CN104845877A - Solid state fermentation reactor and application thereof - Google Patents

Solid state fermentation reactor and application thereof Download PDF

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Publication number
CN104845877A
CN104845877A CN201510186386.4A CN201510186386A CN104845877A CN 104845877 A CN104845877 A CN 104845877A CN 201510186386 A CN201510186386 A CN 201510186386A CN 104845877 A CN104845877 A CN 104845877A
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China
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pipeline
coil pipe
fermentor tank
state fermentation
solid
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CN201510186386.4A
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Chinese (zh)
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冯小海
汤宝
梁金丰
徐虹
许宗奇
姜永翔
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南京工业大学
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Abstract

The invention discloses a solid state fermentation reactor and an application thereof. The reactor comprises a horizontal cylindrical stainless steel fermentation tank body, a stainless steel jacket is welded on the outer wall of the above tank, and the tank is also provided with a gas outlet, probes, a steam inlet, a steam outlet, a material outlet, movable top covers and a two-fluid spray device; and coil pipes and trays are arranged in the inner cavity of the tank body. The invention also discloses an application of the reactor in cultivation of aerobic, anaerobic and facultative anaerobic microbes. The reactor has the advantages of simple structure, convenience and practicality, can solve the problems of difficult material disinfection, easy bacterial contamination, large labor intensity and low working efficiency of traditional solid state fermentation, realizes automatic control of the solid state fermentation process, and facilitates industrial application of the solid state fermentation.

Description

A kind of solid-state fermentation reactor and application thereof

Technical field

The invention belongs to fermentation engineering and process engineering technical field, relate to a kind of solid-state fermentation reactor and application thereof.

Background technology

Solid state fermentation refers under not having or almost not having free water, the bioprocesses that one or more microorganisms carry out on solid substrate, there is the advantages such as simple to operate, energy consumption is low, fermenting process is easy to control, agriculture residues, trade waste particularly can be utilized as solid substrate.At present in biological manufacturing industry, liquid submerged fermentation is still in dominant position, and tracing it to its cause is that shortage one is multi-functional, the reaction unit of fermentation stability.Existing solid fermentation device mainly contains shallow tray fermentation, aerobic fermentation pond, drum-type fermentation etc., but the shortcomings such as usually shallow tray fermentation condition is rough, ventilation and heat performance is poor, fermenting process is wayward, there is the drawbacks such as charging discharging is loaded down with trivial details, labour intensity is high, easy microbiological contamination in fermenting tank for fermentation, serious, the huge shearing force of drum-type fermentation energy consumption affects quality product, and these reactor function singlenesses, cannot the process such as the sterilizing of complete independently raw material, fermentation, drying, fermenting process is also not easy to control.Defect on visible equipment causes solid state fermentation to complete industrial large-scale production, also causes the application of solid-state fermentation technology almost to stay cool, and therefore, develops a kind of solid-state fermentation reactor that is multiple functional, simple and practical, fermentation stability extremely urgent!

Summary of the invention

Technical problem to be solved by this invention be overcome exist in prior art function singleness, complex structure and the problem such as impracticable, the solid-state fermentation reactor of functions such as providing a kind of and can wet to the sterilizing needed for solid materials fermentation, cooling, inoculation, fermentation, temperature control, control, ventilate, be dry.

The technical problem that the present invention also will solve is to provide above-mentioned solid-state fermentation installation and is cultivating the application in aerobic microbiological, anaerobion and amphimicrobe.

For solving the problems of the technologies described above, the technical solution used in the present invention is as follows:

A kind of solid-state fermentation reactor, it comprises fermentor tank, coil pipe, tray and second fluid nozzle system;

Described fermentor tank is horizontal fermentor tank, and fermentor tank periphery is provided with chuck, and both sides, fermentation tank level direction are provided with the first removable top cover and the second removable top cover;

Fermentor tank inside is provided with 1 ~ 10 layer of coil pipe from top to bottom, movable storing tray on every layer of coil pipe; Every layer of coil pipe is hollow structure, and one end of coil pipe is water-in, and the other end is water outlet;

Fermentor tank top is provided with production well, is provided with temp probe, moisture probe and pressure probe near production well;

Position near removable top cover in fermentor tank vertically arranges second fluid nozzle system, described second fluid nozzle system comprises 1 ~ 10 nozzle and second fluid nozzle pipeline, nozzle is connected by the outside gas-liquid pipeline of second fluid nozzle pipeline and fermentor tank, and the outside gas-liquid pipeline of described fermentor tank is sterilized water pipeline, seed liquor pipeline, vapour line and gas piping parallel with one another.

Wherein, described fermentor tank is stainless steel, and fermentor tank bottom is provided with discharge port, and fermentor tank is fixed on base by the first support of bottom and the second support.

Wherein, described chuck is provided with steam inlet and vapour outlet, and steam inlet is located at chuck top, and vapour outlet is located at chuck lowermost end, and steam inlet and vapour outlet position in the horizontal direction away from, improve steam to the heats of tank body.

