CN114586844B - Durian yoghourt capable of reducing blood fat and relaxing bowel and preparation method thereof - Google Patents

Durian yoghourt capable of reducing blood fat and relaxing bowel and preparation method thereof Download PDF

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CN114586844B
CN114586844B CN202210310973.XA CN202210310973A CN114586844B CN 114586844 B CN114586844 B CN 114586844B CN 202210310973 A CN202210310973 A CN 202210310973A CN 114586844 B CN114586844 B CN 114586844B
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durian
fermentation
filtrate
milk
blood fat
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CN114586844A (en
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袁飞连
杨林
张小云
胡超亭
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Jiangxi Sunshine Dairy Co ltd
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Jiangxi Sunshine Dairy Co ltd
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    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/123—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt
    • A23C9/1234—Fermented milk preparations; Treatment using microorganisms or enzymes using only microorganisms of the genus lactobacteriaceae; Yoghurt characterised by using a Lactobacillus sp. other than Lactobacillus Bulgaricus, including Bificlobacterium sp.
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
    • A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
    • A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
    • A23C9/1307—Milk products or derivatives; Fruit or vegetable juices; Sugars, sugar alcohols, sweeteners; Oligosaccharides; Organic acids or salts thereof or acidifying agents; Flavours, dyes or pigments; Inert or aerosol gases; Carbonation methods
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/123—Bulgaricus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/165—Paracasei
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/11—Lactobacillus
    • A23V2400/175—Rhamnosus
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
    • A23V2400/00—Lactic or propionic acid bacteria
    • A23V2400/21—Streptococcus, lactococcus
    • A23V2400/249—Thermophilus

Abstract

The invention discloses durian yoghourt capable of reducing blood fat and relaxing bowel and a preparation method thereof, and the durian yoghourt comprises the following raw materials in parts by weight: 10 to 15 parts of durian pulp, 70 to 80 parts of raw milk, 5 parts of fructo-oligosaccharide powder, 2 parts of probiotics fermentation product filtrate, 0.01 to 0.05 part of common starter and 0.1 to 0.3 part of stabilizer; the probiotics fermentation product filtrate comprises 1 part of lactobacillus rhamnosus BBEK4 fermentation filtrate and 1 part of lactobacillus paracasei JN-1 fermentation filtrate. Compared with common probiotics, the durian yoghourt prepared by fermenting the added probiotics lactobacillus rhamnosus BBEK4 and lactobacillus paracasei JN-1 with durian and raw milk has very remarkable blood fat reducing and bowel relaxing effects, and has remarkable synergistic effects among the lactobacillus rhamnosus BBEK4, lactobacillus paracasei JN-1 and durian, and the blood fat reducing and bowel relaxing effects are remarkable.

Description

Durian yoghourt capable of reducing blood fat and relaxing bowel and preparation method thereof
Technical Field
The invention relates to the technical processing field of dairy products, in particular to durian yoghourt capable of reducing blood fat and relaxing bowel and a preparation method thereof.
Background
Durian belongs to tropical fruits, the pulp consists of meat bags with fake seed coats, the meat color is light yellow, and the viscous juice is a fruit with great economic value. The durian has extremely high nutritive value and the reputation of 'king of tropical fruits', has huge market demand potential, can strengthen the body, invigorate the spleen and tonify qi, tonify the kidney and strengthen yang, warm the body and belongs to the nourishing and beneficial fruits, and can be made into the yogurt cloth Ding Lei nutritional and leisure food with special taste and texture and rich nutrition. Although durian is rich in nutrition and has medicinal effects, the durian is not suitable for people, and is a strong heat fruit, which is easy to cause dryness-heat of the body and cause symptoms such as excessive internal heat, dry throat, dry tongue, sore throat and the like.
At present, although the durian has extremely high nutritive value, the medicinal value of the durian is not deeply developed in the development process of the durian product, and the yogurt pudding type product developed by the durian only has the performance of common yogurt, so that the development team screens out the probiotics which can cooperate with the durian, thereby developing durian yogurt, and having the effects of reducing blood fat and relaxing bowel.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides durian yoghourt capable of reducing blood fat and relaxing bowel and a preparation method thereof, so as to solve the problems in the technical background.
