CN1958780B - Solid-state fermentation bag, and application - Google Patents
Solid-state fermentation bag, and application Download PDFInfo
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- CN1958780B CN1958780B CN2006100543183A CN200610054318A CN1958780B CN 1958780 B CN1958780 B CN 1958780B CN 2006100543183 A CN2006100543183 A CN 2006100543183A CN 200610054318 A CN200610054318 A CN 200610054318A CN 1958780 B CN1958780 B CN 1958780B
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Abstract
This invention relates to a fermentation bag suitable for solid-state aerobic fermentation of a variety of microbes and its applications. The fermentation bag is made of microporous gas-permeable material, and the diameters of the micropores are 0.01-5.0 mum. The fermentation bag can satisfy solid-state aerobic fermentation of a variety of microbes, and can effectively prevent alien bacteria pollution. The fermentation process has high controllability. Besides, the fermentation bag has such advantages as high loading efficiency of fermentation equipment and saved space, and is suitable for mass production.
Description
Technical field
The present invention relates to a kind of solid microbe fermentation technology, particularly a kind of fermentation bag and application thereof that is fit to multiple microorganism such as fungi, bacterium, yeast and actinomycetic solid-state aerobic fermentation.
Background technology
Solid state fermentation is the technology that microorganism such as fungi, yeast, bacterium and actinomycetes utilize solid substrate to ferment and grow under certain temperature, humidity, oxygen condition.Be widely used in field production zymin, single cell protein, organic acid, alcohol and biotic pesticide etc. such as foodstuffs industry, medicine, agricultural at present.Because solid state fermentation compares with liquid fermenting, have that management expectancy is not high, technical process is simple, characteristics such as raw material sources are wide, cheap, and the low and cost of energy consumption is low, the current attention that is subjected to countries in the world gradually.
But there is heat passage difficulty height in solid state fermentation, stirs difficulty and produces shortcomings such as the site space utilization ratio is low, is difficult to reach the requirement of large-scale industrial production.In recent years, various countries drop into great amount of manpower and material resources solid state fermentation equipment are researched and developed, make solid fermentation industry developed, successively develop multiple solid-state fermentation installation, use more have disc type bio-reactor, air solid fluidized bed bio reactor, filling bed type bio-reactor, revolving drum type bioreactor and stirring type bioreactor etc. at present.These solid-state reactors exist that charging ratio is low, apparatus expensive, energy consumption are big, narrow application range and can not mass-producing etc. defective, make these fermentation equipment be difficult to wide popularization and application.As: air solid fluidized bed bio apparatus expensive, little, the power consumption of charging ratio; The bottom of filling bed type bio-reactor bed produces thermograde to the top, adopts forced ventilation to spread the microorganisms heat, makes dry materials, influences microorganism growth; Revolving drum type bioreactor and stirred reactor dispel the heat by shell, and heat transfer efficiencies descends when scale operation, and shearing force can be destroyed the mould structure of growing in rotation or whipping process simultaneously, is not suitable for the fermentation of mould etc.
The domestic industry solid state fermentation generally adopts low legacy equipment tray or the cultivation pool of input to ferment at present.Tray or cultivation pool fermentation have heat and mass efficient height, can effectively regulate the temperature of fermentation, wet, gas by envrionment conditions, can satisfy most aerobic solid ferment process in theory.But tray or cultivation pool fermentation are open system, easily pollute, and have influenced the purity of tunning.For the contaminated solution problem, people adopt the polypropylene plastics pocket sealing and fermenting that has transom to replace tray or cultivation pool fermentation, have reduced the assorted probability of fermentation sense to a certain extent.But because small area air ventilation membrane air flow is limited, the supply of oxygen is limited, and the charging volume of plastics bag is between 1～4 liter, be difficult to expand the scale of production, in addition, moisture content distributes difficulty, the water of condensation that produces between yeast phase influences thalli growth, and discharging also can increase the assorted chance of sense when dry.
Summary of the invention
The object of the present invention is to provide a kind of assorted bacterium of effectively avoiding infection, be fit to the fermentation bag of multiple microbe in solid state aerobic fermentation.Adopt this fermentation bag to ferment, can reach the requirement of large-scale industrial production.
Another object of the present invention is to provide new purposes, i.e. new application in solid state fermentation with above-mentioned solid-state fermentation bag.
The present invention is achieved in that a kind of solid-state fermentation bag, it is characterized in that: it is made by the micro-porous permeable material, and its micropore size is 0.01～5.0 μ m.