Wherein, the first removable top cover and fermentor tank sidewall junction are provided with standing bolt, and the second removable top cover and ferment pot sidewall junction are provided with standing bolt.

Wherein, fermentor tank inside is provided with three layers of coil pipe from top to bottom, be respectively the first coil pipe, the second coil pipe and the 3rd coil pipe, coil pipe is fixed on fermentor tank inside by stiffened steel plates, movable storing the first tray on first coil pipe, movable storing the second tray on second coil pipe, movable storing the 3rd tray on the 3rd coil pipe.

Wherein, described tray is iron pallet, and tray bottom even is drilled with several aperture; Wherein, the aperture of preferred aperture is 2mm, and pitch of holes is 2cm.

Wherein, described coil pipe is stainless steel pipes, and plan view shape is snakelike, and the water-in of coil pipe is communicated with outlet conduit respectively at the inlet channel outside fermentor tank with water outlet, and passing into cold water can cool tank body, and passing into hot water can carry out drying to material.

Wherein, described second fluid nozzle optimum system choosing comprises 4 nozzles, the first second fluid nozzle pipeline and the second second fluid nozzle pipeline, 4 nozzles adopt mode in parallel simultaneously with the first second fluid nozzle pipeline and the second second fluid nozzle pipeline connection; Sterilized water pipeline and vapour line parallel with one another, then to connect with the first second fluid nozzle pipeline; Seed liquor pipeline and gas piping parallel with one another, then to connect with the second second fluid nozzle pipeline; Sterilized water pipeline is provided with first liquid strainer and second liquid strainer, and vapour line is provided with vapour generator, the external inoculation bottle of seed liquor pipeline, and gas piping is provided with the first gas filter and the second gas filter; Sterilized water pipeline and seed liquor pipeline are respectively equipped with gas meter, vapour line and gas piping are respectively equipped with gas meter.

Wherein, 4 nozzles can provide fan spray pattern in fermentor tank.

Above-mentioned solid-state fermentation reactor carries out the application in solid state fermentation at cultivation aerobic microbiological, anaerobion and amphimicrobe.

Wherein, the using method of above-mentioned solid-state fermentation reactor is:

Charging: open tank body left end removable top cover, takes out after solid materials mixes by tray and is distributed in equably on tray.

Sterilizing: pass into steam arrival tank interior by second fluid nozzle after entering steam heating tank body by chuck and carry out sterilizing, control sterilising temp, pressure, time as required; After sterilizing terminates, simultaneously from chuck, coil pipe input water coolant, reduce tank body and temperature of charge, and pass into gas pressurize is carried out to tank body;

Fermentation: temperature, humidity access seed liquor by negative pressure after adapting to, opens inoculation pipeline valve, seed liquor is sprayed on material surface, completes inoculation.In fermenting process, control temperature, humidity, air flow are stable.

Dry: to need dried material according to practical application, hot water drying material can be passed into suitable moisture from coil pipe.

Discharging: open removable top cover, takes out tray, unloads material.

Beneficial effect:

Of the present inventionly a kind ofly collect in-situ sterilization, fermentation, drying integrated solid-state fermentation reactor, sterilizing is thorough, fermenting process is stable, simple to operate, can be applicable to the fields such as agricultural fertilizer, microbiobacterial agent, animal-feed, no matter be which kind of application, this solid fermentation reactor all has the following advantages or effect:

1, simple to operate, multiple functional: the solid-state fermentation reactor in the present invention can directly complete sterilizing, fermentation, drying process, greatly reduces the labour intensity in production process.

2, mass transfer radiating efficiency is high: general fermentor tank internal gas is difficult to be evenly distributed, the nutrition of material lower floor is caused to be difficult to be utilized by microorganism, and a large amount of heat produced in fermenting process, be difficult to distribute, cause that leavening temperature is too high is unfavorable for microorganisms producing.Second fluid nozzle in the present invention can form air-flow at every one deck of tray, and have aperture bottom tray, can effectively flow through gas, through material, tray lower floor material is contacted with oxygen fully, be conducive to fermentation, the gas simultaneously flowed can take away the heat given out in fermenting process preferably, reduces the ability consumption cooling and bring.

3, heat and moisture preserving is effective: the chuck in the present invention, coil pipe can start recirculated water during the fermentation simultaneously, can regulate temperature rapidly, keeps the stable of temperature; The sterilized water that second fluid nozzle sprays becomes atomization shape, can be sprayed onto material equably surperficial and inner, play the effect of humidification.

4, second fluid nozzle is powerful: the second fluid nozzle in the present invention integrates humidification, inoculation, ventilation, steam-pipe function, eliminates multiple pipeline and brings the loaded down with trivial details of tank structure.

5, in addition, the sterilized water that in the present invention, humidification is used is through prepared by liquid filter, and more traditional sterilising method is simple, save the energy spent by sterilizing simultaneously.