Chinese patent 201810465929.X (university of south of the river) discloses a lactobacillus rhamnosus (Lactobacillus rhamnosus) BBE K4, the lactobacillus rhamnosus (Lactobacillus rhamnosus) has been preserved in the chinese collection of typical cultures at 12 months 19 of 2017 with a preservation number of cctccc No. M2017807. The lactobacillus rhamnosus BBE K4 used in the invention is presented by researchers at the university of Jiangnan.
202111386682.0 (university of Jiangnan) discloses Lactobacillus paracasei (Lactobacillus paracasei) JN-1 which is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms) for 2021, 6 months and 21 days, wherein the preservation address is Hospital No. 3 of North Chen West Lu 1 in the Korean region of Beijing city, and the preservation number is CGMCC No.22745; lactobacillus paracasei JN-1 has strong gastrointestinal tract digestive system resistance and bile salt tolerance, can be used for preventing and treating spontaneous enteritis, and has good functions and flavor. The Lactobacillus paracasei JN-1 used in the present invention was given from researchers at the university of south China.
In the research and practice process, the invention discovers that the yogurt prepared by fermenting the main components of the combined lactobacillus rhamnosus BBE K4 and the lactobacillus paracasei JN-1 together with durian and raw milk can effectively enhance the treatment effect on high cholesterol and constipation of mice, the yogurt prepared by fermenting the main components of the lactobacillus rhamnosus BBE K4 or the lactobacillus paracasei JN-1 together with durian and raw milk does not have obvious treatment effect on high cholesterol and constipation of mice, or the yogurt prepared by fermenting the main components of the raw milk without adding durian has obvious effect on high cholesterol and constipation of mice, which means that the lactobacillus rhamnosus BBE K4, the lactobacillus paracasei JN-1 and the durian have very obvious blood lipid reducing and bowel relaxing effects, and 3 substances have obvious synergistic effects. Lactobacillus rhamnosus BBE K4 and lactobacillus paracasei JN-1 are safe probiotics, and have good prospects as oral agents.
In order to achieve the above object, the present invention is realized by the following technical scheme:
durian yoghourt capable of reducing blood fat and relaxing bowel comprises the following raw materials in parts by weight: 10 to 15 parts of durian pulp, 70 to 80 parts of raw milk, 5 parts of fructo-oligosaccharide powder, 2 parts of probiotics fermentation product filtrate, 0.01 to 0.05 part of common starter and 0.1 to 0.3 part of stabilizer.
Further, the probiotics fermentation product filtrate comprises 1 part of lactobacillus rhamnosus BBE K4 fermentation filtrate and 1 part of lactobacillus paracasei JN-1 fermentation filtrate.
Further, the preparation method of the lactobacillus rhamnosus BBE K4 fermentation filtrate comprises the following steps:
(1) Inoculating the preserved lactobacillus rhamnosus BBE K4 into an MRS culture medium under the aseptic condition, and performing activation culture to obtain seed liquid for fermentation culture;
(2) Inoculating the lactobacillus rhamnosus BBE K4 seed solution obtained in the step (1) into an MRS culture medium according to the inoculum size of 3-5 v/v%, and culturing for 22-26 h at 36-38 ℃; wherein, the MRS culture medium configuration: 1L distilled water is dissolved with 10g peptone, 8g beef extract, 4g yeast extract, 20g glucose, 801mL Tween-801, 2g dipotassium hydrogen phosphate, 5g sodium acetate trihydrate, 2g ammonium citrate, 0.2g magnesium sulfate heptahydrate and 0.05g potassium manganese sulfate tetrahydrate.
(3) Centrifuging the fermentation broth obtained in step (2) at 8000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain filtrate with concentration of 3×10 8 CFU/mL of sterile lactobacillus rhamnosus BBE K4 fermentation filtrate.
Further, the preparation method of the lactobacillus paracasei JN-1 fermentation filtrate comprises the following steps:
(1) Inoculating preserved lactobacillus paracasei JN-1 into an MRS culture medium under the aseptic condition, and performing activation culture for 24-48 hours at the temperature of 35-38 ℃ to obtain a culture serving as seed liquid for fermentation culture;
(2) Inoculating lactobacillus paracasei JN-1 seed liquid obtained in the step (1) into an MRS culture medium according to an inoculum size of 2-4 v/v percent, and performing anaerobic culture for 20-48 hours at the temperature of 35-38 ℃ to obtain a fermentation liquor;
(3) Centrifuging the fermentation broth obtained in step (2) at 6000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain filtrate with concentration of 3×10 8 CFU/mL sterile Lactobacillus paracasei JN-1 fermentation filtrate.
