present invention also provides a product for treating dry eyes, wherein the active ingredient comprises the wolfberry fermentation product.

[0019] Furthermore, the product for treating dry eyes is a medicine or a health product.

[0020] The present invention discloses the following technical effects:

[0021] The present invention has found that a new wolfberry deep-processed product can be developed by applying the snow lotus fungus agent to wolfberry fermentation. Experiments have confirmed that compared with wolfberry puree, the wolfberry deep-processed product can significantly improve the symptoms of dry eye, specifically by increasing tear secretion, prolonging tear film breakup time, and reducing corneal epithelial damage.

[0022] The invention adopts snow lotus fungus as the fermentation strain to ferment wolfberry pulp, opens up a new wolfberry deep processing method, and provides technical support for further improving the commercial application value of wolfberry. DETAILED DESCRIPTION

[0023] Various exemplary embodiments of the present invention will now be described in detail. This detailed description should not be considered as limiting the present invention, but should be understood as a more detailed description of certain aspects, features, and embodiments of the present invention.

[0024] It should be understood that the terms described in the present invention are only for describing a particular embodiment and are not intended to limit the present invention. In addition, for the numerical range in the present invention, it should be understood that each intermediate value between the upper and lower limits of the scope is also specifically disclosed. The intermediate value in any stated value or stated range, and each smaller range between any other stated value or intermediate value in the described range is also included in the present invention. The upper and lower limits of these smaller ranges can be independently included or excluded in the scope.

[0025] Unless otherwise indicated, all technical and scientific terms used herein have the same meanings as those generally understood by those skilled in the art. Although the present invention describes only preferred methods and materials, any methods and materials similar or equivalent to those described herein may also be used in the implementation or testing of the present invention. All documents mentioned in this specification are incorporated by reference to disclose and describe the methods and / or materials associated with the documents. In the event of a conflict with any incorporated document, the content of this specification shall prevail.

[0026] It will be apparent to those skilled in the art that various modifications and variations may be made to the specific embodiments of the present invention description without departing from the scope or spirit of the present invention. Other embodiments derived from the present invention description will be apparent to those skilled in the art. The present invention description and examples are exemplary only.

[0027] The words “include,” “including,” “have,” “contain,” etc. used in this document are open-ended terms, meaning including but not limited to.

[0028] The snow lotus fungus used in the present invention was donated by Zhejiang University and has been disclosed in the document "Li Yun. Research on Identification and Probiotic Properties of Snow Lotus Fungus Yeast". The present inventor promises to release it to the public 20 years from the filing date of the present invention.

[0029] Example 1

[0030] (1) Wash fresh wolfberries, dry them, and grind them with a grinder to obtain wolfberry puree. During grinding, water 4 times the weight of the wolfberries is added.

[0031] (2) The wolfberry puree was sterilized at 121° C. for 15 min. After cooling, the sterilized wolfberry puree was inoculated with Saussurea spp. at an inoculum amount of 5% (w / v). The mixture was fermented at 28° C. and 120 rpm for 48 h. The mixture was then heated to 100° C. and sterilized for 5 min to obtain a fermentation liquid.

[0032] (3) The fermentation liquid is centrifuged to obtain a supernatant, and then the supernatant is concentrated by a rotary evaporator to a solid content of 50 wt %, to obtain a wolfberry extract.

[0033] Example 2

[0034] (1) Fresh wolfberries were washed and dried, and then ground with a grinder to obtain wolfberry puree. During the grinding, 4.5 times the weight of water relative to the weight of the wolfberries was added.

[0035] (2) The wolfberry puree was sterilized at 121° C. for 15 min. After cooling, the sterilized wolfberry puree was inoculated with Saussurea spp. at an inoculum amount of 4% (w / v). The mixture was fermented at 27° C. and 130 rpm for 42 h. The mixture was then heated to 100° C. and sterilized for 5 min to obtain a fermentation liquid.

[0036] (3) The fermentation liquid is centrifuged to obtain a supernatant, and then the supernatant is concentrated by a rotary evaporator to a solid content of 50 wt %, to obtain a wolfberry extract.