Above-mentioned micro-porous permeable material is bilayer or the three-decker that is composited by micro-porous permeable rete and ventilative carrier layer.When being three-decker, the micro-porous permeable rete is arranged between the two ventilative carrier layer.
Above-mentioned micro-porous permeable rete or be PVDF membrane layer (PVDF) or for tetrafluoroethylene rete (PTFE) or for polyethersulfone rete (PES) or for CN-CA layer (CN-CA) or for polypropylene millipore filtration layer, its aperture is 0.01～5.0 μ m.
Above-mentioned ventilative carrier layer or be the woven layer of cloth of polyester or for polyester acupuncture carpet veneer or for the woven layer of cloth of glass fibre.In fact, key of the present invention is the selection of micro-porous permeable rete, and for ventilative carrier layer, what it played only is the effect of carrier, so long as the gas permeable material that possesses certain intensity all can, the aperture of this carrier layer also can be greater than the micropore size of microporous breathable film.
When adopting solid-state fermentation bag fermentation materials of the present invention, under aseptic condition, pack into through the described solid-state fermentation bag and the sealing of sterilising treatment connecing bacterial classification after the solid culture material sterilising treatment earlier, be placed on then in proving room or the fermentation unit, by temperature, relative humidity and the air capacity control fermentation of regulating proving room or fermentation unit.
The present invention also provides the new application of the used micro-porous permeable material of above-mentioned solid-state fermentation bag, i.e. its application in the fermentation unit of conduct production muscardine spore, beauveria bassiana spore, Bt Biotrol BTV, tropina.
Adopt solid-state fermentation bag of the present invention, the thickness of the material of adorning can be regulated according to material properties and strain characteristic etc. in its bag, because the micro-porous permeable material has good venting capability, extraneous oxygen can effectively see through each layer of gas permeable material and enter in the fermentation bag, satisfy the demand of fermentation to oxygen, simultaneously heat of fermentation discharges in the fermentation bag, is equivalent to the thin layer fermentation, can effectively control fermentation by environment temperature, wet, gas.In addition, the aperture of the microporous breathable film of described micro-porous permeable material is 0.01～5 μ m, has water-proof function, can effectively stop the microorganism and the tunning that enter and control in the fermentation bag of extraneous assorted bacterium to leak in the immobilized solid ferment process.The fermentation bag sealing of being made by the micro-porous permeable material has heat and mass efficient height, easy to control and resistant to pollution advantage when carrying out solid state fermentation, can satisfy the aerobic solid ferment process of microorganism.The fermentation later stage needs drying as tunning, the characteristics hermetically drying that the fermentation bag permeability that can utilize the micro-porous permeable material to make is good, thus prevent in the open drying process that leavened prod is subjected to polluting and tunning is diffused in the environment.
All things considered, the present invention has the following advantages:
1, applied widely.The fermentation bag that is made into because of the micro-porous permeable material carries out solid state fermentation to have every bacterium, ventilating performance, and heat and mass efficient height, easy to control and resistant to pollution advantage can satisfy the aerobic solid ferment process of multiple microorganism in the fermenting process.
2, effectively prevent living contaminants.Microorganism is static fermentation in airtight fermentation bag, and the micro-pore diameter of microporous breathable film is 0.01～5.0 μ m, can effectively prevent the pollution of assorted bacterium, and anti-miscellaneous operation is convenient, reduces the requirement to yeasting.
3, fermenting process has good controllability.Fermentation bag is made for the micro-porous permeable material, can effectively with the external world gaseous interchange take place, and by temperature, humidity, the oxygen of dynamic adjustments yeasting, can effectively control fermentation.
4, adopt this fermentation bag to ferment, the charging ratio height of fermentation unit, conserve space easily realizes large-scale industrial production.
5, adopt fermentation bag of the present invention to carry out sealing and fermenting, the air seasoning after the fermentation can be carried out in position, avoids the stirring operation of traditional technology, reduces labour intensity.
Description of drawings
Below in conjunction with drawings and Examples the present invention is further described, but the present invention is not limited to this.
The bilayer structure synoptic diagram of the micro-porous permeable material that Fig. 1 embodiment of the invention 1 described solid-state fermentation bag adopts;
The three-decker synoptic diagram of the micro-porous permeable material that Fig. 2 embodiment of the invention 3 described solid-state fermentation bags adopt.
Embodiment
Embodiment 1: the solid-state fermentation bag fermentative production muscardine spore that adopts the micro-porous permeable material to make
Green muscardine fungus (Metarhizium) is one of disinsection fungal that is used for the earliest pest control, finds that now the whole world has 200 various insects to be caused death by this fungi infestation approximately, also is one of most important disinsection fungal.