Furthermore, the common starter is a 1:1 mixture of lactobacillus bulgaricus and streptococcus thermophilus.
In the invention, fructo-oligosaccharide is a soluble dietary fiber, and can reach the large intestine to be selectively utilized by beneficial flora in the intestinal tract because the fructo-oligosaccharide is not digested and absorbed after being ingested by a human body, thereby effectively regulating the intestinal flora. Meanwhile, the stability of the fructo-oligosaccharide under the acidic condition is better than that of sucrose, and compared with the sucrose, the fructo-oligosaccharide has fresh and cool sweet taste, is pure in taste, does not have any aftertaste, does not change sweetness, and has no influence on the taste of durian yoghourt.
On the other hand, the invention also provides a preparation method of the durian yoghourt capable of reducing blood fat and relaxing bowel, which comprises the following steps:
(1) Pretreatment of raw milk: raw milk is cooled to 2-6 ℃ for storage after being subjected to milk cleaning and pre-sterilization in sequence, then is subjected to secondary pre-sterilization before burdening, and is cooled to 2-6 ℃ for storage after sterilization;
(2) Adding durian pulp, fructooligosaccharide powder and a stabilizer into the sterilized raw milk, uniformly mixing, homogenizing for 2 times under 15-20 MPa, and sterilizing at 80-90 ℃ for 15min to obtain durian milk;
(3) And (3) adding the common starter and the probiotics fermentation product filtrate into the mixed milk when the durian milk is cooled to about 42-45 ℃, uniformly mixing, and culturing at constant temperature for 6-8 hours at 40-42 ℃ to obtain the durian yoghourt capable of reducing blood fat and relaxing bowel after being matured for 24 hours.
In the technical scheme, the preparation process of the durian pulp comprises the following steps: washing peeled and cored durian pulp, cutting into pieces, pulping by a pulping machine, heating the pulp to 80-120 ℃, preserving heat for 10-25 min for sterilization treatment, cooling to obtain durian pulp, and storing for standby.
Compared with the prior art, the invention has the beneficial effects that:
compared with common probiotics, the durian yoghourt prepared by fermenting the added probiotics lactobacillus rhamnosus BBE K4 and lactobacillus paracasei JN-1 with durian and raw milk has very remarkable blood fat reducing and bowel relaxing effects, and has remarkable synergistic effects among the lactobacillus rhamnosus BBE K4, lactobacillus paracasei JN-1 and durian, and has remarkable blood fat reducing and bowel relaxing effects.
Drawings
FIG. 1 is a bar graph showing the effect of different yoghurt groups on rat serum TC after modeling and after dosing in example 3;
FIG. 2 is a bar graph showing the effect of different yoghurt groups on rat serum TG, LDL-C, HDL-C after administration in example 3.
Detailed Description
Other advantages and effects of the present invention will become apparent to those skilled in the art from the following disclosure, which describes the embodiments of the present invention with reference to specific examples. The invention may be practiced or carried out in other embodiments that depart from the specific details, and the details of the present description may be modified or varied from the spirit and scope of the present invention. It should be noted that the following embodiments and features in the embodiments may be combined with each other without conflict.
It should be noted that the illustrations provided in the following embodiments merely illustrate the basic concept of the present invention by way of illustration, and only the components related to the present invention are shown in the drawings and are not drawn according to the number, shape and size of the components in actual implementation, and the form, number and proportion of the components in actual implementation may be arbitrarily changed, and the layout of the components may be more complicated.
Example 1
Durian yoghourt capable of reducing blood fat and relaxing bowel comprises the following raw materials in parts by weight: 12 parts of durian pulp, 80 parts of raw milk, 5 parts of fructooligosaccharide powder, 2 parts of probiotics fermentation product filtrate and 0.04 part of common starter (the mixed concentration of lactobacillus bulgaricus and streptococcus thermophilus microbial inoculum is not less than 10) 10 CFU/g) and 0.1 to 0.3 part of stabilizer. Wherein the stabilizer is gelatin and/or low-ester pectin.
