CN102433287B - Direct-vat bacillus subtilis starter and preparation method thereof - Google Patents
Direct-vat bacillus subtilis starter and preparation method thereof Download PDFInfo
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Abstract
The invention relates to a direct-vat bacillus subtilis starter and a preparation method thereof. The method comprises the following steps of: performing seed culture on bacillus subtilis, performing high-density scale-up culture on the bacillus subtilis, and preparing a bacterium starter by a freeze-drying method to finally obtain the direct-vat bacillus subtilis starter. A culture medium for the seed culture and the high-density scale-up culture of the bacillus subtilis comprises the following components in percentage by weight: 2 percent of sucrose, 1 percent of bean pulp, 0.1 percent of monopotassium phosphate, 0.05 percent of magnesium sulfate and the balance of water, wherein the pH value is 7.0 to 7.2. Concentration conditions for preparing the bacterium starter by the freeze-drying method are that: the centrifugal rotating speed is 5,000r/min, the centrifugal time is 10min, the centrifugal temperature is 4 DEG C, and a cryoprotectant contains 5 percent of maltodextrin, 4 percent of sucrose, 0.8 percent of trehalose and 0.2 percent of tween. The survival rate of the direct-vat bacillus subtilis starter produced by the method is 95.3 percent, the viable count is 2.69*10<10>cfu/g, and ideal liquid and solid fermentation effects can be achieved according to one thousandth of inoculation amount. The preparation method for developing the novel high-activity direct-vat bacillus subtilis starter on the basis of the strain is applied to the liquid-state fermentation and solid-state fermentation of bacillus subtilis-related products.
Description
Technical field
The invention belongs to the biotechnological formulation field, especially a kind of direct-vat bacillus subtilis starter and preparation method thereof.
Background technology
Throw type leaven refers to the concentrated and standardized lyophilize starter culture of a series of height, can directly add heat treated raw material and ferment, and need not other pre-treatment work such as it activate, spreads cultivation.The at present application of direct-vat bacillus subtilis starter has no report both at home and abroad.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art part, provide a kind of simple to operate, viable bacteria content is very high, vigor is high, direct-vat bacillus subtilis starter that inoculum size is little and preparation method thereof.
The present invention realizes that the technical scheme of purpose is as follows:
A kind of preparation method of direct-vat bacillus subtilis starter; comprise the subtilis seed culture; subtilis high-density amplification culture; concentration; add protective material; freeze-drying prepares the thalline starter; obtain at last direct-vat bacillus subtilis starter; the substratum weight percent of described subtilis seed culture and high-density amplification culture is: sucrose 2; dregs of beans 1; potassium primary phosphate 0.1; sal epsom 0.05; add water to 100%; pH7.0-7.2; freeze-drying prepares the concentrated condition of thalline starter: centrifugal rotational speed 5000r/min; centrifugal 10min; centrifuging temperature is 4 ℃, and the protective material that adopts is for containing 5% maltodextrin; 4% sucrose; the aqueous solution of 0.8% trehalose and 0.2% tween.
And, the name of described subtilis is called TK-1, specific name Bacillus subtilis, deposit number is: CGMCC No.4731, preservation date: on April 2nd, 2011, the preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center.
And described seed culture condition is: cultivate for 37 ± 1 ℃, incubation time is 20h.
And the survival rate of subtilis is 95.3% in the described direct-vat bacillus subtilis starter, and viable count is 2.69 * 10
10Cfu/g.
And described direct-vat bacillus subtilis starter is applied to liquid state fermentation by 1 ‰ inoculum size, and the highest viable count is 1.03 * 10
10Cfu/mL.
And direct-vat bacillus subtilis starter is applied to solid state fermentation by 1 ‰ inoculum size, and the highest viable count is 0.985 * 10
10Cfu/g.
A kind of direct-vat bacillus subtilis starter is characterized in that: comprise lyophilized vaccine and subtilis, described lyophilized vaccine is 5% maltodextrin, 4% sucrose, 0.8% trehalose, 0.2% tween.
And, the name of described subtilis is called TK-1, specific name Bacillus subtilis, deposit number is: CGMCC No.4731, preservation date: on April 2nd, 2011, the preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center.
Advantage of the present invention and positively effect are:
1, after subtilis TK-1 bacterial strain passes through fed-batch fermentation high-density amplification culture among the present invention, adopt freeze-drying to prepare the thalline starter, the survival rate of the direct-vat bacillus subtilis starter of producing with present method is 95.3%, and viable count is 2.69 * 10
10Cfu/g, the inoculum size according to 1 ‰ just can reach desirable liquid state and solid state fermentation effect.
