CN114586844A - Durian yogurt capable of reducing blood fat and relaxing bowel and preparation method thereof - Google Patents

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

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CN114586844A
CN114586844A CN202210310973.XA CN202210310973A CN114586844A CN 114586844 A CN114586844 A CN 114586844A CN 202210310973 A CN202210310973 A CN 202210310973A CN 114586844 A CN114586844 A CN 114586844A
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durian
fermentation
blood fat
reducing blood
filtrate
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CN114586844B (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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    • 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

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Abstract

The invention discloses durian yoghourt capable of reducing blood fat and relaxing bowel and a preparation method thereof, wherein the durian yoghourt comprises 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 probiotic fermentation product filtrate, 0.01-0.05 part of common leavening agent and 0.1-0.3 part of stabilizing agent; the probiotic fermentation product filtrate comprises 1 part of lactobacillus rhamnosus BBEK4 fermentation filtrate and 1 part of lactobacillus paracasei JN-1 fermentation filtrate. Different from common probiotics, the added probiotics Lactobacillus rhamnosus BBEK4 and Lactobacillus paracasei JN-1 are matched with durian and raw milk to prepare durian yoghourt which has very obvious functions of reducing blood fat and relaxing bowel, the Lactobacillus rhamnosus BBEK4, Lactobacillus paracasei JN-1 and durian have obvious synergistic effects, and the effects of reducing blood fat and relaxing bowel are obvious.

Description

Durian yogurt 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 yogurt capable of reducing blood fat and relaxing bowel and a preparation method thereof.
Background
The durian belongs to tropical fruits, the pulp of the durian consists of meat bags with aril, the meat is light yellow, and the durian is succulent and is a fruit with high economic value. The durian has extremely high nutritive value, is reputed by the king of tropical fruits, has huge market demand potential, can build the body, invigorate the spleen and tonify qi, tonify the kidney and strengthen yang, warm the body, belongs to fruits which are beneficial to nourishing, and can be made into yoghourt pudding type nutritive and leisure food which has special taste and texture and rich nutrition. Although durian is rich in nutrition and has medicinal efficacy, durian is not suitable for people, because durian is a fruit with strong heat, the durian is easy to cause body dryness-heat, and symptoms such as excessive internal heat, dry throat, dry tongue, sore throat and the like are caused.
At present, although the durian is extremely high in nutritive value, the medicinal value of the durian is not deeply developed in the durian product development process, and the yoghourt pudding products developed by the durian only have the performance of common yoghourt.
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 (Jiangnan university) discloses a Lactobacillus rhamnosus BBE K4, wherein the Lactobacillus rhamnosus rod (Lactobacillus rhamnosus) is preserved in the China center for type culture collection in 12-19.2017 with the preservation number of CCTCC NO: M2017807. The lactobacillus rhamnosus BBE K4 used in the invention is given by researchers of south China university.
202111386682.0 (university of Jiangnan) discloses a Lactobacillus paracasei JN-1, which is preserved in the China general microbiological culture Collection center at 21.6.21.2021, with the preservation address of No. 3 Xilu 1 Hospital of the sunward area of Beijing, and the preservation number of CGMCC No. 22745; the lactobacillus paracasei JN-1 has strong resistance of gastrointestinal digestive system and bile salt tolerance, can be applied to preventing and treating spontaneous enteritis, and has good effect on both functions and flavor. The lactobacillus paracasei JN-1 used in the invention is given by researchers at the university of Jiangnan.
In the research and practice process of the invention, the team discovers that the yogurt prepared by combining lactobacillus rhamnosus BBE K4, lactobacillus paracasei JN-1, durian and raw milk as main components and fermenting can effectively enhance the treatment effect on high cholesterol and constipation of mice, the yogurt prepared by singly using lactobacillus rhamnosus BBE K4 or lactobacillus paracasei JN-1 and fermenting by using durian and raw milk as main components has no remarkable treatment effect on high cholesterol and constipation of mice, or the yogurt prepared by fermenting raw milk without durian has no remarkable effect on high cholesterol and constipation of mice, which shows that lactobacillus rhamnosus BBE K4, lactobacillus paracasei JN-1 and durian have very remarkable functions of reducing blood fat and relaxing bowel, and 3 substances have remarkable synergistic effect. The lactobacillus rhamnosus BBE K4 and the lactobacillus paracasei JN-1 are safe probiotics and have a bright prospect as an oral reagent.