In this example, the probiotic fermented product filtrate contained 1 part of lactobacillus rhamnosus BBE K4 fermented filtrate and 1 part of lactobacillus paracasei JN-1 fermented filtrate.
Further, the preparation method of the lactobacillus rhamnosus BBE K4 fermentation filtrate comprises the following steps:
(1) Inoculating the preserved lactobacillus rhamnosus BBE K4 into an MRS culture medium under the aseptic condition, and performing activation culture to obtain seed liquid for fermentation culture;
(2) Inoculating the lactobacillus rhamnosus BBE K4 seed solution obtained in the step (1) into an MRS culture medium according to the inoculum size of 3-5 v/v%, and culturing for 22-26 h at 36-38 ℃; wherein, the MRS culture medium configuration: 1L distilled water is dissolved with 10g peptone, 8g beef extract, 4g yeast extract, 20g glucose, 801mL Tween-801, 2g dipotassium hydrogen phosphate, 5g sodium acetate trihydrate, 2g ammonium citrate, 0.2g magnesium sulfate heptahydrate and 0.05g potassium manganese sulfate tetrahydrate.
(3) Centrifuging the fermentation broth obtained in step (2) at 8000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain filtrate with concentration of 3×10 8 CFU/mL of sterile lactobacillus rhamnosus BBE K4 fermentation filtrate.
The preparation method of the lactobacillus paracasei JN-1 fermentation filtrate comprises the following steps:
(1) Inoculating preserved lactobacillus paracasei JN-1 into an MRS culture medium under the aseptic condition, and performing activation culture for 24-48 hours at the temperature of 35-38 ℃ to obtain a culture serving as seed liquid for fermentation culture;
(2) Inoculating lactobacillus paracasei JN-1 seed liquid obtained in the step (1) into an MRS culture medium according to an inoculum size of 2-4 v/v percent, and performing anaerobic culture for 20-48 hours at the temperature of 35-38 ℃ to obtain a fermentation liquor;
(3) Centrifuging the fermentation broth obtained in step (2) at 6000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain filtrate with concentration of 3×10 8 CFU/mL sterile Lactobacillus paracasei JN-1 fermentation filtrate.
In this example, the general starter is a 1:1 mixture of Lactobacillus bulgaricus and Streptococcus thermophilus. For example, the common starter is Streptococcus thermophilus lyophilized powder (10) 10 CFU/g) 0.02 parts of Lactobacillus bulgaricus freeze-dried powder (10) 10 CFU/g) 0.02 parts of mixed bacterial powder (food grade).
The preparation method of durian yoghourt capable of reducing blood fat and relaxing bowel provided by the embodiment comprises the following steps of:
(1) Pretreatment of raw milk: raw milk is cooled to 4 ℃ for storage after being subjected to milk cleaning and pre-sterilization in sequence, then is subjected to secondary pre-sterilization before burdening, and is cooled to 4 ℃ for storage after sterilization;
(2) Adding durian pulp, fructooligosaccharide powder and a stabilizer into the sterilized raw milk, uniformly mixing, homogenizing for 2 times under 15-20 MPa, and sterilizing at 80-90 ℃ for 15min to obtain durian milk;
(3) And (3) adding the common starter and the probiotics fermentation product filtrate into the mixed milk when the durian milk is cooled to about 45 ℃, uniformly mixing, and then culturing for 6 hours at a constant temperature under the condition of 42 ℃ to obtain the durian yoghourt capable of reducing blood fat and relaxing bowel after being matured for 24 hours.