2, the present invention adopts freeze-drying to prepare direct-vat bacillus subtilis starter first, compares with former liquid seed inoculation, and the starter inoculum size is little, directly uses, and labor savings shortens the production cycle, can prevent effectively that again bacterial classification from polluting and degeneration; Preserve convenient management, the advantages such as product with stable quality.
3, the present invention has determined that subtilis adopts centrifugal rotational speed 5000r/min in concentration process, 4 ℃ of lower centrifugal 10min, and centrifugal yield is 98.83%.
4, the present invention has determined that protective material is 5% maltodextrin, 4% sucrose, 0.8% trehalose, 0.2% tween in the thalline freezing dry process, and final subtilis freeze-drying survival rate is 93%, and viable count is 2.69 * 10
10Cfu/g.
5, the direct-vat bacillus subtilis starter that adopts the inventive method preparation is applied to liquid state fermentation by 1 ‰ inoculum size, and the highest viable count is 1.03 * 10
10Cfu/mL, the direct-vat bacillus subtilis starter of employing the inventive method preparation is applied to solid state fermentation by 1 ‰ inoculum size, and the highest viable count is 0.985 * 10
10Cfu/g.
6, the fermentation of bacillus subtilis agent of the present invention's preparation has the following advantages: viable bacteria content is very high, and vigor is high, thereby inoculum size is little; Can directly use, avoid lengthy and tedious enlarged culturing process, both labor savings shortens the production cycle, can prevent effectively that again bacterial classification from polluting and degeneration; Preserve convenient management; Be easy to production control technique, product with stable quality; After processing, simple rehydration can directly use as the seed bacterial strain.
Description of drawings
Fig. 1 is that the present invention carries out cell collecting time to the impact of starter viable count;
Fig. 2 be lyophilized vaccine on the impact of starter viable count, annotate: No. 1 10% maltodextrin; No. 2 10% sucrose; No. 3 10% trehaloses; No. 4 10% tweens; No. 5 5% maltose+5% sucrose; No. 6 5% maltodextrin+4% sucrose+0.8% trehalose+0.2% tweens; No. 7 5% maltodextrin+4% sucrose+1% tweens; No. 8 5% maltodextrin+4% sucrose+1% trehaloses;
Fig. 3 is the effect comparison that fermentation of bacillus subtilis agent and fermented liquid are seeded in liquid culture medium.
Fig. 4 is the effect comparison that fermentation of bacillus subtilis agent and fermented liquid are seeded in solid medium.
Embodiment
Below in conjunction with embodiment, the present invention is further described, and following embodiment is illustrative, is not determinate, can not limit protection scope of the present invention with following embodiment.
1. the seed liquor of thalline preparation
The bacterial classification of glycerine pipe cold storage is received in the test tube that liquid seed culture medium is housed, and is that 37 ℃, rotating speed are to cultivate under the condition of 150r/min in temperature.
Wherein liquid seed culture medium (g/L) is: peptone 10g, and Nacl 10g, yeast soak powder 5g, and cultivate in shaking table pH7.0～7.2, and its temperature is 37 ℃, and rotating speed is 150r/min.
3. the high-density amplification culture of thalline
Inoculum size with 5% is inoculated in the 200L tank with subtilis and carries out fermentation culture, and liquid amount 160L cultivates 20h in 37 ± 1 ℃.The employing peristaltic pump adds the NaOH solution of 2mol/L and regulates pH, makes it to maintain about 6.8, and fermentation is adopted and left standstill fed-batch fermentation, carries out gentle agitation after the feed supplement, and the soya-bean oil of interpolation 0.3% is made defoamer.
4. bacterial classification is collected
It is a step of whole technology key that bacterial classification is collected, comparatively suitable method is to adopt supercentrifuge to carry out centrifugation then to carry out vacuum lyophilization at present, it can adapt to industrialization production requirements fully, so mostly adopt centrifugal and the cryodesiccated way of vacuum-drying.
5. the protectant selection of thalline
After having collected a large amount of subtilis thalline, must add the bacterial classification protective material so that subtilis avoids the damage of extraneous adverse factor.Adding protective material can effectively improve survival rate, the survival time of subtilis and keep the original characteristic of subtilis.
6. lyophilize
With the thalline collected with carry out pre-freeze after protective material fully mixes; the pre-freeze temperature is controlled at-20 ℃～-30 ℃; the pre-freeze time decides with the biomass surface-area, and mostly between-20 ℃～-30 ℃, dried starter moisture content is lower than 3% and is advisable drying temperature.
