CN1742543A - Commercial production method for edible mushroom liquid culture bacteria, apparatus and automatic sterilized inoculator - Google Patents

Abstract

The present invention discloses an edible fungus liquid culture strain commercial production method, its equipment and automatic sterile inoculating machine. It is characterized by that the edible fungus production is divided into two portions. One portion is edible fungus strain-making industry and the other portion is edible fungus cultivation industry.

Description

Commercial production method for edible mushroom liquid culture bacteria, equipment and automatic sterilized inoculator

Technical field:

The automatic sterilized inoculator tool of indispensability when production method, sterile vacuum filling apparatus and this product that the present invention relates to a kind of edible mushroom liquid culture bacteria product that can popularization and application used especially can allow the production of edible mushroom liquid culture bacteria and the equipment that popularization and application reaches standardization, automation, scale;

Technical background:

At present, in the edible fungi liquid strain application facet, just, produce liquid culture bacteria voluntarily by the mushroom farming so that the production equipment of making liquid spawn to be provided, this mode has restricted the popularization and application of liquid spawn and the realization of edible mushroom suitability for industrialized production target, because:

1, the production of hybrid seeds equipment investment of edible fungi liquid strain is huge; For example, the production process of one 10,000 bag cultivating kind culture bag, the fund that needs input to buy the production of hybrid seeds equipment of corresponding production of hybrid seeds amount 270L is 55800,00 yuan, and the production of solid spawn only need drop into 50000 yuans, the production cycle 7-15 of liquid spawn days, though than production cycle 30-40 days of solid spawn, reduced 3-4.8 doubly, but fund input amount 1116 times have really been increased; Ratio between investments is seriously lost perseverance; Even increase the production cycle, also just a production cycle is successively decreased 1 times, and the cost recovery time is long to making;

2, the production risk of liquid spawn is big; The production of liquid spawn needs the culture environment of cleaning and good level professional technology, could guarantee that bacterial classification does not pollute; Present vast mushroom farming is a first-selection with the conventional solid bacterial classification still;

3, liquid production of hybrid seeds equipment in actual applications can not be harmonious with actual production; The as above routine production of hybrid seeds equipment of buying, its output is constant 270L/10000 bag, but expand the scale of production in the mushroom farming, a production cycle, this complete equipment just can not satisfy production requirement when increasing by 20,000 or 30,000 bag cultivating kind culture bag, can only increase the production of hybrid seeds and produce 2-3 time, each 7-15 days, so just prolonged the cultivation and production process,, greatly reduced the application advantage of liquid spawn to making cell age be difficult to be consistent; As select the production of hybrid seeds equipment of corresponding production of hybrid seeds amount for use, will increase investment again;

4, existing automation inoculating facility is difficult to popularization and application; As the automation inoculation device that the full standing grain bacterium in Dalian already produces, its price is more than 50,000 yuans, and inoculum concentration is 2160 bags/hour, and needs to be equipped with the liquid fungus seed jar; The automatic vaccination machine price of TianXing, Cao County, Shandong liquid fungus seed instrument factory production is 2,980,000 yuans in addition, is that present home-made machine is cheapest, but its inoculum concentration only is 1000 bags/hour; And these two kinds of types all need use in gnotobasis, so cost an arm and a leg, factor affecting such as complex structure, inoculation environment are strict, existing type is difficult to popularization and application;

5, the can production process of existing edible fungi liquid strain is easily polluted, and the storage time of liquid spawn is short;

Summary of the invention:

The present invention seeks to disclose a kind of commercial production method for edible mushroom liquid culture bacteria, equipment and automatic sterilized inoculator; Overcome the deficiency that edible mushroom liquid culture bacteria that prior art causes can not popularization and application, can not only provide the liquid culture bacteria of high-quality cheapness, green safety and supporting with it simple, practical, the first bacterium inoculating mechanism of high-efficiency automatic for vast mushroom farming, but also the production of hybrid seeds operation of liquid spawn is separated from the whole process of production of edible fungus culturing, professional technique unit can be engaged in production by the present invention, the liquid culture bacteria product of high-quality and high-efficient automatic sterilized inoculator product, be supplied to the mushroom farming to be engaged in the later stage cultivation and production and use; Just Edible Fungi is divided into two, promptly formed the new production model of edible mashroom cultivating industry and cultivation two independences of industry and associated, fundamentally simplified the Edible Fungi chain, changed the traditional mode of production pattern of edible mushroom, and can make production of hybrid seeds industry and the cultivation industry of edible mushroom all possess the advantages for development that become the industrialization industry;

A kind of commercial production method for edible mushroom liquid culture bacteria of the present invention is:

1-1. the preparation of seed

The quality of seed quality is very big to the fermentation influence; Be decided by bacterial classification itself and seed culture condition;

The seed requirement that edible fungi submerged fermentation is used

(1) thalli growth is vigorous, and is energetic

The cell age of seed normally is controlled at the exponential phase later stage of thalline; The seed cell age with 48-96 hour for well;

(2) the bacterium bulb diameter is little, quantity is many

The nutrition composition of medium has certain influence to the shape of bacterium ball; Usually, contain the many medium of sugar, the bacterium bulb diameter of turning out is little, and the medium viscosity is big, and the bacterium ball is also less; In addition, than using reciprocating type shaking table, the bacterium bulb diameter is less with rotary shaking table; In shaking bottle, add several beades or a little spring carries out shaken cultivation, the bacterium bulb diameter is diminished;

(3) purity height

The seed that edible fungi submerged fermentation is used requires high-purity, and any pollution can not be arranged;

1-2, slant strains preparation

The original strain that is used for submerged fermentation need be through the purification and the evaluation of strictness;

Medium: glucose 2.0%, yeast extract 0.5%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.1%, agar 2.0%, pH nature;

Sterilization: under 120-126 ℃ condition, sterilized 30 minutes;

Condition of culture: 25 ℃, 7-10 days;

1-3, shake-flask seed preparation

Medium: potato 20%, brown sugar 1.0-1.5%, glucose 1.0-1.5%, wheat bran 3%, peptone 0.15-0.3%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.1%, vitamin V B110 mg/litre, surplus is a water, pH value 6.5-7.0.

