CN101361506A - Fermentation yogurt capable of reducing cholesterol and preparation method thereof - Google Patents

Fermentation yogurt capable of reducing cholesterol and preparation method thereof Download PDF

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Publication number
CN101361506A
CN101361506A CNA2008100134918A CN200810013491A CN101361506A CN 101361506 A CN101361506 A CN 101361506A CN A2008100134918 A CNA2008100134918 A CN A2008100134918A CN 200810013491 A CN200810013491 A CN 200810013491A CN 101361506 A CN101361506 A CN 101361506A
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milk
cholesterol
lactic acid
acid bacteria
norcholesterol
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侯红漫
郭东启
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Dalian Polytechnic University
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Dalian Polytechnic University
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Abstract

The invention discloses a cholesterol-lowering ferment cultured milk and a preparation method thereof. The material milk is prepared by fermenting 3 mixing strains of the lactobacillus GL-03, bulgaria bacillus and streptococcus thermophilus, and a hawthorn extract; the technique comprises the following steps: the material milk is needed for milk-test and purifying, and then can be prepared for use, sucrose and a stabilizing agent are dissolved by little solution, and then added in the milk liquid after purifying; the temperature is enhancedto 60 DEG C for carrying out homogenate, sterilization, the temperature is lowered down to 43 DEG C, inoculation is carried out in a greenhouse of 40-45 DEG C for fermenting 4-5 hours, the temperature is cooled to under 10 DEG C after the pH value is 4.6-4.8, then the ferment cultured milk is done after being replaced to a refrigeratory of 0-4 DEG C for refrigeration. Shown by GL-03 culture in vitro test, the ferment cultured milk has strong acid and cholate resistant ability, good adhesive, obvious inhibit to common pathogenic entero becteria, without drug tolerance factors, is a lactobacillus applicable to permanent planting of human body intestinal tract, and simultaneously the deterioration ratio of the cholesterol in vitro can be up to 56 percent; proved by the animal-test, the cholesterol-lowering cultured milk has the function of lowering serum cholesterol.

Description

Fermentation yogurt of a kind of norcholesterol and preparation method thereof
Technical field
The present invention relates to the fermented milk preparation, is with lactobacillus-fermented and to be equipped with the wholefood extract be that lactic acid bacteria growth promoter is made.The technical field that also relates to norcholesterol function yogurt.
Background technology
Along with the progress of the development of society, science and technology, people's living standard improves constantly, and daily diet nutritional also increases day by day, but that thing followed negative effect is the incidence of cardiovascular and cerebrovascular diseases such as atherosclerotic, coronary heart disease is constantly soaring.1996 annual health statistics yearbooks according to World Health Organization's announcement, the whole world is because of dead 1,530 ten thousand people of angiocardiopathy, account for 29% of total death toll, therefore angiocardiopathy is in first of the world's five big causes of the death, and their common pathologic basis all are artery sclerosis.Atherosclerotic is facilitated by many factors, and wherein most important hazards are that hyperlipidemia is the rising of cholesterolemia and blood triglyceride.Thereby, keep in the blood cholesterol concentration in normal range (NR), atherosis and cardiovascular and cerebrovascular disease is vital for prevention of arterial [1]
The intake standard that nutrition that the U.S., China are domestic and medical expert recommend the food cholesterol is 250-300mg/d/ people.But along with the increase with diet nutritional of improving constantly of living standards of the people, the intake of cholesterol is easy to exceed standard.According to Shanghai Inst. of Cardio-Vascular Diseases's investigation to 3467 Shanghai residents in the period of 1997-1999, because the nutrient intake of every day is than being significantly increased the seventies in 20th century, fat increases to 31.8% by 24.1%, cholesterol rises to 388mg by 282mg, carbohydrate then reduces to 53.7% by 65.7%, and Shanghai people's blood fat then obviously increases.Therefore research does not influence flavour of food products, and the economical and practical reduction food and the method for human serum cholesterol have become current food quality, the raising people's living standard important research project improved [2]
The research to lactobacillus for degrading cholesterol abroad starts to walk early.1977, Gilliland to enteron aisle lactobacillus norcholesterol effect study, propose lactobacillus and in growth course, promote the catabolism of cholesterol, thereby reduce the viewpoint of cholesterol level by the degraded cholate [3]Rasic etc. has also confirmed the effect that removes to cholesterol of lactobacillus acidophilus and two Bifidobacteriums subsequently [4]Human body and zoopery up to now also proved the norcholesterol effect of lactic acid bacteria [5]
The domestic research about lactic acid bacteria norcholesterol aspect of China is started late, and most level that all rests in vitro study, yet there are no the report and the appearance of the lactobacillus for degrading cholesterol series products of success.