Example 2
The yoghurt prepared in comparative examples 1 to 6 is similar to example 1 except that part of the ingredients are different, and the ingredients of comparative examples 1 to 6 are shown in Table 1:
table 1 example 1, comparative examples 1 to 6 ingredients table of different yogurts produced
Note that:the microbial inoculum in the lactobacillus paracasei fermenting filtrate in comparative example 6 is purchased and preserved in China general microbiological culture Collection center (CGMCC) 1.9089, the preparation method of the lactobacillus paracasei fermenting filtrate is similar to that of the lactobacillus paracasei JN-1 fermenting filtrate in example 1, and the concentration of the lactobacillus paracasei CGMCC 1.9089 microbial inoculum in the fermenting filtrate is 3×10 8 CFU/mL。
Example 3
Rat grouping: 100 male rats weighing 200-220 g were divided into 10 groups, and there was no statistical difference between the groups. The 10 groups were a blank control group (normal rats), a positive control group (lovastatin group), a negative control group, example 1 and comparative examples 1 to 6 treated groups, each group of 10 rats. Wherein, the normal rat group is used as a blank control, and the positive control group, the negative control group and the treatment group are all rats successfully molded by using hyperlipidemia.
The method for molding the hyperlipoidemia rats comprises the following steps: 100 male rats were adaptively fed for 1 week, 10 rats were randomly selected and continued to be given basal feed (78.8% basal feed, 10% lard, 10% egg yolk powder, 1% cholesterol, 0.2% bile salt) as a blank group, and the remaining 90 were given high-fat feed as a high-fat model group. After modeling, orbital blood was collected from rats, and serum Total Cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) levels were measured, and when the model group was compared with the blank group, the model was established and the test results are shown in table 2, when TC or TG was significantly increased in serum of high fat rats in the model group (P < 0.05).
After modeling is successful, the blank control group and the negative control group are not dosed, and the equal volume of distilled water is filled into the stomach every day; the positive control group is subjected to gastric lavage after being given with lovastatin suspension, and the administration amount of the lovastatin suspension is converted into the dosage of rats according to the drug administration instruction, and the stomach is irrigated at 30 mg/kg.d; the treatment groups (example 1, comparative example 1 to comparative example 6) were fed at 2ml/100 g.d; all groups were continuously treated for 30 days, after last administration, orbital blood was collected from rats after 12h of fasting, and serum Total Cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) levels were determined. The test results are shown in tables 2 and 3.
TABLE 2 Effect of different yogurt groups on rat serum TC, TG
Note that: a: p <0.05 (compared to blank); b: p <0.05 (compared to negative control group); c: p <0.05 (compared to positive control).
TABLE 3 Effect of different groups of yogurt on rat serum LDL-C, HDL-C
Note that: a: p <0.05 (compared to blank); b: p <0.05 (compared to negative control).
As can be seen from table 2 and fig. 1, the TC level of the rats in the negative control group is significantly increased compared with that in the blank control group after modeling, which indicates that the modeling of the hyperlipidemia rats is successful. Following dosing, the TCs of the example 1, comparative examples 1 to 6 showed a decreasing trend compared to modeling, and the serum TCs of the positive control, example 1, comparative examples 4 to 6 were significantly different compared to the negative control; while example 1 rat serum TC was significantly reduced compared to the positive control group. After modeling from table 2 and fig. 2, TG levels were significantly increased in the negative control compared to the blank control, indicating successful modeling of the hyperlipidemic rats. The TG values of the rats in the blank control group and the negative control group did not change significantly during the test. The positive control, example 1, comparative examples 4 to 6 all had significantly lower serum TG than the negative control. The experiment shows that the yoghurt provided by the invention can not only reduce the serum total cholesterol of a high-fat rat, but also effectively reduce the serum triglyceride.
As can be seen from table 3 and fig. 2, the positive control group, example 1, and comparative examples 4 to 6 all significantly reduced serum LDL-C compared to the negative control group. The yogurt provided by the invention can effectively reduce the serum LDL-C level of rats. After modeling, the serum HDL-C values of rats were significantly different from group to group, and the serum HDL-C levels of the rats of the blank control group, the positive control group, example 1, and comparative examples 1 to 6 were all on an ascending trend during the experiment, and the negative control group was significantly lower than the blank control group, and by the end of the experiment, the serum LDL-C of the positive control group, example 1, comparative examples 4 to 6 was significantly elevated compared to the negative control group. Proved by the invention, the yoghurt provided by the invention can effectively improve the HDL-C level of rat serum.
