CN102994380B - The preparation technology of concentrated lactic acid bacterium leavening agent and special equipment thereof - Google Patents
The preparation technology of concentrated lactic acid bacterium leavening agent and special equipment thereof Download PDFInfo
- Publication number
- CN102994380B CN102994380B CN201210594501.8A CN201210594501A CN102994380B CN 102994380 B CN102994380 B CN 102994380B CN 201210594501 A CN201210594501 A CN 201210594501A CN 102994380 B CN102994380 B CN 102994380B
- Authority
- CN
- China
- Prior art keywords
- lactic acid
- fermentation
- micro filter
- ceramic micro
- tank
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Separation Using Semi-Permeable Membranes (AREA)
Abstract
The invention discloses preparation technology and the special equipment thereof of a kind of concentrated lactic acid bacterium leavening agent.The special equipment of this concentrated lactic acid bacterium leavening agent preparation technology, it includes a charging, fermentation system I and ceramic membrane filter system II；Connect between this system I and system II.The preparation technology of this concentrated lactic acid bacterium leavening agent, including: use special equipment as the aforementioned, during lactobacillus-fermented cultivation to exponential phase, ceramic micro filter film is used to be filtered to remove metabolite inhibitor and Partial fermentation liquid, supplement new fermentation culture subsequently, repeat aforementioned operation 1 ~ 3 time afterwards, then fermentation liquor ceramic micro filter membrane filtration is i.e. obtained lactic acid bacteria fermenting agent.The preparation technology of the present invention and the setting especially of special equipment thereof can effectively realize the continuous High Density Cultivation of lactic acid bacteria, and thalline survival rate is high, and ensure that bacterium solution lengthy fermentation, improves lactic acid bacteria thalline accumulation.
Description
Technical field
The present invention relates to preparation technology and the special equipment thereof of a kind of concentrated lactic acid bacterium leavening agent.
Background technology
In recent years, along with raising and the enhancing of health perception of living standards of the people, China's fermented dairy product
Yield with every year on average 25% speed increment, the consumption per head of cheese product is also improving year by year.Send out
Ferment agent effect in Yoghourt and cheese manufacturing process is extremely important, choosing and making thus for leavening
One of standby deciding factor becoming dairy industry development.As far back as developed countries such as 19 end of the centurys, America and Europes
Starting to develop starter culture concentrates, China's lactic acid bacteria fermenting agent research is fewer, has independent intellectual property right
The research and development of lactic acid bacteria fermenting agent almost blank.
In addition to having independent intellectual property right strain breeding thereof aspect, the production equipment of leavening industrialization is also
Another important aspect.In existing preparation technology and equipment, carry out lactic acid bacteria cultivation by fermentation tank,
Obtaining Lactobacillus Survival the highest, culture efficiency is relatively low.Prepared by efficient concentration type leavening it is crucial that want
Lactic acid bacteria is carried out High Density Cultivation, therefore, develops a kind of low cost input, concentration with short production cycle
Preparation technology and the special equipment thereof of lactic acid bacteria fermenting agent are imperative.
Summary of the invention
The technical problem to be solved is, it is provided that the preparation work of a kind of concentrated lactic acid bacterium leavening agent
Skill and special equipment thereof.The preparation technology of the present invention and arranging especially and can effectively realizing of special equipment thereof
The continuous High Density Cultivation of lactic acid bacteria, thalline survival rate is high, and ensure that bacterium solution lengthy fermentation, carries
High lactic acid bacteria thalline accumulation.Present invention incorporates ceramic membrane filter method, chemistry neutralisation, fed-batch
Cultivation etc., and in ceramic membrane filter technique, use different pore size ceramic membrane series system, it is greatly improved
Lactic acid bacteria culture density, improves leavening production efficiency.
The invention provides the special equipment of a kind of concentrated lactic acid bacterium leavening agent preparation technology, it include into
Material, fermentation system I and ceramic membrane filter system II；Connect between this system I and system II；
Described charging, fermentation system I include a head tank, a fermentation tank and a pH-meter；Should
Head tank is connected with the entrance of this charging, the fermentation tank of fermentation system, and the outlet manifold of this fermentation tank is two
Road, a road is discharging opening, and another road is through the entrance of a filter and a feeding engine with this filtration system II
Connecting, this fermentation tank and a pH-meter are connected；The outlet conduit of this head tank, this fermentation tank
It is equipped with valve between outlet conduit, pH-meter and fermentation tank, the inlet duct of this fermentation tank sets
There is a feeding engine, this fermentation tank is additionally provided with a warning inspection including a pH electrode and a temperature probe
Measurement equipment；
Described ceramic membrane filter system II includes two ceramic micro filter film equipments, two ceramic micro filter film equipments
Series connection forms ceramic micro filter film equipment group, and the entrance of this ceramic micro filter film equipment group passes through a feeding engine and
Filter is connected with the outlet of described charging, the fermentation tank of fermentation system I, this ceramic micro filter film equipment group
Outlet be connected with the entrance of described charging, the fermentation tank of fermentation system I, this ceramic micro filter film equipment group
In each ceramic micro filter film equipment be provided with a filter liquor discharging opening, the inlet tube of this ceramic micro filter film equipment group
Road, outlet conduit are equipped with valve, the pottery in this ceramic micro filter film equipment group, close to feeding engine
The ceramic micro filter membrane aperture of microfiltration membranes equipment is more than the ceramic micro filter membrane aperture of another ceramic micro filter film equipment.