Prepared culture medium is sub-packed in the triangular flask, and puts the 3-4 bead in triangular flask, has filled in tampon, waterproof paper on the bundle; Autoclaving, when being cooled to below 30 ℃, inserts the slant strains 2-4 piece of 0.5 centimetre of 2 size at 1.4 kg/cm, 2 pressure, sterilization 30 minutes, is sandwiched in then and carries out shaken cultivation on the shaking table, and its Frequency Design is 80-120 time/minute; Cultivation temperature was cultivated 2-7 days at 25 ℃;

The preparation method of one-level shake-flask seed, secondary shake-flask seed is identical;

1-4. preparation one-level, secondary seed preparation

Broadcast bottle seed further expanding propagation become first order seed, first order seed more expanding propagation becomes two seeds; First order seed and secondary seed are cultivated in seeding tank, and the first order seed medium is selected the shake-flask culture base for use, and the medium of secondary seed is selected the shake-flask culture base for use, and the medium of secondary seed is selected fermentation medium for use;

2-1. ferment tank technological parameter control

Raw material and culture medium preparation

Medium: potato 10-20%, wheat bran 2-5%, brown sugar 0.5-2.0%, peptone 0.1-0.3%, yeast extract 0.1-0.3%, potassium dihydrogen phosphate 0.1-0.2%, magnesium sulfate 0.05-0.15%, VB11 0-20 mg/litre, polyoxypropylene glycerine 0.03-0.035%, surplus are water, pH value 4.0-8.0%; Culture medium preparation: in the following order

(1) make potato sweat and wheat bran juice: fresh potato digs the bud peeling, is cut into the sheet or the bar of 0.5-0.4 cm thick, adds 5 times of poach and use 6-8 layer filtered through gauze when shortcake does not rot, and it is standby to get juice; The wheat bran that fresh nothing is gone mouldy adds 10 times of water boils to 20 minute, and it is standby to cross leaching juice;

(2) composite soil fermented bean drink and wheat bran juice;

(3) will add behind potassium dihydrogen phosphate, the molten water of magnesium sulfate in the mixed juice;

(4) will add behind brown sugar, glucose, the molten water of peptone in the mixed juice again;

(5) vitamin B1 being pulverized the back adds in the mixed juice again;

(6) be that 10% hydrochloric acid or sodium hydroxide are adjusted pH value with concentration;

(7) sterilization: medium is packed in the culture tank, when temperature reaches 121-126 ℃, kept 30-35 minute;

(8) cooling: after the sterilization, feed the cold water cooling immediately, make the medium temperature drop to 23-30 ℃;

2-2, cell age

The shake-flask seed cell age was controlled at 4-10 days, and the cell age of first order seed and secondary seed is 48-96 hour;

2-3, inoculum concentration

Inoculum concentration when food has deep-fermentation is 10%-20%;

2-4, temperature

The temperature of edible fungi submerged fermentation is at 22-30 ℃;

The throughput of 2-5, edible fungi submerged fermentation is with 0.2-1.5 ventilation volume/culture fluid volume/minute be advisable;

2-6, mixing speed

The mixing speed of edible fungi submerged fermentation generally is 180-500 rev/min;

2-7, acid-base value

Optimal pH is 5.0-7.2, and flat mushroom is 5.0-6.5; Need timing sampling during the fermentation, measure acid-base value, and adjust;

2-8, tank pressure

To keep certain tank pressure during the fermentation, be generally 0.2-0.7 kg/cm 2;

2-9, foam control

3. mechanical defoaming

Dependence is loaded on jar interior froth breaking and starches the surface tension that the variation of intense mechanical vibration and pressure destroys bubble; Reduce the stability of foam, impel lather collapse;

4. chemical froth breaking

Defoamer commonly used has natural oil and synthetic defoamer two classes; The natural oil that is used for froth breaking has: peanut oil, soya-bean oil, cotton oil etc.; Synthetic defoamer commonly used has the bubble enemy; Great majority are to use the bubble enemy to do defoamer at present, and addition is about 0.006%;

5. fermentation termination

Reasonably putting jar time should could determine with reference to factors such as the outward appearance of production concentration, the rate of filtration, amino nitrogen content, residual sugar amount, mycelia form, pH, zymotic fluid and viscositys; Measure during the fermentation;

6. mycelia quantitative determination

Assay method commonly used has two kinds:

A) centrifugal process

Measure the sample liquid of certain volume, through 3000 rev/mins centrifugal 10 minutes, the tipping supernatant claims bacterium mud weight;

B) cross elimination

According to the size of mycelium pellet, select suitable specification sieve for use, sample liquid is filtered with sieve, with the flushing of 20-30 times of volume water, the culture fluid of carrying secretly is formed flushing again, filter mycelium dry down at 80 ℃, dry to constant weight with 105 ℃ again, weigh;

7. the mycelium pellet size is measured

Sample earlier after filtration, rinse well, take a morsel mycelium pellet in culture dish, add a little distillation water and the bacterium ball is scattered and fully stretch, measure facing to mycelium pellet at the bottom of culture dish with slide measure;

8. purity test

Inspection is to stagnate living contaminants is arranged; Inspection method can be cultivated or the cultivation of phenol red meat soup with dull and stereotyped, and checking after 24-28 hour has not germ contamination; Also can be by microexamination, just having seen clearly not with oily mirror, bacterium exists;

9. pH measurement

Change by the pH that check to find ferment liquid, the process of passable hydrolysis and fermentation is also understood pollution condition indirectly; In general, along with the carrying out of fermentation, phenomenon appears ging up again in pH, and this is because the result of thalline self-dissolving;

10. sugar content

1. the mensuration of zymotic fluid total reducing sugar;

2. the mensuration of zymotic fluid reducing sugar;

11. amino acid whose mensuration;

12. the zymotic fluid smell is checked

The gas checking of zymotic fluid can be in the exhaust outlet inspection of fermentation tank;

13. canned production

When the liquid culture bacteria in the fermentation tank reaches fermentation termination, after qualified Upon Strict Inspection, be transported to by aseptic pipeline in the separating box of sterile vacuum filling production lines, by the sterile vacuum filling mechanism liquid spawn under vacuum state, can and seals the bung of PET polyester bucket under vacuum state with the disposable plastic intelligent lid in PET polyester bucket, then, it is pre-chilled to below 5 ℃, in 1-5 ℃ refrigerated room, preserves and sell by the form of chain direct selling;

The sterile vacuum filling production lines is by connecting gear, and vacuum filling mechanism manages and covers cover supplier, vavuum pump, motor, speed changer, clutch composition;

It is characterized in that:

Connecting gear comprises chain scraper conveyor, thumb wheel, and chain scraper conveyor also is listed in the annular rotary platform outside in the vacuum filling machine structure, vacuum filling mechanism below;