In order to achieve the purpose, the invention is realized by the following technical scheme:
the durian yoghourt capable of reducing blood fat and relaxing bowel comprises 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 probiotic fermentation product filtrate, 0.01-0.05 part of common leavening agent and 0.1-0.3 part of stabilizing agent.
Further, the probiotic 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) under the aseptic condition, inoculating the preserved lactobacillus rhamnosus BBE K4 in an MRS culture medium for activation culture and then taking the activated lactobacillus rhamnosus BBE K4 as a seed solution for fermentation culture;
(2) inoculating the lactobacillus rhamnosus BBE K4 seed solution obtained in the step (1) in an MRS culture medium by an inoculation amount of 3-5 v/v%, and culturing for 22-26 h at 36-38 ℃; wherein the MRS medium configuration is: 10g of peptone, 8g of beef extract, 4g of yeast extract, 20g of glucose, 801mL of tween-801, 2g of dipotassium phosphate, 5g of sodium acetate trihydrate, 2g of ammonium citrate, 0.2g of magnesium sulfate heptahydrate and 0.05g of potassium manganese sulfate tetrahydrate are dissolved in 1L of distilled water.
(3) Centrifuging the fermentation liquid obtained in step (2) at 8000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain fermentation liquid with concentration of 3 × 108CFU/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 the preserved lactobacillus paracasei JN-1 into an MRS culture medium under an aseptic condition, and performing activated culture at 35-38 ℃ for 24-48 h to obtain a culture as a seed solution for fermentation culture;
(2) inoculating the lactobacillus paracasei JN-1 seed solution obtained in the step (1) into an MRS culture medium in an inoculation amount of 2-4 v/v%, and carrying out anaerobic culture at 35-38 ℃ for 20-48 h to obtain a fermentation liquid;
(3) centrifuging the fermentation liquid obtained in step (2) at 6000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain 3 × 108CFU/mL sterile Lactobacillus paracasei JN-1 fermentation filtrate.
Further, the common type leaven is a mixture of lactobacillus bulgaricus and streptococcus thermophilus 1: 1.
In the invention, the fructo-oligosaccharide is soluble dietary fiber, and after being ingested by a human body, the fructo-oligosaccharide can reach the large intestine and be selectively utilized by beneficial flora in the intestinal tract because the fructo-oligosaccharide is not digested and absorbed, thereby effectively regulating the intestinal flora. Meanwhile, the stability of the fructo-oligosaccharide under the acidic condition is superior to that of cane sugar, and compared with the cane sugar, the fructo-oligosaccharide is fresh and cool in sweetness, pure in taste, free of any aftertaste, and free of change in sweetness, and does not affect the taste of durian yogurt.
On the other hand, the invention also provides a preparation method of the durian yoghourt capable of reducing blood fat, moistening intestines and relaxing bowels, which comprises the following steps:
(1) pretreatment of raw milk: cleaning and pre-sterilizing raw milk in sequence, cooling to 2-6 ℃ for storage, then performing secondary pre-sterilization treatment before batching, and cooling to 2-6 ℃ for storage after sterilization for later use;
(2) adding the durian pulp, the fructo-oligosaccharide powder and the 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 adding a common starter and probiotic fermentation product filtrate into the mixed milk when the durian milk is cooled to about 42-45 ℃, uniformly mixing, culturing at constant temperature of 40-42 ℃ for 6-8 h, and ripening for 24h to obtain the durian yoghourt capable of reducing blood fat and relaxing bowel.
In the technical scheme, the preparation process of the durian pulp comprises the following steps: cleaning peeled and shelled durian pulp, cutting into blocks, pulping by using a pulping machine, heating the pulp obtained by pulping to 80-120 ℃, preserving heat for 10-25 min, sterilizing, and cooling to obtain the durian pulp, and storing for later use.
