阅读说明：本技术 一种功能性蓝莓羊干酪及其制备方法和应用 (Functional blueberry sheep cheese and preparation method and application thereof ) 是由 孙锐 孔悦 杨明冠 韩燕苓 于 2020-12-30 设计创作，主要内容包括：本发明提供一种功能性蓝莓羊干酪及其制备方法和应用,所述制备方法以蓝莓干果作为固定干酪益生菌的载体。本发明的方法能够最大程度的实现对益生菌细胞群体的固定化,能够在羊干酪产品的长期贮存过程中始终保持益生菌的有效数量在10～6-10～7CFU/g,同时,本发明的方法实现了高的酸化率,确保了羊干酪的高品质。(The invention provides functional blueberry sheep cheese and a preparation method and application thereof. The method can realize immobilization of probiotic cell populations to the maximum extent, and can always keep the effective quantity of probiotics at 10 in the long-term storage process of the sheep cheese product 6 ‑10 7 CFU/g, and meanwhile, the method realizes high acidification rate and ensures high quality of the sheep cheese.)

1. A preparation method of functional blueberry sheep cheese comprises the step of taking dried blueberry fruits as carriers for fixing probiotics of cheese.

2. The method of manufacturing according to claim 1, comprising: preparing a cheese probiotic wet biomass, mixing the cheese probiotic wet biomass with dried blueberry fruits, and fermenting to prepare a carrier fixed with cheese probiotics for cheese production; heating goat milk, cooling, adding a seed starter, incubating, and adding rennin to obtain a coagulum; adding a carrier fixed with cheese probiotics into the coagulum to prepare curdled milk, and curing the curdled milk to obtain the sheep cheese product.

3. The method of manufacturing according to claim 1, comprising: preparing a cheese probiotic wet biomass for cheese production; heating goat milk, cooling, adding a seed starter, incubating, and adding rennin to obtain a coagulum; and adding cheese probiotic wet biomass and dried blueberries into the coagulum to prepare curds, and curing the curds to obtain the sheep cheese product.

4. The preparation method according to claim 2 or 3, wherein when the carrier immobilized with the cheese probiotics is prepared, the mass relation between the blueberry dried fruit and the cheese probiotics wet biomass is 5-10: 0.5-3.

5. The method according to claim 2, wherein the method for preparing the carrier comprises: mixing dried blueberry fruit, probiotic wet biomass and MRS, fermenting at 30-40 deg.C without stirring, pouring out liquid, and washing with sterile water.

6. The method of claim 2 or 3, wherein the curing comprises: the curd is stored at 20-25 ℃ with an initial saline content of 8-15% (w/v), and when the pH is lowered to 3.4-5.2, and when the saline becomes 4-6% w/v, the sample is stored at 3-5 ℃ for further maturation for 30-80 days.

7. The preparation method according to claim 2 or 3, wherein the goat milk is a mixture of ewe milk and goat milk, and the mixing volume ratio of the ewe milk to the goat milk is 6-7: 4-3;

preferably, the goat milk is mixed, heated at 65-70 deg.C for 30-60 min, and then cooled at 37 deg.C.

8. The method according to claim 2 or 3, wherein the black bean extract and/or the tannic acid are added to the sheep cheese during the preparation of the sheep milk coagulation, wherein the black bean extract is added in an amount of 2.5-5mg/100g of sheep milk, and the tannic acid is added in an amount of 5-7mg/100g of sheep milk.

9. The functional blueberry sheep cheese prepared by the preparation method of any one of claims 1 to 8.

10. Use of the functional blueberry sheep cheese of claim 9 in the preparation of a food product having antioxidant function or preventing atherosclerosis.

Technical Field

The invention relates to the field of microorganisms and fermentation, and particularly relates to functional blueberry sheep cheese and a preparation method and application thereof.

Background

The information in this background section is only for enhancement of understanding of the general background of the invention and is not necessarily to be construed as an admission or any form of suggestion that this information forms the prior art that is already known to a person of ordinary skill in the art.

The global food market is constantly demanding functional foods that contain new ingredients developed by technology and have beneficial health effects and a wide range of nutritional effects. The incorporation of natural compounds with beneficial effects has been used in the production of functional dairy products. In particular, a variety of antioxidant supplements have been used in the cheese making process, whether single phenolic compounds such as tea polyphenols or natural plant extracts (e.g. grape or green tea extracts, cranberry powder, etc.), and novel natural preservatives (e.g. pomegranate rind extract, thyme l. essential oil, etc.) have been widely incorporated into cheese to improve microbial safety and organoleptic characteristics.