List of references
[1] Li Ruijie translates. reduce cholesterol levels prevention coronary cardiopathy. and JAMA Chinese edition [M] .2000,19 (6): 297-303.
[2] Zhang Jiacheng, Luo Chengxiang. lactic acid bacteria removes the research [J] of effect, Food Science .1998 (3): 20-23 to cholesterol in the food
[3]Maire?B,Colin?H,and?Cormac?G.M.Bile?Salt?Hydrolase?Activity?inProbiotics[J].Applied?and?Evironmental?Microbiology,2006,72(3):1729-1738
[4]He?F,Ouwehand?AC,Isolauri?E,et?al.Comparison?of?mucosal?adhesionand?species?identification?of?bifidobacteria?isolated?fromhealthy?and?allergic?infants[J].FEMS?Immunol?Med?Microbiol,200l,30(1):43-47
[5] Liu Hanling. lactobacillus norcholesterol The Characteristic Study [D] in the tradition dairy products of pastoral area, the Inner Mongol. Agricultural University of the Inner Mongol, 2006
Summary of the invention
The object of the present invention is to provide a kind of yogurt production method with norcholesterol effect.Wherein mainly comprise the separation screening of efficient lactobacillus for degrading cholesterol, the prescription of norcholesterol yogurt fermentation, the technological condition for fermentation of norcholesterol yogurt.
1, the screening of lactobacillus for degrading cholesterol GL-03 and cholesterol degradation are induced
The used bacterial classification of the present invention is accredited as Lactobacillus casei cheese subspecies (Lactobacillus casei subsp.casei) for the lactic acid bacteria GL-03 of the efficient norcholesterol that filters out through the 16SrDNA order-checking from natural fermented cheese.Lactobacillus for degrading cholesterol GL-03 tests through substrate for induction, its cholesterol degradation rate 50% is increased to 56% before induce, simultaneously its have preferably stomach juice-resistant, bile tolerance and to the intestinal epithelial cell adhesiveness, suppress characteristic such as pathogenic bacteria, be a kind of probio that is suitable for the field planting human body intestinal canal.
Separating screening method: take by weighing an amount of natural fermented cheese, after the sterilized water dilution, sterile working is drawn 2ml and is added in the skimmed milk of sterilizing, and 37 ℃ leave standstill enrichment culture 48h.Sterile working is got the above-mentioned enrichment culture liquid adding of 1ml and is contained in the aseptic improvement KENJI selective medium of 9ml (containing 0.04% bromine thyme phenol orchid), 37 ℃ shaken cultivation 1-2 days, get nutrient solution 100 μ L then and be added in the same culture medium and cultivate, need three tubes to cultivate altogether.Ratio turbidity and indicator change in color according to culture medium, get for the third time the MRS fluid nutrient medium that bacterium liquid 200 μ L after cultivating join pH2 and cultivate 6h, getting its 100 μ L nutrient solution is transferred to contain in the 0.30% pig cholate MRS fluid nutrient medium and cultivates 24h, that gets 200 μ L nutrient solutions then is coated on the modified MRS flat board 37 ℃ of constant temperature culture 48~72h.Picking produces the colonies typical of acid, line separation repeatedly, purifying, and Gram, microscopy are till pure bacterium colony.Then, identify through 16SrDNA order-checking carrying out bacterial classification.
Directional induction: after the lactic acid bacteria GL-03 activation with seed selection, quantitatively be added in the KENJI fermentation medium that contains 0.25% cholesterol and cultivate, the concentration of a cholesterol of every cultivation all will increase by 0.15%, five domestication cultivate after, make that content of cholesterol reaches 1mg/mL in the culture medium.Select well-grown lactic acid bacteria GL-03, in containing the MRS fluid nutrient medium of cholesterol 0.1mg/mL, measure the variation of its cholesterol degradation rate before and after inducing and acclimating, therefrom filter out the norcholesterol ability and improve bigger bacterial strain.
2, the fermentation condition of norcholesterol yogurt and technology
The present invention is a kind of norcholesterol fermentation yogurt, not only adds the lactic acid bacteria GL-03 of efficient norcholesterol in acidified milk, has added haw thorn extract simultaneously.Test shows that haw thorn extract is that lactobacter growth promotes the factor, and an amount of interpolation can improve in the yogurt about 100 times of viable lactic acid bacteria quantity, and haw thorn extract can reduce body's cholesterol with the lactic acid bacteria synergy simultaneously.