In the present invention, described special equipment the most also includes purging system (CIP system) III on the spot；
Entirety after described system I and the series connection of system II is connected with system III；This on the spot purging system III include
An acid tank, an alkali tank and a deionization water pot, this acid tank, this alkali tank and this deionized water parallel with one another
It is equipped with valve on the inlet duct of tank and outlet conduit, this acid tank, this alkali tank and this deionization water pot
It is additionally provided with total valve, this acid tank, this alkali tank and the outlet conduit of this deionization water pot after inlet duct parallel connection
Further respectively have pump.
In the present invention, described fermentation tank is preferably also provided with the interlayer of circulation cycle water.
In the present invention, described pH electrode is preferably with the alarm detection equipment precision of described temperature probe
± 0.1 unit.
In the present invention, at the entrance and exit of described ceramic filtration membrane equipment group, flow detector can be provided with respectively
And pressure detector, its precision is preferably ± 0.1 unit.
In the present invention, in described ceramic micro filter film equipment, the material of ceramic micro filter film is preferably Al2O3,
Or Al2O3With ZrO2Composite.The shell of described ceramic micro filter film equipment is preferably ALSL304
Type or ALSL316 type stainless steel, be more preferably ALSL316 type stainless steel.
A ceramic micro filter film dress in the present invention, in described ceramic micro filter film equipment group, close to feeding engine
Standby ceramic micro filter film pore size is preferably 0.20 μm～0.45 μm, the hole of another ceramic micro filter film
Footpath is preferably sized to 0.02 μm～0.20 μm.
In the present invention, in described pH-meter, be preferably equipped with the ammoniacal liquor of 4～7mol/L, more preferably equipped with
The ammoniacal liquor of 5mol/L.
In the present invention, the pH value in described fermentation tank is preferably stablized by described pH-meter
In the range of pH5.5 ~ 6.2, preferably pH5.5 ~ 6.0 or pH6.0 ~ 6.2.
Present invention also offers the preparation technology of a kind of concentrated lactic acid bacterium leavening agent, it comprises the steps:
Use special equipment as the aforementioned, during lactobacillus-fermented cultivation to exponential phase, use described pottery
Porcelain membrane filtration system removes metabolite inhibitor and original volume percentage 40% ~ 60% zymotic fluid, subsequently
Supplement new fermentation culture, repeat aforementioned operation 1 ~ 3 time afterwards, then by fermentation liquor ceramic membrane filter system
System filters and i.e. obtains lactic acid bacteria fermenting agent.
In the present invention, described lactic acid bacteria can be for the lactic acid bacteria described in the routine of this area or its mixed lactic
Bacterium.The preferred streptococcus thermophilus of described mixing lactic acid bacteria (S.thermophilus) and moral formula lactobacillus Bulgaria
Subspecies (L.delbrueckii subsp.bulgaricus) mixing lactic acid bacteria, or streptococcus thermophilus (S.
Thermophilus), moral formula lactobacillus subspecies bulgaricus (L.delbrueckii subsp.bulgaricus) and addicted to
Lactobacillus lactis (L.acidophilus) mixing lactic acid bacteria.
In the present invention, lactobacillus-fermented is preferably spread cultivation through twice by lactic acid bacteria strains before cultivating and is followed by
Plant and cultivate.The inoculum concentration of fermented and cultured is preferably 2mL/100mL～3mL/100mL.
In the present invention, the fermentation culture conditions of described lactic acid bacteria can be the lactobacillus-fermented that this area is conventional
Condition of culture, is specifically adjusted zymotic fluid and condition of culture according to the concrete bacterial classification cultivated.
In the present invention, the described mensuration during exponential phase is obtained between pH value preferably by measuring
, described pH value preferably pH5.0～pH6.2 is exponential phase.
In the present invention, the temperature of described fermented and cultured preferably 35 DEG C～42 DEG C, more preferably 37 DEG C～40 DEG C.
In the present invention, described metabolite inhibitor refers to that produce in lactobacillus-fermented incubation has
Machine acid and other meeting lactic acid bacteria inhibiting continue the metabolite of fermented and cultured.
In the present invention, described fermentation culture can be the nutrient solution through sterilization, preferably through Pasteur
Sterilizing.Wherein, described pasteurize condition is preferably 115 DEG C～121 DEG C sterilization 4s～8s.Described
Fermentation culture preferred MRS culture medium.The concentration preferred mass mark of described MRS culture medium
The MRS culture medium of 6%.
In the present invention, the fluid treatment mode that described ceramic micro filter membrane filtration system filters preferably mistake
Flow operation.Wherein, the fluid treatment mode filtered when described ceramic micro filter membrane filtration system is cross-flow behaviour
When making, the tangential velocity of described zymotic fluid is preferably 0.5m/s～1m/s.
In the present invention, the mistake film pressure that described ceramic micro filter membrane filtration system filters is preferably
0.50MPa～0.90MPa.
In the present invention, in described ceramic micro filter membrane filtration system, the size of ceramic micro filter film is with m2
Meter is preferably for filtering fermentating liquid volume numerical value 0.005 in terms of L～0.050 times.
In the present invention, for keeping the suitable viscosity of zymotic fluid, when described ceramic micro filter membrane filtration system filters
Broth temperature is preferably 20 DEG C～35 DEG C, is more preferably 25 DEG C～30 DEG C.