[0037] Example 3

[0038] (1) Wash fresh wolfberries, dry them, and grind them with a grinder to obtain wolfberry puree. During grinding, add 5 times the weight of water relative to the weight of the wolfberries.

[0039] (2) The wolfberry puree was sterilized at 121° C. for 15 min. After cooling, the sterilized wolfberry puree was inoculated with Saussurea spp. at an inoculum amount of 3% (w / v). The mixture was fermented at 25° C. and 140 rpm for 36 h. The temperature was then raised to 100° C. and sterilized for 5 min to obtain a fermentation liquid.

[0040] (3) The fermentation liquid is centrifuged to obtain a supernatant, and then the supernatant is concentrated by a rotary evaporator to a solid content of 40 wt % to obtain a wolfberry extract.

[0041] Comparative Example 1

[0042] (1) Wash fresh wolfberries, dry them, and grind them with a grinder to obtain wolfberry puree. During grinding, water 4 times the weight of the wolfberries is added.

[0043] (2) The wolfberry pulp was sterilized at 121° C. for 15 min, cooled, fermented and cultured at 28° C. and 120 rpm for 48 h, then heated to 100° C. and sterilized for 5 min, and then centrifuged to obtain a supernatant, which was concentrated to a solid content of 60 wt % using a rotary evaporator to obtain a wolfberry extract.

[0044] Effect verification example

[0045] 1. Materials and Methods

[0046] 1.1 Experimental Materials

[0047] Experimental animals: 8-week-old healthy female C57BL / 6 mice, weighing 20±2 g.

[0048] 1.2 Establishment of dry eye mouse model

[0049] 5 μL of benzalkonium chloride solution (concentration 2 mg / mL) was dripped into the conjunctival sac of both eyes of mice twice a day for 14 days to prepare a dry eye mouse model.

[0050] 1.3 Experimental groups and drug administration

[0051] The experiment was divided into a control group, a model group, a positive drug group, an experimental group 1, and an experimental group 2; wherein, healthy female C57BL / 6 mice were used as the control group, and dry eye mice were used as the model group, and these two groups were not subjected to any drug administration; the positive drug group was to drip sodium hyaluronate eye drops into both eyes of the dry eye mice, with an eye drop volume of 5 μL per eye, twice a day; experimental group 1 and experimental group 2 were to apply the wolfberry extract prepared in Example 1 and comparative example 1 to both eyes of the dry eye mice, 0.1 g per eye, twice a day. Each group was administered for 2 consecutive weeks.

[0052] 1.4 Index detection

[0053] Tear secretion detection: When the mouse is not anesthetized, put the phenol red cotton thread into the conjunctival sac of the mouse, loosen the eyelids to allow the eyelids to close naturally, time for 1 minute, take out the phenol red cotton thread, measure the length of the red part of the phenol red cotton thread with a ruler, and record it.

[0054] Tear film breakup time detection: 5 μL of 1% sodium fluorescein solution was dropped into the conjunctival sac of mice. The mice were allowed to blink freely to allow the sodium fluorescein solution to be evenly distributed on the corneal surface. After 1 minute, the mice were observed under cobalt blue light under a slit lamp and the time when the first black spot appeared on the cornea was recorded. The average value was taken after 3 consecutive times.

[0055] Corneal epithelial injury score: Sodium fluorescein was instilled into the ocular surface to assess the degree of corneal epithelial damage. A 1% sodium fluorescein saline solution was instilled into the mouse conjunctival sac, and the mouse conjunctival sac was rinsed with 0.9% NaCl, and excess moisture around the eye was wiped off with gauze. The mouse corneal staining was observed under the cobalt blue light of a slit lamp microscope, and photographed and recorded. The ocular surface was divided into 4 quadrants according to different parts, and the corneal fluorescein sodium staining degree and area were divided into 0 to 3 points. The larger the staining area and the more severe the degree, the higher the score. The total score of the 4 quadrants was 0 to 12 points.

[0056] 2. Results

[0057] 2.1 Tear secretion test results

[0058] The test results of tear secretion are shown in Table 1. As can be seen from Table 1, the tear secretion of dry eye mice is significantly reduced compared with that of healthy mice. The wolfberry extract prepared by fermentation of snow lotus fungus in the present invention can significantly increase the tear secretion.