Embodiment 1-1: a kind of solid-state fermentation bag is characterized in that: it is made by the micro-porous permeable material; Micro-porous permeable material in this example is the bilayer structure (referring to Fig. 1) that is composited by micro-porous permeable rete 1 and ventilative carrier layer 2; Its micro-porous permeable rete 1 is PVDF membrane layer (PVDF), and its aperture is 0.1 μ m, and its ventilative carrier layer 2 is the woven layer of cloth of polyester.The micro-porous permeable material is made the circle bag of diameter 20cm, long 40cm.
Embodiment 1-2: a kind of solid-state fermentation bag is characterized in that: it is made by the micro-porous permeable material; Micro-porous permeable material in this example is the bilayer structure (referring to Fig. 1) that is composited by micro-porous permeable rete 1 and ventilative carrier layer 2; Its micro-porous permeable rete 1 is tetrafluoroethylene rete (PTFE), and its aperture is 0.2 μ m, and its ventilative carrier layer 2 is the woven layer of cloth of polyester.The micro-porous permeable material is made the circle bag of diameter 20cm, long 40cm.
The contriver adopts described solid-state fermentation bag of the foregoing description 1-1,1-2 and traditional polypropylene fermentation bag respectively as the fermentation unit of producing muscardine spore:
The solid materials rice soaks 2～3h in 28 ℃ of water, 121 ℃ of moist heat sterilization 30min, and liquid green muscardine fungus bacterial classification is inserted in the cooling back, pours under aseptic condition in sterilization embodiment 1-1, the described fermentation circle of the 1-2 bag, loads coefficient 95%, the sealing sack; Is contrast with two ends with the aseptic polypropylene fermentation bag that seals the diameter 20cm of film phonograph seal, long 40cm, loading coefficient and be 30% (is its maximal packing coefficient, increase the filling coefficient again and will be difficult to heat radiation), insert the solid fermentation workshop, keep 25～28 ℃ of room temps, relative humidity 90%～100%.Ferment after 8 days, the beginning fan-drying, control group is poured material in the plastic tub into, expects thick 10cm (control group must be poured out material could be dry), and material is still stayed dried in place in the bag in embodiment 1-1, the 1-2 fermentation bag.Adopting cyclonic separator to collect spore behind fermentation, the dry end calculates spore production rate and measures spore water content, living contaminants rate (the results are shown in Table 1).
Table 1 solid-state fermentation bag and polypropylene fermentation bag are produced the correlation data of muscardine spore
As can be seen from Table 1: solid-state fermentation bag of the present invention is higher than under the condition of 3～5 times in conventional polypropylene bag at the material charging ratio, spore production rate and spore content reach or a little more than conventional polypropylene bag production level, illustrate that fermentation bag of the present invention is suitable for the solid-state aerobic fermentation of fungi; Water content and the Polypropylene Bag of fermentation bag tunning muscardine spore of the present invention after fan-drying, collection do not have significant difference, this is because solid-state fermentation bag of the present invention is fermenting the later stage, through powerful air draft the form that moisture passes through to evaporate in the fermentation bag is scattered and disappeared, illustrate that solid-state fermentation bag of the present invention still has good drying property under the condition of sealing; It is about 0.001% that solid-state fermentation bag of the present invention is produced muscardine spore living contaminants rate, only 1/10th of the spore living contaminants rate of producing for Polypropylene Bag, illustrate that solid-state fermentation bag of the present invention is suitable for producing the solid state fermentation strict to living contaminants.Therefore, adopt the solid-state fermentation bag of the present invention clear superiorities such as (being more than 3 times of Polypropylene Bag) of producing that the muscardine spore powder has easy, saving of work and time, pollution rate is low, throughput is high.
Embodiment 2: the solid-state fermentation bag fermentative production beauveria bassiana spore that adopts the micro-porous permeable material to make
Embodiment 2-1: a kind of solid-state fermentation bag is characterized in that: it is made by the micro-porous permeable material; Micro-porous permeable material in this example is the bilayer structure (referring to Fig. 1) that is composited by micro-porous permeable rete 1 and ventilative carrier layer 2; Its micro-porous permeable rete 1 is polyethersulfone rete (PES), and its aperture is 0.8 μ m, and its ventilative carrier layer 2 is a polyester acupuncture carpet veneer.The micro-porous permeable material is made the circle bag of diameter 16cm, long 80cm.