In the present invention, in obtained lactic acid bacteria fermenting agent, cell concentration is preferably
1.0×1011Cfu/mL～8.5 × 1011cfu/mL。
In the present invention, after described lactic acid bacteria fermenting agent preparation, the most also to ceramic micro filter film with send out
Ferment tank cleans on the spot.Wherein, described cleaning on the spot is preferably:
1. deionized water pre-flush 5～8min；
2. 75 DEG C～85 DEG C of alkaline cleaner circulations 15～20min；
Described alkaline cleaner preferably mass percent be 1.5%～2.0% NaOH water-soluble
Liquid；
3. deionized water rinsing 5～8min；
4. 65 DEG C～70 DEG C of acidic cleaner circulations 15～20min；
Described acidic cleaner preferably mass percent is the aqueous solution of nitric acid of 1.0%～1.5%；
5. deionized water rinsing 5min；
Wherein, described deionized water can be the deionized water that this area is conventional, preferably deionized water
Index be content of organics ＜ 8mg/kg, pollutant index ＜ 3, silica ＜ 5mg/kg, iron ＜
0.5mg/kg, and manganese ＜ 0.5mg/kg.
In the present invention, described cleaning on the spot is preferably additionally operable to the sterilization before preparing lactic acid bacteria fermenting agent and disappears
Poison, described sterilizing condition is 85 DEG C～90 DEG C of deionization hot water circuit 25min～30min.
Agents useful for same of the present invention and raw material the most commercially, and are intended to do aseptic process.
Without prejudice to the field on the basis of common sense, above-mentioned in the present invention each technical characteristic can any group
Conjunction obtains preferred embodiments.
The most progressive effect of the present invention is: the invention provides the system of a kind of concentrated lactic acid bacterium leavening agent
Standby technique and special equipment thereof.This preparation technology and arranging especially of special equipment thereof can effectively realize breast
The continuous High Density Cultivation of acid bacterium, thalline survival rate is high.The pottery of two of which different pore size size especially
Microfiltration membranes, this filtration system can effectively get rid of lactobacillus-fermented metabolite, it is ensured that bacterium solution is sent out for a long time
Ferment, improves lactic acid bacteria thalline accumulation；Additionally this equipment introduces a pH-meter and also can regulate breast in real time
PH in acid bacteria fermentation tank so that it is stable in the range of certain pH, neutralizes the lactic acid that metabolism produces, reduces
The sour environment inhibitory action to lactobacter growth, thus promote lactic acid bacteria raised growth.
Accompanying drawing explanation
Fig. 1 is the special equipment knot of the concentrated lactic acid bacterium leavening agent preparation technology of a preferred embodiment of the present invention
Composition.
Detailed description of the invention
Further illustrate the present invention below by the mode of embodiment, but the most therefore limit the present invention to
Among described scope of embodiments.The experimental technique of unreceipted actual conditions in the following example, according to often
Rule method and condition, or select according to catalogue.
Embodiment 1 efficiently prepares the process special equipment of concentrated lactic acid bacterium leavening agent
As it is shown in figure 1, the present invention provides a kind of special equipment efficiently preparing lactic acid bacteria fermenting agent, it is main
Including charging, fermentation system I and the ceramic membrane filter system II being connected with this charging, fermentation system I；
This charging, fermentation system I include head tank S1 and fermentation tank S2；This head tank S1 with this
The entrance of ferment tank S2 connects, and the outlet manifold of this fermentation tank S2 is two-way, and a road is discharging opening, another
Road is connected with the entrance of this ceramic filter system II through filter P8, feeding engine P2；Fermentation tank S2
Connect a pH-meter S5；The outlet conduit of this head tank S1 is provided with valve 13, this fermentation tank S2
Two-way outlet pipeline on, discharging opening one tunnel is provided with valve 15, and another road is provided with valve 12, this fermentation
The connection pipeline of tank S2 and pH-meter S5 is provided with valve 14, the inlet duct of this fermentation tank S2
It is provided with feed pump P1, this fermentation tank S2 is additionally provided with and includes pH electrode P6's and temperature probe P7
Alarm detection equipment.
This ceramic membrane filter system II includes two ceramic micro filter film equipments, ceramic micro filter film S3 and pottery
Microfiltration membranes S4 is serially connected each pottery in formation ceramic micro filter film equipment group, and ceramic micro filter film equipment group
The pore size of microfiltration membranes is different and S3 aperture is relatively big, the entrance of ceramic micro filter film S3 and fermentation tank S2
Outlet connect, the outlet of ceramic micro filter film S4 is connected with the entrance of fermentation tank S2, ceramic micro filter film
Second outlet of S3 and ceramic micro filter film S4 is filter liquor discharging opening, and is provided with valve 24,25, pottery
Porcelain microfiltration membranes S3 inlet duct is provided with valve 21 and feeding engine P2；Ceramic micro filter film S3 is micro-with pottery
Filter membrane S4 two ends further respectively have flow, pressure-detecting device S11 and S10.
This is efficiently prepared lactic acid bacteria fermenting agent special equipment and also includes purging system III on the spot, cleans system on the spot
System III is connected with system I after whole series connection and II.This on the spot purging system include sour tank parallel with one another
S7, alkali tank S8 and deionization water pot S6, the inlet duct of acid tank S7 is provided with valve 34, outlet
Road is provided with valve 31, and the inlet duct of this alkali tank S8 is provided with valve 35, outlet conduit is provided with valve
Door 32, the inlet duct of this deionization water pot S6 is provided with valve 36, outlet conduit is provided with valve
33, it is additionally provided with total after the inlet duct parallel connection of this acid tank S7, this alkali tank S8 and this deionization water pot S6
Valve 23, the outlet conduit of this acid tank S7, this alkali tank S8 and this deionization water pot S6 sets the most respectively
There are pump P3, pump P4 and pump P5.