(1) actication of culture and leavening are made
Lactic acid bacteria GL-03, bacillus bulgaricus, the streptococcus thermophilus of refrigerator preservation are transferred in the liquid MRS culture medium by the MRS inclined-plane, cultivate 12h, activate three times for 37 ℃; The bacterial classification of activation is inoculated in the skimmed milk power liquid that contains 10% (W/V) of sterilization by 2%, and 37 ℃ are cultured to curdled milk, transfer and make leavening three times.
(2) preparation of haw thorn extract
Get hawthorn 50 grams, earlier material is cut into broken end or powder, add 3~8 times water, soak 2~4h, boil the back and continue 30~40min with the line fire; Leach decocting liquid, add 3~5 times water, carry out two frying in shallow oil (20~30min) leach decocting liquid, add 2~4 times water, carry out three frying in shallow oil (20~30min), merging filtrate is concentrated into 100mL, gets supernatant, and 115 ℃, sterilization 15min, 4 ℃ of refrigerators are preserved standby.
(3) acidified milk composition of raw materials
The raw material quality per distribution ratio of functional norcholesterol sour milk is:
Fresh milk or recombined milk 80.0~85.0
Milk powder 0.0~2.0
Sucrose 5.0~7.0
Hawthorn extract 4.0~6.0
Three kinds of mixed bacterias 2.0~5.0
Stabilizing agent CMC-Na 0.1
Wherein three kinds of mixed bacterias are that lactic acid bacteria GL-03 (Lactobacillus casei subsp.casei), bacillus bulgaricus (Lactobacillus bulgaricus) and 3 kinds of bacterium of streptococcus thermophilus (Streptococcusthermophilus) mix, its ratio is 1:0.5~1.0:0.5~1.0, fermentation temperature 40-45 ℃.
(4) technological process
Raw milk → clean breast → standardization → preheating (60 ℃) → homogeneous (15~17Mpa) → sterilization → cooling → interpolation leavening → packing → fermentation → refrigeration (0~5 ℃) → check → transmission
The concrete operations step:
1) fresh milk (or recombined milk) is examined, purifies, is waited to join through milk.
2) sucrose, stabilizing agent are dissolved with a small amount of emulsion (50-60 ℃), add in the milk liquid after purifying, stir, be warming up to and carry out homogeneous (pressure is 15-17Mpa) about 58~62 ℃.
3) the milk liquid behind the homogeneous is heated to 90 ℃, 5min or ultra high temperature short time sterilization (135 ℃ 3-5s), are cooled to 43 ℃, inoculation fermentation then.
4) feed liquid quick branch in inoculation back installs to sell and uses container, and delivers to 40-45 ℃ of thermostatic chamber fermentation 4-5h, to pH4.6-4.8.Be cooled to rapidly below 10 ℃, and move to 0-4 ℃ of freezer refrigeration after-ripening.
3, norcholesterol yogurt quality standard
(1) organoleptic indicator:
Color and luster: the color and luster uniformity is creamy white or is with little yellow slightly.
Flavour smell: have distinctive flavour of cultured milk and smell that the homolactic fermentation agent is made, alcohol-free fermented flavour, musty and other bad smell.
Structural state: grumeleuse uniform and smooth, no bubble, permission small amount of whey are separated out.
(2) physical and chemical index:
Butter oil 〉=3.00wt%, real milk solids 〉=10wt%, 70~110 ° of T of acidity, granulated sugar 〉=5.0wt%, mercury≤0.01%ppm.
Every 100g norcholesterol yogurt contains heat 60-70Kcal, non-fat solid 〉=8.1g, protein 〉=3.0g, granulated sugar 〉=5.0g, viable lactic acid bacteria number 〉=1 * 10 6Cfu/g.
(3) microbiological indicator: coliform≤90/milliliter, pathogenic bacteria must not detect.
4, the zoopery of norcholesterol acidified milk
To being the norcholesterol yogurt that main fermenting agent is made, carried out reducing in the body zoopery of serum cholesterol with lactic acid bacteria GL-03.
With 30 male white mouses (available from Dalian Medical Univ's Experimental Animal Center), 20~25 ℃ of temperature, the feed of freely drinking water in the environment of relative humidity 50 ± 5%, normal diet is fed behind the 3d, weigh one by one, do not have apparent difference by average weight between each group and be divided into 3 groups again, 10 every group.A group: basal diet group (normal group), the basal feed of feeding; B group: hyperlipidemia model group, the high lipid food of feeding; C irritates the stomach group: high lipid food+norcholesterol yogurt, irritated stomach 1 time in per 2 days, and to carry out at early about 9 o'clock, A, B group gavages 0.9% physiological saline, and the C group gavages the norcholesterol yogurt, only irritates stomach amount 1mL/.