Taken together, the blood lipid lowering effect of example 1 was better than that of the positive control group. The liquid beverage does not contain durian pulp, but only contains one fermentation filtrate (comparative example 1 and comparative example 2) with the lowest blood fat reducing effect; the lipid-lowering effect was slightly higher, but not significantly higher, than that of the one containing only one fermentation filtrate (comparative example 1 and comparative example 2) without durian pulp but with the two fermentation filtrates (comparative example 3); the durian pulp and one of the fermented filtrate (comparative examples 4, 5 and 6) can reduce blood lipid, and the effect is equivalent to that of a positive control group, which shows that the durian pulp has obvious effect of improving the blood lipid reducing effect on the filtrate; when the two filtrates are matched with durian pulp to prepare the yoghurt, the blood lipid reducing effect is highest, even higher than that of lovastatin solution, which indicates that the two filtrates have good synergistic effect with each other, and the blood lipid reducing effect can be greatly improved.
Example 4
Grouping mice: 160 male BALB/c mice with the weight of 18-22 g are taken, the mice are randomly divided into 2 large groups, namely a small intestine movement experimental group and a defecation experimental group, each large group is divided into 10 small groups, and the mice in the groups have no statistical difference. The 10 groups in each large group were a blank control group (normal mice), a positive control group (xylooligosaccharide group), a negative control group (constipation model group), example 1 and comparative examples 1 to 6 treatment groups, respectively. 8 mice per group. Wherein, the normal mouse group is used as blank control, and the positive control group, the negative control group and the treatment group are all successful mice with constipation modeling.
The method for establishing the constipation model comprises the following steps: after mice are grouped, each treatment group (the treatment group comprising 2 large groups) is fed with yoghurt at regular time with 2 ml/(100 g.BW) each day, a blank control group and a negative control group are filled with volumes of purified water, a positive control group is filled with a gastric xylooligosaccharide suspension at regular time each day, the gastric volume is 1.5 g/(kg.BW), the mice are fed with yoghurt and continuously fed for 14 days, during the period, each group of mice is fed with yoghurt, drinking water and activity normally, after the yoghurt and xylooligosaccharide suspension are fed for the last time, all the mice of the two large groups are not fed with water forbidden for 20 hours, the mice of the negative control group, the positive control group and the treatment group are filled with a gastric hydrochloric loperamide aqueous solution, the gastric volume of the blank control group is 10 mg/(kg.BW), and the mice of the blank control group are filled with the same volumes of purified water.
1. Small intestine exercise experiment
After the loperamide hydrochloride is filled in the stomach for 30min, the treatment group is respectively fed with yoghourt containing corresponding dosage and prepared by edible ink, the positive control group is filled with xylose oligosaccharide suspension containing corresponding dosage and prepared by ink, the blank control group and the model control group are filled with ink with the same volume, after 25min, all mice in the small intestine movement experiment group are immediately killed by pulling the neck, the intestinal membranes are opened, the intestinal tracts are separated, the intestinal tracts from the pylorus and the lower end to the back blind intestines are cut off, the small intestines are gently pulled into a straight line and measured, the length of the intestinal tracts is recorded as 'total length of the small intestines', the ink propelling rate is calculated according to the following formula and is recorded as 'ink propelling length' from the pylorus to the length of the leading edge part of the ink, and the test results are shown in table 4.
Ink advance (%) = (ink advance length/total length of small intestine) ×100%
2. Stool test
After the loperamide hydrochloride is filled in the stomach for 30min, the treatment group is respectively fed with yoghourt containing corresponding dosage and prepared by edible ink, the xylose oligosaccharide suspension containing corresponding dosage and prepared by the ink for the stomach filling of the positive control group, the ink with the same volume and the same volume as the gastric control group and the model control group are filled with the ink, the mice of the defecation experiment group are fed in a single cage, normally ingest, drink water and move, the black stool time of the first grains of each mouse is observed and recorded from the time of filling the gastric ink, the number and the weight of the black stool grains are calculated within 5h, and the water content of the excrement is calculated according to the following formula to judge the influence on the defecation of the constipation mice, and the test results are shown in table 4.
Fecal moisture content (%) = [ (fecal wet weight-fecal dry weight)/fecal wet weight ] ×100%
TABLE 4 Effect of different yogurt groups on mouse intestinal motility and defecation
Note that: a: p <0.05 (compared to blank); b: p <0.05 (compared to negative control group); c: p <0.05 (compared to positive control).