Below as a example by preparing lactic acid bacteria fermenting agent, specifically describe concentrated lactic acid bacterium leavening agent of the present invention and prepare
The process special equipment course of work is as follows:
Step one: use purging system (CIP system) on the spot to clean whole path.Closedown valve 11,
Valve 14, valve 15, valve 24, valve 25, valve 32, valve 31, valve 35, valve 34
With pump P3, pump P4, open other all valves and pump P5, pump P1, pump P2, use deionized water
After pre-flush 5～8min, close valve 11, valve 14, valve 15, valve 24, valve 25, valve
Door 31, valve 34, valve 36, valve 33 and pump P3, pump P5, open other all valves and pumps
P4, pump P1, pump P2, (described alkaline cleaner is preferably to use 75 DEG C～85 DEG C of alkaline cleaners
Sodium hydrate aqueous solution for mass percent is 1.5%～2.0%) circulate 15～20min after close valve closing
Door 11, valve 14, valve 15, valve 24, valve 25, valve 32, valve 31, valve 35,
Valve 34 and pump P3, pump P4, open other all valves and pump P5, pump P1, pump P2, and employing is gone
After ionized water pre-flush 5～8min, close valve 11, valve 14, valve 15, valve 24, valve
25, valve 32, valve 35, valve 36, valve 33 and pump P4, pump P5, opens other all valves
Door and pump P3, pump P1, pump P2, use 65 DEG C～70 DEG C of acidic cleaner (described acidic cleaners
Preferably for aqueous solution of nitric acid that mass percent is 1.0～1.5%) circulate 15～20min after, close
Valve 11, valve 14, valve 15, valve 24, valve 25, valve 32, valve 31, valve 35,
Valve 34 and pump P3, pump P4, open other all valves and pump P5, pump P1, pump P2, and employing is gone
Ionized water pre-flush 5～8min.
Wherein, the described concrete conditions in the establishment of a specific crime of cleaning tool on the spot is:
1. deionized water pre-flush 5～8min；
2. 75 DEG C～85 DEG C of alkaline cleaner circulations 15～20min；
Described alkaline cleaner is preferably water-soluble for NaOH that mass percent is 1.5%～2.0%
Liquid；
3. deionized water rinsing 5～8min；
4. 65 DEG C～70 DEG C of acidic cleaner circulations 15～20min；
Described acidic cleaner is preferably for aqueous solution of nitric acid that mass percent is 1.0～1.5%；
5. deionized water rinsing 5min；
Wherein, described deionized water is the deionized water that this area is conventional, is preferably deionized water
Index is content of organics ＜ 8mg/kg, pollutant index ＜ 3, silica ＜ 5mg/kg, iron ＜
0.5mg/kg, and manganese ＜ 0.5mg/kg.
In the present invention, described cleaning on the spot is preferably additionally operable to the sterilization before preparing lactic acid bacteria fermenting agent and disappears
Poison, described sterilizing condition is 85 DEG C～90 DEG C of deionization hot water circuit 25min～30min.
Step 2: access exponential phase lactic acid bacteria strains by head tank S1 and carry out in fermentation tank S2
Cultivate, utilize pH electrode P6 and temperature probe P7 detection growth situation that fermentation tank S2 connects,
When pH is down to 6.0, open valve 14, the 4 ~ 7mol/L ammoniacal liquor in pH-meter S5 is injected
In fermentation tank S2, within regulation pH maintains 5.5～6.2 number ranges, to neutralize lactobacillus-fermented product
The metabolites such as raw lactic acid, thus extend the exponential phase of lactobacter growth.
Step 3: the lactic acid bacteria strains in fermentation tank S2 is cultivated, visits through pH electrode P6 and temperature
When the alarm detection device detector detection lactic acid bacteria of pin P7 is cultivated to exponential phase later stage pH≤5.2,
Open valve 12, valve 21, other valves of valve 22(are turned off) and feeding engine P2, use
Ceramic micro filter film S3, S4 of series connection is filtered to remove metabolite inhibitor (such as lactic acid, acetic acid) and former
The zymotic fluid of percentage by volume 40% ~ 60%, when less than the 1/3 of filtrate flow prompt drop to initial flow rate,
Close valve 12, valve 21, valve 22 and feeding engine P2, stop micro-filtration operation.
Step 4: Open valve 13 and feeding engine P1 carry out feed supplement, then lactic acid bacteria to fermentation tank S2
In fermentation tank S1, proceed fermented and cultured, remaining step with two, step 3.So it is repeated
1～3 time.
Step 5: utilize fermentation tank S2 be provided with the interlayer of circulation cycle water zymotic fluid is cooled to 20 DEG C～
35℃；Hereafter Open valve 12, valve 21, valve 22 and other valves of feeding engine P2(all close
Close), the lactic acid bacteria thalline zymotic fluid in fermentation tank S2 is carried out micro-filtration filtration, filters after terminating, close
Valve 12, valve 21, valve 22 and feeding engine P2, and Open valve 15 i.e. can be from discharging
Concentrated lactic acid bacterium leavening agent is collected at Kou.