Fed mouse after 30 days, fasting 12h, eye socket blood sampling, 4 ℃ of refrigerations, TC, TG, HDL-C, the LDL-C of mensuration blood fat.After the blood sampling, the disconnected neck of mouse is put to death, and gets liver,kidney,spleen after the dissection respectively and weighs.
After feeding 30 days, it is as shown in table 1 that each organizes the mouse blood lipid level.B group serum TC and TG content have shown and have been higher than A group (P<0.05), illustrate that the 30 days hyperlipidemia models of mouse high lipid food of feeding successfully build up.The serum TC of C group and TG content are showing reduction (P<0.05) than the B group, and the LDL-C level of the mouse of B group and C group obviously descends, and all feeds prescription is little to the content influence of mouse HDL-C; Atherogenic index (AI) is the index that the reflection angiocardiopathy is easily suffered from degree.The AI value of B that high lipid food is fed group mouse is 0.365, and is the highest in each group, and the mouse AI value of irritating stomach norcholesterol yogurt descends to some extent, and the AI value is 0.211, and organizes with B and to compare difference and showing (P<0.05); The HDL-C/TC value of the mouse of B group of feeding behind the high lipid food is showing and is reducing, be 73.3%, the HDL-C/TC value of irritating the C group mouse of hydrochloric acid in gastric juice breast is showing and is raising, be 86.2%, and that the HDL-C/TC ratio of normal control group A group mouse remains is the highest, illustrates that this norcholesterol yogurt has the function that reduces serum cholesterol.
Table 1 norcholesterol yogurt is to the influence of mouse blood fat
Annotate: AI=TC-HDL-C/HDL-C
In the mouse 30 days fed, each is organized, and the mouse hair color is bright and clean, and appetite is normal, and natural death does not take place.Body weight and liver,kidney,spleen and body weight percentage are shown in table 2 and 3.The body weight of B group mouse of high lipid food of feeding obviously increases (P<0.05), and C group mouse body weight and A organize not showing property difference (P〉0.05).Feed the liver weight/body weight of B group mouse of high lipid food apparently higher than the mouse of basal diet group, and irritate stomach group (C group) be lower than hyperlipidemia model group (B group), the effect owing to the norcholesterol yogurt is described, make the fatty amount of stockpiling minimizing in mouse liver.Kidney weight/body weight and spleen weight/body weight are not seen apparent difference between each group, show during to mouse stomach norcholesterol yogurt and do not see obvious toxic-side effects.
Table 2 norcholesterol yogurt is to the influence (G) of mouse body weight
Figure A200810013491D00101
Table 3 norcholesterol yogurt is to the influence of the dirty body ratio of mouse
Figure A200810013491D00102
In sum, efficient lactobacillus for degrading cholesterol-Lactobacillus casei cheese subspecies that the present invention uses, separation is from natural fermented dairy products, the in vitro culture test shows that it has very strong acidproof, bile tolerance ability, adhesiveness is good, common pathogenic entero becteria there is the obvious suppression effect, do not carry r factor, it is a kind of lactic acid bacteria that is suitable for the human body intestinal canal field planting, external cholesterol degradation rate can reach 56% simultaneously, and the zoopery of norcholesterol shows that also this yogurt can obviously reduce the mice serum cholesterol in the body.
It is the Yoghourt fermentation microbial inoculum that the present invention adopts the Lactobacillus casei cheese subspecies of efficient norcholesterol first, is lactic acid bacteria growth promoter with the haw thorn extract, produces efficient norcholesterol yogurt.Experiment in vitro, interior animal experiment show that all this norcholesterol yogurt has the function that reduces serum cholesterol, and effect obviously is better than the existing literature report.After adding haw thorn extract in the raw milk, fermentation yogurt viable lactic acid bacteria number can improve 10 2Doubly, can obviously strengthen the functional of the prebiotic effect of yogurt lactic acid bacteria and norcholesterol.