In addition, various methods of cheese making have been developed to meet consumer demand for healthier, safer and high quality dairy products. Various studies have used probiotic bacteria, in particular strains of lactobacilli and bifidobacteria, which have shown that they are present in appropriate amounts (10)⁶-10⁷CFU/g) when present in cheese products can produce health benefits to the human body. The existing methods such as gel immobilization or microencapsulation can immobilize the strains, but when applied to food, especially fermented food, the method often affects the quality and flavor of the product, and the method is difficult to maintain the proper probiotic quantity during long-term storage.

Disclosure of Invention

In order to overcome the defects of the prior art, the invention provides a functional blueberry sheep cheese and a preparation method and application thereof. The method can realize immobilization of probiotic cell populations to the maximum extent, and can always keep the effective quantity of probiotics at 10 in the long-term storage process of the sheep cheese product⁶-10⁷CFU/g, and meanwhile, the method realizes high acidification rate and ensures high quality of the sheep cheese.

In a first aspect of the invention, the invention provides a preparation method of functional blueberry sheep cheese, which comprises the step of taking dried blueberry fruits as carriers for fixing probiotics in cheese.

The dried blueberry fruit provided by the invention is dried blueberry fruit with the water content within 12%.

In some embodiments of the present invention, the probiotic cheese bacteria are streptococcus thermophilus and lactobacillus bulgaricus, which may be formulated conventionally in the art, and in preferred embodiments of the present invention, the preferred ratio of the two is 1: 1.

In some embodiments of the invention, the method for preparing the sheep cheese comprises the following steps: preparing a cheese probiotic wet biomass, mixing the cheese probiotic wet biomass with dried blueberry fruits, and fermenting to prepare a carrier fixed with cheese probiotics for cheese production; heating goat milk, cooling, adding a seed starter, incubating, and adding rennin to obtain a coagulum; adding a carrier fixed with cheese probiotics into the coagulum to prepare curdled milk, and curing the curdled milk to obtain the sheep cheese product.

In still other embodiments of the present invention, the method for preparing the sheep cheese of the present invention comprises: preparing a cheese probiotic wet biomass for cheese production; heating goat milk, cooling, adding a seed starter, incubating, and adding rennin to obtain a coagulum; and adding cheese probiotic wet biomass and dried blueberries into the coagulum to prepare curds, and curing the curds to obtain the sheep cheese product.

In some embodiments of the invention, when the carrier immobilized with the cheese probiotics is prepared, the mass relation between the blueberry dried fruit and the cheese probiotics wet biomass is 5-10: 0.5-3.

In the research process, the inventor finds that the amount of the dried blueberry fruit and probiotic wet biomass can influence the fixed amount of the strain, and in the embodiment of the invention, when the mass relation between the dried blueberry fruit and the cheese probiotic wet biomass is 5-10: 0.5-3, high viable cell fixation can be obtained, especially when the content is 5-10: 2-3, the fixation effect is better. In one or more embodiments of the invention, the inventors found that when the dried blueberry fruit is matched with the probiotic wet biomass at a ratio of 10:2, the fixed amount of somatic cells is not increased more significantly than when the dried blueberry fruit is matched with the probiotic wet biomass at a ratio of 5:2, and therefore, in the embodiments of the invention, 5:2 is a relatively superior dosage relationship.

In some embodiments of the invention, the method of preparing the carrier comprises: mixing dried blueberry fruit, probiotic wet biomass and MRS culture medium, fermenting at 30-40 deg.C preferably 37 deg.C for 24 hr without stirring, pouring out liquid, and washing with sterile water.

In an embodiment of the invention, said curing comprises: the curd is stored at 20-25 ℃ with an initial saline content of 8-15% (w/v), and when the pH is lowered to 3.4-5.2, and when the saline becomes 4-6% w/v, the sample is stored at 3-5 ℃ for further maturation for 30-80 days.

In some embodiments, the curing comprises: the curd was stored at 22 ℃ with an initial saline content of 8-15% (w/v), and when the pH was lowered to 3.4-5.2, the samples were stored at 4 ℃ for further maturation for 60 days when the saline became 4-6% w/v.