Accompanying drawing explanation

Fig. 1 is the front view of solid-state fermentation reactor device.

Fig. 2 is the side-view of solid-state fermentation reactor device.

Fig. 3 is the coil pipe floor map in solid-state fermentation reactor device.

Fig. 4 is the second fluid nozzle system schematic in solid-state fermentation reactor device.

Drawing illustrates: fermentor tank-1; Stiffened steel plates-1-2; First removable top cover-2-1; Second removable top cover-2-2; First support-3-1; Second support-3-2; Base-3; Chuck-4-1; Steam inlet 4-2; Vapour outlet 4-3; First coil pipe-5-1; Second coil pipe-5-2; 3rd coil pipe-5-3; Water-in-5-4; Water outlet-5-5; Second fluid nozzle system-6; Nozzle-6-1, the first second fluid nozzle pipeline-6-2; Second second fluid nozzle pipeline-6-3; Vapour generator-6-4; First liquid strainer-6-5; Second liquid strainer-6-6; First gas filter-6-7; Second gas filter-6-8; Inoculation bottle-6-9; Temp probe-7-1; Moisture probe-7-2; With pressure probe-7-3; Discharge port-8; Air outlet-9; First tray 10-1; Second tray 10-2; 3rd tray 10-3.

Embodiment

According to following embodiment, the present invention may be better understood.But those skilled in the art will readily understand, concrete material proportion, processing condition and result thereof described by embodiment only for illustration of the present invention, and should can not limit the present invention described in detail in claims yet.

Embodiment 1

Solid-state fermentation reactor as shown in Figure 1, it comprises fermentor tank 1, coil pipe, tray and second fluid nozzle system 6.

Fermentor tank 1 is horizontal stainless steel fermentor tank, fixing on the base 3 by the first support 3-1 of bottom and the second support 3-2.Fermentor tank periphery is provided with chuck 4-1, chuck 4-1 is provided with steam inlet 4-2 and vapour outlet 4-3, steam inlet 4-2 is located at chuck 4-1 top, and vapour outlet 4-3 is located at chuck 4-1 lowermost end, and steam inlet 4-2 and vapour outlet 4-3 position in the horizontal direction away from.

Fermentor tank 1 horizontal direction both sides are provided with the first removable top cover 2-1 and the second removable top cover 2-2; First removable top cover 2-1 and fermentor tank 1 sidewall junction are provided with standing bolt 2-3, and the second removable top cover 2-2 and ferment tank 1 sidewall junction are provided with standing bolt 2-3.Fermentor tank 1 top is provided with production well 9, is provided with temp probe 7-1, moisture probe 7-2 and pressure probe 7-3 near production well 9; Fermentor tank 1 bottom is provided with discharge port 8.

As shown in Figure 2, fermentor tank 1 inside is provided with three layers of coil pipe from top to bottom, be respectively the first coil pipe 5-1, the second coil pipe 5-2 and the 3rd coil pipe 5-3, every layer of coil pipe is hollow structure, one end of coil pipe is water-in 5-4, the other end is water outlet 5-5, and it is inner that coil pipe is fixed on fermentor tank 1 by stiffened steel plates 1-2.As shown in Figure 3, coil pipe is stainless steel, and plan view shape is snakelike, also can be beneficial to the shape of holding tray for other.The water-in 5-4 of coil pipe is communicated with outlet conduit respectively at the inlet channel outside fermentor tank with water outlet 5-5.

Wherein, movable storing the first tray 10-1 on the first coil pipe 5-1, movable storing the second tray 10-2 on the second coil pipe 5-2, on the 3rd coil pipe 5-2, the 3rd tray 10-3 is put in activity; Wherein, described tray is iron pallet, and tray bottom even is drilled with several aperture, and the aperture of aperture is 2mm, and pitch of holes is 2cm.By opening the removable top cover of fermentor tank both sides, tray can be pushed or take out.

Position near removable top cover 2-2 in fermentor tank 1 vertically arranges second fluid nozzle system 6.As shown in Figure 4, described second fluid nozzle system 6 comprises 4 nozzle 6-1, the first second fluid nozzle pipeline 6-2 and the second second fluid nozzle pipeline 6-3, and 4 nozzle 6-1 adopt mode in parallel to be communicated with the second second fluid nozzle pipeline 6-3 with the first second fluid nozzle pipeline 6-2 simultaneously; Sterilized water pipeline and vapour line parallel with one another, then to connect with the first second fluid nozzle pipeline 6-2; Seed liquor pipeline and gas piping parallel with one another, then to connect with the second second fluid nozzle pipeline 6-3; Sterilized water pipeline is provided with first liquid strainer 6-5 and second liquid strainer 6-6, and vapour line is provided with vapour generator 6-4, the external inoculation bottle 6-9 of seed liquor pipeline, and gas piping is provided with the first gas filter 6-7 and the second gas filter 6-8; Sterilized water pipeline and seed liquor pipeline are respectively equipped with gas meter, vapour line and gas piping are respectively equipped with gas meter.This second fluid nozzle system can spray any one or a few in this of sterilized water, steam, seed liquor and gas with the flow velocity of specifying in fermentor tank.Can carry out disinfection in fermentor tank by spraying into steam as required before fermentation, or spray into the conduct protection gas such as nitrogen by gas piping.