Step 6: after described concentrated lactic acid bacterium leavening agent preparation, the most also to ceramic micro filter film and
Fermentation tank cleans on the spot.Wherein, the described concrete operations of cleaning on the spot are with step one.
In following embodiment, percentage is mass percent.
In the present invention, described thalline counting mode uses flat board casting, will zymotic fluid or thalline mistake
Filtrate carries out the gradient dilution of 1:10 with SPSS (NaCl solution of 0.85wt%), to closing
Suitable dilution gradient, draws 1mL mycelium dilution liquid and is transferred on aseptic flat board, treating that sterile letheen is cultivated
When base (depending on the kind of culture medium is according to concrete bacterial classification) is cooled to 45 DEG C～50 DEG C, topple over 15mL～
25mL, on the flat board containing mycelium dilution liquid, mixes and static cooling.Each sample carries out 3 times puts down
Row test.Cooling, the flat board of solidification are positioned over permanent ability of swimming incubator and carry out cultivating (cultivation temperature and time
According to depending on bacterial classification), cultivate and count after terminating.
(streptococcus thermophilus, moral formula lactobacillus subspecies bulgaricus are mixed for embodiment 2 lactic acid bacteria mixed culture fermentation agent
Close leavening)
Formula for a product:
| Raw material | Ingredient requirement | Addition |
| Fermented by mixed bacterium agent | Spread cultivation twice | 10kg |
| MRS culture medium | Biochemical reagents | 60kg |
| Water | Food-grade | 930kg |
The preparation method of lactic acid bacteria fermenting agent includes the following: the mixed fermentation liquid that spreads cultivation through twice with 2
The ratio of mL/100mL is added to 5000L in the MRS fluid nutrient medium of the 6% of pasteurize, rises
Beginning Mixed Microbes bulk concentration is 9.5 × 109cfu/mL.Ferment under the conditions of 40 DEG C, utilize pH-meter to make
Mixed bacteria environment in fermentation tank maintains pH6.0 ~ 6.2, and after maintaining fermentation 6h, closes valve 14,
When mixed bacteria grows to the logarithmic phase later stage, pH is down to when 5.2 proceed by ceramic micro filter membrane filtration, phase
Between, fermented by mixed bacterium liquid is through a ceramic micro filter film feed operation.Through twice feed supplement and ceramic membrane repeatedly
After filtration, final collection obtains zymotic fluid 480L, and cell concentration is 4.0 × 1011Cfu/mL is thermophilic chain
Coccus, moral formula lactobacillus subspecies bulgaricus mixed culture fermentation agent.
Wherein, described culture medium pasteurize condition is: temperature 121 DEG C, 4 seconds time.
Wherein, the material of this ceramic micro filter film is Al2O3；The pore size of ceramic micro filter film S3 is
0.45 μm, the pore size of ceramic micro filter film S4 are 0.20 μm.The material of this ceramic micro filter film shell is
ALSL316 type stainless steel.
Wherein, described ceramic micro filter film filter condition is: pressure 0.20MPa, and tangential velocity is 0.5m/s,
Filtration temperature is 25 DEG C.
(streptococcus thermophilus, moral formula lactobacillus subspecies bulgaricus are mixed for embodiment 3 lactic acid bacteria mixed culture fermentation agent
Close leavening)
Formula for a product:
| Raw material | Ingredient requirement | Addition |
| Fermented by mixed bacterium agent | Spread cultivation twice | 10kg |
| MRS culture medium | Biochemical reagents | 60kg |
| Water | Food-grade | 930kg |
The preparation method of lactic acid bacteria fermenting agent includes the following: the mixed fermentation liquid that spreads cultivation through twice with 2
The ratio of mL/100mL is added to 5000L in the MRS fluid nutrient medium of the 6% of pasteurize, rises
Beginning Mixed Microbes bulk concentration is 9.5 × 109cfu/mL.Ferment under the conditions of 40 DEG C, utilize pH-meter to make
Mixed bacteria environment in fermentation tank maintains pH6.0 ~ 6.2, and after maintaining fermentation 6h, closes valve 14,
When mixed bacteria grows to the logarithmic phase later stage, pH is down to when 5.2 proceed by ceramic micro filter membrane filtration, phase
Between, fermented by mixed bacterium liquid is through a ceramic micro filter film feed operation.Through twice feed supplement and ceramic membrane repeatedly
After filtration, final collection obtains zymotic fluid 550L, and cell concentration is 1.0 × 1011Cfu/mL is thermophilic chain
Coccus, moral formula lactobacillus subspecies bulgaricus mixed culture fermentation agent.
Wherein, described culture medium pasteurize condition is: temperature 121 DEG C, 4 seconds time.
Wherein, the pore size of ceramic micro filter film S3 be 0.20 μm, ceramic micro filter film S4 aperture big
Little is 0.02 μm.
Wherein, described ceramic micro filter film filter condition is: pressure 0.15MPa, and tangential velocity is 1m/s,
Filtration temperature is 25 DEG C.