During the course of the research, the inventors have found that acidification of the curd is critical to ensure a high quality and proper maturation process of the sheep cheese. An important key to the success of making brine cheese is the high acidification rate achieved by the starter culture, which can exhibit a lower pH at the end of the maturation of the sheep cheese. The maturation operation of the present invention ensures that this key is achieved, and in some embodiments of the invention, the pH of the matured sheep cheese is kept low at 4.45-4.55, and the acidity, expressed as lactic acid content, can reach 0.84-0.87g/100 g cheese, with a lower pH and higher acidification rate compared to the prior art.

In some embodiments of the invention, the goat milk is a mixture of ewe's milk and goat milk in a mixing volume ratio of 6-7: 4-3; in some preferred embodiments, the mixing volume ratio is 2:1, which allows better quality assurance of the goat milk from the milk ingredients.

In some embodiments of the invention, the goat milk is mixed, heated at 65-70 ℃ for 30min-60min, and then cooled at 37 ℃.

In some embodiments of the present invention, the black bean extract and/or the tannic acid may be added to the sheep cheese during the preparation of the sheep milk coagulation, wherein the black bean extract is added in an amount of 2.5-5mg/100g of sheep milk, and the tannic acid is added in an amount of 5-7mg/100g of sheep milk.

The black bean extract is an ethanol extract of black beans, is rich in polyphenols, and can effectively prevent atherosclerosis by being mixed with tannic acid. In some embodiments of the present invention, the black soybean extract and/or tannic acid can also assist chymosin to coagulate milk protein and accelerate the formation of coagulum.

In a second aspect of the invention, the invention provides the functional blueberry sheep cheese prepared by the first method; the invention also provides application of the functional blueberry sheep cheese in preparing food with an antioxidant function or preventing atherosclerosis.

Compared with the prior art, the invention has the advantages that:

the invention uses the dried blueberry fruit as the lactobacillus casei immobilized biocatalyst to generate the highest immobilized cell population (>10⁹cfu/g). The method of the invention ensures that the sheep cheese has a lower pH value at the end of maturation, which can be as low as about 4.4, and realizes a high acidification rate, and the acidity can reach more than 0.85 by the representation of the lactic acid content, thereby ensuring the high quality of the sheep cheese. The process of the invention increases the number of cheese probiotics and the sheep cheese product of the invention is stored in a suitable amount (10) for a storage time of at least 100 days⁶-10⁷CFU/g) keeps the quantity of probiotics, and overcomes the defect that the long-term stable maintenance of the quantity of probiotics is difficult to realize in the prior art. In addition, the sheep cheese prepared by taking the dried blueberry fruits as the carrier has fruit aroma attributes, such as fruity aroma, flowery aroma and brandy aroma, only exists in the process of taking the dried blueberry fruits as the fixed carrier, and the content of the sheep cheese is increased in the curing process, so that a plurality of terpenes and carbonyl compounds are enriched; the feta of the present invention tastes soft and pleasant, while the tartness of the berries is masked in the environment of the feta type cheese. The biotechnological method for developing the fortified sheep cheese by using the blueberries as natural and biodegradable carriers not only increases the nutritional value of cheese foods and popular foods, but also makes a virtuous circle by using a green technical method, and has high positive significance on human health.

Detailed Description

The invention will be further illustrated with reference to the following specific examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention. The experimental procedures, in which specific conditions are not noted in the following examples, are generally carried out according to conventional conditions or according to conditions recommended by the manufacturers.

Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. The reagents or starting materials used in the present invention can be purchased from conventional sources, and unless otherwise specified, the reagents or starting materials used in the present invention can be used in a conventional manner in the art or in accordance with the product specifications. In addition, any methods and materials similar or equivalent to those described herein can be used in the methods of the present invention. The preferred embodiments and materials described herein are intended to be exemplary only.

Example 1

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Dried blueberry fruits after being dried in the air are taken as a fixed carrier of probiotic strains of cheese. 10g of dried blueberry fruit, 2g of probiotic wet cheese biomass (2g) and 500mL of MRS were mixed and fermented without stirring at 37 ℃ for 24 hours. Then, the liquid is poured out and washed twice with a proper amount of sterile aqueous solution to obtain the carrier fixed with the cheese probiotics for the cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Example 2

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Dried blueberry fruits after being dried in the air are taken as a fixed carrier of probiotic strains of cheese. 5g of dried blueberry fruit, 2g of probiotic wet biomass cheese and 500mL of MRS were mixed and fermented without stirring at 37 ℃ for 24 hours. Then, the liquid is poured out and washed twice with a proper amount of sterile aqueous solution to obtain the carrier fixed with the cheese probiotics for the cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Example 3

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point 2g of cheese probiotic wet biomass and 5g of blueberry air dried fruit were added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Example 4