Embodiment 2: aerobic microbiological (producing gamma-polyglutamic acid-agricultural fertilizer for subtilis solid state fermentation)

Gamma-polyglutamic acid-(γ-polyglutamic acid, γ-PGA) be by a class homogeneous polyamino acid of Microbe synthesis, basic framework is the straight-chain molecule be formed by connecting by γ-amido linkage by Pidolidone and D-Glu, and relative molecular mass can reach 200 × 10 4da.Owing to there is a large amount of hydrogen bonds and wetting ability free carboxy in its structure, there is excellent performance of keeping humidity, be widely used in agricultural fertilizer.

The microorganism of fermenting in the present embodiment is that ((culture presevation number: CGMCC 0833), directly uses microbiobacterial agent (5 × 10 to the subtilis Bacillus subtilisNX-2 that obtained the authorization of this laboratory 10cfu/g) inoculate, inoculum size is 25g/kg.

Solid-state fermentation culture medium: the bacterium slag 600g/kg after mushroom cultivation, msg meal 400g/kg, industry glycerol 40g/kg.

With reference to figure 1, make a solid-state fermentation reactor device of the present invention, and on the device of the present embodiment making, carry out solid fermentation production gamma-polyglutamic acid-agricultural fertilizer, below by embodiment, the present invention is further described.Fermentor tank main body 1 adopts stainless material to make, tank body internal diameter 50cm, length 80cm, thickness 6mm, two ends, left and right are removable top cover 2-1,2-2, are cast with chuck 4-1 in the middle part of tank wall, and chuck left upper end is connected with steam inlet 4-2, bottom righthand side has vapour outlet 4-3.Air outlet 9, temp probe 7-1, moisture probe 7-2, tensimeter 7-3 are positioned at tank body left upper end, and discharge port 8 is positioned at tank body lower-left end.Coil pipe 5-1,5-2,5-3 are that stainless steel is made, and are positioned at tank interior upper, middle and lower-ranking, and two ends are respectively water-in 5-4 and water outlet 5-5, and this coil pipe can cool tank body, carry out drying to the material after fermentation.Each placement iron pallet above coil pipe, be tray 10-1,10-2,10-3, specification is length 70cm, and width is 40cm.It is 2cm that this tray bottom is evenly drilled with pitch of holes, and aperture is the aperture of 2mm, puts 2 layers of gauze, prevent material from falling before loaded material bottom tray.The right-hand member of fermentor tank main body 1 is connected with second fluid nozzle system 6, and this system is made up of gas pipeline and fluid pipeline in tank body outside, and described gas pipeline is provided with gas prefilter 6-8, fine filter 6-9, and by under meter coutroi velocity; Fluid pipeline is provided with liquid prefilter 6-5, fine filter 6-6, and by under meter coutroi velocity, this pipeline is humidifier pipeline, for tank interior material keeps the skin wet; Steam conveying pipe is positioned at both sides, and is provided with vapor filter 6-4, is separately provided with inoculation pipeline 6-9 in gas pipeline side, external inoculation bottle.

Production operation is as follows: take 3kg Lentinus Edodes fungus slag, 2kg msg meal, 200g industry glycerol, after tank body mixes outward, evenly be laid on pallet after regulating pH to about 7.0, thickness is about 5cm, is advanced by pallet tank body internal fixtion good, open vapour generator and first again sterilizing is carried out to tank body through chuck heating, maintain 121 DEG C, 0.1Mpa 20min, pass into sterile air after sterilizing terminates and carry out pressurize, ON cycle water is cooled to 35 DEG C to tank body simultaneously, controls material moisture about 65%.Taking 125g B.subtilis NX-2 microbial inoculum is dissolved in 100ml sterilized water, is sprayed on material surface inoculates by second fluid nozzle, by PLC system holding temperature 35 DEG C, humidity 65% in fermenting process, and air flow 100ml/min.We carry out tranquillization stuffiness cultivation as a comparison with 5L filter flask simultaneously, investigate the performance of apparatus of the present invention.

Result shows: in reactor of the present invention, and meta-bolites gamma-polyglutamic acid-concentration can reach 168g kg -1, microbe colony number reaches 60 × 10 8cfu g -1, improve 31.5%, 33.3% respectively than in 5L filter flask, visible reactor of the present invention is feasible and is efficient.Finally, passing into hot water by coil pipe, to make culture be dried to contained humidity be 10%.

