CN107772477A - A kind of compound probiotic powder preparation rich in selenium and preparation method thereof - Google Patents
A kind of compound probiotic powder preparation rich in selenium and preparation method thereof Download PDFInfo
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- CN107772477A CN107772477A CN201711206009.8A CN201711206009A CN107772477A CN 107772477 A CN107772477 A CN 107772477A CN 201711206009 A CN201711206009 A CN 201711206009A CN 107772477 A CN107772477 A CN 107772477A
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- bifidobacterium
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Abstract
The invention belongs to compound probiotic powder preparation field, and in particular to a kind of compound probiotic powder preparation rich in selenium and preparation method thereof, the compound probiotic powder preparation, include following compositions：Selenium-rich Bifidobacterium V9, maltodextrin, glucose, citric acid, FOS, rich in Organic Selenium, it is the good food source for supplementing Organic Selenium, there is good stability in cryopreservation.
Description
Technical field
The invention belongs to compound probiotic powder preparation field, and in particular to a kind of compound probiotic powder preparation rich in selenium and
Its preparation method.
Background technology
The utilization of new resource for food result in new probiotics strain and continue to bring out, and lactic acid bacteria is that a kind of can utilize can
The general name of the gram-positive bacteria of a large amount of lactic acid is produced in fermentable carbohydrate fermentation, and it is well-known industrial prebiotic
Bacterium, it has very high healthy nutritive value, and caused organic acid can improve the utilization rate of the elements such as phosphorus, iron after fermentation, promotes
The absorption of iron and VD, cholesterol is reduced, stimulating immune system, strengthens host immunity.Bifidobacterium can absorb inorganic selenium simultaneously
It is largely converted into Organic Selenium；Albumen selenium is the main occurrence patterns of selenium in selenium-rich Bifidobacterium, Organic Selenium protein,
Distribution in polysaccharide and nucleic acid is followed successively by：Albumen selenium>Polysaccharide selenium>Nucleic acid selenium；In albumen selenium, selenium is mainly distributed on molecular weight not
In albumen or peptide chain more than 20ka.In albumen selenium, selenium is generally attached in protein in the form of selenoaminoacid, and selenium is covalently tied
Close and find there are 2 kinds of forms of selenocysteine (SeCys) and selenomethionine (SeMet) in protein.
Selenium-rich Bifidobacterium V9, it is that to filter out one plant of selenium rich ability in a kind of strain excellent from Resource of lactic bacteria database storehouse stronger
Bacterial strain, contrast test identification is carried out to it, it is as a result most strong for animal bifidobacteria V9 selenium rich ability, and it is named as selenium-rich
Bifidobacterium V9, show that the selenium-rich content of the bacterial strain is 10 μ g/ml by the selenium-rich experiment of bacterial strain.To obtained strains continuous passage
Cultivated for 10 generations, selenium rich ability is stable, and genetic stability is good.
Selenium element in the present invention in selenium-rich Bifidobacterium V9 is incorporated on protein, low and have symphysis for development cost
The selenium-rich bifidobacterium preparations of first effect.Bifidobacterium V9 has prebiotic effect, is widely used in various food, such as acid
Breast, milk, formulated infant milk, cereals, cheese and dietary supplements etc..On the one hand, Bifidobacterium V9 is solid to lactose, courage
The metabolism of many kinds of substance such as alcohol and oxalic acid has facilitation.On the other hand, Bifidobacterium V9 also has anti-oxidant, antiulcer, resisted
Tumour, the germ reproduction that suppresses, regulation gut flora balance and the effect for maintaining immune system balance.Bifidobacterium V9 can be by nothing
Machine selenium is converted into organic selenoprotein, and these selenoproteins have good anti-oxidation function.Therefore, using Bifidobacterium V9 selenium-rich, both
Can meet the needs of people are to probiotics, and can increase product organic selenium content.Therefore, using Bifidobacterium V9 to inorganic selenium
It is enriched with, selenium-rich bacterial strain, which is applied to, can reach multiple health care in probiotics preparation, have very high practical application
Value.