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Dried blueberry fruits after being dried in the air are taken as a fixed carrier of probiotic strains of cheese. 5g of dried blueberry fruit, 2g of probiotic wet cheese biomass (2g) and 500mL of MRS were mixed and fermented without stirring at 37 ℃ for 24 hours. Then, the liquid is poured out and washed twice with a proper amount of sterile aqueous solution to obtain the carrier fixed with the cheese probiotics for the cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, mixing with dried powder of ethanol solution of semen Sojae Atricolor (added amount of 2.5mg/100g milk) and tannin (added amount of 5mg/100g milk). Adding chymosin (added in an amount of 100kg milk and 20ml chymosin solution) to obtain coagulum.

The preparation method of the black bean ethanol solution dry powder comprises the following steps: 10g of ground air-dried black beans were immersed in 50ml of ethanol, stirred at room temperature (40 cycles/min) for 2-4h, and then water was added to make the ratio of ethanol: the water ratio is 80: 20, and the mixture is kept still for 7 days. The extract was filtered through a Xinhua No. 1 filter paper. Then, the solvent is removed under reduced pressure at 55-60 ℃ in a rotary evaporator, and the black bean ethanol solution is dried by vacuum freeze-drying to obtain dry powder of the black bean ethanol solution.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Comparative example 1

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point 2g of cheese probiotic wet biomass was added, mixed thoroughly and left to stand for 20min to give curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Comparative example 2

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Dried blueberry fruits after being dried in the air are taken as a fixed carrier of probiotic strains of cheese. 5g of dried blueberry fruit, 5g of probiotic wet biomass cheese and 500mL of MRS were mixed and fermented without stirring at 37 ℃ for 24 hours. Then, the liquid is poured out and washed twice with a proper amount of sterile aqueous solution to obtain the carrier fixed with the cheese probiotics for the cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Comparative example3

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Dried blueberry fruits after being dried in the air are taken as a fixed carrier of probiotic strains of cheese. 5g of dried blueberry fruit, 2g of probiotic wet biomass cheese and 500mL of MRS were mixed and fermented without stirring at 37 ℃ for 24 hours. Then, the liquid is poured out and washed twice with a proper amount of sterile aqueous solution to obtain the carrier fixed with the cheese probiotics for the cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

And finally, putting the standing curd into 10% (w/v) saline water for curing for 60 days to obtain the cheese product.

Comparative example 4

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Dried blueberry fruits after being dried in the air are taken as a fixed carrier of probiotic strains of cheese. 5g of dried blueberry fruit, 2g of probiotic wet cheese biomass (2g) and 500mL of MRS were mixed and fermented without stirring at 37 ℃ for 24 hours. Then, the liquid is poured out and washed twice with a proper amount of sterile aqueous solution to obtain the carrier fixed with the cheese probiotics for the cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v) and further aged for 60 days when the pH dropped to 4. + -. 0.5. And (5) obtaining the cheese product after the ripening is finished.

Comparative example 5

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Respectively removing impurities from pericarpium Citri Tangerinae, lemon peel and okra peel, sterilizing at 120 deg.C for 6min, oven drying at 100 deg.C for 0.4h, and pulverizing to obtain pericarpium Citri Tangerinae, lemon peel and okra peel powder with fineness of 200 mesh; adding 20g of mixed powder of orange peel powder, 20g of lemon peel powder and 30g of okra peel powder into 2600mL of sterilized deionized water with the pH value of 6.0, carrying out vortex oscillation at 300rpm for 20min, mixing uniformly, carrying out micro-jet treatment for 5 times at 300MPa, adjusting the pH value of the solution to 6.0, incubating for 4h at the temperature of 30 ℃ and the rotation speed of 30rpm/min, then centrifuging for 12min at 10 ℃ and 10000rpm/min to remove precipitates, and carrying out vacuum rotary evaporation concentration at 50 ℃ to obtain 500mL of concentrated solution; and (3) taking 2g of cheese probiotic wet biomass and the concentrated solution, carrying out vortex oscillation at 150rpm for 5min, uniformly mixing, adjusting the pH value of the mixed solution to 6.0, and incubating for 4h at 26 ℃ and the rotation speed of 80rpm/min to obtain the carrier fixed with the cheese probiotics for cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Comparative example 6

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Taking the dried apple slices after air drying as a fixed carrier of the probiotic strains of cheese. 5g of dried apple, 2g of cheese probiotic wet biomass (2g) and 500mL of MRS were mixed and fermented without stirring at 37 ℃ for 24 hours. Then, the liquid is poured out and washed twice with a proper amount of sterile aqueous solution to obtain the carrier fixed with the cheese probiotics for the cheese production.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point the prepared cheese probiotic immobilized carrier was added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Comparative example 7

Probiotic casei (Streptococcus thermophilus and Lactobacillus bulgaricus, 1:1) were grown in MRS liquid medium at 37 ℃ for 72 hours. The cheese probiotic wet biomass was collected by centrifugation at 5000rpm for 10 minutes and stored at 5 ℃.