Embodiment 4 lactic acid bacteria mixed culture fermentation agent (streptococcus thermophilus, moral formula lactobacillus subspecies bulgaricus with
Lactobacillus acidophilus mixed culture fermentation agent)
Formula for a product (in terms of final products per ton):
| Raw material | Ingredient requirement | Addition |
| Fermented by mixed bacterium liquid | Spread cultivation twice | 15kg |
| MRS culture medium | Biochemical reagents | 60kg |
| Water | Food-grade | 925kg |
The preparation method of lactic acid bacteria fermenting agent includes the following: the fermented by mixed bacterium liquid that will spread cultivation through twice
Add to 5000L in the MRS fluid nutrient medium of the 6% of pasteurize with the ratio of 3mL/100mL,
Initial Mixed Microbes bulk concentration is 9.0 × 109cfu/mL.Ferment under the conditions of 37 DEG C, utilize pH-meter
Make the mixed bacteria environment in fermentation tank maintain pH5.5 ~ 6.0, and after maintaining fermentation 6h, close valve
14, when mixed bacteria grows to the logarithmic phase later stage, pH is down to 5.2 and proceeds by ceramic micro filter membrane filtration,
Period, fermented by mixed bacterium liquid is through a ceramic micro filter film feed operation.Through twice feed supplement and pottery repeatedly
After membrane filtration, final collection obtains zymotic fluid 480L, and cell concentration is 8.5 × 1011Cfu/mL is thermophilic
Streptococcus, moral formula lactobacillus subspecies bulgaricus and lactobacillus acidophilus mixed culture fermentation agent.
Wherein, described culture medium pasteurize condition is: temperature 121 DEG C, 4 seconds time.
Wherein, the material of this ceramic micro filter film is Al2O3With ZrO2Composite；Ceramic micro filter film
The pore size of S3 be 0.45 μm, the pore size of ceramic micro filter film S4 be 0.20 μm.
Wherein, described ceramic micro filter film filter condition is: pressure 0.20MPa, and tangential velocity is 0.5m/s,
Filtration temperature is 25 DEG C.
Embodiment 5 lactic acid bacteria mixed culture fermentation agent (streptococcus thermophilus, moral formula lactobacillus subspecies bulgaricus with
Lactobacillus acidophilus mixed culture fermentation agent)
Formula for a product (in terms of final products per ton):
| Raw material | Ingredient requirement | Addition |
| Fermented by mixed bacterium liquid | Spread cultivation twice | 15kg |
| MRS culture medium | Biochemical reagents | 60kg |
| Water | Food-grade | 925kg |
The preparation method of lactic acid bacteria fermenting agent includes the following: the fermented by mixed bacterium liquid that will spread cultivation through twice
Add to 5000L in the MRS fluid nutrient medium of the 6% of pasteurize with the ratio of 3mL/100mL,
Initial Mixed Microbes bulk concentration is 9.0 × 109cfu/mL.Ferment under the conditions of 37 DEG C, utilize pH-meter
Make the mixed bacteria environment in fermentation tank maintain pH5.5 ~ 6.0, and after maintaining fermentation 6h, close valve
14, when mixed bacteria grows to the logarithmic phase later stage, pH is down to 5.2 and proceeds by ceramic micro filter membrane filtration,
Period, fermented by mixed bacterium liquid is through a ceramic micro filter film feed operation.Through twice feed supplement and pottery repeatedly
After membrane filtration, final collection obtains zymotic fluid 505L, and cell concentration is 1.2 × 1011Cfu/mL is thermophilic
Streptococcus, moral formula lactobacillus subspecies bulgaricus and lactobacillus acidophilus mixed culture fermentation agent.
Wherein, described culture medium pasteurize condition is: temperature 121 DEG C, 4 seconds time.
Wherein, the material of this ceramic micro filter film is Al2O3With ZrO2Composite；Ceramic micro filter film
The pore size of S3 be 0.20 μm, the pore size of ceramic micro filter film S4 be 0.02 μm.
Wherein, described ceramic micro filter film filter condition is: pressure 0.15MPa, and tangential velocity is 1m/s,
Filtration temperature is 25 DEG C.
Claims (12)
1. the special equipment of a concentrated lactic acid bacterium leavening agent preparation technology, it is characterised in that include into
Material, fermentation system I and ceramic membrane filter system II；This charging, fermentation system I and ceramic membrane filter
Connect between system II；
Described charging, fermentation system I include a head tank, a fermentation tank and a pH-meter；Should
Head tank is connected with the entrance of this charging, the fermentation tank of fermentation system I, and the outlet manifold of this fermentation tank is
Two-way, a road is discharging opening, and another road is through a filter and a feeding engine and this ceramic membrane filter system
The entrance of II connects, and this fermentation tank and a pH-meter are connected；The outlet conduit of this head tank, should
It is equipped with valve, the entrance of this fermentation tank between the outlet conduit of fermentation tank, pH-meter and fermentation tank
Pipeline is provided with a feeding engine, this fermentation tank is additionally provided with one and includes a pH electrode and a temperature probe
Alarm detection equipment；
Described ceramic membrane filter system II includes two ceramic micro filter film equipments, two ceramic micro filter film equipments
Series connection forms ceramic micro filter film equipment group, and the entrance of this ceramic micro filter film equipment group passes through a feeding engine and
Filter is connected with the outlet of described charging, the fermentation tank of fermentation system I, this ceramic micro filter film equipment group
Outlet be connected with the entrance of described charging, the fermentation tank of fermentation system I, this ceramic micro filter film equipment group
In each ceramic micro filter film equipment be provided with a filter liquor discharging opening, the inlet tube of this ceramic micro filter film equipment group
Road, outlet conduit are equipped with valve, the pottery in this ceramic micro filter film equipment group, close to feeding engine
The ceramic micro filter membrane aperture of microfiltration membranes equipment is more than the ceramic micro filter membrane aperture of another ceramic micro filter film equipment；
In described ceramic micro filter film equipment group, the pottery of the ceramic micro filter film equipment close to feeding engine is micro-
Filter sizes size is 0.20 μm～0.45 μm, and the pore size of another ceramic micro filter film is
0.02 μm～0.20 μm.