In sterile vacuum filling mechanism casing, the lower right is square double acting hydraulic cylinder, on the carrier bar on the annular rotary platform beaming roller below its piston is fixed on, grinding tooth wheel in the carrier bar right-hand member is fixed with, interior grinding tooth wheel is meshed with spur gear, and spur gear is connected with motor, speed changer, the clutch of below by power transmission shaft, angular wheel again; The piston left side of square hydraulic cylinder is provided with impact switch, hydraulic cylinder body then is arranged in the cabinet, its top be fuel tank with the three-position electromagnetic reversal valve that links to each other of fluid cylinder cylinder body and pressure switch, pipeline, the left side is the device that declutches, the device that declutches is connected with semicircle sealing tile fragment, be fixed with second half circular seal tile fragment on the cabinet wall of left side on the other side, be provided with impact switch in this tile fragment; The fuel tank left side is an electric oil pump, its export pipeline place is provided with pressure switch, the oil pump top is two three-position electromagnetic reversal valves, be connected with the hydraulic cylinder that is vertically fixed on cabinet left side cavity inner top with the hydraulic cylinder of crosswise fixed respectively, be connected with the pushing cover sealing baffle on the piston of top, right side hydraulic cylinder at top, cabinet right side; Be connected with on the piston of left side top hydraulic cylinder and cover groove, lid groove right openings, built-in jump ring, with on the lid channel opening corresponding cavity onesize opening is arranged also, open lower end, horizontal direction are provided with the bung slide plate, the slide plate top has the bung inlet on the cabinet casing; Be provided with the slide plate chamber in the sealing link stopper; Arranged outside has near switch below the device left side casing that declutches;

Declutch above the device, in the cavity that can seal in oil pump left side, the right side is provided with linear electric motors, and its mover top is connected with flow controller and limit switch and perfusing hole;

Flow controller is connected by the outer catheter of bellows and cavity, and the catheter other end links to each other with separating box by metal tube; Vacuum tube and catheter also are listed in cavity outside upper center, and opening is to communicate with cavity at cavity wall, the low-vacuum load-tripping device of the other end and upper end, casing left side, magnetic valve is connected, magnetic valve links to each other with gettering container by metal catheter again, separating box is fixed on gettering container upper end, and the gettering container lower end is fixed on the rolling bearing on the support; Air intake duct links to each other with vavuum pump by the rotatory sealing loose joint; The separating box top is connected with the sealing rotary guide pipe and links to each other with receiver;

Send and cover capping machine and remove head and be provided with near switch;

The automatic sterilized inoculator of the supporting special use of a kind of commercial production method for edible mushroom liquid culture bacteria;

By the vaccination mechanism of frame inner and upper and below indexing transfer mechanism and motor form; It is characterized in that: in vaccination mechanism, anion purifier is arranged on the lower end and links to each other with body, slide-bar is the left and right sides above it, and the lower end is fixed on the frame beam, and the upper end links to each other with screw rod, the screw rod upper end is fixed on the frame top, be connected with support and slide plate by slide block on the slide-bar, continuous syringe is horizontally arranged in the support, and its cylindrical shell is fixed on the rack beam, the pipette of lower end links to each other with the bellows of hopper mechanism, and its piston then is fixed on the slide plate below;

Inoculation syringe needle upper end links to each other with the syringe lower end, the lower end is arranged in anion purifier utmost point grid gap, the cylinder stent top is connected with the chain bar at grade with the slide plate top, and cylinder stent lifting chain bar and power transmission shaft branch are listed in the left and right sides of the rotten and power transmission shaft of slide plate lifting chain; Power transmission shaft all is fixed on the screw rod and can regulates on the pedestal of shift position with nut up and down according to rule, syringe rack lifting chain bar and power transmission shaft are fixed on the pedestal below, piston slide plate lifting chain bar and power transmission shaft are fixed on the top, they use synchronous cog belt, toothed belt wheel is connected, slide plate lifting chain bar power transmission shaft passes through synchronous cog belt again, toothed belt wheel and the upper fixed final drive shaft on housiung separator, regulating wheel is connected, the final drive shaft rear end is a toothed belt wheel, by the speed change power transmission shaft of synchronous cog belt and below, synchronous cog belt, toothed wheel and motor are linked;

In the indexing transfer mechanism, directed beaming roller is positioned at two ends, the inboard left and right sides of frame, the axle of its rear and front end is fixed in the rolling bearing on the frame, the back shaft roll row is listed in the frame top, its upper end-face edge and directed beaming roller upper end-face edge are at grade, attached moving axis roller is positioned at and supports the beaming roller below, and its lower edge and directed beaming roller lower edge are at grade; The conveyer belt locating wheel is between the attached moving axis roller and directed beaming roller on right side, front and back two rows, and be fixed on the frame by axle bed, conveyor belt loop is around directed beaming roller, support beaming roller, the outside of attached moving axis roller and locating wheel, the conveyer belt surface is provided with movable tooth grid, conveyer belt is fixed with jump ring on the edge, the right side is sliding sleeve and slide block below conveyer belt, and sliding sleeve is fixed on the housiung separator, and slide block is in sliding sleeve, the slide block left end links to each other with support, two groups of front and back are connected with crossbeam, and chain bar one end is fixed on the crossbeam middle part, the other end is connected on the speed change power transmission shaft, links to each other with motor by synchronous cog belt and belt wheel;

Hopper mechanism is arranged on vaccination mechanism frame upper right side, links to each other with frame by support; The support upper end is antibacterial smart seat, built-in anion purifier, antibacterial smart seat is connected with following fluid storage tank by the loose joint ball valve, and fluid storage tank is provided with diffluence pass, terminate on the bellows on the diffluence pass of fluid storage tank, the lower end is connected on the continuous syringe pipette in the vaccination mechanism; Motor and anion purifier switch are fixed on the frame of middle part;

Description of drawings

Fig. 1 is edible mushroom liquid culture bacteria commercialization technological process of production figure of the present invention;

Fig. 2 is a sterile vacuum liquid filling machine structural representation of the present invention;

Fig. 3 is a sterile vacuum filling production lines structure schematic top plan view of the present invention;

Fig. 4 is the present invention's device sealed structural representation that declutches;

Fig. 5 is that the present invention is covered groove and advanced the sealing baffle structural representation;

Fig. 6 is sterile vacuum filling mechanism control circuit figure of the present invention;

Fig. 7 is an automatic sterile inoculation device structural representation of the present invention;

Fig. 8 is a hopper structural scheme of mechanism of the present invention;

Fig. 9 is the end view of Fig. 7;

Figure 10 is the vertical view of Fig. 7;

Figure 11 is second embodiment of the invention figure;

Figure 12 is the accurate chain bar of a present invention structural representation;

Embodiment:

As shown in Figure 1: edible mushroom liquid culture bacteria commercialization production technology

The preparation of seed

The purpose of seed preparation is to breed sufficient amount, healthy and strong, highly purified seed greatly most; The quality of seed quality is very big to the fermentation influence; Seed quality is decided by bacterial classification itself on the one hand, is decided by the seed culture condition on the other hand to a great extent;

1, the used seed requirement of edible fungi submerged fermentation

(1) thalli growth is vigorous, and is energetic

The vigor of seed is relevant with the nutrition condition of medium and cell age; Should select nutritious seed culture medium for use, preferably without synthetic medium; The cell age of seed normally is controlled at the exponential phase later stage of thalline; As Asparagus, the seed cell age with 48-96 hour for well;