Mixing the ewe milk and goat milk at a volume ratio of 2:1, heating at 65 deg.C for 30min, and cooling at 37 deg.C.

Adding seed starter at 1% (v/v) level, incubating for 20min, and adding chymosin (20 ml chymosin solution added to 100kg milk) to obtain coagulum.

Subsequently, 1kg of the coagulum was cut into cubes (diameter less than 1cm) and at this point 2g of cheese probiotic wet biomass and 5g of tea polyphenols were added, mixed thoroughly and left to stand for 20min to obtain curd.

The curd is then gradually transferred to a circular mold and periodically stirred to facilitate whey drainage. The curd was then removed from the mold and allowed to stand for 10 min.

Finally subjecting the standing curd to a maturation operation comprising: the curd was stored at 22 ℃ with an initial saline content of 10% (w/v), and when the pH was lowered to 4. + -. 0.5, the sample was stored at 4 ℃ for further maturation for 60 days when the saline content became 5% w/v. And (5) obtaining the cheese product after the ripening is finished.

Examples of the experiments

1. The number of viable bacteria immobilized on the immobilized carrier and the number of viable bacteria after 100 days of storage of the cheese product were measured according to the method of GB 4789.35-2016 (lactic acid bacteria test for food microbiology), and the results are shown in Table 1.

TABLE 1

2. The pH and acidity of the cheese product after maturation was measured, wherein the pH was measured directly at room temperature using a pH meter and the acidity was measured according to the method of GB 5009.239-2016, "determination of acidity of food", wherein the acidity of the sample is expressed as lactic acid content and the results are reported in table 2.

TABLE 2

3. DPPH clearance test

The samples (example 1 and comparative example 7) and the positive control (Vc) solutions were prepared with double distilled water at a concentration of 0.5mg/ml, respectively, for further use. The solution was pipetted 200. mu.l each with a pipette gun and diluted to 8 serial dilutions of 100. mu.l in 96-fold gradient, while 100. mu.l of double distilled water was pipetted instead of the test solution as a blank control. Adding 100 mul of 750 mu M DPPH ethanol solution into each hole, fully mixing, reacting for 30 minutes at room temperature under the condition of dark black, and measuring the light absorption value of the hole solution at 517nm on a microplate reader according to the formula: clearance (%) - (A)_Control-A_{To be measured})/A_Control]x100 calculation of the respective clearance, wherein A_{To be measured}Is the light absorption value of DPPH.solution when the solution to be detected is added; a. the_ControlThe absorbance value of DPPH & solution when the solution to be measured is not added. DPPH free radical scavenging ability of sample to be detectedIC for strength and weakness of₅₀This means that a smaller value indicates a higher scavenging ability.

TABLE 3 antioxidant Performance study

	DPPH clearance (IC)50)
		Example 1	14.7
Comparative example 7	15.3
		Vc	16.1

Note: IC (integrated circuit)₅₀Defined as the concentration of antioxidant required at a DPPH.clearance of 50%.

4. Sensory evaluation of cheese products

15 selected evaluations were made, and sensory evaluation criteria are shown in table 4, and the results are recorded in table 5.

TABLE 4 sensory evaluation criteria Table

TABLE 5 evaluation of cheese quality

	Aroma (average score)	Mouthfeel (average score)	Texture (average score)
				Example 1	6.8	6.8	6.4
Example 2	6.7	6.8	6.5
				Example 3	6.6	6.7	6.5
Example 4	6.7	6.6	6.5
				Comparative example 1	6.6	2.0	5.4
Comparative example 2	6.4	2.5	3.8
				Comparative example 3	5.4	6.1	5.4
Comparative example 4	5.6	6.2	5.0
				Comparative example 5	4.5	6.1	5.1
Comparative example 6	4.8	6.8	6.2
				Comparative example 7	6.2	5.7	5.1

Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that changes may be made in the embodiments and/or equivalents thereof without departing from the spirit and scope of the invention. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.