The specific embodiment
Embodiment 1
The preparation of full-cream yogurt
(1) lactic acid bacteria GL-03 makes with the following method:
Sterile working: natural fermented cheese is drawn 2ml and is added in the skimmed milk of sterilizing after the sterilized water dilution, and 37 ℃ leave standstill enrichment culture 48h; Get 1ml enrichment culture liquid and add and to contain in the aseptic improvement KENJI selective medium of the blue 9ml of 0.04% bromine thyme phenol, 37 ℃ shaken cultivation 1-2 days; Get nutrient solution 100 μ L and be added in the same culture medium and cultivate, need three tubes to cultivate altogether; Ratio turbidity and indicator change in color according to culture medium, get for the third time the MRS fluid nutrient medium that bacterium liquid 200 μ L after cultivating join pH2 and cultivate 6h, getting its 100 μ L nutrient solution is transferred to contain in the 0.30wt% pig cholate MRS fluid nutrient medium and cultivates 24h, that gets 200 μ L nutrient solutions then is coated on the modified MRS flat board 37 ℃ of constant temperature culture 48~72h; Picking produces the colonies typical of acid, line separation repeatedly, purifying, and Gram, microscopy are till pure bacterium colony; Then, identify through 16SrDNA order-checking carrying out bacterial classification;
Directional induction: after the lactic acid bacteria GL-03 activation with seed selection, quantitatively be added in the KENJI fermentation medium that contains 0.25% cholesterol and cultivate, the concentration of a cholesterol of every cultivation all will increase 0.15wt%, after five domestications are cultivated, makes that content of cholesterol reaches 1mg/mL in the culture medium; Select well-grown lactic acid bacteria GL-03, in containing the MRS fluid nutrient medium of cholesterol 0.1mg/mL, measure the variation of its cholesterol degradation rate before and after inducing and acclimating, therefrom filter out the bigger bacterial strain of norcholesterol ability raising and form.
(2) actication of culture and leavening are made:
Lactobacillus casei GL-03, bacillus bulgaricus, streptococcus thermophilus are transferred in the liquid MRS culture medium by the MRS inclined-plane respectively, cultivate 12h, activate three times for 37 ℃; Three bacterial classifications after the activation are inoculated in the skimmed milk power liquid that contains 10% (W/V) of sterilization by 1:0.5:0.5, and 37 ℃ are cultured to curdled milk, transfer and make leavening three times.
(3) preparation of haw thorn extract
Get hawthorn 50 grams, earlier material is cut into broken end or powder, add 3 times water, soak 4h, boil the back and continue 40min with the line fire; Leach decocting liquid, filter residue adds 5 times water, carries out two and fries in shallow oil 30min, leaches decocting liquid, and filter residue adds 3 times water, carries out three and fries in shallow oil 20min, and merging filtrate is concentrated into 100mL, gets supernatant, and 115 ℃, sterilization 15min, 4 ℃ of refrigerators are preserved standby.
(4) preparation of composition of raw materials and fermentation yogurt: raw material quality per distribution ratio is:
Fresh milk 82.0%, milk powder 1.9%, sucrose concentration 5.0%, hawthorn extract 6.0%, lactic acid bacteria GL-03, bacillus bulgaricus and 3 kinds of bacterium mixed proportions of streptococcus thermophilus are 1:0.5:0.5, inoculum concentration is 5.0%, stabilizing agent CMC-Na0.1%.
Operating procedure:
Sucrose, stabilizing agent with a small amount of emulsion (50-60 ℃) dissolving, are added in the milk liquid after purifying, add the hawthorn extract in proportion simultaneously and stir, be warming up to 60 ℃ and carry out homogeneous (pressure is 15-17Mpa).Milk liquid behind the homogeneous is heated to 90 ℃, 5min or ultra high temperature short time sterilization (135 ℃ 3-5s), are cooled to 43 ℃, then with lactic acid bacteria GL-03, bacillus bulgaricus and the inoculation of streptococcus thermophilus mixed bacteria.Inoculation back feed liquid branch fast installs in the container, and delivers to 43 ℃ of thermostatic chambers fermentation 4h, to pH4.6-4.8.Be cooled to rapidly below 10 ℃, and move to 0-4 ℃ of freezer refrigeration after-ripening.
(4) product specification and physical and chemical index:
Butter oil 〉=3.00wt%, real milk solids 〉=10wt%, 70~110 ° of T of acidity, granulated sugar 〉=5.0wt%, mercury≤0.01%ppm.
Every 100g norcholesterol yogurt contains heat 65Kcal, protein 4.0g, lipid 3.1g, granulated sugar 5.1g, viable lactic acid bacteria number 〉=1 * 10 6Cfu/g.
Embodiment 2
The preparation of partially skimmed yogurt
(1) lactic acid bacteria GL-03 makes with embodiment 1 method.
(2) actication of culture and leavening are made:
Lactobacillus casei GL-03, bacillus bulgaricus, streptococcus thermophilus are transferred in the liquid MRS culture medium by the MRS inclined-plane respectively, cultivate 12h, activate three times for 37 ℃; Three bacterial classifications after the activation are inoculated in the skimmed milk power liquid that contains 10% (W/V) of sterilization by 1:0.5:0.5, and 37 ℃ are cultured to curdled milk, transfer and make leavening three times.