2. special equipment as claimed in claim 1, it is characterised in that: described special equipment also includes
One purging system III on the spot；Described charging, fermentation system I and ceramic membrane filter system II series connection after whole
Body is connected with purging system III on the spot；This on the spot purging system III include a parallel with one another acid tank, one
Alkali tank and a deionization water pot, this acid tank, this alkali tank and the inlet duct of this deionization water pot and outlet
It is equipped with valve on road, is additionally provided with after the inlet duct parallel connection of this acid tank, this alkali tank and this deionization water pot
Total valve, the outlet conduit of this acid tank, this alkali tank and this deionization water pot further respectively has pump.
3. special equipment as claimed in claim 1, it is characterised in that: described fermentation tank is additionally provided with stream
The interlayer of logical recirculated water；
And/or, it is respectively equipped with flow detector and pressure at the entrance and exit of described ceramic micro filter film equipment group
Force detector；
And/or, in described ceramic micro filter film equipment, the material of ceramic micro filter film is Al2O3, or Al2O3
With ZrO2Composite；
And/or, the shell of described ceramic micro filter film equipment is ALSL304 type or ALSL316 type stainless steel；
And/or, described pH-meter built with 4～7mol/L ammoniacal liquor.
4. special equipment as claimed in claim 1, it is characterised in that: the pH value in described fermentation tank
Stable in the range of pH5.5～6.2 by described pH-meter.
5. special equipment as claimed in claim 4, it is characterised in that: the pH value in described fermentation tank
Stable in the range of pH5.5～6.0 or pH6.0～6.2 by described pH-meter.
6. the preparation technology of a concentrated lactic acid bacterium leavening agent, it is characterised in that comprise the steps: to make
With the special equipment as described in any one of Claims 1 to 5, lactobacillus-fermented is cultivated to exponential phase
Period, described ceramic membrane filter system II is used to remove metabolite inhibitor and original volume percentage
40%～60% zymotic fluid, supplements new fermentation culture subsequently, repeats aforementioned operation 1～3 times afterwards, then will
Fermentation liquor ceramic membrane filter system II filters and i.e. obtains lactic acid bacteria fermenting agent.
7. preparation technology as claimed in claim 6, it is characterised in that: described lactic acid bacteria is ability
Lactic acid bacteria described in the routine of territory or its mixing lactic acid bacteria；Described mixing lactic acid bacteria is streptococcus thermophilus (S.
Thermophilus) mix with moral formula lactobacillus subspecies bulgaricus (L.delbrueckii subsp.bulgaricus)
Close lactic acid bacteria, or streptococcus thermophilus (S.thermophilus), moral formula lactobacillus subspecies bulgaricus (L.
Delbrueckii subsp.bulgaricus) and lactobacillus acidophilus (L.acidophilus) mixing lactic acid bacteria.
8. preparation technology as claimed in claim 6, it is characterised in that: before lactobacillus-fermented is cultivated
By lactic acid bacteria strains inoculated and cultured after twice spreads cultivation；The inoculum concentration of fermented and cultured be 2mL/100mL～
3mL/100mL；
And/or, the described mensuration during exponential phase obtains indirectly by measuring pH value, described
PH value is that pH5.0～pH6.2 is exponential phase；
And/or, the temperature of described fermented and cultured is 35 DEG C～42 DEG C；
And/or, described fermentation culture is the nutrient solution through sterilization.
9. preparation technology as claimed in claim 8, it is characterised in that: the temperature of described fermented and cultured
Degree is 37 DEG C～40 DEG C；
And/or, described fermentation culture is the nutrient solution through pasteurize.
10. preparation technology as claimed in claim 9, it is characterised in that: the bar of described pasteurize
Part is 115 DEG C～121 DEG C sterilization 4s～8s.
11. preparation technologies as claimed in claim 6, it is characterised in that: described fermentation culture is
MRS culture medium；
And/or, the fluid treatment mode that described ceramic membrane filter system II filters is cross-flow operation；
And/or, the mistake film pressure that described ceramic membrane filter system II filters is 0.50MPa～0.90MPa；
And/or, in described ceramic membrane filter system II, the size of ceramic micro filter film is with m2Meter, for
Filtering fermentating liquid volume numerical value 0.005 in terms of L～0.050 times；
And/or, when described ceramic membrane filter system II filters, broth temperature is 20 DEG C～35 DEG C；
And/or, in obtained lactic acid bacteria fermenting agent, cell concentration is 1.0 × 1011Cfu/mL～
8.5×1011cfu/mL；
And/or, after described lactic acid bacteria fermenting agent preparation, also the most clear to ceramic micro filter film and fermentation tank
Wash.