(2) the bacterium bulb diameter is little, quantity is many

The nutrition composition of medium has certain influence to the shape of bacterium ball; Usually, contain the many medium of sugar, the bacterium bulb diameter of turning out is little, and the medium viscosity is big, and the bacterium ball is also less; In addition, than using reciprocating type shaking table, the bacterium bulb diameter is less with rotary shaking table; In shaking bottle, add several beades or a little spring carries out shaken cultivation, the bacterium bulb diameter is diminished;

(3) purity height

The seed that edible fungi submerged fermentation is used requires high-purity, and any pollution, particularly germ contamination can not be arranged; The edible fungi submerged fermentation failure, the overwhelming majority is owing to germ contamination causes;

2, slant strains preparation

The original strain that is used for submerged fermentation need be through the purification and the evaluation of strictness; Medium: glucose 2.0%, yeast extract 0.5%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.1%, agar 2.0%, pH nature;

Sterilization: under 120-126 ℃ condition, sterilized 30 minutes;

Condition of culture: 25 ℃, 7-10 days;

3, shake-flask seed preparation

Medium: potato 20%, brown sugar 1.0-1.5%, glucose 1.0-1.5%, wheat bran 3%, peptone 0.15-0.3%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.1%, vitamin V B110 mg/litre, surplus is a water, pH value 6.5-7.0.

Prepared culture medium is sub-packed in the triangular flask, the bottled 100-150 milliliter of common 500 milliliters triangle medium, the medium of the bottled 200-250 milliliter of 750 milliliters triangle; And in triangular flask, put 3-4 bead (also can not putting), filled in tampon, waterproof paper on the bundle (or brown paper); Autoclaving (1.4 kg/cm, 2 pressure, sterilization 30 minutes), when being cooled to below 30 ℃, insert the slant strains 2-4 piece of 0.5 centimetre of 2 size, be sandwiched in then and carry out shaken cultivation on the shaking table, its Frequency Design is 80-120 time/minute; Cultivation temperature was cultivated 2-7 days at 25 ℃;

The preparation method of one-level shake-flask seed, secondary shake-flask seed is identical;

4, preparation one-level, secondary seed preparation

Except that the gemma of white fungus, general growth is all slower in edible mushroom, in order to shorten fermentation time, needs to improve inoculum concentration in the production, and the inoculum concentration of general edible fungi submerged fermentation is 10%-20%, even higher; One 10 tons fermentation tank, with 7 tons of calculating of loading amount, required seed will the 700-1400 kilogram, obviously, and such grain weight, it is unappeasable far away depending merely on shake-flask seed; For this reason, broadcast bottle seed further expanding propagation become first order seed, first order seed more expanding propagation becomes two seeds; First order seed and secondary seed are cultivated in seeding tank, and first class seed pot is generally the 50-100 liter, and the secondary seed jar is the 500-1000 liter; The first order seed medium can be selected the shake-flask culture base for use, and the medium of secondary seed in order to reduce cost, can be selected the shake-flask culture base for use, and the medium of secondary seed in order to reduce cost, can be selected fermentation medium for use; The culture technique of seeding tank sees also following ferment tank process portion;

Three, ferment tank technological parameter control

1, raw material and culture medium preparation

Medium: potato 10-20%, wheat bran 2-5%, brown sugar 0.5-2.0%, peptone 0.1-0.3%, yeast extract 0.1-0.3%, potassium dihydrogen phosphate 0.1-0.2%, magnesium sulfate 0.05-0.15%, VB11 0-20 mg/litre, polyoxypropylene glycerine 0.03-0.035%, surplus are water, pH value 4.0-8.0%; Culture medium preparation: in the following order

(1) make potato sweat and wheat bran juice: fresh potato digs the bud peeling, is cut into the sheet or the bar of 0.5-0.4 cm thick, adds 5 times of poach and use 6-8 layer filtered through gauze when shortcake does not rot, and it is standby to get juice; The wheat bran that fresh nothing is gone mouldy adds 10 times of water boils to 20 minute, and it is standby to cross leaching juice;

(2) composite soil fermented bean drink and wheat bran juice;

(3) will add behind potassium dihydrogen phosphate, the molten water of magnesium sulfate in the mixed juice;

(4) will add behind brown sugar, glucose, the molten water of peptone in the mixed juice again;

(5) vitamin B1 being pulverized the back adds in the mixed juice again;

(6) be that 10% hydrochloric acid or sodium hydroxide are adjusted pH value with concentration;

(7) sterilization: medium is packed in the culture tank, kept 30-35 minute when degree reaches 121-126 ℃ overflowing;

(8) cooling: after the sterilization, feed the cold water cooling immediately, make the medium temperature drop to 23-30 ℃;

2, cell age

The vigor of cell age and seed is closely related; If cell age is too little, mycelial concentration is low in the seed liquor, prolongs fermentation period; If cell age is too big, bacterial classification is aging, the self-dissolving of part thalline, and the vigor of seed is low, reduces fermentation yield; Different edible mushrooms, the cell age difference that it is suitable; The seed of different stage, its cell age are also different; Usually, the shake-flask seed cell age was controlled at 4-10 days, and the cell age of first order seed and secondary seed is 48-96 hour;

3, inoculum concentration

Inoculum concentration when usually, food has deep-fermentation is 10%-20%; The size of inoculum concentration directly has influence on the speed of this bacterial classification growth and breeding in fermentation tank; Inoculum concentration is big, can shorten fermentation period; This is because contain the outer hydrolase of a large amount of born of the same parents in the seed liquor, helps the hydrolysis utilization to raw material; The mycelia amount is big in the seed liquor, and the thalline accumulation is also fast in the fermentation; Simultaneously, the rapid grown bacterial classification of culture environment is occupied, and living contaminants is controlled;

4, temperature

The temperature of edible fungi submerged fermentation is different because of kind; Usually the fastest 22-30 ℃ of growth, recovery rate (output) is the highest; Woodear is best with 28 ℃, and Asparagus is 25 ℃;

5, the size of throughput influences the dissolved oxygen concentration in the culture fluid, thereby influences growth rate and the mycelia yield of mycelia; In addition, the size of throughput also influences the form of thalline, and when throughput was sufficient, the sphere of thalline was easily filtered as hickory chick, but when hypoventilation, mycelia does the disperse state growth, and zymotic fluid is sticky as a result, filtration difficulty, and harvest subtracts greatly; Yet throughput can not be too big, otherwise the culture fluid moisture loss is serious, and the throughput of edible fungi submerged fermentation is with 0.2-1.5 ventilation volume/culture fluid volume/minute be advisable; For example: preceding 24 hours of the woodear submerged fermentation with 0.25 ventilation volume/culture fluid volume/minute throughput, back 24 hours with 0.5 ventilation long-pending/culture fluid volume/minute be advisable; Asparagus then be preceding 48 hours with 1.0 ventilation volume/culture fluid volume/minute, back 48 hours with 1.5 ventilation volume/culture fluid volume/minute be advisable;