Embodiment 3: aerobic microbiological (for solid state fermentation subtilis phosphate solubilizing bacteria microbial inoculum)

Phosphorus element is one of required nutritive element of growth and development of plants, and soil lacks growth and the grain yield that phosphorus directly affects farm crop, lacks phosphorus cultivated area reach 74% in China, and in soil, the phosphorus of more than 95% is invalid state, is difficult to directly be absorbed by plant.There is many phosphate solubilizing microorganisms in soil, the phosphoric acid salt of these insolublies can be degraded into free form, so that plant absorption utilizes, therefore, carry out phosphate solubilizing microorganism exploitation, reduce production of phosphate fertilizer cost, solve phosphate fertilizer and to pollute and to improve the research of P use efficiency aspect very necessary.

Phosphate solubilizing bacteria Bacillus subtilis JT-1 in the present embodiment is screened by this laboratory and obtains from the phosphorus ore in Chinese fir a strip of land between hills phosphorus ore area, Yichang, Hubei Province, be preserved in China typical culture collection center (being called for short CCTCC), preserving number is CCTCCM 2014367.

Seed culture medium: glucose 10g/L, NaCl 5g/L, peptone 10g/L;

Solid fermentation substratum: corn cob meal and feces of livestock and poultry mass ratio are 3:1; Supplement cane molasses 10%, (NH 4) 2sO 40.3%, MnSO 44H 2o 0.015%, pH 6.8 ~ 7.0.

With the production equipment that embodiment 2 makes, carry out solid state fermentation and produce phosphate solubilizing bacteria active bacteria formulation, its step is with the step of embodiment 2, just concrete fermentation condition is distinguished to some extent, below provide concrete fermentation condition: access a full ring Bacillus subtilis JT-1 from the inclined-plane of preservation in 500mL seed culture fluid, under 35 DEG C of 150rpm, cultivate 15h.Inoculum size is 4.8%, and solid fermentation matrix quality is 5kg, and sterilising conditions is 121 DEG C, 20min.Leavening temperature controls at 35 ~ 37 DEG C, and humidity is 65 ~ 70%, air flow 80ml/min, and fermentation total time 30h, passes into hot water drying substratum moisture to 10% by coil pipe after fermentation ends, obtain phosphate solubilizing bacteria microbial inoculum.Simultaneously with 5L filter flask for experiment contrast group.

Phosphate solubilizing bacteria microbial inoculum ability measures: load 100mL inorganic phosphorus liquid nutrient medium in 500mL triangular flask, medium component is glucose 10g/L, (NH 4) 2sO 41g/L, MgSO 47H 2o 0.3g/L, NaCl 0.3g/L, FeSO 47H 2o 0.03g/L, MnSO 44H 2o 0.03g/L, Ca 3(PO 4) 25.0g/L.Add 5g phosphate solubilizing bacteria microbial inoculum again, do simultaneously and do not inoculate contrast (only adding 5g solid medium), all process are carried out 3 times and are repeated.At 30 DEG C, 150rpm is shaking culture 7d respectively, by nutrient solution centrifugal 20min under 6000rpm, measures phosphorus decomposing amount (water-soluble phosphorus content) by molybdenum antimony resistance colorimetric method.

Result shows, compared with contrast filter flask, in solid-state fermentation reactor of the present invention, Bacillus subtilis JT-1 can grow well, and after single Batch fermentation 30h, colony number reaches 55.6 × 10 10cfu g -1, compare 35 × 10 of photograph 10cfu g -1, improve 37%.Dissolving P capacity measurement result shows, and in the aqueous solution, phosphorus content is 750mg L -1, phosphorus decomposing rate reaches 75%, and this shows that the phosphate solubilizing bacteria microbial inoculum of this reactor solid state fermentation has higher dissolving P capacity.

Embodiment 4: aerobic microbiological (for solid state fermentation subtilis potassium solubilizing bacteria)

Potassium is the required nutritive element of crop growth, accounts for dry matter content 0.2 ~ 4.1%, and in plant growth and development process, potassium can strengthen the disease and insect resistance of crop, resistant to lodging, drought resisting and the anti-adversity ability such as cold-resistant, thus improves the yield and quality of crop.According to soil survey information display, the arable land potassium deficiency of China about 70%, the arable land severe potassium deficiency of about 45%.And the potash fertilizer turnout Zhi Zhan world 0.34% of China, and consumption accounts for 14.7%, although annual import 2,000,000 tons of potash fertilizer, still can not meet the demand of agriculture production to potash fertilizer.These have become the important factor of restriction crop yield and quality raising.Silicate bacteria, also known as potassium bacterium, refers to the mineral such as the silico-aluminate that can decompose in soil, and the potassium of insoluble is transformed into the quasi-microorganism that soluble status utilizes for plant absorption, it can also secrete various plants hormone Promoting plant growth in addition.Therefore, the potassium solubilizing bacteria resource that, ability of dissolving potassium effective by potassium decomposing is strong excavates the not available potassium element in soil, and to developing the economy, ecological agriculture tool is of great significance.