[0059] Table 1 Test results of tear secretion

[0060]

[0061]

[0062] Note: The data in the table are the discoloration length of phenol red cotton thread; compared with the model group, * P<0.05, ** P<0.01, *** P<0.001.

[0063] 2.2 Tear film breakup time test results

[0064] The test results of tear film breakup time are shown in Table 2. As can be seen from Table 2, the tear film breakup time of dry eye mice is significantly reduced compared with that of healthy mice. The wolfberry extract prepared by fermentation of snow lotus fungus in the present invention can significantly prolong the tear film breakup time.

[0065] Table 2 Test results of tear film breakup time

[0066] Group Before administration(s) After administration(s) Control group 12.04±0.84 <![CDATA[13.17±0.59 *** ]]> Model Group 5.67±0.61 5.73±0.48 Positive drug group 5.24±0.84 <![CDATA[7.19±0.32 * ]]> Experimental Group 1 5.21±0.57 <![CDATA[8.34±0.27 * ]]> Experimental Group 2 5.68±0.49 6.51±0.43

[0067] Note: The data in the table are tear film breakup time, in seconds; compared with the model group, * P<0.05, *** P<0.001.

[0068] 2.3 Corneal epithelial injury scoring results

[0069] The corneal epithelial injury scoring results are shown in Table 3. The corneal epithelial injury was evaluated according to the degree of sodium fluorescein staining, and the higher the score, the more severe the corneal injury. According to Table 3, it can be seen that there was no corneal fluorescence staining in the control group, and the corneal epithelial injury score of the model group was significantly increased. The wolfberry extract prepared by the fermentation of snow lotus fungus in the present invention can significantly reduce the corneal epithelial injury score, that is, reduce corneal epithelial injury.

[0070] Table 3 Corneal epithelial injury scores

[0071] Group Before administration After administration Control group 0 0 Model Group 8.5±0.61 8.6±0.47 Positive drug group 8.4±0.53 <![CDATA[1.9±0.32 *** ]]> Experimental Group 1 8.5±0.27 <![CDATA[1.5±0.19 *** ]]> Experimental Group 2 8.6±0.35 <![CDATA[4.1±0.50 ** ]]>

[0072] Note: Compared with the model group, * P<0.05, *** P<0.001.

[0073] The embodiments described above are only descriptions of the preferred modes of the present invention, and are not intended to limit the scope of the present invention. Without departing from the design spirit of the present invention, various modifications and improvements made to the technical solutions of the present invention by ordinary technicians in this field should all fall within the protection scope determined by the claims of the present invention.

Claims

1. A method for preparing a wolfberry fermented product, It is characterized in that The following steps are involved: Inoculating snow lotus fungus into wolfberry puree, and subjecting it to fermentation treatment to obtain fermentation liquid; The fermentation liquid is centrifuged to obtain a supernatant, which is then concentrated to obtain a wolfberry extract, namely the wolfberry fermentation product.

2. The preparation method according to claim 1, It is characterized in that The inoculation amount of the snow lotus fungus is 3-5%.

3. The preparation method according to claim 1, It is characterized in that The fermentation treatment is carried out at a temperature of 25-28° C., a rotation speed of 120-140 rpm, and a time of 36-48 hours.

4. The preparation method according to claim 1, It is characterized in that The solid content of the wolfberry extract is 40-60wt%.

5. A wolfberry fermented product prepared according to the preparation method according to any one of claims 1 to 4.

6. Application of snow lotus fungus in wolfberry fermentation, It is characterized in that The wolfberry fermentation is used to prepare a product for treating dry eye.

7. Use of the wolfberry fermented product according to claim 5 in preparing a product for treating dry eye.

8. The use according to claim 7, It is characterized in that The product for treating dry eye is a medicine or a health product.

9. A product for treating dry eyes. It is characterized in that The active ingredient comprises the wolfberry fermented product according to claim 5.

10. The product according to claim 9, It is characterized in that The product for treating dry eye is a medicine or a health product.

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