6, mixing speed

Stirring can promote the dissolving of oxygen in culture fluid in the air; The mixing speed of edible fungi submerged fermentation generally is 180-500 rev/min; The used mixing speed of woodear submerged fermentation is 500 rev/mins, and Asparagus is 220-240 rev/min;

7, acid-base value

Different types of edible mushroom all has an Optimum pH, and in suitable acid-base value environment, mycelial growth is fast; For example: the optimal pH of woodear is 6.0-6.5, and Asparagus is 6.3, and mushroom is 4.5-6.5, and straw mushroom is 6.8-7.2, and flat mushroom is 5.0-6.5; Need timing sampling during the fermentation, measure acid-base value, and adjust;

8, tank pressure

In order to increase the solvability of oxygen in zymotic fluid, and produce negative pressure outside air is polluted in entering jar by the aperture because of stirring, will keep certain tank pressure during the fermentation, be generally 0.2-0.7 kg/cm 2 for preventing;

9, foam control

(3) mechanical defoaming

Dependence is loaded on jar interior froth breaking and starches the surface tension that the variation of intense mechanical vibration and pressure destroys bubble; Reduce the stability of foam, impel lather collapse; This debubbling method is very not thorough, also needs sometimes by chemical froth breaking method;

(4) chemical froth breaking

Mainly be to borrow defoamer to reduce the mechanical dado of foam and the surface tension that reduces foam, foam is broken because of unbalance stress; Defoamer commonly used has natural oil and synthetic defoamer two classes; The natural oil that is used for froth breaking has: peanut oil, soya-bean oil, cotton oil etc.; Synthetic defoamer commonly used has the bubble enemy; Great majority are to use the bubble enemy to do defoamer at present, and addition is about 0.006%;

9, fermentation termination

Whether fermentation termination is accurate, and the output (units per ml cycle) that improves product is had extremely important meaning; Reasonably putting jar time should could determine with reference to factors such as the outward appearance of production concentration, the rate of filtration, amino nitrogen content, residual sugar amount, mycelia form, pH, zymotic fluid and viscositys; Whether measure during the fermentation, can understand course of fermentation, it is normal to have understood ferment, have not contaminated, for fermentation termination determine give information;

8. mycelia quantitative determination

Assay method commonly used has two kinds:

A) centrifugal process

Measure the sample liquid of certain volume, through 3000 rev/mins centrifugal 10 minutes, the tipping supernatant claims bacterium mud weight; The accuracy of this method is accurately according to culture fluid nutrition; If trophic component partly is insoluble, as using things such as wheat bran, yellow thing powder, prepared culture is also got off with mycelia one coprecipitation after centrifugal because of also have certain insoluble residue after fermenting, and makes measurement result higher; If centrifugal with sucrose, the result is accurately so, but technical sophistication, difficult the grasp;

B) cross elimination

According to the size of mycelium pellet, select suitable specification sieve (as soil sample sieve) for use, sample liquid is filtered with sieve, with 20-30 times of volume water flushing, the culture fluid of carrying secretly is formed flushing again, filter mycelium dry down at 80 ℃, dry to constant weight with 105 ℃ again, weigh;

9. the mycelium pellet size is measured

Sample earlier after filtration, rinse well, take a morsel mycelium pellet in culture dish, add a little distillation water and the bacterium ball is scattered and fully stretch (attention adds on a small quantity, and is unsuitable too much or very few), with slide measure at the bottom of culture dish facing to the mycelium pellet measurement;

(3) purity test

Mainly be to check it is to stagnate living contaminants is arranged; Inspection method can be cultivated or the cultivation of phenol red meat soup with dull and stereotyped, and checking after 24-28 hour has not germ contamination; This method required time of obtaining a result is longer; Also can be by microexamination, just having seen clearly not with oily mirror, bacterium exists;

10. pH measurement

Change by the pH that check to find ferment liquid, the process of passable hydrolysis and fermentation is also understood pollution condition indirectly; In general, along with the carrying out of fermentation, phenomenon appears ging up again in pH, and this is because the result of thalline self-dissolving; In addition, if germ contamination occurs,, the pH of zymotic fluid is quickly fallen to below 5.0 because the reproduction speed of bacterium is very fast, and produces many machine acid;

(5) saccharinity determining

1. the assay method of the introduction of the 22 to 24 page of the assay method of the mensuration zymotic fluid total reducing sugar of zymotic fluid total reducing sugar " biochemical technology introduction " (People's Education Publishing House) that can write with reference to biochemical microorganism teaching and research room of biology department of Zhongshan University carries out;

2. the mensuration zymotic fluid of zymotic fluid reducing sugar through 4000 rev/mins centrifugal 10 minutes, get supernatant and measure content of reducing sugar with the DNS method;

(6) method of the 61 to the 63 page of introduction of " Biochemistry Experiment " (Shanghai science tech publishing house) that amino acid whose assay method can be write with reference to Zhu Jian in the amino acid whose mensuration zymotic fluid is measured;

(7) the zymotic fluid smell is checked

The gas checking of zymotic fluid can be in the exhaust outlet inspection of fermentation tank; If fermentation is normal, can smell one mushroom perfume (or spice) or the original smell of culture fluid in exhaust ports, in the fermentation later stage, the transmission of smell possibility is tart flavour slightly; If germ contamination is arranged, ferment after 12 hours, then can smell acid smell in exhaust ports;

Five, canned production

When the liquid culture bacteria in the fermentation tank reaches fermentation termination, after qualified Upon Strict Inspection, be transported to by aseptic pipeline in the separating box of sterile vacuum filling production lines, by the sterile vacuum filling mechanism liquid spawn under vacuum state, can and seals the bung of PET polyester bucket under vacuum state with the disposable plastic intelligent lid in PET polyester bucket, then, it is pre-chilled to below 5 ℃, in 1-5 ℃ refrigerated room, preserves and sell by the form of chain direct selling; Its storage life is the longest to reach 6 months, and its product quality, performance are constant;

As Fig. 2-shown in Figure 6: edible mushroom liquid culture bacteria commercialization production equipment motion mode is:

Turn on the power switch (DK), introduce power supply, electromotor (1-46) and vavuum pump (1-55) energising work, motor is moving to be dropped to rotating speed in the setting range by speed changer, and grinding tooth wheels (1-41) in by angular wheel (1-45), power transmission shaft (1-44) kinetic energy being passed to, make grinding tooth wheel in being fixed on go up and rotation platform (1-42) in carrier bar on vacuum filling mechanism and conveyer on the left of thumb wheel (1-1) together move;

Run to the head below of cover supplier (1-25) when filling mechanism, when the sensor on the head (1-26) detects punctuate (1-27) on bung inlet (1-28) limit, point driving switch (1-25ZJ) closure, cover supplier (1-25) crawl operation is once sent into bung in the bung inlet (1-28) in the vacuum filling mechanism; Drop on the slide plate (1-30) left side of pushing cover sealed baffle (1-17);