7. mensuration viable count
The general colony counting method that adopts, operation steps is: take by weighing the starter 1g for preparing, be dissolved in the stroke-physiological saline solution of 9ml, shake up the rear bacterium liquid of getting 1ml with liquid-transfering gun and add in the stroke-physiological saline solution of another 9ml, dilute, take turns doing ten times of gradient dilutions.After having diluted, get respectively suitable extent of dilution 1ml with liquid-transfering gun and throw flat board into, pour the substratum mixing into, take turns doing three parallel, put 38 ℃ of incubators into and be inverted and leave standstill cultivation.
8. the packing of starter
The starter viable count needs greater than 10
10Cfu/g, final powder formulation product must carry out the vacuum seal packing, and in cryopreservation, the quality guaranteed period is approximately 1a.
Below be the determining of some parameters that the direct-throwing subtilis is prepared the production method of agent, and the optimization of some culture condition.
The screening of direct-vat bacillus subtilis starter preparation condition
1. the Bacillus subtilis strain collection time determines
Bacterial growth has experienced respectively lag phase, logarithmic phase, balance period and paracme, and at different times, growing state is different, respectively live bacterial count is carried out in its sampling.
Screening example one
Liquid state fermentation substratum: sucrose 2g, dregs of beans 1g, dipotassium hydrogen phosphate 0.1g, sal epsom 0.05g, pH7.0～7.2 inoculation and culture condition: inoculum size 5% (v/w), 37 ± 1 ℃ of culture temperature, incubation time 18h.The viable count that records is 3.01 * 10
10Cfu/ml.
Screening example two
Liquid state fermentation substratum: sucrose 2g, dregs of beans 1g, potassium primary phosphate 0.1g, sal epsom 0.05g, pH7.0～7.2 inoculation and culture condition: inoculum size 5% (v/w), 37 ± 1 ℃ of culture temperature, incubation time 20h.The viable count that records is 3.43 * 10
10Cfu/ml.
Screening example three
Liquid state fermentation substratum: sucrose 2g, dregs of beans 1g, potassium primary phosphate 0.1g, sal epsom 0.05g, pH7.0～7.2
Inoculation and culture condition: inoculum size 5% (v/w), 37 ± 1 ℃ of culture temperature, incubation time 22h.The viable count that records is 3.36 * 10
10Cfu/ml.
The result
The suitableeest incubation time of liquid seed fermentation liquid is 20h (seeing accompanying drawing 1).
2. determining of the centrifugal rotational speed when liquid bacterial agent is collected and time
To increase the liquid bacterial agent that the subtilis after bacterium is cultivated is collected by whizzer, the revolution when centrifugal and time are larger on the impact of thalline, and under the equal conditions, along with the increase of centrifugal revolution, the survival rate of centrifugal rear bacterium reduces,
Screening example 1
With the thalline liquid that ferments, under the condition of 4000r/min, centrifugal 10min, centrifugal front viable count 3.43 * 10
10Cfu/ml, viable count is 3.25 * 10 in the centrifugal rear sedimentation liquid
10Cfu/ml, centrifugal rear sedimentation liquid thalline survival rate is 94.75%.
Screening example 2
With the thalline liquid that ferments, under the condition of 4000r/min, centrifugal 15min, centrifugal front viable count 3.43 * 10
10Cfu/ml, viable count is 3.34 * 10 in the centrifugal rear sedimentation liquid
10Cfu/ml, centrifugal rear sedimentation liquid thalline survival rate is 97.38%.
Screening example 3
With the thalline liquid that ferments, under the condition of 5000r/min, centrifugal 10min, centrifugal front viable count 3.43 * 10
10Cfu/ml, viable count is 3.39 * 10 in the centrifugal rear sedimentation liquid
10Cfu/ml, centrifugal rear sedimentation liquid thalline survival rate is 98.83%.
The result: centrifugal optimum condition is 5000r/min, and centrifugation time is 10min (seeing Table 1).
3. lyophilized vaccine determines
Protectant concentration of below mentioning is the weight percent concentration of the aqueous solution of tie substance.
Behind thalline and the good lyophilized vaccine mixing that goes out, make its fully charge, then carry out vacuum lyophilization.Must carry out pre-freeze to sample, the temperature of this research pre-freeze-25 ℃～-30 ℃, pre-freeze time 4hr, freezing thickness is 0.5mm, carries out lyophilize again, condition is: cold well temperature-30 ℃, 50 ℃ of plate temperature, freeze-drying time 20h.