Embodiment 2-2: a kind of solid-state fermentation bag is characterized in that: it is made by the micro-porous permeable material; Micro-porous permeable material in this example is the bilayer structure (referring to Fig. 1) that is composited by micro-porous permeable rete 1 and ventilative carrier layer 2; Its micro-porous permeable rete 1 is CN-CA layer (CN-CA), and its aperture is 0.8 μ m, and its ventilative carrier layer 2 is a polyester acupuncture carpet veneer.The micro-porous permeable material is made the circle bag of diameter 16cm, long 80cm.
Embodiment 2-3: a kind of solid-state fermentation bag is characterized in that: it is made by the micro-porous permeable material; Micro-porous permeable material in this example is the bilayer structure (referring to Fig. 1) that is composited by micro-porous permeable rete 1 and ventilative carrier layer 2; Its micro-porous permeable rete 1 is a polypropylene millipore filtration layer, and its aperture is 0.8 μ m, and its ventilative carrier layer 2 is a polyester acupuncture carpet veneer.The micro-porous permeable material is made the circle bag of diameter 16cm, long 80cm.
Be control group still, except that the bacterium of adopting respectively is that the mode of production is with embodiment 1 muscardine with the polypropylene fermentation bag.Fermentation, dry back are adopted cyclonic separator to collect spore and are calculated spore production rate and measure the living contaminants rate and spore content (the results are shown in Table 2).
Table 2 solid-state fermentation bag and polypropylene fermentation bag are produced the correlation data of beauveria bassiana spore
As can be seen from Table 2: embodiment 2-1 of the present invention, 2-2,2-3 solid-state fermentation bag, can obviously improve fermented quality and productive rate, have good anti-polygamy energy simultaneously.
Embodiment 3: the solid-state fermentation bag fermentative production Biotrol BTV (BT) that adopts the micro-porous permeable material to make
Biotrol BTV (BT) is present most popular a kind of bacterium living beings agricultural chemicals, this sterilant all has in various degree toxic action to 526 kinds of insects of 10 orders, to lepidopterous kind more than 100, for example the insect of fruit, cotton, forest, tobaccos etc. such as cabbage caterpillar, cotton instar, Pyrausta nubilalis (Hubern)., oriental tobacco budworm all has toxic action, be the sterilant of wide spectrum, kill pests has broad application prospects in agricultural, health, environmental protection.
Embodiment 3-1: a kind of solid-state fermentation bag is characterized in that: it is made by the micro-porous permeable material; Micro-porous permeable material in this example is the three-decker (referring to Fig. 2) that is composited by micro-porous permeable rete 1 and ventilative carrier layer 2; Its micro-porous permeable rete 1 is tetrafluoroethylene rete (PTFE), and its aperture is 2 μ m, and its ventilative carrier layer 2 is the woven layer of cloth of polyester, and the tetrafluoroethylene rete is the middle layer.Microporous compound film is become the bar shaped bag of long 1m, wide 0.5m, thick 3cm ± 0.5cm.
Utilize the wheat bran solid medium to produce BT.Wheat bran, lime powder and water were mixed thoroughly by 5: 0.5: 4.5, and this routine described fermentation bag rear enclosed of packing into steams material 30min, and 0.5% commodity BT bacterium powder is inserted in the cooling back, puts on the fermentation frame of solid fermentation chamber, keeps 25～30 ℃ of temperature, relative humidity more than 90%.With the shallow tray fermentation is control group, and the wheat bran solid medium steams 30min cooling back and inserts 0.5% commodity BT bacterium powder, puts into the tray of proving room, expects thick 2cm ± 0.5cm.Ferment after 2 days analytical unit gemma number, toxicity evaluation (the results are shown in Table 3) after assorted situation of inspection sense and the drying and crushing.
Table 3 solid-state fermentation bag and shallow tray fermentation are produced the correlation data of Bacillus thuringiensis (BT)
As can be seen from Table 3: adopt solid-state fermentation bag fermentative production Bacillus thuringiensis of the present invention obviously to be better than shallow tray fermentation, the assorted rate of sense was zero when solid-state fermentation bag of the present invention was produced Bacillus thuringiensis, and the same period shallow tray fermentation the assorted rate of sense be 5.2%; Solid-state fermentation bag can directly obtain to contain several 280～34,000,000,000/g of gemma, and the Bacillus thuringiensis product that its poisoning is tired and reached 19000～20000IU/mg is significantly higher than shallow tray fermentation.
Embodiment 4: the solid-state fermentation bag fermentative production tropina that adopts the micro-porous permeable material to make
Embodiment 4-1: a kind of solid-state fermentation bag is characterized in that: it is made by the micro-porous permeable material; Micro-porous permeable material in this example is the bilayer structure (referring to Fig. 1) that is composited by micro-porous permeable rete 1 and ventilative carrier layer 2; Its micro-porous permeable rete 1 is tetrafluoroethylene rete (PTFE), and its aperture is 4 μ m, and its ventilative carrier layer 2 is the woven layer of cloth of glass fibre.The micro-porous permeable material is become the bar shaped bag of long 2m, wide 0.5m, thick 4cm ± 1cm.
With the waste residue behind rice, the Starch Production glucose is raw material, adopts cereuisiae fermentum manufacturing feedstuff protein by solid state fermentation (tropina).Waste residue, batching are added water and an amount of inorganic salt mixes thoroughly and makes that the substratum initial water content is 65%, initial pH6.2, above-mentioned solid-state fermentation bag sealing back steaming 30min packs into, it is the cereuisiae fermentum of 1 * 108/ml that concentration is inserted in the cooling back, inoculum size is 5%, put on the fermentation frame of solid fermentation chamber, leavening temperature is that 28～30 ℃, relative humidity are more than 90%.With the fermenting tank for fermentation is control group.Cultivate after 3 days, dry to constant weight for 65 ℃ and measure crude protein with Kjeldahl determination.The result shows: utilize the thick tropina of solid-state fermentation bag production to reach more than 30%, average out to 34.5% improves 30% than control group, illustrates that solid-state fermentation bag of the present invention is suitable for solid fermentation and produces animal feeding-stuff containing somatic protein.
Claims (8)
1. solid-state fermentation bag, it is characterized in that: it is made by the micro-porous permeable material, and its micropore size is 0.01～5.0 μ m; Described micro-porous permeable material is bilayer or the three-decker that is composited by micro-porous permeable rete (1) and ventilative carrier layer (2).
2. solid-state fermentation bag as claimed in claim 1 is characterized in that: the micro-porous permeable rete (1) of described three-decker is arranged between the two ventilative carrier layer (2).
3. solid-state fermentation bag as claimed in claim 1 or 2, it is characterized in that: described micro-porous permeable rete (1) or for the PVDF membrane layer or for the tetrafluoroethylene rete or for the polyethersulfone rete or for the CN-CA layer or for polypropylene millipore filtration layer, its aperture is 0.01～5.0 μ m; Described ventilative carrier layer (2) or be the woven layer of cloth of polyester or for polyester acupuncture carpet veneer or for the woven layer of cloth of glass fibre.
4. method with the described solid-state fermentation bag fermentation materials of the arbitrary claim of claim 1～3, it is characterized in that: during fermentation materials, under aseptic condition, pack into through the described solid-state fermentation bag and the sealing of sterilising treatment connecing bacterial classification after the solid culture material sterilising treatment earlier, be placed on then in proving room or the fermentation unit, by temperature, relative humidity and the air capacity control fermentation of regulating proving room or fermentation unit.
5. as the application of the described solid-state fermentation bag of 1～3 arbitrary claim in the fermentation unit of conduct production muscardine spore.
6. as the application of the described solid-state fermentation bag of 1～3 arbitrary claim in the fermentation unit of conduct production beauveria bassiana spore.
7. as the application of the described solid-state fermentation bag of 1～3 arbitrary claim in the fermentation unit of conduct production Biotrol BTV.
8. as the application of the described solid-state fermentation bag of 1～3 arbitrary claim in the fermentation unit of conduct production tropina.
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| CN106234669A (en) * | 2016-06-24 | 2016-12-21 | 中国土产畜产进出口总公司 | Grow dim and use fermentation bag in a kind of Fu-brick tea surface |
| CN110305801B (en) * | 2019-07-31 | 2021-04-09 | 四川省农业科学院经济作物育种栽培研究所 | Method for culturing beauveria bassiana spore powder by using fungus bags |
| CN112063537A (en) * | 2020-09-24 | 2020-12-11 | 瑞楚生物科技(江苏)有限公司 | Method capable of maintaining spore viability in discothrix bacterium liquid |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1328864A (en) * | 2001-06-06 | 2002-01-02 | 周宗娴 | Composite filtering film with ground filled with porous material and its production method |
| CN1342512A (en) * | 2001-09-04 | 2002-04-03 | 华东理工大学 | Millipore filter film for fermentation industry and its preparing process |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN1328864A (en) * | 2001-06-06 | 2002-01-02 | 周宗娴 | Composite filtering film with ground filled with porous material and its production method |
| CN1342512A (en) * | 2001-09-04 | 2002-04-03 | 华东理工大学 | Millipore filter film for fermentation industry and its preparing process |
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| CN 1328864 A,摘要和权利要求书. |
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