12. preparation technologies as claimed in claim 11, it is characterised in that: described MRS culture medium
The MRS culture medium that concentration is mass fraction 6%；
And/or, when the fluid treatment mode that described ceramic membrane filter system II filters is cross-flow operation,
The tangential velocity of described zymotic fluid is 0.5m/s～1m/s；
And/or, when described ceramic membrane filter system II filters, broth temperature is 25 DEG C～30 DEG C；
And/or, described cleaning on the spot is: 1. deionized water pre-flush 5～8min；2. 75 DEG C～85 DEG C
Alkaline cleaner circulation 15～20min；3. deionized water rinsing 5～8min；4. 65 DEG C～70 DEG C of acidity
Cleaning agent circulation 15～20min；5. deionized water rinsing 5min；Described alkaline cleaner is quality hundred
Proportion by subtraction is the sodium hydrate aqueous solution of 1.5%～2.0%；Described acidic cleaner is that mass percent is
The aqueous solution of nitric acid of 1.0%～1.5%；The index of described deionized water is content of organics ＜ 8mg/kg,
Pollutant index ＜ 3, silica ＜ 5mg/kg, iron ＜ 0.5mg/kg, and manganese ＜ 0.5mg/kg；
And/or, described cleaning on the spot is additionally operable to the sterilizing before preparing lactic acid bacteria fermenting agent, described
Sterilizing condition is 85 DEG C～90 DEG C of deionization hot water circuit 25min～30min.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210594501.8A CN102994380B (en) | 2012-12-31 | 2012-12-31 | The preparation technology of concentrated lactic acid bacterium leavening agent and special equipment thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210594501.8A CN102994380B (en) | 2012-12-31 | 2012-12-31 | The preparation technology of concentrated lactic acid bacterium leavening agent and special equipment thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102994380A CN102994380A (en) | 2013-03-27 |
| CN102994380B true CN102994380B (en) | 2016-08-31 |
Family
ID=47923494
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210594501.8A Active CN102994380B (en) | 2012-12-31 | 2012-12-31 | The preparation technology of concentrated lactic acid bacterium leavening agent and special equipment thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102994380B (en) |
Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103449853A (en) * | 2013-09-16 | 2013-12-18 | 云南宝鑫实业集团有限公司 | Full-automatic system for continuously producing microbial fertilizer |
| CN103710279B (en) * | 2013-10-18 | 2015-07-15 | 中国农业大学 | Staphylococcus simulans L-RG18 fermentation medium for use and high density culture method |
| FI126781B (en) | 2015-05-25 | 2017-05-31 | Neste Corp | Methods for continuous production of microorganisms |
| CN113789253B (en) * | 2021-09-30 | 2024-04-09 | 四川省食品发酵工业研究设计院有限公司 | Lactic acid bacteria high-density fermentation system and fermentation method |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101037247A (en) * | 2007-02-05 | 2007-09-19 | 山东齐东方投资有限公司 | Method for preparing biologic polymers flocculant by micrococcus |
| CN201850276U (en) * | 2010-05-07 | 2011-06-01 | 江苏大学 | Micro-filtration membrane coupling high-density culture device for lactic acid bacteria |
| CN102311912B (en) * | 2010-07-06 | 2013-07-31 | 光明乳业股份有限公司 | Preparation process of lactic acid bacteria starter and special equipment thereof |
-
2012
- 2012-12-31 CN CN201210594501.8A patent/CN102994380B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN102994380A (en) | 2013-03-27 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102311912B (en) | Preparation process of lactic acid bacteria starter and special equipment thereof | |
| Desmasures et al. | Microbiological composition of raw milk from selected farms in the Camembert region of Normandy | |
| CN102994380B (en) | The preparation technology of concentrated lactic acid bacterium leavening agent and special equipment thereof | |
| CN109536406B (en) | Weak post-acidification streptococcus thermophilus JMCC16, separation and purification method and application | |
| CN106754470A (en) | The Lactobacillus rhamnosus in one plant of lacto source and its application | |
| CN104940968B (en) | biological air freshener and preparation method thereof | |
| CN104818230A (en) | Lactobacillus plantarum L01 having cholesterol degrading function and application thereof | |
| CN107125745A (en) | Adjust ferment of gut flora and bowel relaxing functions and preparation method thereof | |
| CN110004096A (en) | One lactobacillus plantarum and its application | |
| CN102113556A (en) | Preparation method for leavening special for goat milk yoghourt | |
| CN101717813B (en) | Method for detecting total number of bacterial colonies in activated lactobacillus drink | |
| CN102250806A (en) | Method for screening lactic acid bacteria capable of producing bacteriocin from plant source materials | |
| CN101153316A (en) | Method for detecting lactobacillus casei in probiotic bacteria milk product | |
| CN105748534B (en) | A kind of compound lactobacillus nipple cleaning solution improving mammilla of milk cattle microecosystem | |
| CN101935688B (en) | Method for detecting and counting live microbes | |
| CN110250396A (en) | Pediococcus acidilactici CGMCC 1.4 is inhibiting the application in aspergillus flavus | |
| JP2007524417A5 (en) | ||
| JP2007524417A (en) | Method for producing a liquid concentrate of bacteria acclimatized to food grade and viable | |
| CN104894037A (en) | Lactococcus lactis subsp.lactis and use thereof for cheese production | |
| CN102369274B (en) | Method for culturing lactic acid bacterium and method for producing fermented milk | |
| CN210419820U (en) | Extraction element of nisin | |
| CN107043715A (en) | A kind of active probiotic freeze-dried powder and preparation method thereof | |
| CN111670815A (en) | Yoghourt processing production device and processing method thereof | |
| CN105349467A (en) | Time-sharing fermentation culture method of plant extract compound probiotics | |
| CN110279056A (en) | Bacillus coagulans ACCC10229 is inhibiting the application in aspergillus flavus |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |