CN110403011A - A kind of bacillus coagulans acidified milk and preparation method thereof - Google Patents

A kind of bacillus coagulans acidified milk and preparation method thereof Download PDF

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CN110403011A
CN110403011A CN201910645531.9A CN201910645531A CN110403011A CN 110403011 A CN110403011 A CN 110403011A CN 201910645531 A CN201910645531 A CN 201910645531A CN 110403011 A CN110403011 A CN 110403011A
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milk
bacillus coagulans
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acidified milk
acidified
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刘冬梅
赵珊
陈晓琦
周钦育
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South China University of Technology SCUT
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    • A23C9/00—Milk preparations; Milk powder or milk powder preparations
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    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
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Abstract

The invention belongs to food processing technology fields, disclose a kind of bacillus coagulans acidified milk and preparation method thereof, this method is to carry out fermentation milk fermentation for main leavening with bacillus coagulans (Bacillus sp.) 13002, and bacillus coagulans acidified milk is made.The present invention compounds the fermentation for carrying out acidified milk using probiotics with Traditional Fermented Milk fermenting microbe, it highly shortened fermentation time, simple process, obtained acidified milk mild acidity, viable count are high, no whey is precipitated, have good rheological properties and retention ability, fragrance component strong, and viable count is still 10 after 4 DEG C of storage 28d9CFU/mL or more can also play the prebiotic effect of probiotics, treat indigestion and constipation, enhance gastrointestinal tract immunity, adjust intestinal flora balance, have great market potential.

Description

A kind of bacillus coagulans acidified milk and preparation method thereof
Technical field
The invention belongs to fermented dairy product fields, and in particular to a kind of bacillus coagulans acidified milk and preparation method thereof.
Background technique
In recent years, currently used anti-with the rising of various bacterial infection disease disease incidence and the enhancing of bacterial drug resistance Raw element drug efficiency is low, expensive, it is also possible to which, there are side effect, researcher may provide the significant steering of attention to improvement The probiotics of the dietary ingredient of health.
Currently, due to the good characteristic of gemma Pseudomonas, the bacillus of extensive evaluation have bacillus subtilis, gram Lloyd's bacillus, Bacillus cercus, bacillus coagulans and bacillus licheniformis.With non-sporogenesis agent (such as lactic acid bar Pseudomonas) it compares, bacterial spore has high temperature resistant, allows to store in a dry form at room temperature and do not have any nocuousness to viability It influences；Spore can reach small intestine, the low ph value of resistance to stomach.Bacillus coagulans belong to enteron aisle lactic acid bacteria, referred to as " have spore newborn Sour bacterium " has the healthcare function as the intestinal beneficial bacteriums such as Bifidobacterium, and belongs to bacillus, has stomach juice-resistant, resistance to height Warm high pressure has no drug resistance, is easy the advantages that culture and preservation.In May, 2004, No. 1126 public affairs of The Ministry of Agriculture of the People's Republic of China, MOA It accuses in " catalogue of feed additive varieties " and bacillus coagulans is included in new feedstuff and new feed additive workable for safety, Applicable range is broiler and growing-finishing pig, illustrates that its safety, validity, economy and the influence to environment have been led to National feed evaluation committee evaluation has been crossed, and has obtained Ministry of Agriculture's approval and is produced by Tianjin nova animal pharmaceutical factory；2016, country defended Bacillus coagulans are included in " the strain list that can be used for food " by planning commission.It is reported that bacillus coagulans are in animal experiment With the function as probiotics, which has important potentiality as mankind probiotics.
Main carriers of the acidified milk as probiotics, i.e., using fresh milk as raw material, again to ox after pasteurize Fermenting microbe is added in milk, cooled down again after everfermentation it is filling or first filling after ferment again refrigeration.Epidemiology is ground with zoology Study carefully the generation for showing that often edible fermented dairy product can inhibit the pathogenic bacteria in gastrointestinal tract, improve gastrointestinal bacterial flora, reduce tumour Rate, the variation compound in reduction gastrointestinal tract and stimulating mucosal immunity reaction.Blueness of the fermented dairy product by consumer in recent years It looks at, there is very big market effect.Therefore, application of the fermenting microbe with good prebiotic effect in fermented dairy product is probed into There is potential economic value.
Summary of the invention
The present invention is directed to the deficiency of existing bacillus coagulans application study, it is therefore an objective to provide a kind of bacillus coagulans hair Kefir milk and preparation method thereof.Bacillus coagulans acidified milk manufacturing process prepared by the present invention is simple, and L-lactic acid is high, system Standard specified in the taste and flavor symbol " GB19302-2010 acidified milk " of product.
In order to achieve the object of the present invention, present invention employs following technical solutions:
A kind of preparation method of bacillus coagulans acidified milk is with bacillus coagulans (Bacillus sp.) 13002 Main fermentation agent carries out fermentation milk fermentation, bacillus coagulans acidified milk is made.
Preferably, the leavening includes bacillus coagulans (Bacillus sp.) 13002 and streptococcus thermophilus (Streptococcus thermophilus) and German-style lactobacillus subspecies bulgaricus (Lactobacillus Delbrueckii subsp bulgericus) compounding, total inoculum concentration of leavening is 0.1~2wt%.
Preferably, steps are as follows for specific fermenting and producing:
(1) milk that will ferment and round carry out pasteurize, are then quickly cooled to 37~45 DEG C；
(2) inoculating starter is acidified in 37~43 DEG C of 3.5~8h of fermentation, then after, and acidified milk is made.
Preferably, the fermentation milk composition are as follows: 10~14wt% skimmed milk power and the recovery milk of dilute cream, whole-fat milk powder Recovery milk or fresh milk, 5~10wt% of sucrose, 0~1.0wt% of stabilizer, 0~5wt% of oligofructose, surplus be water；It is described Pasteurize is 85~95 DEG C of 5~30min of wet heat sterilization；Fermentation time is 4~6h；The condition being acidified after described are as follows: after at 4 DEG C It is acidified 8~15h.The stabilizer is sodium carboxymethylcellulose etc..
Preferably, the preparation method of the leavening includes the following steps:
(1) activation of bacillus coagulans carried out using YPD fluid nutrient medium, expand culture, using MRS fluid nutrient medium Carry out streptococcus thermophilus, the activation of German-style lactobacillus subspecies bulgaricus, expansion culture；
(2) each bacterium solution after will be enlarged by culture is centrifuged and is cleaned 2~3 times with PBS buffer solution, by bacterium mud: protectant matter After amount volume ratio 1:1~1:3 adds freeze drying protectant to mix, vacuum freeze drying is carried out, the formula of the freeze drying protectant is 5~10wt% of trehalose, 5~10wt% of skimmed milk power are dissolved in 0.9% physiological saline.
Preferably, in the leavening bacillus coagulans, streptococcus thermophilus, German-style lactobacillus subspecies bulgaricus jelly The mass ratio of dry powder is 2:(0.01~1): (0.01~1).It is highly preferred that the bacillus coagulans, streptococcus thermophilus, German-style The mass ratio of the freeze-dried powder of lactobacillus subspecies bulgaricus is 2:(0.1~0.5): (0.1~0.5).
Preferably, the viable count of leavening are as follows: bacillus coagulans 1~7 × 1010CFU/g, streptococcus thermophilus 1~2 × 1010CFU/g, German-style lactobacillus subspecies bulgaricus 1~2 × 1010CFU/g。
Preferably, oligofructose additive amount is 1~5wt% in milk of fermenting, and leavening inoculum concentration is 0.5~1.5wt%.
Preferably, the leavening includes bacillus coagulans (Bacillus sp.) CGMCC NO.7431, thermophilus Bacterium (Streptococcus thermophilus) CGMCC 1.2741 and German-style lactobacillus subspecies bulgaricus (Lactobacillus delbrueckii subsp bulgericus)CGMCC 1.2902。
The titratable acidity of the bacillus coagulans acidified milk of above method preparation is 70~90 ° of T；The condensation gemma bar The L-lactic acid of bacterium acidified milk is 70~120mmol/L.Viable count in the bacillus coagulans acidified milk reaches 109CFU/mL or more.
The bacillus coagulans show through mouse toxicology test inspection result to female male KM mouse oral toxicity LD50> 10.0g/kg weight, the true nontoxic grade in border.
The present invention compared with prior art, has the advantages that
(1) starter culture bacillus coagulans used in the present invention can not only generate the organic acids such as lactic acid, acetic acid Inhibit the growth of pathogenic bacteria, moreover it is possible to secrete the antibacterial materials such as coagulin, penetrating cell film, to streptococcus, staphylococcus aureus Equal pathogenic bacteria have antagonism, so as to improve body expression environment, improve the premunition of body, reduce the generation of disease.
(2) present invention compounds the fermentation for carrying out milk using probiotics with Traditional Fermented Milk fermenting microbe, greatly shortens Fermentation time (shortening to 4h or so), at low cost, simple process, obtained acidified milk are suitable compared to traditional zymotic lactic acid degree In, viable count is high, no whey is precipitated, has good rheological properties and retention ability, fragrance component strong, after 4 DEG C of storage 28d Viable count is still 109CFU/mL or more, great market potential.
(3) present invention is coagulated from the understanding for enriching the mankind on theoretic and applying in dairy products to bacillus coagulans It ties bacillus and monosaccharide and disaccharide, the oligosaccharides in milk is degraded to lactic acid and other organic acids, increase the nutriture value of acidified milk Value, and viable count is high in acidified milk, as the carrier of probiotics, can also give full play to its prebiotic effect, such as promotes point of gastric juice It secretes, the wriggling of gastrointestinal tract, treats indigestion and constipation, enhance gastrointestinal tract immunity, improve antioxidant ability of organism, adjust intestines The balance of road flora.
The bacillus coagulans (Bacillus sp.), were obtained by separation screening in fermentation pickled vegetable, and in April, 2013 It is preserved within 8th China Committee for Culture Collection of Microorganisms's common micro-organisms center (referred to as: CGMCC, address: court, Beijing The institute 3 of positive area's North Star West Road 1, Institute of Microorganism, Academia Sinica), deposit number is CGMCC NO.7431.The bacterial strain is It is disclosed in Chinese patent CN105154358B, belongs to the prior art.
Detailed description of the invention
Fig. 1 is that Box-Behnken Optimum Experiment fits response surface design interaction influence figure in embodiment 2, and wherein a is A (inoculating proportion) and B (oligofructose) response surface figure, b are A (inoculating proportion) and B (oligofructose) contour map, c are A (strain Proportion) and C (inoculum concentration) response surface figure, d are A (inoculating proportion) and C (inoculum concentration) contour map, e are B (oligofructose) and C (inoculum concentration) response surface figure, f are B (oligofructose) and C (inoculum concentration) contour map.
Fig. 2 is that different strain leavening compounds the acidified milk Pfansteihl chromatogram than under in embodiment 3, and wherein a is condensation Bacillus: streptococcus thermophilus: the acidified milk Pfansteihl color when inoculating proportion of German-style lactobacillus subspecies bulgaricus is 1:0:0 Spectrogram (peak 10 is Pfansteihl), b are bacillus coagulans: streptococcus thermophilus: the strain of German-style lactobacillus subspecies bulgaricus is matched Acidified milk Pfansteihl chromatogram when than for 3:0.5:0.5 (peak 9 is Pfansteihl).
Fig. 3 is the acidified milk Pfansteihl chromatogram of fermentation 4h in embodiment 4 (peak 9 is Pfansteihl).
Fig. 4 is 3 medium viscosity of embodiment with shear rate change figure.
Fig. 5 is 4 medium viscosity of embodiment with shear rate change figure.
Specific embodiment
Below with reference to embodiment, invention is further described in detail, but the scope of protection of present invention not office It is limited to this.
Embodiment 1
Bacillus coagulans (Bacillus sp.) CGMCC NO.7431 acute toxinology experiment
(1) SPF grades KM mouse 20,18~22g of weight are provided, experimental animal by Guangdong Medical Lab Animal Center Production licence number: SCXK (Guangdong) 2013-0002, the certification of fitness number: No.44007200056215.Feed is cured by Guangdong Province It learns Experimental Animal Center to provide, production licence number: SCXK (Guangdong) 2013-0002, the certification of fitness number: No.44200300018665. it is used after animal house quarantine in the court's is qualified after animal is bought, experimental animal uses credit number: SYXK (Guangdong) 2014-0137, animal house temperature: 20~26 DEG C, relative humidity: 40~70%.
(2) bacillus coagulans freeze-dried powder 5.0g is weighed, is formulated into 10.0mL with pure water.Using limitation method, if 10.0g/ The dosage group of kg weight learns from else's experience and quarantines qualified mice 20, half male and half female.Preceding animal overnight fasting is tested, not water restriction. Empty stomach stomach-filling 1 time, each stomach-filling capacity is 20.0mL/kg weight.Observe and record immediately the poisoning manifestations of animal, death toll and Death time, observation period 14d.Toxicity grading is judged by acute oral toxicity test 4.3.2.4 and annex G evaluation regulation.
(3) mouse is found no within the observation period and poisoning manifestations and death occurs, after the test, to surviving animals solution It cuts open, naked eyes no abnormality seen.See Table 1 for details.
1 tested material of table is to chmice acute Oral toxicity test result
(4) oral solution of BCS containing bacillus coagulans 13002 is to male and female KM chmice acute Oral toxicity LD50 > 10.0g/kg weight, the true nontoxic grade in border.
Embodiment 2
A kind of bacillus coagulans acidified milk and preparation method thereof comprising following steps:
(1) preparation for milk of fermenting: whole milk powder 12wt%, white granulated sugar 8wt%, oligofructose 1.5wt%, stabilizer carboxylic Sodium carboxymethylcellulose pyce 0.2wt%, surplus are water, and mixing slowly shakes up；
(2) pasteurize of fermentation milk and round: 85 DEG C of wet heat sterilization 30min are quickly cooled to 42 DEG C；
(3) by inoculum concentration inoculation compounding leavening (bacillus coagulans, streptococcus thermophilus, the German-style lactobacillus of 1.5wt% It is 3:0.5:0.5 that subspecies bulgaricus strain freeze-dried powder, which compounds mass ratio,；Wherein each strain freeze-dried powder viable count are as follows: condensation gemma Bacillus 1 × 1010CFU/g, streptococcus thermophilus 1 × 1010CFU/g, German-style lactobacillus subspecies bulgaricus 1 × 1010CFU/g), in 42 DEG C of fermentation 4.5h, at 4 DEG C after be acidified 12h.
Zymotechnique in embodiment 2 is optimized by single factor experiment, including starter culture compounding ratio, leavening inoculation Amount and oligofructose additive amount, to make acidified milk mild acidity, there is fast good retentiveness, curdled milk time, color and mouthfeel Well, it gives off a strong fragrance.Therefore, fermenting microbe bacillus coagulans, streptococcus thermophilus, German-style lactobacillus subspecies bulgaricus strain freeze The optimum compound proportion of dry powder be (2~4): 0.5:0.5, oligofructose additive amount be 1~5wt%, leavening inoculum concentration be 0.5~ 1.5wt%.According to the center combination principle that Box-Behnken is designed, 3 levels (table 3) of 3 factors response are chosen, are used The experiment of 3 factor of Design-Expert8.0.6 software design, 3 horizontal respone face measures titratable acidity and sensory evaluation scores, and with drop Determining acidity is the best fermentation parameter that index determines bacillus coagulans acidified milk.Zymotechnique the results are shown in Table 4.
Titratable acidity measurement: titratable acidity is measured referring to the measurement of total acid in 12456-2008 food of GB/T, weighs 10g Yoghurt example carries out 3 times of dilution processing, then prepares the standard solution of sodium hydroxide of 0.1mol/L, carries out titration processing, records The NaOH solution volume V mL of consumption, according to × V × 10 acidity calculation formula ° T=[c (NaOH)/0.1].
Subjective appreciation: selected subjective appreciation group person (10 people), and related training is carried out to it, to the outer of acidified milk Sight, flavor, quality and mouthfeel are given a mark, and subjective appreciation standards of grading are as shown in table 2.
2 subjective appreciation standards of grading of table
3 Box-Behnken experimental design of table
The response surface analysis experimental design and result of 4 acidified milk of table
Each response surface group organoleptic quality evaluations of 5 acidified milk of table
As shown in Table 6, influence size of each factor to the titratable acidity of acidified milk be successively are as follows: C > B > A, i.e. inoculum concentration > Oligofructose > inoculating proportion.The P < 0.0001 of regression model, the P value for losing quasi- item is 0.2994, illustrates that the regression equation is extremely aobvious It writes.It is not significant to lose quasi- inspection, illustrates that X factor interferes very little to test result, does not need the item for introducing more high reps, model Appropriately.By software analysis it is found that titratable acidity (Y) compounds than (A), oligofructose additive amount independent variable starter culture (B), the multiple regression equation of leavening inoculum concentration (C):
Y=92.19484-0.20872A-0.9879B+3.44866C-0.027956AB+0.43322A C+1.97485BC- 1.10407A2-1.04077B2-2.99387C2
It is significant, i.e., this method that the equation, which expresses the relationship between the titratable acidity of acidified milk and 3 independents variable, It is reliable.Regression equation Monomial coefficient is bigger, and the interaction coefficient of BC is bigger in interaction term coefficient, illustrates oligofructose Interaction between additive amount and inoculum concentration is big, and the interaction coefficient of AC, AB are relatively small, illustrates that starter culture compounds ratio And the interaction of inoculum concentration, oligofructose additive amount and strain compounding than between is smaller.
By software analysis it is found that the coefficient of determination R of the model2It is 0.9760, Adj is 0.9451 after adjustment, illustrates this Equation and actual conditions fit well preferably reflect acidity and inoculating proportion, oligofructose additive amount, inoculum concentration pass System.Illustrate that the model dependency is good, stable experiment confidence level is higher.
In order to examine the validity of equation, variance point is carried out to the mathematical model of the titratable acidity in the acidified milk of measurement Analysis, and the partial regression coefficient of each factor is detected.The regression coefficient highly significant of C, P < 0.0001, explanation connect in first order Kind amount has highly significant effect to the titratable acidity of acidified milk.The regression coefficient of B is only second to C, illustrates the additive amount of oligofructose Influence to the titratable acidity of acidified milk is only second to leavening inoculum concentration.Specific response surface design is as shown in Figure 1.
After handling test data using Design-Expert 8.0.6 software, the optimum process condition of acidified milk is predicted are as follows: Starter culture freeze-dried powder quality is compounded than (bacillus coagulans: streptococcus thermophilus: German-style lactobacillus subspecies bulgaricus) 3.22:0.5:0.5, oligofructose additive amount (B) 1.56%, leavening inoculum concentration 1.315%, theoretical titratable acidity are 94.6689.Theoretical titratable acidity is 93.2468.Consider experiment actual conditions, fermentation parameter is adjusted are as follows: strain compounding Than (bacillus coagulans: streptococcus thermophilus: lactobacillus bulgaricus) (A) 3:0.5:0.5, oligofructose additive amount (B) 1.5%, inoculum concentration (C) 1.5%.Predicted value is further verified, under best fermentation parameter, carries out parallel test three times, The average value for calculating acidified milk titratable acidity is 90.59, fits rate up to 97.15% with predicted value, illustrates that the Optimized model has Higher feasibility.
6 regression model variance analysis of table
Acidified milk viable count is 8.0 × 10 under optimal zymotechnique in embodiment 29CFU/mL, L-lactic acid are 101.33mmol/L；Viable count is 1.1 × 10 after 4 DEG C of storage 28d9CFU/mL。
Embodiment 3
A kind of bacillus coagulans acidified milk and preparation method thereof comprising following steps:
(1) preparation for milk of fermenting: whole milk powder 12wt%, white granulated sugar 8wt%, oligofructose 1.5wt%, stabilizer carboxylic Sodium carboxymethylcellulose pyce 0.2wt% is mixed with water, is slowly shaken up；
(2) pasteurize of fermentation milk and round: 85 DEG C of wet heat sterilization 30min are quickly cooled to 42 DEG C；
(3) by the inoculum concentration inoculation compounding leavening of 1.5wt%, at 42 DEG C of fermentation 6h, 4 DEG C after acidification 12h.
The compounding leavening bacillus coagulans, streptococcus thermophilus, German-style lactobacillus subspecies bulgaricus strain freeze It includes 1:0:0 and 3:0.5:0.5 that dry powder, which compounds mass ratio,；Wherein each strain freeze-dried powder viable count are as follows: bacillus coagulans 1 × 1010CFU/g, streptococcus thermophilus 1 × 1010CFU/g, German-style lactobacillus subspecies bulgaricus 1 × 1010CFU/g。
Acidified milk described in the embodiment of the present invention 3 is subjected to subjective appreciation, is shown in Table 7.Leavening condenses as can be seen from Table 7 When bacillus, streptococcus thermophilus, German-style lactobacillus subspecies bulgaricus strain freeze-dried powder compounding mass ratio are 3:0.5:0.5, Flavour, the structural state of acidified milk are preferable.
7 acidified milk subjective appreciation of table
The yoghurt example of 10mL is taken out in corresponding small beaker, after calibrating pH meter, starts the pH measurement of yoghurt example, to Registration reads and carries out respective record after stablizing, as shown in table 8；It is surveyed referring to the measurement of total acid in 12456-2008 food of GB/T Determine titratable acidity, weigh 10g yoghurt example and carry out 3 times of dilution processing, then prepare the standard solution of sodium hydroxide of 0.1mol/L, Titration processing is carried out, the NaOH solution volume V mL of consumption is recorded, according to acidity calculation formula ° T=[c (NaOH)/0.1] × V × 10, the results are shown in Table 8；It takes yoghurt example 10mL to carry out 3000r/min centrifugation 10min processing, isolates corresponding whey, It weighs and records Yoghourt and go before whey and go the quality after whey, the two ratio is the retention ability of yoghurt example, such as 8 institute of table Show；Referring to the measurement refractometer method of 12295-1990 soluble solid content of GB, measured with hand held refractometer soluble Solid content, as shown in table 8；L-lactic acid, (Liu Dongmei, Wu Hui, Yu Yigang wait high performance liquid chromatography to bubble to reference literature The chiral separation and measurement [J] modern food science and technology .2007 (08): 74-76. of Pfansteihl and D-ALPHA-Hydroxypropionic acid in dish) method into Row, using U.S.'s Waters high performance liquid chromatograph, the chiral separation pillar used be Phenomenex company Chire × 3126 (D)-penicillamir, 4.60mm ID × 250mm L, flow velocity 1mL/min, Detection wavelength 254.0nm, sample introduction Amount is 5 μ L, and mobile phase is 0.2mmol/L copper-bath (aqueous isopropanol that solvent is 5%), and column temperature is 40 DEG C, as a result such as Shown in Fig. 2, table 8, the respectively Pfansteihl chromatogram and L-lactic acid of acidified milk described in the embodiment of the present invention 3.
The basic physical and chemical index of acidified milk as described in the above method measurement embodiment of the present invention 3, as shown in table 8.It can by table 8 The acidified milk Pfansteihl yield for knowing that leavening of three kinds of strain compoundings than under is fermented is higher, retention ability, soluble solid water It is flat suitable, and pH, titratable acidity of the 3:0.5:0.5 compounding than under are more excellent.
The basic physical and chemical index of 8 acidified milk of table
Rheometer can be used for measuring the rheological characteristics such as macromolecule melt, Polymer Solution, gel, lotion, suspension, food Matter is a kind of quick, easy, effective measurement method.Rotational rheometer generates simple by the relative motion of a pair of of fixture Shear flow can be used to the rheological property for quickly determining the various aspects such as viscosity, the elasticity of material, be the important set of modern rheometer At part.Using the viscoelastic sexual behaviour of TA DHR Stress Control rotational rheometer measurement acidified milk.It is first that acidified milk sample is suitable Hour hands are beaten 10 times, rear to beat 10 times counterclockwise.The stainless steel flat plate of diameter 40mm is selected to pop one's head in, the gap of plate and bottom surface is 1mm, test temperature are controlled in (20 ± 0.5) DEG C.Strain sweep parameter are as follows: frequency 1rad/s, range of strain be 0.1~ 100%.As a result as shown in figure 4, being acidified milk viscosity described in the embodiment of the present invention 3 with shear rate change figure.It is found that with The increase of shear rate, apparent viscosity first increases to be gradually reduced afterwards, and when shear rate is greater than 100s-1When, 3:0.5:0.5 compounding Apparent viscosity than under is increasingly greater than the compounding ratio of 1:0:0, illustrates that bacillus coagulans are compounded with traditional yogurt leavening When, it can produce more exocellular polysaccharide, improve the viscosity of acidified milk.
Embodiment 4
A kind of bacillus coagulans acidified milk and preparation method thereof comprising following steps:
(1) preparation for milk of fermenting: whole milk powder 12wt%, white granulated sugar 8wt%, oligofructose 1.5wt%, stabilizer carboxylic Sodium carboxymethylcellulose pyce 0.2wt% is mixed with water, is slowly shaken up；
(2) pasteurize of fermentation milk and round: 85 DEG C of wet heat sterilization 30min are quickly cooled to 42 DEG C；
(3) by inoculum concentration inoculation compounding leavening (bacillus coagulans, streptococcus thermophilus, the German-style lactobacillus of 1.5wt% It is 3:0.5:0.5 that subspecies bulgaricus strain freeze-dried powder, which compounds mass ratio,；Wherein each strain freeze-dried powder viable count are as follows: condensation gemma Bacillus 1 × 1010CFU/g, streptococcus thermophilus 1 × 1010CFU/g, German-style lactobacillus subspecies bulgaricus 1 × 1010CFU/g), in 42 DEG C of fermentation 4h, at 4 DEG C after be acidified 12h.
Acidified milk described in the embodiment of the present invention 3 is subjected to subjective appreciation, and is compared with the acidified milk of fermentation 6h, is seen Table 9.
9 acidified milk subjective appreciation of table
The yoghurt example of 10mL is taken out in corresponding small beaker, after calibrating pH meter, starts the pH measurement of yoghurt example, to Registration reads and carries out respective record after stablizing, as shown in table 10；Referring to the measurement of total acid in 12456-2008 food of GB/T Titratable acidity is measured, 10g yoghurt example is weighed and carries out 3 times of dilution processing, the sodium hydroxide standard for then preparing 0.1mol/L is molten Liquid carries out titration processing, records the NaOH solution volume V mL of consumption, according to acidity calculation formula ° T=[c (NaOH)/0.1] × V × 10, the results are shown in Table 10；It takes yoghurt example 10mL to carry out 3000r/min centrifugation 10min processing, isolates corresponding Whey weighs and records Yoghourt and goes before whey and go the quality after whey, and the two ratio is the retention ability of yoghurt example, such as table Shown in 10；It, can with hand held refractometer measurement referring to the measurement refractometer method of 12295-1990 soluble solid content of GB Dissolubility solid content, as shown in table 10；L-lactic acid, with embodiment 3, as a result as shown in Fig. 3, table 10, respectively the present invention is real Apply the Pfansteihl chromatogram and L-lactic acid of acidified milk described in example 4.
Acidified milk described in the embodiment of the present invention 4 is carried out to the measurement of basic physical and chemical index, and the acidified milk with fermentation 6h It compares, is shown in Table 10.
The basic physical and chemical index of acidified milk described in the embodiment of the present invention 4 is as shown in table 10.As shown in Table 10 when fermentation 4h, Acidity relatively fermentation 6h is suitable for, and other indices also comply with standard specified in " GB19302-2010 acidified milk ", Pfansteihl Content is up to 101.33mmol/L.
The basic physical and chemical index of 10 acidified milk of table
Rheometer can be used for measuring the rheological characteristics such as macromolecule melt, Polymer Solution, gel, lotion, suspension, food Matter is a kind of quick, easy, effective measurement method.Rotational rheometer generates simple by the relative motion of a pair of of fixture Shear flow can be used to the rheological property for quickly determining the various aspects such as viscosity, the elasticity of material, be the important set of modern rheometer At part.Using the viscoelastic sexual behaviour of TA DHR Stress Control rotational rheometer measurement acidified milk.It is first that acidified milk sample is suitable Hour hands are beaten 10 times, rear to beat 10 times counterclockwise.The stainless steel flat plate of diameter 40mm is selected to pop one's head in, the gap of plate and bottom surface is 1mm, test temperature are controlled in (20 ± 0.5) DEG C.Strain sweep parameter are as follows: frequency 1rad/s, range of strain be 0.1~ 100%.As a result as shown in figure 5, being acidified milk viscosity described in the embodiment of the present invention 4 with shear rate change figure.It is found that with The increase of shear rate, apparent viscosity first increases to be gradually reduced afterwards, and fermentation 4h differs smaller with the apparent viscosity for the 6h that ferments, that is, is existed It is save the cost in production, fermentation time can be shortened.

Claims (10)

1. a kind of preparation method of bacillus coagulans acidified milk, which is characterized in that with bacillus coagulans (Bacillus Sp.) 13002 be main leavening, carries out fermentation milk fermentation, bacillus coagulans acidified milk is made.
2. the method according to claim 1, wherein the leavening includes bacillus coagulans (Bacillus Sp.) 13002 with streptococcus thermophilus (Streptococcus thermophilus) and German-style lactobacillus subspecies bulgaricus The compounding of (Lactobacillus delbrueckii subsp bulgericus), total inoculum concentration of leavening is 0.1~ 2wt%.
3. method according to claim 1 or 2, which is characterized in that steps are as follows for specific fermenting and producing:
(1) milk that will ferment and round carry out pasteurize, are then quickly cooled to 37~45 DEG C；
(2) inoculating starter is acidified in 37~43 DEG C of 3.5~8h of fermentation, then after, and acidified milk is made.
4. according to the method described in claim 3, it is characterized in that, the fermentation milk forms are as follows: 10~14wt% defatted milk It is powder and the recovery milk of dilute cream, the recovery milk of whole-fat milk powder or fresh milk, 5~10wt% of sucrose, 0~1.0wt% of stabilizer, oligomeric 0~5wt% of fructose, surplus are water；The pasteurize is 85~95 DEG C of 5~30min of wet heat sterilization；Fermentation time is 4~6h； The condition being acidified after described are as follows: 8~15h is acidified after at 4 DEG C.
5. method according to claim 1 or 2, which is characterized in that the preparation method of the leavening includes the following steps:
(1) activation of bacillus coagulans carried out using YPD fluid nutrient medium, expand culture, carried out using MRS fluid nutrient medium Streptococcus thermophilus, the activation of German-style lactobacillus subspecies bulgaricus, expansion culture；
(2) each bacterium solution after will be enlarged by culture is centrifuged and is cleaned 2~3 times with PBS buffer solution, by bacterium mud: protectant mass body After product adds freeze drying protectant to mix than 1:1~1:3, vacuum freeze drying is carried out, the formula of the freeze drying protectant is seaweed Sugar 5~10wt%, 5~10wt% of skimmed milk power are dissolved in 0.9% physiological saline.
6. according to the method described in claim 5, it is characterized in that, bacillus coagulans in the leavening, streptococcus thermophilus, The mass ratio of the freeze-dried powder of German-style lactobacillus subspecies bulgaricus is 2:(0.01~1): (0.01~1)；The viable count of leavening Are as follows: bacillus coagulans 1~7 × 1010CFU/g, streptococcus thermophilus 1~2 × 1010CFU/g, German-style lactobacillus Bulgaria Subspecies 1~2 × 1010 CFU/g。
7. according to the method described in claim 6, it is characterized in that, the bacillus coagulans, streptococcus thermophilus, German-style cream bar The mass ratio of the freeze-dried powder of bacterium subspecies bulgaricus is 2:(0.1~0.5): (0.1~0.5)；Oligofructose adds in fermentation milk Dosage is 1~5wt%, and leavening inoculum concentration is 0.5~1.5wt%.
8. method according to claim 1 or 2, which is characterized in that the leavening includes bacillus coagulans (Bacillus sp.) CGMCC NO.7431, streptococcus thermophilus (Streptococcus thermophilus) CGMCC1.2741 With German-style lactobacillus subspecies bulgaricus (Lactobacillus delbrueckiisubsp bulgericus) CGMCC1.2902。
9. a kind of bacillus coagulans acidified milk prepared by claim 1~8 any one method.
10. according to the method described in claim 9, it is characterized in that, the titratable acidity of the bacillus coagulans acidified milk For 70~90 ° of T；The L-lactic acid of the bacillus coagulans acidified milk is 70~120mmol/L.
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CN115353995A (en) * 2022-08-22 2022-11-18 华南理工大学 Compound leavening agent and preparation method and application thereof
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