Compared with the prior art, the invention has the beneficial effects that:
different from common probiotics, the added probiotics lactobacillus rhamnosus BBE K4 and lactobacillus paracasei JN-1 are matched with durian and raw milk to prepare durian yoghourt which has very obvious functions of reducing blood fat and relaxing bowel, the lactobacillus rhamnosus BBE K4, the lactobacillus paracasei JN-1 and the durian have obvious synergistic effect, and the effects of reducing blood fat and relaxing bowel are obvious.
Drawings
FIG. 1 is a bar graph of the effect of different groups of yogurt on rat serum TC after modeling and after dosing in example 3;
FIG. 2 is a bar graph of the effect of different groups of yogurt on rat serum TG, LDL-C, HDL-C after administration in example 3.
Detailed Description
The embodiments of the present invention are described below with reference to specific embodiments, and other advantages and effects of the present invention will be easily understood by those skilled in the art from the disclosure of the present specification. The invention is capable of other and different embodiments and of being practiced or of being carried out in various ways, and its several details are capable of modification in various respects, all without departing from the spirit and scope of the present invention. It is to be noted that the features in the following embodiments and examples may be combined with each other without conflict.
It should be noted that the drawings provided in the following embodiments are only for illustrating the basic idea of the present invention, and the drawings only show the components related to the present invention rather than being drawn according to the number, shape and size of the components in actual implementation, and the type, amount and proportion of each component in actual implementation can be changed freely, and the layout of the components can be more complicated.
Example 1
The 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 fructo-oligosaccharide powder, 2 parts of probiotic fermentation product filtrate and 0.04 part of common leavening agent (the mixed concentration of lactobacillus bulgaricus and streptococcus thermophilus inoculants is not less than 10)10CFU/g) and 0.1-0.3 part of stabilizer. Wherein the stabilizer is gelatin and/or low ester pectin.
In this embodiment, the probiotic 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) under the aseptic condition, inoculating the preserved lactobacillus rhamnosus BBE K4 in an MRS culture medium for activation culture and then taking the activated lactobacillus rhamnosus BBE K4 as a seed solution for fermentation culture;
(2) inoculating the lactobacillus rhamnosus BBE K4 seed solution obtained in the step (1) in an MRS culture medium by an inoculation amount of 3-5 v/v%, and culturing for 22-26 h at 36-38 ℃; wherein the MRS medium configuration is: 10g of peptone, 8g of beef extract, 4g of yeast extract, 20g of glucose, 801mL of tween-801, 2g of dipotassium phosphate, 5g of sodium acetate trihydrate, 2g of ammonium citrate, 0.2g of magnesium sulfate heptahydrate and 0.05g of potassium manganese sulfate tetrahydrate are dissolved in 1L of distilled water.
(3) Centrifuging the fermentation liquor obtained in the step (2) at a centrifugal rotation speed of 8000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain 3 × 108CFU/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 the preserved lactobacillus paracasei JN-1 into an MRS culture medium under an aseptic condition, and performing activated culture at 35-38 ℃ for 24-48 h to obtain a culture as a seed solution for fermentation culture;
(2) inoculating the lactobacillus paracasei JN-1 seed solution obtained in the step (1) into an MRS culture medium in an inoculation amount of 2-4 v/v%, and carrying out anaerobic culture at 35-38 ℃ for 20-48 h to obtain a fermentation liquid;
(3) centrifuging the fermentation liquid obtained in step (2) at 6000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain 3 × 108CFU/mL sterile Lactobacillus paracasei JN-1 fermentation filtrate.
In this example, the common starter culture is a mixture of Lactobacillus bulgaricus and Streptococcus thermophilus 1: 1. For example, a common type of starter is lyophilized Streptococcus thermophilus powder (10)10CFU/g)0.02 parts and lactobacillus bulgaricus freeze-dried powder (10)10CFU/g)0.02 parts of mixed bacterial powder (food grade).
The preparation method of the durian yoghourt capable of reducing blood fat and relaxing bowel provided by the embodiment comprises the following steps:
(1) pretreatment of raw milk: cleaning and pre-sterilizing raw milk in sequence, cooling to 4 ℃ for storage, then performing secondary pre-sterilization treatment before batching, and cooling to 4 ℃ for storage after sterilization for later use;
(2) adding the durian pulp, the fructo-oligosaccharide powder and the 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 adding a common starter and the filtrate of the probiotic fermentation product into the mixed milk when the durian milk is cooled to about 45 ℃, uniformly mixing, culturing at the constant temperature of 42 ℃ for 6h, and ripening for 24h to obtain the durian yoghourt capable of reducing blood fat and relaxing bowel.
Example 2
The yoghurts prepared in comparative examples 1 to 6 are similar to those prepared in example 1, except that part of the ingredients are different, and the ingredients in comparative examples 1 to 6 are shown in table 1:
TABLE 1 ingredient Table for different yoghurts obtained in example 1, comparative examples 1 to 6
Figure BDA0003567111230000051
Note: the microbial inoculum in the lactobacillus paracasei fermentation filtrate in the comparative example 6 is purchased and preserved in China general microbiological culture collection center with the preservation number of CGMCC 1.9089, the preparation method of the lactobacillus paracasei fermentation filtrate is similar to the preparation method of the lactobacillus paracasei JN-1 fermentation filtrate in the example 1, and the microbial inoculum concentration of the lactobacillus paracasei in the fermentation filtrate is 3 multiplied by 10 CGMCC 1.90898CFU/mL。
Example 3
Grouping rats: 100 male rats with the weight of 200-220 g are divided into 10 groups, and the rats in each group have no statistical difference. The 10 groups were blank control group (normal rats), positive control group (lovastatin group), negative control group, example 1 and treatment groups of comparative examples 1 to 6, respectively, and 10 rats per group. Wherein, the normal rat group is used as a blank control, and the positive control group, the negative control group and the treatment group use the rats successfully modeled by the hyperlipidemia.
The modeling method of the hyperlipemia rat comprises the following steps: 100 male rats were adaptively fed for 1 week, 10 rats were randomly selected to continue to be given basal diet (78.8% basal diet, 10% lard, 10% egg yolk powder, 1% cholesterol, 0.2% bile salt) as a blank control group, and the remaining 90 rats were given high fat diet as a high fat model group. After modeling, orbital blood is collected from rats, serum Total Cholesterol (TC), Triglyceride (TG), high-density lipoprotein cholesterol (HDL-C) and low-density lipoprotein cholesterol (LDL-C) levels are measured, TC or TG in serum of high-fat rats in a model group is remarkably increased (P is less than 0.05) compared with a blank control group, the model is determined to be established, and the test result is shown in table 2.
After the modeling is successful, the blank control group and the negative control group are not administrated, and the same volume of distilled water is infused into the stomach every day; feeding lovastatin suspension to the positive control group for intragastric administration, converting into dosage for rats according to the medication guidance of the drug, and performing intragastric administration at the dosage of 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 treated continuously for 30 days, and after the last administration and after fasting for 12h, orbital blood was collected from the rats and serum Total Cholesterol (TC), Triglyceride (TG), high density lipoprotein cholesterol (HDL-C) and low density lipoprotein cholesterol (LDL-C) levels were measured. The test results are shown in tables 2 and 3.
TABLE 2 Effect of different groups of yoghurt on TC, TG in rat serum
Figure BDA0003567111230000061
Note: a: p <0.05 (compared to blank); b: p <0.05 (compared to negative control); c: p <0.05 (compared to positive control).
TABLE 3 Effect of different groups of yoghurts on rat serum LDL-C, HDL-C
Figure BDA0003567111230000062
Note: 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 hyperlipidemic rats is successful. After administration, TC in the groups of example 1 and comparative examples 1 to 6 is in a descending trend during modeling, and the TC in the serum of the rats in the positive control group, example 1 and comparative examples 4 to 6 is significantly different from that in the negative control group; meanwhile, the serum TC of the rat in the example 1 is obviously reduced compared with that of the positive control group. After modeling is carried out by the table 2 and the figure 2, compared with a blank control group, the TG level of the negative control group is obviously increased, which indicates that the modeling of the hyperlipoidemia rat is successful. The TG values of the rats in the blank control group and the negative control group are not changed significantly in the test process. Compared with the negative control group, the positive control group, the example 1 and the comparative examples 4 to 6 have obviously reduced serum TG. The tests show that the yoghourt provided by the invention not only can reduce the serum total cholesterol of high-fat rats, but also can effectively reduce the serum triglyceride of the high-fat rats.