At the volley, sensor (1-3) measure that thumb wheel (1-1) is delivered to the brake lining (1-5) of vacuum filling mechanism to the bacterium liquid bucket (1-2) on the conveyer belt and the bung inlet that (1-7) forms under the time, simultaneously the switch (1-33ZJ) of closed electric oil pump (1-33), electromagnetism three position directional valve (1-31) left bit switch (JCI) (1-38) are (JC2) and the switch (HKI) of the device that declutches (1-4), the device that declutches (1-4) is opened, and brake lining (1-5) (1-7) separately; The pushing cover sealed baffle (12-17) that the piston of hydraulic cylinder (1-29) drives its top moves left, bung (1-19) is prolonged slide plate (1-30) to be sent to and to cover in the groove (1-18), opening on lid groove (1-18) the right side cavity (1-13) is sealed, when the pressure in the hydraulic cylinder (1-29) reaches setting pressure, pressure switch (YKI) disconnects, and makes the pressurize of electromagnetism three position directional valve (1-31) outage meta; Hydraulic cylinder (1-39) drives entire mechanism and descends, make the bung end of bacterium liquid bucket (1-2) enter into brake lining (1-5) (1-7) between, when hydraulic cylinder (1-39) when dropping to extreme lower position, impact switch (1-35) HK1 commutation just makes the device that declutches (1-4) outage, and the bung of bacterium liquid bucket (1-2) is sealed in the left side cavity (1-13) in the vacuum filling mechanism; Magnetic valve (1-22) energising on vacuum tube (1-14) pipeline is opened, and (1-13) inhales vacuum to cavity; Low-vacuum load-tripping device (1-24) obtains electricity work; When reaching in the cavity (1-13) when setting vacuum (3000-4000Pa), low-vacuum load-tripping device (1-24) is gone up gauge tap (KI) closure, make flow controller (1-10) obtain to establish by cable and open, the liquid bacterium in perfusing hole (1-8) is filled into bacterium liquid bucket (1-2); Switch (HK2) commutation when reaching the setting flow is closed flow controller (1-10) outage; And the right bit switch (JC7) of the switch (FC) of the coil that falls back (FC) of connection linear electric motors (1-11) and electromagnetism three position directional valve (1-16) moves right linear electric motors (1-11) drive flow controllers (1-10); When arriving desired location, limit switch (FC) outage stops linear electric motors; The piston of hydraulic cylinder (1-22) and lid groove (1-18) move downward, and the bung (1-19) that will cover in the groove (1-18) is pressed on the bung of bacterium liquid bucket (1-2), and bacterium liquid bucket (1-2) is sealed; So far, the piston of fluid cylinder (1-22) reaches range, makes impact switch (1-6) K2 closure just, allows the device (1-4) that declutches obtain to establish by cable by delay switch (JC3) and open simultaneously; Solenoid directional control valve (1-16) (1-31) (1-38) obtains electricity left side position, and hydraulic cylinder (1-22) (1-29) turns back to initial position, and lid groove (1-18) is with the K3 closure just of the impact switch (1-21) on the link stopper (1-20); The forward of connecting linear electric motors (1-11) feeds coil (XC), and linear electric motors (1-11) are delivered to assigned address left with flow controller (1-10), limit switch (XC) outage, and linear electric motors stop; The cylinder body of hydraulic cylinder (1-39) rises, switch (1-40) HKI magnetic valve (1-23) outage that resets just; Delay switch (JC3) outage simultaneously, the device that declutches obtains; Hydraulic cylinder (1-35) continues to rise, vacuum filling mechanism is separated with bacterium liquid bucket (1-2), bacterium liquid bucket (1-2) prolongs conveyer (1-44) and goes to next procedure, when hydraulic cylinder (1-22) (1-29) reaches setting pressure in (1-39), pressure switch (YK2) disconnects, and makes (1-31) (1-38) outage meta of electromagnetism three position directional valve (1-16); Overflow valve (1-36) is opened under pressure and is made oil flow back into oil cylinder; Finish the expendable process to this whole system;

As shown in Figure 7, the automatic vaccination machine kind of drive of the present invention is:

At first open the switch (1-36) of anion purifier (1-2), purify the inoculation space, and culture bag vertically is arranged in the teeth groove on the conveyer belt (2-25), open the switch (2-38) of motor (2-34) then, entire mechanism is moved, (2-35) drops to 1.5r/min with rotating speed by toothed belt wheel, and through synchronous cog belt, toothed belt wheel (2-16) (2-14) (2-10) (2-35) respectively kinetic energy is sent to chain bar (2-33) (2-18) (2-9), the chain bar is clockwise rotated with power transmission shaft; Chain bar (2-9) in vaccination mechanism, the support (2-5) of continuous syringe when (2-18) forwarding peak to (2-4) and the slide plate (2-8) of piston (2-7) slide block (2-6) by its edge prolongs slide-bar (2-3) and is raised to the extreme higher position, continuous syringe (2-4) is finished by pipette (2-24), in the bacterium liquid bucket (2-19) of bellows (2-23) inspiration from the antibacterial smart seat (2-20), 5mL liquid spawn in loose joint ball valve (2-21) flows into fluid storage tank, when chain bar (2-9) (2-18) forwards the right side horizontal level to, support (2-5) piston (2-7) slide plate (2-8) of continuous syringe (2-4) descends thereupon, chain bar (2-33) moves right, promote support (2-32), the jump ring (2-29) that makes slide block (2-31) prolong sliding sleeve (2-30) promotion conveyer belt (2-25) edge prolongs locating wheel (2-28) moving linearly to the right; Make conveyer belt (2-25) prolong beaming roller (2-26) (2-27) (2-37) turn left;

One group of 4 bags of culture bag is delivered to the below of inoculating syringe needle (2-1), and chain bar (2-33) continues to move downward is moved to the left slide block (2-31), and conveyer belt (2-25) is parked in the below of inoculation syringe needle (2-1) under the effect of directed beaming roller (2-27); At this moment, chain bar (2-9) (2-18) is continued to rotate by horizontal level, the support (2-5) of continuous syringe (2-4) and the slide plate (2-8) of piston (2-7) also descend thereupon, make inoculation syringe needle (2-1) thrust culture bag and start injection bacterial classification, the slide plate (2-8) that (2-18) drives the support (2-5) of continuous syringe (2-4) and piston (2-7) when chain bar (2-9) is injected when moving to minimum point and is finished, chain bar (2-9) (2-18) continues to move to the left side during horizontal level, the support (2-2) of continuous syringe (2-4) and the slide plate (2-8) of piston (2-7) rise inoculation syringe needle (2-1) are transferred in culture bag fully, at this moment chain bar (2-33) has been transported to the left lateral journey position of left end, so far, entire mechanism has been finished seeded process one time, reciprocal in this way, reached continuous, the purpose of aseptic inoculation, inoculum concentration is per hour 3600 bags;