As can be seen from table 4, the small intestine ink push rate of the mice in the negative control group was significantly reduced compared to the blank group, indicating that the model construction was successful. Compared with the negative control group, the positive control group has obviously increased small intestine ink propulsion rate (P < 0.05), which proves that the experimental system is feasible; the small intestine ink advancing rate of example 1, comparative examples 4 to 6 was significantly increased compared to the negative control group; meanwhile, the small intestine ink propelling rate of the mice in the example 1 is obviously increased compared with that of the positive control group.
As can be seen from table 4, the first-grain defecation time of the negative control group was significantly increased, the average number of black bowel movements was significantly reduced within 5 hours, and the average weight of black bowel movements was reduced within 5 hours, compared with the blank control group, indicating that the model establishment was successful. Compared with the negative control group, the positive control group has obviously reduced first-granule defecation time, obviously increased average defecation granule number in 5h and average defecation weight in 5h, and shows that the experimental system is feasible. The first granule defecation time of example 1 and comparative examples 4 to 6 was significantly reduced compared with the negative control group, and the average number of black discharge granules in 5 hours and the average weight of black discharge in 5 hours were significantly increased; meanwhile, compared with a positive control group, the first granule defecation time of the example 1 is obviously reduced, and the average number of black excrement grains in 5 hours and the average weight of black excrement in 5 hours are obviously increased. In addition, the water content of the feces of the mice in the negative control group is the lowest, and compared with the feces of other mice in each group, the wet degree of the feces is reduced, and the feces properties are drier and harder.
In summary, compared with the positive control group, the embodiment 1 has remarkable difference in bowel relaxing effect and better effect, which indicates that when the two filtrates (lactobacillus rhamnosus BBE K4 fermentation filtrate and lactobacillus paracasei JN-1 fermentation filtrate) are matched with durian pulp, the bowel relaxing effect can be greatly improved. Whereas the bowel relaxing effect with only one fermentation filtrate (comparative example 1 and comparative example 2) was the lowest in the treatment group without durian pulp; the effect of relaxing bowel is slightly higher than that of only one fermentation filtrate (comparative example 1 and comparative example 2), but is not significantly higher, without durian pulp, but with two fermentation filtrates (comparative example 3); the durian pulp and one of the fermentation filtrate (comparative examples 4, 5 and 6) have the effect of relaxing bowel, and the effect is equivalent to that of a positive control group, which shows that the durian pulp has obvious effect of improving the effect of relaxing bowel on the filtrate; when the two filtrates are matched with durian pulp to prepare the yoghurt, the effect of relaxing bowel is highest, even higher than that of a positive control group, which shows that the two filtrates have good synergistic effect with each other, and the effect of relaxing bowel can be greatly improved.
In conclusion, in the research and practice process, the invention discovers that the yogurt prepared by fermenting the main components of the combined lactobacillus rhamnosus BBE K4 and the lactobacillus paracasei JN-1 together with durian and raw milk can effectively enhance the treatment effects on high cholesterol and constipation of mice, and the yogurt prepared by fermenting the main components of the independent lactobacillus rhamnosus BBE K4 or the lactobacillus paracasei JN-1 together with durian and raw milk has no obvious treatment effects on high cholesterol and constipation of mice, or the yogurt prepared by fermenting the main components of the raw milk does not add durian into the raw milk has no obvious effects on high cholesterol and constipation of mice, so that the yogurt has very obvious effects on reducing blood fat and relaxing bowel, and 3 substances have obvious synergistic effects. Lactobacillus rhamnosus BBE K4 and lactobacillus paracasei JN-1 are safe probiotics, and have good prospects as oral agents.
The above examples merely illustrate specific embodiments of the invention, which are described in more detail and are not to be construed as limiting the scope of the invention. It should be noted that it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the spirit of the invention, which are all within the scope of the invention.