As can be seen from table 3 and fig. 2, the serum LDL-C of the rats of the positive control group, example 1 and comparative examples 4 to 6 was significantly reduced compared to that of the negative control group. The yoghourt provided by the invention is proved to be capable of effectively reducing the LDL-C level of rat serum. After modeling, the values of the rat serum HDL-C between the groups are significantly different, the levels of the rat serum HDL-C of the blank control group, the positive control group, the rat serum HDL-C of the example 1 and the rat serum of the comparative examples 1 to 6 are all in an ascending trend in the experimental process, the negative control group is significantly lower than the blank control group, and the rat serum LDL-C of the positive control group, the rat serum of the example 1 and the rat serum of the comparative examples 4 to 6 are significantly increased compared with the negative control group by the end of the experiment. The yoghourt provided by the invention is proved to be capable of effectively improving the level of HDL-C in the serum of a rat.
As a result, the blood lipid lowering effect of example 1 was better compared with that of the positive control group in example 1. But not containing durian pulp, only one fermentation filtrate (comparative example 1 and comparative example 2) had the lowest effect of reducing blood lipid; the blood lipid lowering effect with two fermentation filtrates (comparative example 3) without durian pulp was slightly higher but not significantly higher than with one fermentation filtrate (comparative example 1 and comparative example 2); the durian pulp and one of the fermentation filtrates (comparative examples 4, 5 and 6) can reduce blood fat, 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 fat reducing effect on the filtrate; when the two filtrates are matched with durian pulp to prepare the yoghourt, the effect of reducing blood fat is the highest, even higher than that of a lovastatin solution, which shows that the two filtrates have good synergistic effect with each other and can greatly improve the effect of reducing blood fat.
Example 4
Grouping mice: 160 male BALB/c mice weighing 18-22 g are taken, the mice are randomly divided into 2 large groups, namely a small intestine movement experiment group and a defecation experiment group, each large group comprises 10 small groups, and the mice in each group have no statistical difference. 10 groups in each large group were a blank control group (normal mice), a positive control group (xylo-oligosaccharide group), a negative control group (constipation model group), example 1, and treatment groups of comparative examples 1 to 6, respectively. Each group had 8 mice. Wherein, the normal mouse group is used as a blank control, and the positive control group, the negative control group and the treatment group use the mice which are successfully modeled by constipation.
The method for establishing the constipation model comprises the following steps: after the mice are grouped, feeding yoghourt to each treatment group (comprising 2 treatment groups of the big groups) at the time of 2ml/(100g BW) every day, feeding pure water with the gastric administration amount of volume to the blank control group and the negative control group, feeding xylo-oligosaccharide suspension to the positive control group at the time of every day with the gastric administration amount of 1.5g/(kg BW) every day, feeding yoghourt for 14 days continuously, wherein the mice of each group eat, drink water and move normally, after the last time of feeding the yoghourt and the xylo-oligosaccharide suspension, all the mice of the two big groups are fasted for 20 hours without water prohibition, feeding the mice of the negative control group, the positive control group and the treatment groups with the aqueous solution of loperamide hydrochloride, and feeding the gastric administration amount of 10mg/(kg BW) to the mice of the blank control group with the same volume of pure water.
1. Small intestine exercise experiment
After 30min of the loperamide hydrochloride for gastric lavage, the treatment components are respectively fed with yoghourt containing corresponding dose prepared by edible ink, xylo-oligosaccharide suspension containing corresponding dose prepared by the ink for gastric lavage of a positive control group, ink with the same volume as the gavage of a blank control group and a model control group, after 25min, immediately pulling the neck to kill all mice in a small intestine movement experiment group, dissecting, opening the abdominal cavity, separating mesentery, cutting the intestinal canal part from the pylorus, the lower end to the ileoceria, placing on a tray, slightly pulling the small intestine into a straight line, measuring the length of the intestinal canal, recording as the total length of the small intestine, recording the length of the ink from the pylorus to the front edge as the length of the ink, calculating the ink propulsion rate according to the following formula, judging the influence on the intestinal peristalsis of the constipation mice, and showing the experiment result in Table 4.