Potassium solubilizing bacteria Bacillus subtilis JT-2 in the present embodiment is by this laboratory from seed selection ore deposit, Zijinshan Area, the Linxian County, Shanxi Province soil being rich in potassium ore, be preserved in China typical culture collection center (being called for short CCTCC), preserving number is CCTCCM 2014368.

Seed culture medium: glucose 10g/L, NaCl 5g/L, peptone 10g/L;

Solid fermentation substratum: Lentinus Edodes fungus slag and straw mass ratio are 2.5:1.5, industry glycerol 5%, yeast powder 2%, NaCl 2%, pH 6.8 ~ 7.0.

With the production equipment that embodiment 2 makes, carry out solid state fermentation and produce potassium solubilizing bacteria active bacteria formulation, its step is with the step of embodiment 2, just concrete fermentation condition is distinguished to some extent, below provide concrete fermentation condition: access a full ring Bacillus subtilis JT-2 from the inclined-plane of preservation in 500mL seed culture fluid, under 30 DEG C of 200rpm, cultivate 8h.Inoculum size is 8%, and solid fermentation matrix quality is 5kg, and sterilising conditions is 121 DEG C, 20min.Leavening temperature controls at 28 ~ 30 DEG C, and humidity is 60 ~ 70%, air flow 150ml/min, and fermentation total time 24h, passes into hot water drying substratum moisture to 10%, obtain potassium solubilizing bacteria microbial inoculum.With the fermentation of 5L filter flask for experiment contrast group.

Potassium solubilizing bacteria microbial inoculum ability of dissolving potassium measures: preparation potassium decomposing substratum (without water-soluble potassium ion) is by 95mL packing and add potassium breeze that 0.5g accurately takes in 250mL triangular flask, 121 DEG C of sterilizing 30min.Add 5g Bacillus subtilisJT-2 microbial inoculum, to add 5g substratum for contrast, 37 DEG C of shaking baths terminate to cultivate after cultivating 25d, nutrient solution is through 4 DEG C of centrifugal 5min of 1000r/min, fully mix with pipette, extract 4mL centrifuged supernatant and 4mL 6mmoL/LiCl, utilize flame spectrophotometer method to survey potassium.

Result shows: compared with control group, Bacillus subtilis JT-2 grows better in solid-state fermentation reactor of the present invention, and after single Batch fermentation 24h, colony number reaches 88.5 × 10 10cfu g -1, improve 28.2% than contrast; Microbial inoculum ability of dissolving potassium measurement result: water-soluble potassium ion content is 175mg/L, and potassium decomposing rate reaches 67%, illustrates that this microbial inoculum has stronger ability of dissolving potassium.

Embodiment 5: anaerobion (producing clostridium butyricum active bacteria preparation for solid state fermentation)

Along with antibiotic widespread use, its shortcoming and drawback also gradually find by people and be familiar with, cause global common concern, Chinese scholars for can the expansion also like a raging fire of the Study and Development of novel product of substitute antibiotics, and also starts as the research of the probiotic bacterium that can overcome above-mentioned drawback the attention causing scholars.

Clostridium butylicum is a kind of Gram-positive sporeformer of anaerobism, in direct rod shape or slightly bending, be extensively present in the enteron aisle of humans and animals body, effectively can suppress the breeding of pathogen enterobacteria, adjustment intestinal microecology balance, reduces the generation of the objectionable impurities such as Ammonia, amine simultaneously.In clostridium butylicum preparation, viable bacteria exists with spore form, has very strong stability to factors such as the high temperature in various severe environment, storage and processing transportation, high humidity, acidity, mechanical friction and extruding.Therefore be added in animal and fowl fodder the high temperature can resisted in feed manufacturing process, Gao Re and gi tract sour environment thereof, there is the very strong market competitiveness.Clostridium butylicum is as a kind of probiotic bacterium, there is following effect: (1), maintenance intestinal microflora balance: clostridium butylicum is not poisoned human body and animal, the growth of the husky bacterium of staphylococcus, candidiasis, klebsiella, Pseudomonas aeruginosa, intestinal bacteria, dysentery bacterium and typhoid fever and spoilage organism can be suppressed, thus decrease the objectionable impuritiess such as amine, indoles and hydrogen sulfide, the breeding of intestinal beneficial bacterium can be promoted again simultaneously.(2), strengthen immunologic function, prophylaxis of tumours occurs: test the growth proving the exocellular polysaccharide Galactose Glucose sugar energy inhibition tumor cell that the cell wall constituent of clostridium butylicum and it produce.(3), produce and should produce thing: clostridium butylicum can produce vitamin B group, vitamin K, amylase, proteolytic enzyme, cellulase and butyric acid etc., and these should produce thing for Growth of Cells and intestinal health is all required.(4), stable resistance is better: clostridium butylicum belongs to gemma Pseudomonas, endogenous spore can be produced, so viability is comparatively strong, possibility heat-resisting, acidproof, resistance to bile, antibiotics resistant, can be widely used in the additive of medicine, functional health-care food and animal-feed.