By the selenium-rich culturing of Bifidobacterium, Bifidobacterium physiological metabolism process can be utilized by the inorganic selenium in nutrient solution
Organic Selenium in thalline is converted into, so as to obtain the selenium-rich product of more high bioavilability, Bifidobacterium selenium-rich active bacteria formulation is made.
Because Bifidobacterium is existing flora under human normal physiological status, therefore take selenium-rich bifidobacteria viable bacteria product on the one hand
The health-care effect of selenium can be utilized, on the other hand can also increase the quantity of Bifidobacterium in human body, so as to improve body health water
It is flat.The system research carried out to selenium-rich Bifidobacterium V9, its characteristic are as follows：After 3h being digested in pH3.0 simulated gastric fluid,
Survival rate still reaches more than 90%；After selenium-rich Bifidobacterium V9 can digest 5h in pH8.0 simulated intestinal fluid, survival rate still reaches
To more than 95%；It is insensitive to 0.3% bovine bile concentration；To enteron aisle lead to diarrhea bacterial strain Shigella shigae, mouse typhus sramana
Salmonella, Escherichia coli, Pseudomonas aeruginosa have different degrees of antagonism；DNA is not present in bacterial strain, also just not
In the presence of the risk that Drug Resistance Plasmidss are propagated in enteron aisle；With immunoregulation effect.After pilot plant test, the bacterial strain is advised in 150L
Mould fermentation thalli density is up to 6 × 109Cfu/ml, zymotic fluid are freeze-dried after thalline is collected by centrifugation and can obtain average viable count
1.0×1011Cfu/ml bacterium powder, and the bacterium powder has good stability.
By the use of microorganism Bifidobacterium V9 as carrier Rich in Trace Element selenium, research heat has at home and abroad been formed at present
Point, microorganism enrichment inorganic selenium have that accumulation ability is strong, high conversion rate, low cost and other advantages, and passing through microbial conversion will
Inorganic selenium is converted into Organic Selenium, can reduce selenium toxicity, improves the physiologically active and absorptivity of selenium.Therefore, selenium-rich bifid bar is studied
Bacterium V9, the selenium rich ability of this kind of probiotics of lactic acid bacteria is especially improved, develop Organic Selenium functional food to provide selenium to body
Source, and the focus studied at present.Exploitation is with China's independent intellectual property right, tallying with the national condition and can be further
The standardization commodity leavening for lifting selenium-enriched health care function has been not coilable trend.
Current domestic selenium-enriched probioticses preparation is especially rare, and really has the selenium-enriched probioticses product of independent intellectual property right
Industrialization it is almost nil.On a large scale using selenium-rich rice, Se rich tea, selenium-enriched edible mushroom etc., not only production cost is high, more important
Be that will not merge normalizing with technique of enriched selenium the effect of probiotics.The product that current external probiotics is combined with selenium is rarely seen
Britain, its trade name Selenomax, Selenium (selenium yeast product).At present, domestic existing a variety of cities of product release containing selenium
.Such as selenium-rich health, natural selenium dietetic liquid, west weir (selenium yeast piece), selenous acid selling sheet, selenium health difficult to understand etc..But probiotics and selenium
With reference to product there is presently no listing.And the bifidobacterium preparations containing selenium, the inorganic selenium in nutrient solution is converted into nontoxic
Thalline includes Organic Selenium, and the Tiny ecosystem regulatory function of selenium and the antitumaous effect of Bifidobacterium and bifid bar mattress is superimposed, is cancer
The treatment of disease drug provides new developing direction.
Selenium-rich probiotic lactobacillus strain and preparation correlation technique and product of the research and development with independent intellectual property right, build
Stand with selenium-rich probiotic lactobacillus production technology and its production demonstration in China with extensive market, make the country that there is characteristic
Double effects effect selenium-enriched probioticses launch.In product development process, main technological difficulties are to selenium-rich
The control optimization of condition of culture, selenium content detection method determine research and prepare suitable selenium-enriched probioticses preparation.It is but overall
For, it is a very potential, significant research to develop cheap selenium-rich Bifidobacterium V9 preparations.