(3) preparation of haw thorn extract
Get hawthorn 50 grams, earlier material is cut into broken end or powder, add 3~8 times water, soak 2~4h, boil the back and continue 30~40min with the line fire; Leach decocting liquid, add 3~5 times water, carry out two frying in shallow oil (20~30min) leach decocting liquid, add 2~4 times water, carry out three frying in shallow oil (20~30min), merging filtrate is concentrated into 100mL, gets supernatant, and 115 ℃, sterilization 15min, 4 ℃ of refrigerators are preserved standby.
(4) preparation of composition of raw materials and fermentation yogurt: raw material quality per distribution ratio is:
Partially skimmed fresh milk 85.0%, sucrose concentration 6.0%, hawthorn extract 4.9%, lactic acid bacteria GL-03, bacillus bulgaricus and 3 kinds of bacterium mixed proportions of streptococcus thermophilus are 1:1:1, inoculum concentration is 4.0%, stabilizing agent CMC-Na0.1%, 43 ℃ of fermentation temperatures.
Operating procedure:
Sucrose, stabilizing agent with a small amount of emulsion (50-60 ℃) dissolving, are added in the milk liquid after purifying, add the hawthorn extract in proportion simultaneously and stir, be warming up to and carry out homogeneous (pressure is 15-17Mpa) about 60 ℃.Milk liquid behind the homogeneous is heated to 90 ℃, and 5min or ultra high temperature short time sterilization (135 ℃, 3-5s), be cooled to 43 ℃ then, lactic acid bacteria GL-03, bacillus bulgaricus and streptococcus thermophilus to be mixed with the 1:1:1 ratio, inoculum concentration is 5wt%.Inoculation back feed liquid branch fast installs in the container, and delivers to 43 ℃ of thermostatic chambers fermentation 4-5h, to pH4.6-4.8.Be cooled to rapidly below 10 ℃, and move to 0-4 ℃ of freezer refrigeration after-ripening.
(5) product specification and physical and chemical index:
Butter oil 〉=0.5~≤ 3.0wt%, real milk solids 〉=10wt%, 70~110 ° of T of acidity, granulated sugar 〉=5.0wt%, mercury≤0.01%ppm.
Every 100g norcholesterol yogurt contains protein 3.0g, lipid 0.5 ~ 3.0g, granulated sugar 5.1g, viable lactic acid bacteria number 〉=1 * 10 6Cfu/g.
Embodiment 3
The preparation of degreasing yogurt
(1) lactic acid bacteria GL-03 makes with embodiment 1 method.
(2) actication of culture and leavening are made:
Lactobacillus casei GL-03, bacillus bulgaricus, streptococcus thermophilus are transferred in the liquid MRS culture medium by the MRS inclined-plane respectively, cultivate 12h, activate three times for 37 ℃; Three bacterial classifications after the activation are inoculated in the skimmed milk power liquid that contains 10% (W/V) of sterilization by 1:0.5:0.5, and 37 ℃ are cultured to curdled milk, transfer and make leavening three times.
(3) preparation of haw thorn extract
Get hawthorn 50 grams, earlier material is cut into broken end or powder, add 3~8 times water, soak 2~4h, boil the back and continue 30~40min with the line fire; Leach decocting liquid, add 3~5 times water, carry out two frying in shallow oil (20~30min) leach decocting liquid, add 2~4 times water, carry out three frying in shallow oil (20~30min), merging filtrate is concentrated into 100mL, gets supernatant, and 115 ℃, sterilization 15min, 4 ℃ of refrigerators are preserved standby.
(4) preparation of composition of raw materials and fermentation yogurt: raw material quality per distribution ratio is:
Defatted fresh milk 85%, sucrose concentration 6.9%, hawthorn extract 6.0%, lactic acid bacteria GL-03, bacillus bulgaricus and 3 kinds of bacterium mixed proportions of streptococcus thermophilus are 1:1:1, inoculum concentration is 2.0%, stabilizing agent CMC-Na0.1%.
Operating procedure:
Sucrose, stabilizing agent with a small amount of emulsion (50-60 ℃) dissolving, are added in the milk liquid after purifying, add the hawthorn extract in proportion simultaneously and stir, be warming up to and carry out homogeneous (pressure is 15-17Mpa) about 60 ℃.Milk liquid behind the homogeneous is heated to 90 ℃, and 5min or ultra high temperature short time sterilization (135 ℃, 3-5s), be cooled to 43 ℃ then, lactic acid bacteria GL-03, bacillus bulgaricus and streptococcus thermophilus are mixed with the 1:1:1 ratio, connect 43 ℃ of kind amounts.Inoculation back feed liquid branch fast installs in the container, and delivers to 43 ℃ of thermostatic chambers fermentation 4-5h, to pH4.6-4.8.Be cooled to rapidly below 10 ℃, and move to 0-4 ℃ of freezer refrigeration after-ripening.
(5) physical and chemical index:
Butter oil≤0.5wt%, real milk solids 〉=10wt%, 70~110 ° of T of acidity, granulated sugar 〉=5.0wt%, mercury≤0.01%ppm.
Every 100g norcholesterol yogurt contains protein 3.0g, lipid 0.5g, granulated sugar 5.1g, viable lactic acid bacteria number 〉=1 * 10 6Cfu/g.
Embodiment 4
The preparation of degreasing yogurt
(1) actication of culture and leavening are made, the preparation of (2) haw thorn extract is with above embodiment,
(3) acidified milk raw material quality per distribution ratio is:
Degreasing recombined milk 80.0%, skimmed milk power 2.0%, sucrose 6.9%, hawthorn extract 6.0%, lactic acid bacteria GL-03, bacillus bulgaricus and 3 kinds of bacterium mixed proportions of streptococcus thermophilus are 1:1:1, inoculum concentration is 5.0%, stabilizing agent CMC-Na0.1%, 43 ℃ of fermentation temperatures.The fermentation operation step is with embodiment 3.
(4) physical and chemical index: with embodiment 3

Claims (5)

1. the fermentation yogurt of a norcholesterol is characterized in that fresh milk or recombined milk make through fermentation with lactic acid bacteria GL-03 (Lactobacillus casei subsp.casei), bacillus bulgaricus (Lactobacillusbulgaricus) and 3 kinds of mixed bacterias of streptococcus thermophilus (Streptococcus thermophilus) and haw thorn extract; Be creamy white or little yellow, grumeleuse uniform and smooth, no bubble allow small amount of whey to separate out; Its quality index is: butter oil 〉=3.00wt%, real milk solids 〉=10wt%, 70~110 ° of T of acidity, granulated sugar 〉=5.0wt%, mercury≤0.01%ppm, every 100g contains heat 60-70Kcal, non-fat solid 〉=8.1g, protein 〉=3.0g, granulated sugar 〉=5.0g, viable lactic acid bacteria several 1 * 10 6Cfu/g, coliform≤90/milliliter, pathogenic bacteria must not detect.
2. the preparation method of the fermentation yogurt of norcholesterol according to claim 1 is characterized in that processing step is:
(1) feedstock production:
(1) actication of culture and leavening are made
With the refrigerator preservation through separation screening, directional induction and lactic acid bacteria GL-03, bacillus bulgaricus and streptococcus thermophilus are transferred to by the MRS inclined-plane in the liquid MRS culture medium, cultivate 12h for 37 ℃, activate three times; The bacterial classification of activation is inoculated in the skimmed milk power liquid that contains 10% (W/V) of sterilization by 2%, and 37 ℃ are cultured to curdled milk, transfer and make leavening three times;
Ratio between lactic acid bacteria GL-03, bacillus bulgaricus and the streptococcus thermophilus is: 1.0:0.5~1.0:0.5~1.0
(2) preparation of haw thorn extract
50 grams are pulverized hawthorn, add 3~8 times water, soak 2~4h, boil the back and continue 30~40min with the line fire; Leach decocting liquid, filter residue adds 3~5 times, 2~4 water then respectively, carries out two of 20~30min with identical method and fries in shallow oil, three fries in shallow oil, and merging filtrate is concentrated into 100mL, gets supernatant, 115 ℃ of sterilization 15min, and 4 ℃ of refrigerators are preserved standby;
(2) preparation of fermentation yogurt:
(3) composition of raw materials
Raw material milk 80.0~85.0
Milk powder 0.0~2.0
Sucrose 5.0~7.0
Hawthorn extract 4.0~6.0
Three kinds of mixed bacterias 2.0~5.0
Stabilizing agent CMC-Na 0.1
Wherein three kinds of mixed bacterias are that lactic acid bacteria GL-03, bacillus bulgaricus and 3 kinds of bacterium of streptococcus thermophilus mix, and its ratio is 1:0.5~1.0:0.5~1.0;
(4) processing step
1) raw material milk is through milk inspection, purification, stand-by;
2) with sucrose, stabilizing agent with a small amount of emulsion in 50-60 ℃ of dissolving, in the milk liquid after add purifying, stir, be warming up to 58~62 ℃, 15-17Mpa carries out homogeneous;
3) the milk liquid behind the homogeneous is heated to 88~92 ℃, the 5min sterilization is cooled to 43 ℃ then, inoculation;
4) the inoculation back, is cooled to rapidly below 10 ℃ to pH4.6~4.8 in 40~45 ℃ of thermostatic chamber fermentation 4~5h, and moves to 0-4 ℃ of freezer refrigeration after-ripening.
3. as the preparation method of the fermentation yogurt of norcholesterol as described in the claim 2, it is characterized in that actication of culture described in (1) of processing step () feedstock production and leavening make through separation screening, directional induction and lactic acid bacteria GL-03 makes with the following method:
Sterile working: natural fermented cheese is drawn 2ml and is added in the skimmed milk of sterilizing after the sterilized water dilution, and 37 ℃ leave standstill enrichment culture 48h; Get 1ml enrichment culture liquid and add and to contain in the aseptic improvement KENJI selective medium of the blue 9ml of 0.04% bromine thyme phenol, 37 ℃ shaken cultivation 1-2 days; Get nutrient solution 100 μ L and be added in the same culture medium and cultivate, need three tubes to cultivate altogether; Ratio turbidity and indicator change in color according to culture medium, get for the third time the MRS fluid nutrient medium that bacterium liquid 200 μ L after cultivating join pH2 and cultivate 6h, getting its 100 μ L nutrient solution is transferred to contain in the 0.30% pig cholate MRS fluid nutrient medium and cultivates 24h, that gets 200 μ L nutrient solutions then is coated on the modified MRS flat board 37 ℃ of constant temperature culture 48~72h; Picking produces the colonies typical of acid, line separation repeatedly, purifying, and Gram, microscopy are till pure bacterium colony; Then, identify through 16SrDNA order-checking carrying out bacterial classification;
Directional induction: after the lactic acid bacteria GL-03 activation with seed selection, quantitatively be added in the KENJI fermentation medium that contains 0.25% cholesterol and cultivate, the concentration of a cholesterol of every cultivation all will increase by 0.15%, five domestication cultivate after, make that content of cholesterol reaches 1mg/mL in the culture medium; Select well-grown lactic acid bacteria GL-03, in containing the MRS fluid nutrient medium of cholesterol 0.1mg/mL, measure the variation of its cholesterol degradation rate before and after inducing and acclimating, therefrom filter out the bigger bacterial strain of norcholesterol ability raising and form.
4. as the preparation method of the fermentation yogurt of norcholesterol as described in the claim 2, it is characterized in that the raw material milk described in the processing step is a recombined milk.
5. as the preparation method of the fermentation yogurt of norcholesterol as described in the claim 2, it is characterized in that described processing step 3), be with 135 ℃ of the heating of the milk liquid behind the homogeneous, carry out superhigh temperature 3-5s sterilization, be cooled to 43 ℃ then, inoculation.
CNA2008100134918A 2008-09-28 2008-09-28 Fermentation yogurt capable of reducing cholesterol and preparation method thereof Pending CN101361506A (en)

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Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102747000A (en) * 2011-04-22 2012-10-24 天津科技大学 Lactobacillus kefiranofaciens and its fermented milk preparation method
CN104430851A (en) * 2013-12-25 2015-03-25 广东燕塘乳业股份有限公司 Fermented milk capable of reducing cholesterol and preparation method of fermented milk
CN105473005A (en) * 2013-08-14 2016-04-06 雀巢产品技术援助有限公司 Dairy composition comprising hawthorn and phytosterols
CN107048401A (en) * 2017-05-03 2017-08-18 贵阳中医学院 Hawthorn plant extraction liquid, fermented yoghourt and preparation method thereof
CN108497062A (en) * 2017-02-28 2018-09-07 株式会社诚元大理 Method for preparing sour milk
CN109938250A (en) * 2017-12-21 2019-06-28 财团法人石材暨资源产业研究发展中心 For adjusting the complex liquid of blood lipid

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102747000A (en) * 2011-04-22 2012-10-24 天津科技大学 Lactobacillus kefiranofaciens and its fermented milk preparation method
CN105473005A (en) * 2013-08-14 2016-04-06 雀巢产品技术援助有限公司 Dairy composition comprising hawthorn and phytosterols
CN104430851A (en) * 2013-12-25 2015-03-25 广东燕塘乳业股份有限公司 Fermented milk capable of reducing cholesterol and preparation method of fermented milk
CN104430851B (en) * 2013-12-25 2017-08-25 广东燕塘乳业股份有限公司 A kind of norcholesterol acidified milk and preparation method thereof
CN108497062A (en) * 2017-02-28 2018-09-07 株式会社诚元大理 Method for preparing sour milk
CN107048401A (en) * 2017-05-03 2017-08-18 贵阳中医学院 Hawthorn plant extraction liquid, fermented yoghourt and preparation method thereof
CN109938250A (en) * 2017-12-21 2019-06-28 财团法人石材暨资源产业研究发展中心 For adjusting the complex liquid of blood lipid

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