Clostridium butylicum (Clostridium butyricum CGMCC 1.336) in the present embodiment is purchased from China General Microbiological culture presevation administrative center.

Seed culture medium: glucose 20g/L, extractum carnis 1.5g/L, peptone 20g/L, calcium carbonate 10g/L, magnesium sulfate heptahydrate 0.2g/L, manganese sulfate monohydrate 0.2g/L, pH 7.0,37 DEG C of static gas wave refrigerator 24h.

Solid-state fermentation culture medium: dregs of beans and wheat bran mass ratio are 1:1, calcium chloride 0.2%, ammonium sulfate 1.5%.

With the production equipment that embodiment 2 makes, carry out solid state fermentation and produce clostridium butyricum active bacteria preparation, its step is with the step of embodiment 2, and just concrete fermentation condition is distinguished to some extent, below provides concrete fermentation condition.

Clostridium butyricum CGMCC 1.336 is anaerobion, and first by actication of culture before inoculation, access seed culture medium, 37 DEG C of static gas wave refrigerator 24h make seed liquor, and inoculum size is 5%.Material matrix quality is 5kg, and sterilising conditions is 115 DEG C, 15min.Need to pass into N after sterilizing terminates 2drain tank interior and ducted air and pressurize is carried out to tank body, and maintaining and pass into N 2speed is 10ml/min.Control pot temperature 37 DEG C in fermenting process, humidity 55-60%, fermentation time is 72h, passes into hot water drying substratum moisture to 15% after fermentation ends by coil pipe.Meanwhile, with 5-L filter flask for tranquillization does not pass into N 2for contrast.

Result shows: in solid-state fermentation reactor of the present invention, and thalli growth is obviously vigorous than contrast (5-L filter flask), and final colony number reaches 2.2 × 10 9cfu g -1, than contrast raising 23.6%.

Embodiment 6 amphimicrobe (for plant lactobacillus solid state fermentation dregs of beans)

Dregs of beans is a kind of protein feed resource of high-quality, and compared with cotton dregs, the dish dregs of rice, peanut meal, its lysine content high (close to Ideal Amino Acid Pattern), digestibility are high, good palatability.Compared with animal derived protein, dregs of beans source is sufficient, not easily by pathogen contamination, not easily oxydative spoilage, few containing harmful toxic matter, safety coefficient is high.Therefore dregs of beans becomes the vegetable protein source of the larger a kind of high-quality of usage quantity in current livestock industry.But containing some antinutritional factor in dregs of beans, mainly contain trypsin inhibitor, phytic acid, soybean lectin, arteries and veins enzyme, oligomericly to ward off, lipoxidase, Soybean antigen protein (sensitizing factor) and goitrin etc.Due to their existence, on the one hand restraining effect played to some digestive ferment in animal body or become stodgy composition etc. with nutritive substance complexing, the utilization ratio of dregs of beans is declined; On the other hand play toxic action to some organ in animal body, cause bad impact to the physiology of animal, growth, health, the impact especially for young animal is more obvious.

Be at present main to the treatment process of antinutritional factor in dregs of beans mainly with chemistry, physical method, have certain removal effect, but it is high and to defects such as dregs of beans Middle nutrition Value Loss are large to there is security, cost.Carry out fermentative processing with plant lactobacillus solid state fermentation means to ordinary student dregs of beans in the present embodiment, research plant lactobacillus is on the impact of antinutritional factor in dregs of beans and nutritive value thereof.

Plant lactobacillus Lactobacillus plantarum CICC 21790 in the present embodiment is purchased from Chinese industrial Microbiological Culture Collection administrative center.

Seed culture medium: glucose 5g/L, peptone 10g/L, extractum carnis 10g/L, yeast extract paste 5g/L, ammonium citrate 2g/L, sodium-acetate 5g/L, Tween80 1mL/L, K 2hPO 42g/L, MgSO 47H 2o 0.2g, MnSO 44H 2o 0.2g/L, pH 6.0 ~ 6.8.Solid-state fermentation culture medium: dregs of beans.

With the solid-state fermentation reactor that embodiment 2 makes, carry out solid fermentation dregs of beans, its step is with the step of embodiment 2, and just concrete fermentation condition is distinguished to some extent, below provides concrete fermentation condition.Lactobacillus plantarum CICC21790 is amphimicrobe, but in view of research experience and experiment convenience, we are according to anaerobic bacteria culture.First connect plant lactobacillus in 500mL seed culture medium, 37 DEG C of static gas wave refrigerator 12h make seed liquor, and inoculum size is 9%.Dregs of beans total mass is 5kg, and sterilising conditions is 121 DEG C, 15min.Need to pass into N after sterilizing terminates 2drain tank interior and ducted air and pressurize is carried out to tank body, and maintaining and pass into N 2speed is 20ml/min.Control pot temperature 37 DEG C in fermenting process, humidity 55-65%, fermentation time is 48h, dried material moisture to 15% after result.Result shows; Lactobacillus plantarum CICC 21790 can degrade trypsin ihhibitor in dregs of beans and lectin effectively, and after 48h, clearance reaches 60.4% respectively, and 99%.