In addition, can be according to the different size of culture bag (bottle), regulate pedestal (2-14) firmly putting on screw rod (2-13) by rule (2-12) with nut (2-17), make inoculation syringe needle (2-1) be in best inoculation position, use screw rod (2-4) to regulate regulating wheel (2-15) simultaneously and make synchronous cog belt remain on work under the certain force of strain state, realize the function of a tractor serves several purposes;

The invention has the beneficial effects as follows:

Can provide that a large amount of storings are convenient, high-quality is cheap, the liquid culture bacteria product of green safety and Simply, practical, cost is low, automaticity is high automatic vaccination machine product, the mushroom farming only needs according to food With bacterium culture bag production scale, buy liquid spawn product and the automatic sterile inoculation device of respective numbers, Can implement to produce, realize the popularization and application of edible fungi liquid strain and the no dirt of inoculation production process Dye automation, scale, thereby improved quality and efficient that edible fungus culturing is produced, and make the mushroom farming Limited fund can take full advantage of on edible mushroom culture bag production link having alleviated truly The financial burden of mushroom farming and labour intensity shorten the Edible Fungi cycle, improve bacterium product quality, realize The purpose of increasing both production and income; Given full play to simultaneously the physiological property of edible fungi liquid strain and produced excellent Gesture promotes edible mashroom cultivating industry and cultivation industry realization scale, standardization, automation, batch production Industry goal;

Description of drawings

In Fig. 2: thumb wheel 1-1; Bacterium liquid bucket 1-2; Sensor 1-3; Device 1-4 declutches; Sealing brake lining 1-5; Impact switch 1-6; Sealing brake lining 1-7; Perfusing hole 1-8; Limit switch 1-9; Flow controller 1-10; Linear electric motors 1-11; Bellows 1-12; Cavity 1-13; Vacuum tube 1-14; Catheter 1-15; Electromagnetism Reversal valve and pipeline 1-16; Pushing cover sealed baffle 1-17; Cover slot 1-18; Bung 1-19; Link stopper 1-20; Impact switch 1-21; Hydraulic cylinder 1-22; Magnetic valve 1-23; Low-vacuum load-tripping device 1-24; Cover supplier 1-25; Sensor 1-26; Punctuate 1-27; Bung entrance 1-28; Hydraulic cylinder 1-29; Slide plate 1-30; Electromagnetic switch Valve and pipeline 1-31; Pressure switch 1-32; Electric oil pump 1-33; Pressure switch 1-34; Fuel tank 1-35; Overflow valve 1-36; Pressure switch 1-37; Solenoid directional control valve and pipeline 1-38; Hydraulic cylinder 1-39; Collision Switch 1-40; Interior grinding tooth wheels 1-41; Rotation platform 1-42; Conveyer 1-43; Power transmission shaft 1-44; Conical tooth and lubrication box 1-45; Motor gear box device clutch 1-46; Woven hose 1-47; Seal rotary Loose joint 1-48; Separating box 1-49; Gettering container 1-50; Rolling bearing 1-51; Rotatory sealing loose joint 1-52; Support 1-53; Air intake duct 1-54; Vavuum pump 1-55; Blast pipe 1-56;

In Fig. 7: inoculation syringe needle 2-1 anion purifier 2-2; Slide-bar 2-3; Continuous syringe 2-4; Support 2-5; Slide block 2-6; Piston 2-7; Slide plate 2-8; Chain bar and power transmission shaft 2-9; Synchronous cog belt reaches Belt wheel 2-10; Take out frame 2-11; Rule 2-12; Screw rod 2-13; Synchronous cog belt and belt wheel 2-14, Regulating wheel 2-15; Spiral shell stalk 2-16; Nut 2-17; Chain bar and power transmission shaft 2-18; Bacterium liquid bucket 2-19; Antibacterial Smart seat and support 2-20; Ball valve and loose joint 2-21; Fluid storage tank 2-22; Bellows 2-23; Imbibition Pipe 2-24; Conveyer belt 2-25; Support beaming roller 2-26; Directed beaming roller 2-27; Locating wheel 2-28; Jump ring 2-29; Fixing sliding sleeve 2-30; Slide block 2-31; Support 2-32; Chain bar and power transmission shaft 2-33; Motor 2-34; Synchronous cog belt and belt wheel 2-35; Clarifier switch 2-36; Attached moving axis rolls 2-37; Motor switch 2-38;

In Figure 12:

Key way 3-1; Propeller boss 3-2, sheet stem body 3-3; Connecting axle 3-4; Rolling bearing 3-5; Power transmission shaft 3-6; Connection Block 3-7; Bolt 3-8;

In another embodiment shown in Figure 11, when power transmission shaft rotates, the chain bar (2-18) of slide plate (2-8) And the chain bar (2-9) of support (2-5) is whole at position and the chain bar (2-33) of A1 firmly putting of CZ Rising of the motion of mechanism makes the position; AZ is the terminal point of support (2-5) rising and the slide plate of piston (2-7) (2-8) rising makes syringe (2-4) be filled the position of liquid spawn; A2, A3, C1, C3 position are Coordinated movement of various economic factors process, be the chain stalk (2-18) that makes slide plate (2-8), support (2-5) chain rotten (2-9) and The relative motion of chain bar (2-33) reaches unanimously, and the C4 position is that chain bar (2-33), slide block (2-31) push away Moving jump ring (2-29), specify the below that makes conveyer belt that the edible mushroom culture bag is delivered to inoculation syringe needle (2-1) The terminal point of inoculation position; It is that inoculation syringe needle (2-1) is finished and lunged culture bag that A4 benevolence is put, the injection bacterial classification Process, A1 is the position of transfering to inoculation syringe needle (2-1), CZ is chain stalk (2-33) band movable slider (2-31) Moving to maximum travel position left, so far is again the next time beginning of seeded process.