Screening example 1
Thalline is mixed by a certain percentage with lyophilized vaccine, condition such as above-mentioned, the lyophilized vaccine composition is 10% maltodextrin, and recording freeze survival rate is 51.3%, and the freeze-drying survival rate is 46.1%, and the lyophilized powder viable count is 0.83 * 10
10Cfu/g.
Screening example 2
Thalline is mixed by a certain percentage with lyophilized vaccine, condition such as above-mentioned, freeze-dried protection composition is: 5% maltodextrin, 4% sucrose, 1% trehalose, recording freeze survival rate is 90.1%, freeze-drying survival rate 88.7%, the lyophilized powder viable count is 1.22 * 10
10Cfu/g.
Screening example 3
Thalline is mixed by a certain percentage with lyophilized vaccine; condition such as above-mentioned, freeze-dried protection composition is: 5% maltodextrin, 4% sucrose, 0.8% trehalose, 0.2% tween, recording freeze survival rate is 89.6%; the freeze-drying survival rate is 88.4%, and the lyophilized powder viable count is 2.69 * 10
10Cfu/g.
The result
The freeze-dried protection composition that should adopt is: 5% maltodextrin, 4% sucrose, 0.8% trehalose, 0.2% tween (seeing accompanying drawing 2).
Conclusion: this experiment adopts this method to produce direct-vat bacillus subtilis starter, and this starter bacteria containing amount is high, and energetic, viable count reaches 2.69 * 10
10Cfu/g.
4. the application of fermentation of bacillus subtilis agent in liquid state fermentation
Screening example 1
The fermentation of bacillus subtilis agent is added in sterilized 0.85% physiological saline, normal temperature rehydration 20min, then access the liquid state fermentation substratum of the bacterium of going out by 0.1% inoculum size, form (%): sucrose 2, dregs of beans 1, dipotassium hydrogen phosphate 0.1, sal epsom 0.05, fermentation culture 18h is carried out in pH7.0～7.2 under 37 ℃ of conditions, survey viable count, record viable count and be: 9.96 * 10
9Cfu/mL.
Screening example 2
Subtilis after liquid activation by the went out liquid state fermentation substratum of bacterium of 5% inoculum size access, is formed (%): sucrose 2, dregs of beans 1, dipotassium hydrogen phosphate 0.1, sal epsom 0.05, pH7.0～7.2, under 37 ℃ of conditions, carry out fermentation culture 18h, survey viable count.
Recording viable count is: 8.53 * 10
9Cfu/mL.
The result
Use direct-throwing bacillus licheniformis starter to contrast in the liquid state fermentation substratum by inoculum size 0.1% and inoculum size 5% liquid inoculation, the highest viable count is respectively 1.03 * 10
10Cfu/mL and 0.903 * 10
9Cfu/mL (accompanying drawing 3).Obviously reduce inoculum size, and can reach better ferment effect.
Five, the application of direct-vat bacillus subtilis starter in solid state fermentation
Screening example 1
The fermentation of bacillus subtilis agent is added in sterilized 0.85% physiological saline, normal temperature rehydration 20min, then by 0.1% inoculation access went out bacterium take dregs of beans as main solid-state fermentation culture medium, form: dregs of beans: wheat bran: Semen Maydis powder=3: 1: 1, leave standstill cultivation 18h at 37 ℃ of constant incubators, survey viable count.
Recording viable count is: 8.21 * 10
9Cfu/g.
Screening example 2
With the subtilis after liquid activation by the inoculum size access of 5% (v/w) went out bacterium take the solid-state fermentation culture medium of dregs of beans as leading, form: dregs of beans: wheat bran: Semen Maydis powder=3: 1: 1, leave standstill cultivation 18h at 37 ℃ of constant incubators, survey viable count.
Recording viable count is: 7.4 * 10
9Cfu/g.
The result
Use direct-vat bacillus subtilis starter to contrast in solid-state fermentation culture medium by inoculum size 0.1% and inoculum size 5% liquid inoculation, the highest viable count is respectively 9.85 * 10
9Cfu/g and 8.43 * 10
9Cfu/g (accompanying drawing 4).There is the lag phase of about 12h in thalline after the starter inoculation, and then growth is quick, and final ferment effect is desirable, can obviously reduce inoculum size.