Claims (9)

1. a solid-state fermentation reactor, is characterized in that, it comprises fermentor tank (1), coil pipe, tray and second fluid nozzle system (6);
Described fermentor tank (1) is horizontal fermentor tank, fermentor tank periphery is provided with chuck (4-1), and fermentor tank (1) horizontal direction both sides are provided with the first removable top cover (2-1) and the second removable top cover (2-2);
Fermentor tank (1) inside is provided with 1 ~ 10 layer of coil pipe from top to bottom, movable storing tray on every layer of coil pipe; Every layer of coil pipe is hollow structure, and one end of coil pipe is water-in (5-4), and the other end is water outlet (5-5);
Fermentor tank (1) top is provided with production well (9), is provided with temp probe (7-1), moisture probe (7-2) and pressure probe (7-3) near production well (9);
Position near removable top cover (2-2) in fermentor tank (1) vertically arranges second fluid nozzle system (6), described second fluid nozzle system (6) comprises 1 ~ 10 nozzle and second fluid nozzle pipeline, nozzle is connected with the outside gas-liquid pipeline of fermentor tank (1) by second fluid nozzle pipeline, and described fermentor tank (1) outside gas-liquid pipeline is sterilized water pipeline, seed liquor pipeline, vapour line and gas piping parallel with one another.
2. solid-state fermentation reactor according to claim 1, it is characterized in that, described fermentor tank (1) is stainless steel, fermentor tank (1) bottom is provided with discharge port (8), and fermentor tank (1) is fixed on base (3) by first support (3-1) of bottom and the second support (3-2).
3. solid-state fermentation reactor according to claim 1, it is characterized in that, described chuck (4-1) is provided with steam inlet (4-2) and vapour outlet (4-3), steam inlet (4-2) is located at chuck (4-1) top, vapour outlet (4-3) is located at chuck (4-1) lowermost end, and steam inlet (4-2) and vapour outlet (4-3) position in the horizontal direction away from.
4. solid-state fermentation reactor according to claim 1, it is characterized in that, first removable top cover (2-1) and fermentor tank (1) sidewall junction are provided with standing bolt (2-3), and the second removable top cover (2-2) and ferment tank (1) sidewall junction are provided with standing bolt (2-3).
5. solid-state fermentation reactor according to claim 1, it is characterized in that, fermentor tank (1) inside is provided with three layers of coil pipe from top to bottom, be respectively the first coil pipe (5-1), the second coil pipe (5-2) and the 3rd coil pipe (5-3), it is inner that coil pipe is fixed on fermentor tank (1) by stiffened steel plates (1-2), first coil pipe (5-1) upper movable storing the first tray (10-1), second coil pipe (5-2) upper movable storing the second tray (10-2), the 3rd tray (10-3) is put in the upper activity of the 3rd coil pipe (5-2).
6. solid-state fermentation reactor according to claim 1 or 5, it is characterized in that, described tray is iron pallet, and tray bottom even is drilled with several aperture.
7. solid-state fermentation reactor according to claim 1 or 5, it is characterized in that, described coil pipe plan view shape is snakelike, and the water-in (5-4) of coil pipe is communicated with outlet conduit respectively at the inlet channel outside fermentor tank with water outlet (5-5).
8. solid-state fermentation reactor according to claim 1, it is characterized in that, described second fluid nozzle system (6) comprises 4 nozzles (6-1), the first second fluid nozzle pipeline (6-2) and the second second fluid nozzle pipeline (6-3), and 4 nozzles (6-1) adopt mode in parallel to be communicated with the second second fluid nozzle pipeline (6-3) with the first second fluid nozzle pipeline (6-2) simultaneously; Sterilized water pipeline and vapour line parallel with one another, then to connect with the first second fluid nozzle pipeline (6-2); Seed liquor pipeline and gas piping parallel with one another, then to connect with the second second fluid nozzle pipeline (6-3); Sterilized water pipeline is provided with first liquid strainer (6-5) and second liquid strainer (6-6), vapour line is provided with vapour generator (6-4), the external inoculation bottle of seed liquor pipeline (6-9), gas piping is provided with the first gas filter (6-7) and the second gas filter (6-8); Sterilized water pipeline and seed liquor pipeline are respectively equipped with gas meter, vapour line and gas piping are respectively equipped with gas meter.
9. solid-state fermentation reactor according to claim 1 carries out the application in solid state fermentation at cultivation aerobic microbiological, anaerobion and amphimicrobe.
CN201510186386.4A 2015-04-17 2015-04-17 Solid state fermentation reactor and application thereof CN104845877A (en)

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