Ink propulsion rate (%) (ink propulsion length/total small intestine length) × 100%
2. Defecation test
After the loperamide hydrochloride for gastric perfusion is fed for 30min, the treatment groups are respectively fed with yoghourt prepared from edible ink, xylo-oligosaccharide suspension prepared from positive control group ink for gastric perfusion and containing corresponding dose, ink with the same gastric perfusion capacity in a blank control group and a model control group, mice in an defecation experimental group are all fed in a single cage, normal food intake, water drinking and activities are carried out, the time for discharging black feces of the first particle of each mouse and the number and weight of the black feces in 5h are observed and recorded from the ink for gastric perfusion, the water content of feces is calculated according to the following formula so as to judge the influence on the defecation of the mice, and the test result is shown in Table 4.
Stool water content (%) (wet stool weight-dry stool weight)/wet stool weight) x 100%
TABLE 4 Effect of different groups of yoghurts on bowel movement and defecation in mice
Figure BDA0003567111230000081
Note: a: p <0.05 (compared to blank); b: p <0.05 (compared to negative control); c: p <0.05 (compared to positive control).
As can be seen from Table 4, the rate of intestinal ink propulsion in the negative control mice was significantly reduced compared to the control blank, indicating that the model was successfully constructed. Compared with the negative control group, the small intestine ink propulsion rate is obviously increased (P is less than 0.05), which indicates that the experimental system is feasible; the small intestine ink propulsion rate of the example 1 and the comparative examples 4 to 6 is obviously increased compared with that of the negative control group; meanwhile, the ink propulsion rate of the small intestine of the mouse in the example 1 is obviously increased compared with that of the positive control group.
As can be seen from Table 4, compared with the blank control group, the first defecation time of the negative control group is significantly increased, the average number of the black defecation granules within 5h is significantly reduced, and the average weight of the black defecation granules within 5h is reduced, which indicates that the model is successfully established. Compared with the negative control group, the positive control group has the advantages that the first defecation time is obviously reduced, the average defecation number within 5h and the average defecation weight within 5h are obviously increased, and the feasibility of the experimental system is proved. Compared with a negative control group, the first defecation time of the example 1 and the comparative examples 4 to 6 is obviously reduced, and the average defecation number within 5h and the average defecation weight within 5h are obviously increased; meanwhile, compared with a positive control group, the first defecation time of the example 1 is obviously reduced, and the average number of the black defecation granules within 5h and the average weight of the black defecation within 5h 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 moisture degree of the feces is reduced, and the feces are dry and hard.
In conclusion, compared with the positive control group, the effect of relaxing bowel of example 1 is significantly different and better, which shows that when two filtrates (lactobacillus rhamnosus BBE K4 fermented filtrate and lactobacillus paracasei JN-1 fermented filtrate) are matched with durian pulp, the synergistic effect can be achieved, and the effect of relaxing bowel is greatly improved. Whereas the effect of relaxing bowel was lowest in the treatment group with no durian pulp, only one fermentation filtrate (comparative example 1 and comparative example 2); the effect of relaxing bowel without durian pulp but with two fermentation filtrates (comparative example 3) was slightly higher but not significantly higher than with only one fermentation filtrate (comparative example 1 and comparative example 2); the durian pulp and one of the fermentation filtrates (comparative examples 4, 5 and 6) have the effect of relaxing bowel, and the effect is equivalent to that of a positive control group, so that the durian pulp has the obvious effect of improving the effect of relaxing bowel on the filtrate; when the two filtrates are matched with durian pulp to prepare the yoghourt, the effect of relaxing bowel is highest and 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 processes of the team, the combination of lactobacillus rhamnosus BBE K4, lactobacillus paracasei JN-1, durian and raw milk is found to be used for fermentation to prepare the yoghourt which can effectively enhance the treatment effect on high cholesterol and constipation of mice, the lactobacillus rhamnosus BBE K4 or lactobacillus paracasei JN-1 is used alone and is matched with the yoghourt prepared by fermentation with durian and raw milk as main components, the treatment effect on high cholesterol and constipation of mice is not obvious, or the effect on high cholesterol and constipation of mice is not obvious by adding durian in raw milk, so that lactobacillus rhamnosus BBE K4, lactobacillus paracasei JN-1 and durian have very obvious functions of reducing blood fat and relaxing bowel, and 3 substances have obvious synergistic effects. The lactobacillus rhamnosus BBE K4 and the lactobacillus paracasei JN-1 are safe probiotics and have a bright prospect as an oral reagent.
The above-mentioned embodiments only express the specific embodiments of the present invention, and the description thereof is specific and detailed, but not construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention.

Claims (7)

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 probiotic fermentation product filtrate, 0.01-0.05 part of common leavening agent and 0.1-0.3 part of stabilizing agent.
2. The durian yogurt capable of reducing blood fat and relaxing bowel according to claim 1, wherein the probiotic fermentation product filtrate comprises 1 part of lactobacillus rhamnosus BBE K4 fermentation filtrate and 1 part of lactobacillus paracasei JN-1 fermentation filtrate.
3. The durian yogurt capable of reducing blood fat and relaxing bowel according to claim 1, wherein the common starter is a 1:1 mixture of lactobacillus bulgaricus lyophilized powder and streptococcus thermophilus lyophilized powder.
4. The durian yogurt capable of reducing blood fat and relaxing bowel according to claim 2, characterized in that the preparation method of the lactobacillus rhamnosus BBE K4 fermentation filtrate comprises the following steps:
(1) under the aseptic condition, inoculating the preserved lactobacillus rhamnosus BBE K4 in an MRS culture medium for activation culture and then taking the activated lactobacillus rhamnosus BBE K4 as a seed solution for fermentation culture;
(2) inoculating the lactobacillus rhamnosus BBE K4 seed solution obtained in the step (1) in an MRS culture medium by an inoculation amount of 3-5 v/v%, and culturing for 22-26 h at 36-38 ℃;
(3) centrifuging the fermentation liquid obtained in step (2) at 8000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain 3 × 108CFU/mL of sterile Lactobacillus rhamnosus BBE K4 fermentation filtrate.
5. The durian yogurt capable of reducing blood fat and relaxing bowel according to claim 2, wherein the preparation method of the lactobacillus paracasei JN-1 fermentation filtrate comprises the following steps:
(1) inoculating the preserved lactobacillus paracasei JN-1 into an MRS culture medium under an aseptic condition, and performing activated culture at 35-38 ℃ for 24-48 h to obtain a culture as a seed solution for fermentation culture;
(2) inoculating the lactobacillus paracasei JN-1 seed solution obtained in the step (1) into an MRS culture medium in an inoculation amount of 2-4 v/v%, and carrying out anaerobic culture at 35-38 ℃ for 20-48 h to obtain a fermentation liquid;
(3) centrifuging the fermentation liquid obtained in step (2) at 6000r/min for 20min, collecting supernatant to obtain fermentation filtrate, and filtering with 0.22um sterilizing filter to obtain 3 × 108CFU/mL sterile Lactobacillus paracasei JN-1 fermentation filtrate.
6. The preparation method of the durian yogurt capable of reducing blood fat and relaxing bowel according to any one of claims 1 to 5, which is characterized by comprising the following steps:
(1) pretreatment of raw milk: cleaning and pre-sterilizing raw milk in sequence, cooling to 2-6 ℃ for storage, then performing secondary pre-sterilization treatment before batching, and cooling to 2-6 ℃ for storage after sterilization for later use;
(2) adding the durian pulp, the fructo-oligosaccharide powder and the 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 adding a common starter and probiotic fermentation product filtrate into the mixed milk when the durian milk is cooled to about 42-45 ℃, uniformly mixing, culturing at constant temperature of 40-42 ℃ for 6-8 h, and ripening for 24h to obtain the durian yoghourt capable of reducing blood fat and relaxing bowel.
7. The method for making the durian yogurt capable of reducing blood fat and relaxing bowel according to claim 6, wherein the preparation process of the durian pulp slurry is as follows: cleaning peeled and shelled durian pulp, cutting into blocks, pulping by using a pulping machine, heating the pulp obtained by pulping to 80-120 ℃, preserving heat for 10-25 min, sterilizing, and cooling to obtain the durian pulp, and storing for later use.
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