Conclusion
By the research to the direct-vat bacillus subtilis starter production technique, subtilis is concentrated adopts final subtilis survival rate to reach 93.2%, and the viable count of starter is 2.69 * 10
10Cfu/g, the inoculum size by 1 ‰ just can reach desirable liquid state or solid state fermentation effect, and the highest viable count is respectively 1.03 * 10
10Cfu/mL, 9.85 * 10
9Cfu/g.
Table 1 centrifugation time of the present invention and rotating speed are on the impact of starter viable count
Claims (6)
1. the preparation method of a direct-vat bacillus subtilis starter, it is characterized in that: comprise the subtilis seed culture, subtilis high-density amplification culture, concentration, add protective material, freeze-drying prepares the thalline starter, obtain at last direct-vat bacillus subtilis starter, the substratum weight percent of described subtilis seed culture and high-density amplification culture is: sucrose 2, dregs of beans 1, potassium primary phosphate 0.1, sal epsom 0.05, add water to 100%, pH 7.0-7.2, the concentration condition is: centrifugal rotational speed 5000r/min, centrifugal 10min, centrifuging temperature is 4 ℃, the lyophilized vaccine that adopts consist of mass percent concentration: 5% maltodextrin, 4% sucrose, the aqueous solution of 0.8% trehalose and 0.2% tween;
The name of described subtilis is called TK-1, specific name: subtilis Bacillus subtilis, deposit number is: CGMCCNo.4731, preservation date: on April 2nd, 2011, the preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center.
2. the preparation method of direct-vat bacillus subtilis starter according to claim 1, it is characterized in that: described seed culture condition is: cultivate for 37 ± 1 ℃, incubation time is 20h.
3. the preparation method of direct-vat bacillus subtilis starter according to claim 1, it is characterized in that: the survival rate of subtilis is 95.3% in the described direct-vat bacillus subtilis starter, viable count is 2.69 * 10
10Cfu/g.
4. the preparation method of direct-vat bacillus subtilis starter according to claim 3, it is characterized in that: described direct-vat bacillus subtilis starter is applied to liquid state fermentation by 1 ‰ inoculum size, and the highest viable count is 1.03 * 10
10Cfu/mL.
5. the preparation method of direct-vat bacillus subtilis starter according to claim 4, it is characterized in that: direct-vat bacillus subtilis starter is applied to solid state fermentation by 1 ‰ inoculum size, and the highest viable count is 0.985 * 10
10Cfu/g.
6. direct-vat bacillus subtilis starter that is prepared by the described preparation method of claim 1; it is characterized in that: comprise lyophilized vaccine and subtilis; described lyophilized vaccine is 5% maltodextrin; 4% sucrose; 0.8% trehalose; 0.2% tween; the name of described subtilis is called TK-1; specific name Bacillus subtilis; deposit number is: CGMCC No.4731; preservation date: on April 2nd, 2011; the preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, depositary institution: China Committee for Culture Collection of Microorganisms common micro-organisms center.
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| CN112980717B (en) * | 2020-12-24 | 2023-07-07 | 云南省微生物发酵工程研究中心有限公司 | Solid bacillus microbial agent and preparation method thereof |
| CN113005040B (en) * | 2021-01-22 | 2023-07-25 | 武汉微康益生菌研究院有限公司 | Bifidobacterium lactis freeze-drying protective agent and application method thereof |
| CN113462573A (en) * | 2021-07-12 | 2021-10-01 | 河北科技大学 | Preservation method of agricultural bacillus liquid microbial inoculum |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102220258A (en) * | 2011-04-20 | 2011-10-19 | 天津科技大学 | Bacillus subtilis generating nattokinase and method for producing nattokinase by fermenting same |
| CN102246969A (en) * | 2011-05-20 | 2011-11-23 | 中国肉类食品综合研究中心 | Starter and preparation method and use thereof |
-
2011
- 2011-12-28 CN CN 201110446721 patent/CN102433287B/en not_active Expired - Fee Related
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102220258A (en) * | 2011-04-20 | 2011-10-19 | 天津科技大学 | Bacillus subtilis generating nattokinase and method for producing nattokinase by fermenting same |
| CN102246969A (en) * | 2011-05-20 | 2011-11-23 | 中国肉类食品综合研究中心 | Starter and preparation method and use thereof |
Non-Patent Citations (2)
| Title |
|---|
| 古绍彬等.直投式酸奶发酵剂高密度培养工艺的研究.《食品工业科技》.2009,第30卷(第11期),摘要. |
| 直投式酸奶发酵剂高密度培养工艺的研究;古绍彬等;《食品工业科技》;20091231;第30卷(第11期);摘要 * |
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