The content of the invention
We use modern analytical technique, respectively by experiment in vitro and zoopery to its prebiotic function, biological characteristics
Property has carried out system research.Further with correlation theories knowledges such as clinical medicine, nutrition, by experimental verification, research is graduated from old-type opera school
Learn formula.Want lyophilized thalline is realized and reached and commercially produce effect, lyophilized survival rate needs to reach higher quantity.
It is a difficulty of research, microcapsule embedded protection is carried out by freeze drying protectant, and to bacterial strain, so as to carry out thalline viable count
Protection.
To achieve the above object, the present invention provides a kind of compound probiotic powder preparation rich in selenium, it is characterised in that it is wrapped
Containing following compositions：Selenium-rich Bifidobacterium V9, maltodextrin, glucose, citric acid, FOS.
The present invention also provides a kind of selenium-enriched probioticses piece, it is characterised in that it includes following compositions：Selenium-rich Bifidobacterium
V9, galactooligosaccharide, maltodextrin, antierythrite, hydroxypropyl cellulose, oligoisomaltose, magnesium stearate, citric acid.
The present invention also provides a kind of food, and it is prebiotic that it includes the compound probiotic powder preparation or selenium rich in selenium described above
Bacterium piece.
The commercially available acquisitions of Bifidobacterium V9 in the present invention.
The present invention also provides a kind of preparation method of the compound probiotic powder preparation rich in selenium, the bifid bar that will be obtained first
Bacterium V9 is fermented, thalline is collected by centrifugation；Bacterium mud freeze-drying after centrifugation is obtained to the viable bacteria bacterium powder of high quality, will be made afterwards
Viable bacteria bacterium powder be used in the production of preparation, Bifidobacterium V9 is inoculated in after optimization in viable count performance highest culture medium,
Its fermentation condition is optimized and (adds selenium time, addition Se content, the mode of addition selenium to study)；It is enlarged again
It is trained two level kind, three-level kind, ferment tank culture；Medium centrifugal concentrates, and collects bacterium mud；Bacterium mud after centrifugation is carried out
Freeze-drying, (skimmed milk powder, ascorbic acid, monosodium glutamate, lactose) protective agent is added, be sufficiently mixed uniformly, carried out vacuum refrigeration and do
It is dry, freeze to moisture it is extremely low when take out, obtain required selenium-rich Bifidobacterium V9 viable bacteria bacterium by the technique such as sieving, crushing
Powder.
Selenium-enriched probioticses preparation prepared by this research, milk is pure, sweet and sour taste, fine and smooth homogeneous, and main physical and chemical index is equal
The relevant regulations met, and Organic Selenium is rich in, it is the good food source for supplementing Organic Selenium.This research is the industrial metaplasia of selenium-rich product
Production provides certain reference.
Unlike the production method of other like products, the production of this product is screening, training from probiotic lactobacillus
Support base optimization, the optimization of condition of culture is started with, determine optimal conditions of fermentation, then to Dialysis culture in process of high-density fermentation,
The fermentation techniques such as coupled fermentation, feedback regulation carry out discussion research, complete the high density fermentation to probiotic lactobacillus.From cell and
Molecular level study protection agent prescription composition and the protection mechanism dried to lactobacillus cell, optimization design is efficient on this basis
Protective agent, the control technology to the thalline death rate in probiotic lactobacillus drying process is completed, reach the relatively low thalline death rate, and
And the bacterium powder prepared using the technology has good stability in cryopreservation.
Brief description of the drawings
Below in conjunction with the accompanying drawings and embodiment the invention will be further described：
Fig. 1 inoculum concentrations and selenium concentration reciprocation response surface；
Fig. 2 selenium concentrations and initial pH value reciprocation response surface；
Fig. 3 150L ferment tank curves；
Fig. 4 bacterial strain PZB-1 production technological process；
The number of active bifid bacteria situation of change in Fig. 5 different hardness tablets；
Bifidobacterium survival rate situation of change in Fig. 6 different hardness tablets.