Claims (3)

1. The durian yoghourt capable of reducing blood fat and relaxing bowel is characterized by comprising the following raw materials in parts by weight: 10-15 parts of durian pulp, 70-80 parts of raw milk, 5 parts of fructo-oligosaccharide powder, 2 parts of probiotics fermentation product filtrate, 0.01-0.05 part of common starter and 0.1-0.3 part of stabilizer; the probiotics fermentation product filtrate comprises 1 part of lactobacillus rhamnosus BBE K4 fermentation filtrate and 1 part of lactobacillus paracasei JN-1 fermentation filtrate;
the common starter is a mixture of lactobacillus bulgaricus freeze-dried powder and streptococcus thermophilus freeze-dried powder in a ratio of 1:1;
the preparation method of the lactobacillus rhamnosus BBE K4 fermentation filtrate comprises the following steps:
(1) Inoculating the preserved lactobacillus rhamnosus BBE K4 into an MRS culture medium under the aseptic condition, and performing activation culture to obtain seed liquid for fermentation culture;
(2) Inoculating the lactobacillus rhamnosus BBE K4 seed solution obtained in the step (1) into an MRS culture medium with an inoculum size of 3-5 v/v%, and culturing for 22-26 h at 36-38 ℃;
(3) Centrifuging the fermentation broth obtained in step (2) at 8000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain filtrate with concentration of 3×10 8 CFU/mL of sterile lactobacillus rhamnosus BBE K4 fermentation filtrate;
the preparation method of the lactobacillus paracasei JN-1 fermentation filtrate comprises the following steps:
(1) Inoculating preserved lactobacillus paracasei JN-1 into an MRS culture medium under the aseptic condition, and performing activation culture for 24-48 hours at the temperature of 35-38 ℃ to obtain a culture serving as seed liquid for fermentation culture;
(2) Inoculating lactobacillus paracasei JN-1 seed liquid obtained in the step (1) into an MRS culture medium according to an inoculum size of 2-4 v/v percent, and performing anaerobic culture for 20-48 hours at the temperature of 35-38 ℃ to obtain a fermentation liquor;
(3) Centrifuging the fermentation broth obtained in step (2) at 6000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain filtrate with concentration of 3×10 8 CFU/mL sterile Lactobacillus paracasei JN-1 fermentation filtrate;
the preparation method of the durian yoghourt capable of reducing blood fat and relaxing bowel comprises the following steps of:
(1) Pretreatment of raw milk: raw milk is cooled to 2-6 ℃ for storage after being subjected to milk cleaning and pre-sterilization in sequence, then is subjected to secondary pre-sterilization before burdening, and is cooled to 2-6 ℃ for storage after sterilization;
(2) Adding durian pulp, fructooligosaccharide powder and a stabilizer into the sterilized raw milk, uniformly mixing, homogenizing for 2 times under 15-20 MPa, and sterilizing at 80-90 ℃ for 15min to obtain durian milk;
(3) And (3) adding the common starter and the probiotics fermentation product filtrate into the mixed milk when the durian milk is cooled to 42-45 ℃, uniformly mixing, and culturing at constant temperature for 6-8 hours at 40-42 ℃ to obtain the durian yoghourt capable of reducing blood fat and relaxing bowel after being matured for 24 hours.
2. The preparation method of durian yoghourt capable of reducing blood fat and relaxing bowel as claimed in claim 1, which is characterized by comprising the following steps:
(1) Pretreatment of raw milk: raw milk is cooled to 2-6 ℃ for storage after being subjected to milk cleaning and pre-sterilization in sequence, then is subjected to secondary pre-sterilization before burdening, and is cooled to 2-6 ℃ for storage after sterilization;
(2) Adding durian pulp, fructooligosaccharide powder and a stabilizer into the sterilized raw milk, uniformly mixing, homogenizing for 2 times under 15-20 MPa, and sterilizing at 80-90 ℃ for 15min to obtain durian milk;
(3) And (3) adding the common starter and the probiotics fermentation product filtrate into the mixed milk when the durian milk is cooled to 42-45 ℃, uniformly mixing, and culturing at constant temperature for 6-8 hours at 40-42 ℃ to obtain the durian yoghourt capable of reducing blood fat and relaxing bowel after being matured for 24 hours.
3. The preparation method of the durian yoghourt capable of reducing blood fat and relaxing bowels as claimed in claim 2, wherein the durian pulp is prepared by the following steps: cleaning peeled and cored durian pulp, cutting, pulping with a pulping machine, heating the pulp to 80-120deg.C, maintaining the temperature for 10-25 min, sterilizing, cooling to obtain durian pulp, and storing for use.
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