Embodiment
In order to further elucidate rather than limit the present invention in more detail, the following example is provided.
The response surface experiments design optimization the condition of selenium-enriched of embodiment 1.
Single factor experiment shows that suitable the condition of selenium-enriched is：Selenium concentration 8ug/mL, inoculum concentration 3%, cultivate to 6h (logarithms
Growth period) when add selenium, incubation time 48h, initial pH are 6.0.
With reference to single factor experiment result, it is major influence factors (being shown in Table 1) to select inoculum concentration, selenium concentration and pH value, with bacterium
Body selenium content is that index carries out Box-Behnken assay optimizations, experimental design and the results are shown in Table 2.Utilize software Design
Expert 7.0 analyzes and processes to test data, as a result as shown in table 3.
Table 1Box-Behnken empirical factors and level
Table 2Box-Behnken experimental designs and result
From table 2 and 3, the factor for influenceing thalline selenium content is followed successively by inoculum concentration ＞ selenium mass concentration ＞ pH from big to small
Value.Obtained regression equation is：Y=543.80+14.00A+10.75B+2.0C+14.00AB
+19.00AC-20.50BC-45.40A2-29.90B2-24.90C2。
Variance analysis (table 9), F are carried out to quadratic polynomial model regression coefficient and result of the testmodel=78.34, probability
Pmodel＜ 0.0001, the regression equation for showing to obtain are notable.Lose and intend item probability Plose=0.2722 ＞ 0.05, i.e., without mistake pseudo-factor
In the presence of.Coefficient of determination R2=0.9902, illustrate that regression equation can preferably reflect selected true pass between parameter and response
System.In addition, the coefficient of variation (CV) is 1.15% ＜ 5%, i.e. regression equation has good reappearance, R2 Pred=0.9014 with
R2 Adj=0.9775 preferably coincide.
The quadratic polynomial model regression coefficient of table 3 estimates and result of the test variance analysis
It is variable to select wherein two factors according to regression equation, and another factor is located at central point and makees Fig. 1 and 2.As seen from Figure 1,
Either with the increase of selenium concentration, or the increase with inoculum concentration, thalline selenium content, which is presented, first increases becoming of reducing afterwards
Gesture.Similar, as initial pH value increases, same trend (Fig. 2) is also presented in thalline selenium content.As shown in Table 9, selenium concentration,
The interaction item of inoculum concentration and the factor of pH value 3 is respectively provided with extremely significantly (P to thalline selenium contentAB＜ 0.01, PBC＜ 0.01, PAC＜
0.01) influence.
It can also be seen that regression equation has maximum of points by Fig. 1 and Fig. 2.Thalline highest is calculated using equation to contain
Selenium amount is 2546.337ug/g, and now inoculum concentration, selenium concentration and pH value are respectively 3.79%, 6.83 μ g/ml and 6.02.Above-mentioned
Under the conditions of carry out confirmatory experiment, bacterial strain selenium content is (2551 ± 4.6) μ g/g, and selenium conversion ratio is (42.2 ± 3.4) %.
The pilot scale fermentation of embodiment 2. is tested
On the basis of bacterial strain High Density Cultivation pilot plant test, the production work that fermentation-scale is amplified to 2t by 150L step by step is completed
Skill amplification research, to determine scale production process.Centrifugation and freezing dry process of the experiment also to thalline after fermentation are to thalline
Influence, zymotic fluid after thalline is collected by centrifugation, freeze-drying can obtain average viable count 2.2 × 1011Cfu/g bacterium powder, energy
Enough meet the requirement of high viable count, thalline survival rate 82%.
2.1 fermentation parameter
By a series of optimal screening early stage, obtain Media Components and condition of culture is as follows：Fermentative medium formula
For：Yeast extract 1.63%, FOS 0.13%, cysteine hydrochloride 0.0076%, casein peptone 1.0%, K2HPO4
0.2%；37 DEG C of cultivation temperature, plants age 6h, inoculum concentration 4%, mixing speed 100r/min, initial pH6.0, ferment 2h when add it is sub-
Sodium selenate, it is 6.83 μ g/ml to make selenium concentration.
2.2 150L fermentation tanks batch fermentations are tested
Fermentation test is carried out according to the culture medium and condition of culture in 150L fermentation tanks, determines bacterial strain in fermentation process
OD600With selenium conversion ratio, Fig. 3 is as a result seen.
As seen from Figure 3, cultivated in 150L fermentation tanks, there is no the obvious delay phase, be directly entered exponential phase；
24h is cultivated, into stationary phase, zymotic fluid OD600Value reaches 0.912, and viable count reaches 6.9 × 109cfu/mL.Selenium conversion ratio with
The increase of thalline quantity and improve, reach maximum 45.1% during 40h.
2.3 2t fermentation tank scale-ups
The success of pre-stage test ensure that the possibility of industrialized production, continue the 2t fermentation tank production technology bars to bacterial strain
Part is amplified experiment.Result of the test shows, is cultivated in 2t fermentation tanks, the growth and selenium conversion ratio variation tendency and life of bacterial strain
Production is horizontal substantially close with the situation in 150L fermentation tanks, and viable count reaches 6.1 × 109Cfu/mL, selenium conversion ratio are 44.9%.
Therefore the zymotechnique that pre-stage test determines completely can be as the technological parameter cultivated in 2t fermentation tanks.
The lyophilized technique of embodiment 3. optimizes
The optimization of 3.1 centrifugal conditions
Zymotic fluid is centrifuged using different centrifuge RPMs, temperature and centrifugation time, then outwelled supernatant, determines sediment
Middle number of viable and cycles of concentration, the results are shown in Table 4.
The centrifugal condition of table 4 is to zymotic fluid centrifugal effect
As can be seen from Table 4, in the case where high score is from revolution, low temperature group is higher than the thalline survival rate of room temperature group 1 times；
Under cryogenic conditions, high score is higher than the bacterium number survival rate of low separation revolution from revolution, thus centrifugal process concentration thalline using low temperature and
High score is from revolution to thalline survival rate than advantageous.But in concentrate, because bacterial strain production acid causes Partial Protein to precipitate, and with
Separated out together with thalline, this largely reduces centrifugal effect, and each group experiment cycles of concentration shown in table is not high.
The optimization of 3.2 lyophilized techniques
It will centrifuge after obtained concentration liquid uniformly mixes with the protective agent of thing based on the skimmed milk selected and be put into ice
Case is stayed overnight.The refrigerating plants such as compressor are opened, 0.5~1h of normal operation, its cold plate is cooled to less than 35 DEG C.Mixing is protected
The concentration thalline 1mL for protecting agent loads cillin bottle, is placed on freeze dryer cold plate.Treat that sample temperature is down to -40 DEG C, open vavuum pump
And heating system.Sample drying bursts apart to outward appearance to be continued to dry 1h (to moisture below 2%), and shutdown stops lyophilized, starting
Pressing device, by cillin bottle vacuum sealing.Obtained freeze-drying bacterium powder is used to determine survival rate and transmission and scanning electron microscopic observation structure.
The early stage of drying vacuum is 0.1bar, and freezing minimum temperature is -38~-40 DEG C.
The screening of 3.3 frost durability agents
Bacterial strain is accessed in seed culture medium and activated, thalline is collected by centrifugation, adds the different frozen-dried protective factors respectively
(2% sucrose, 0.1% sulfur-containing amino acid, 1.5% sodium glutamate, 1% tween, 10% marine algae extract, 0.3% vitamin c),
After fully mixing, it is dispensed into oneself sterilized aluminum dish and is freeze-dried.After lyophilized sterilizing is linked into by 0.6% inoculum concentration
Containing in seleno culture medium afterwards, after 100r/min cultivates 36h, determines the conversion ratio of its selenium by 32 DEG C.It is shown in Table 5.Find out from result, paddy
Propylhomoserin sodium is protected larger to thalline selenium conversion ratio.
The influence that the frost durability agent of table 5 converts to thalline selenium
The screening of 3.4 freeze drying protectants
Influence of the freeze drying protectant of table 6 to Strain survival rate
Selected protective agent has certain protecting effect to PZB-1 it can be seen from the result of table, wherein with adonis amurensis
The effect of alcohol is the most notable, freezes survival rate up to 53.45%, but adonite is a kind of system applied in terms of medical science
Agent, costly, application is restricted price.The protecting effect of marine algae extract is unexpected, and its lyophilized survival rate is reachable
52.40%, it is probably because a certain amount of trehalose serves preferable protecting effect in marine algae extract, separately to analyze its reason
Composition is sufficiently complex in outer marine algae extract, and especially algal polysaccharides content is higher, the guarantor of this kind of polysaccharide and other compositions to thalline
Shield effect requires study.The protecting effect of sodium glutamate is more significant, it is lyophilized after thalline survival rate reach 50.29%, this with it is other
The experimental result of strain is consistent.In addition, sucrose and tween are to apply wider freeze drying protectant, their effect in this experiment
Preferably, thalline survival rate is respectively 38.71% and 40.00% after freezing.β-phosphoric acid tween disodium salt, Span80 protecting effects one
As.Yolk is not easy to freeze, and its structural state is sticky to be difficult to count, and the sterilization of yolk is a more scabrous problem, therefore
Uncomfortable cooperation freeze drying protectant.
The determination of the bacterial strain PZB-1 bacterium powder production technologies of embodiment 4.
The optimal lyophilisation condition that can determine bacterial strain PZB-1 by above-mentioned lyophilized Optimum Experiment is：Centrifugal rotational speed 8000r/
Min, 0~4 DEG C of centrifuging temperature, centrifugation time 5min；- 30 DEG C of pre-freezing temperature, pre-freeze time 2h；Fill liquid thickness 10mm；It is lyophilized to protect
Protecting agent prescription is：The marine alga juice of+1% vitamin c of 5% sucrose+0.8% sodium glutamate of+2% tween+5%.In summary, bacterial strain
PZB-1 bacterium powder preparation technology flow chart is as shown in Figure 4.
Bacterium powder is freezed according to the above-mentioned technique productions of Fig. 4, the bacterium powder indices of acquisition are shown in Table 7.From data in table, obtain
The viable count for obtaining bacterium powder is 2.5 × 1011Cfu/g, Se content are 2145 μ g/g；Water content is 4.3%, long-term less than being suitable for
Store the 6% of bacterium powder condition；Thalline survival rate is 84%.Under the preparation condition of the culture and bacterium powder, in prepared bacterium powder
Miscellaneous bacteria, pathogenic bacteria and mould are not detected, meet the hygienic requirements as leavening.
The bacterium powder index of table 7
The determination of the pressed powder technique of embodiment 5.
The manufacture craft of traditional wet granulation is to put mixed powder in mixer, adds suitable amount of adhesive or moistening
Agent, softwood processed, wet granular processed, wet granular is dry and whole grain, is produced after whole grain by tabletting machine.This preparation method of granules work
Sequence is cumbersome, and labor intensity is big, and adhesive or wetting agent dosage are larger, increases production cost.Direct powder compression technology is the brightest
Aobvious advantage is its economy.
5.1 tablet formulation
Selenium-enriched probioticses piece is positioned at popular 1 ︰ 10 sweet tea acid ratio, and sweetener is with acid with sucrose, lactose, Portugal
Grape sugar, oligoisomaltose, galactooligosaccharide, antierythrite, citric acid carry out Experiment of Compatibility.As a result show, with glucose, lemon
Lemon acid is aided with flavoring apple essence, product sour and sweet palatability as filler, flavouring.Antierythrite is sweet, does not have particle during chewing
Feel, be in the endothermic reaction during dissolving, have cooling feeling in mouth during chewing.Therefore a certain amount of antierythrite is added in formula.
The selection of adhesive：Maltodextrin can be used as filler, increase chewiness, but addition can excessively make tablet into
Shape is difficult, loose pieces easily occurs.And oligoisomaltose and galactooligosaccharide are not only effective to proliferation of probiotics, moreover it is possible to play adhesive
Effect.Add the microcrystalline cellulose that compressibility is good, helps stream, adhesive effect again simultaneously, solve tablet and embark on journey and disintegration
Contradiction.
The selection of glidant：Simultaneous selection magnesium stearate, superfine silica gel powder, talcum powder are selected, the results showed that stearic
Sour magnesium not only helps stream effect best, also improves the finish of tablet.
5.2 tablet forming technique
Dispensing, mixing：Supplementary material is crossed into 16 mesh sieves in advance, according to charge ratio, first by the unclassified stores in addition to magnesium stearate
Mixed 12 minutes on mixing machine, add magnesium stearate and mix 2 minutes.
The material mixed is subjected to tabletting.
The determination of 5.3 tablet hardnesses
Using direct powder compression, the auxiliary material for meeting food hygiene requirement is crossed into 40 mesh sieves, addition bacterial strain PZB-1 respectively
Lyophilized bacterium powder 10% and other auxiliary materials, mixing machine mixing 12min, upper tabletting machine, tablet press machine pressure is controlled, obtains 4 kinds of pieces
Agent hardness is respectively 40N, 60N, 80N, 100N tablet.Overall process must operate under certain cleaning condition.
5.3.1 the viable count in different hardness tablet
Viable count in different hardness tablet is detected using flat board tilt-pour process, and calculates survival rate, as a result sees Fig. 5 and Fig. 6.
As can be seen from Figure 5 when selenium-enriched probioticses tablet hardness is respectively 0N, 40N, 60N, 80N, 100N, bacterial strain is measured
PZB-1 viable count is respectively 6.0 × 109cfu/g、1.9×109cfu/g、1.6×109cfu/g、1.4×109cfu/g、9.0
×108cfu/g.Survival rate is respectively 32.3%, 26.2%, 23.4%, 15.6%.
As can be seen from Figure 6 when selenium-enriched probioticses tablet hardness is respectively 40N, 60N, 80N, 100N, bacterial strain PZB-1 is measured
Survival rate be respectively 32.3%, 26.2%, 23.4%, 15.6%.
5.3.2 the length of a film of different tablet hardnesses and piece are thick
Because final probiotic tablet piece type is olive shape, for the needs of tablet packaging, to the thickness and length of tablet
Degree there are certain requirements.Test and the piece thickness and length of a film of the tablet of 4 kinds of different hardness are determined respectively, the results are shown in Table 7.By table
In the result of tabletting twice can be seen that piece is thick, length of a film is negatively correlated with piece hardness.Meanwhile when being formulated identical, tablet hardness
Bigger, its surface is brighter and cleaner.
The tablet hardness of table 7 and thickness and the relation of length
In summary, with the increase of tablet hardness, the survival rate of Bifidobacterium gradually reduces in tablet, viable count also by
Gradually reduce, bacterial strain proof pressure ability is very weak.When tabletting hardness is 60N and 80N, the survival rate of Bifidobacterium is only 25%
Left and right, though viable count substantially reduces, the order of magnitude remains within 109.At the same time, with the increase of tablet hardness, tablet
Thickness and length be gradually reduced.When tabletting hardness is 80N, piece thickness is 5.891mm, a length of 20.196mm of piece, meets bag
Reload request.It is more to consider thalline survival rate and viable count, smoothness, color, shape, piece thickness, the length of a film of probiotic tablet etc.
Aspect factor, it is most suitable tablet hardness to test finally selected hardness 80N.The viable count of finished tablet is 1.4 × 109Cfu/g, bacterium
Strain PZB-1 survival rate is 23.4%, and Se content is 18.9 μ g/g.
The probiotics tablets that the pressed powder technology of above-mentioned determination prepares addition possess following advantage：(1) because double bacterium are recognized
To there is many wholesome effects to human body, so the addition prebiotic substance such as galactooligosaccharide and oligoisomaltose sugar can promote
The increase of probiotics quantity or its active enhancing.(2) pressed powder technology is used, by the powder of medicine and suitable auxiliary material
After sieving and mix respectively, without pelleting (wet granular or dry particl), direct pressing is in blocks, and this technical process is simple, no
It must pelletize, dry, energy-conservation is time saving, preferably protects the activity of probiotics.(3) in traditional wet granulation, to consider to stick
Mixture addition, particle drying time, the alcohol granulation sieving factor such as time, the machine quantity that produces needs is more, space is big, and by
Need to rely on micro-judgment in some factors, it is thus possible to cause unstable product quality, difference is big etc. between batch；And directly press
The most significant advantage of chip technology is reduction of equipment and the cost of running：Its simple production process, without pelletizing, sieving, drying,
The process such as whole grain and middle sampling Detection, reduce corresponding equipment factory building investment and inspection cost and labor intensity, save
Time and the energy, and do not make end product quality stable, difference is small between batch, can because the experience of worker determines the quality of product
Strong operability, continuous production is guaranteed, is especially suitable for the production management of GMP requirements.
Claims (8)
1. a kind of compound probiotic powder preparation rich in selenium, it is characterised in that it includes following compositions：Selenium-enriched probioticses pulvis：
Selenium-rich Bifidobacterium V9, maltodextrin, glucose, citric acid, FOS
2. a kind of selenium-enriched probioticses piece, it is characterised in that it includes following compositions：Selenium-rich Bifidobacterium V9, galactooligosaccharide, wheat
Bud dextrin, antierythrite, hydroxypropyl cellulose, oligoisomaltose, magnesium stearate, citric acid.
3. the preparation method of the compound probiotic powder preparation rich in selenium in claim 1, it is characterised in that first by Bifidobacterium
V9 is inoculated in culture medium, and its fermentation condition is optimized and (adds selenium time, addition Se content, the mode of addition selenium to grind
Study carefully)；It is enlarged again and is trained two level kind, three-level kind, ferment tank culture；Medium centrifugal concentrates, and collects bacterium mud；
Bacterium mud after centrifugation is freeze-dried, adds (skimmed milk powder, ascorbic acid, monosodium glutamate, lactose) protective agent, is sufficiently mixed
It is even, carry out vacuum freeze drying, freeze to moisture it is extremely low when take out, obtain required richness by the technique such as sieving, crushing
Selenium Bifidobacterium V9 viable bacteria bacterium powders.
4. the preparation method of compound probiotic powder preparation according to claim 3, it is characterised in that fermentation condition is as follows：
Selenium concentration 8ug/mL, inoculum concentration 3%, selenium is added when cultivating to 6h (exponential phase), incubation time 48h, initial pH are 6.0.
5. the preparation method of compound probiotic powder preparation according to claim 3, it is characterised in that fermentation condition is：Connect
Kind amount, selenium concentration and pH value are respectively 3.79%, 6.83 μ g/ml and 6.02.
6. the preparation method of compound probiotic powder preparation according to claim 3, it is characterised in that fermentation condition is as follows,
Fermentative medium formula is：Yeast extract 1.63%, FOS 0.13%, cysteine hydrochloride 0.0076%, casein peptone
1.0%, K2HPO40.2%；37 DEG C of cultivation temperature, plant age 6h, inoculum concentration 4%, mixing speed 100r/min, initial pH6.0, hair
Sodium selenite is added during ferment 2h, it is 6.83 μ g/ml to make selenium concentration.
7. the preparation method of compound probiotic powder preparation according to claim 3, it is characterised in that freeze-drying condition
For：Centrifugal rotational speed 8000r/min, 0~4 DEG C of centrifuging temperature, centrifugation time 5min；- 30 DEG C of pre-freezing temperature, pre-freeze time 2h；Dress
Liquid thickness 10mm；Frozen-dried protective agent prescription is：The marine alga of+1% vitamin c of 5% sucrose+0.8% sodium glutamate of+2% tween+5%
Juice.
8. a kind of food, it includes institute in the compound probiotic powder preparation or claim 2 rich in selenium described in claim 1
The selenium-enriched probioticses piece stated.
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