Claims (3)

1. commercial production method for edible mushroom liquid culture bacteria:

1-1. the preparation of seed

The quality of seed quality is very big to the fermentation influence; Be decided by bacterial classification itself and seed culture condition;

The seed requirement that edible fungi submerged fermentation is used

(1) thalli growth is vigorous, and is energetic

The cell age of seed normally is controlled at the exponential phase later stage of thalline; The seed cell age with 48-96 hour for well;

(2) the bacterium bulb diameter is little, quantity is many

The nutrition composition of medium has certain influence to the shape of bacterium ball; Usually, contain the many medium of sugar, the bacterium bulb diameter of turning out is little, and the medium viscosity is big, and the bacterium ball is also less; In addition, than using reciprocating type shaking table, the bacterium bulb diameter is less with rotary shaking table; In shaking bottle, add several beades or a little spring carries out shaken cultivation, the bacterium bulb diameter is diminished;

(3) purity height

The seed that edible fungi submerged fermentation is used requires high-purity, and any pollution can not be arranged;

1-2, slant strains preparation

The original strain that is used for submerged fermentation need be through the purification and the evaluation of strictness; Medium: glucose 2.0%, yeast extract 0.5%, potassium dihydrogen phosphate 0.1%, magnesium sulfate 0.1%, agar 2.0%, pH nature;

Sterilization: under 120-126 ℃ condition, sterilized 30 minutes;

Condition of culture: 25 ℃, 7-10 days;

1-3, shake-flask seed preparation

Medium: potato 20%, brown sugar 1.0-1.5%, glucose 1.0-1.5%, wheat bran 3%, peptone 0.15-0.3%, potassium dihydrogen phosphate 0.2%, magnesium sulfate 0.1%, vitamin V B110 mg/litre, surplus is a water, pH value 6.5-7.0.

Prepared culture medium is sub-packed in the triangular flask, and puts the 3-4 bead in triangular flask, has filled in tampon, waterproof paper on the bundle; Autoclaving, when being cooled to below 30 ℃, inserts the slant strains 2-4 piece of 0.5 centimetre of 2 size at 1.4 kg/cm, 2 pressure, sterilization 30 minutes, is sandwiched in then and carries out shaken cultivation on the shaking table, and its Frequency Design is 80-120 time/minute; Cultivation temperature was cultivated 2-7 days at 25 ℃;

The preparation method of one-level shake-flask seed, secondary shake-flask seed is identical;

1-4. preparation one-level, secondary seed preparation

Broadcast bottle seed further expanding propagation become first order seed, first order seed more expanding propagation becomes two seeds; First order seed and secondary seed are cultivated in seeding tank, and the first order seed medium is selected the shake-flask culture base for use, and the medium of secondary seed is selected the shake-flask culture base for use, and the medium of secondary seed is selected fermentation medium for use;

2-1. ferment tank technological parameter control

Raw material and culture medium preparation

Medium: potato 10-20%, wheat bran 2-5%, brown sugar 0.5-2.0%, peptone 0.1-0.3%, yeast extract 0.1-0.3%, potassium dihydrogen phosphate 0.1-0.2%, magnesium sulfate 0.05-0.15%, VB11 0-20 mg/litre, polyoxypropylene glycerine 0.03-0.035%, surplus are water, pH value 4.0-8.0%;

Culture medium preparation: in the following order

(1) make potato sweat and wheat bran juice: fresh potato digs the bud peeling, is cut into the sheet or the bar of 0.5-0.4 cm thick, adds 5 times of poach and use 6-8 layer filtered through gauze when shortcake does not rot, and it is standby to get juice; The wheat bran that fresh nothing is gone mouldy adds 10 times of water boils to 20 minute, and it is standby to cross leaching juice;

(2) composite soil fermented bean drink and wheat bran juice;

(3) will add behind potassium dihydrogen phosphate, the molten water of magnesium sulfate in the mixed juice;

(4) will add behind brown sugar, glucose, the molten water of peptone in the mixed juice again;

(5) vitamin B1 being pulverized the back adds in the mixed juice again;

(6) be that 10% hydrochloric acid or sodium hydroxide are adjusted pH value with concentration;

(7) sterilization: medium is packed in the culture tank, when temperature reaches 121-126 ℃, kept 30-35 minute;

(8) cooling: after the sterilization, feed the cold water cooling immediately, make the medium temperature drop to 23-30 ℃;

2-2, cell age

The shake-flask seed cell age was controlled at 4-10 days, and the cell age of first order seed and secondary seed is 48-96 hour;

2-3, inoculum concentration

Inoculum concentration when food has deep-fermentation is 10%-20%;

2-4, temperature

The temperature of edible fungi submerged fermentation is at 22-30 ℃;

The throughput of 2-5, edible fungi submerged fermentation is with 0.2-1.5 ventilation volume/culture fluid volume/minute be advisable;

2-6, mixing speed

The mixing speed of edible fungi submerged fermentation generally is 180-500 rev/min;

2-7, acid-base value

Optimal pH is 5.0--7.2, and flat mushroom is 5.0-6.5; Need timing sampling during the fermentation, measure acid-base value, and adjust;

2-8, tank pressure

To keep certain tank pressure during the fermentation, be generally 0.2-0.7 kg/cm 2;

2-9, foam control

(1) mechanical defoaming

Dependence is loaded on jar interior froth breaking and starches the surface tension that the variation of intense mechanical vibration and pressure destroys bubble; Reduce the stability of foam, impel lather collapse;

(2) chemical froth breaking

Defoamer commonly used has natural oil and synthetic defoamer two classes; The natural oil that is used for froth breaking has: peanut oil, soya-bean oil, cotton oil etc.; Synthetic defoamer commonly used has the bubble enemy; Great majority are to use the bubble enemy to do defoamer at present, and addition is about 0.006%;

3-1. fermentation termination

Reasonably putting jar time should could determine with reference to factors such as the outward appearance of production concentration, the rate of filtration, amino nitrogen content, residual sugar amount, mycelia form, pH, zymotic fluid and viscositys; Measure during the fermentation;

(1) mycelia quantitative determination

Assay method commonly used has two kinds:

1. centrifugal process

Measure the sample liquid of certain volume, through 3000 rev/mins centrifugal 10 minutes, the tipping supernatant claims bacterium mud weight;

2. cross elimination

According to the size of mycelium pellet, select suitable specification sieve for use, sample liquid is filtered with sieve, with the flushing of 20-30 times of volume water, the culture fluid of carrying secretly is formed flushing again, filter mycelium dry down at 80 ℃, dry to constant weight with 105 ℃ again, weigh;

(2) the mycelium pellet size is measured

Sample earlier after filtration, rinse well, take a morsel mycelium pellet in culture dish, add a little distillation water and the bacterium ball is scattered and fully stretch, measure facing to mycelium pellet at the bottom of culture dish with slide measure;

(3) purity test

Inspection is to stagnate living contaminants is arranged; Inspection method can be cultivated or the cultivation of phenol red meat soup with dull and stereotyped, and checking after 24-28 hour has not germ contamination; Also can be by microexamination, just having seen clearly not with oily mirror, bacterium exists;

(4) pH measurement

Change by the pH that check to find ferment liquid, the process of passable hydrolysis and fermentation is also understood pollution condition indirectly; In general, along with the carrying out of fermentation, phenomenon appears ging up again in pH, and this is because the result of thalline self-dissolving;

(5) sugar content

1. the mensuration of zymotic fluid total reducing sugar;

2. the mensuration of zymotic fluid reducing sugar;

(6) amino acid whose mensuration;

(7) the zymotic fluid smell is checked

The gas checking of zymotic fluid can be in the exhaust outlet inspection of fermentation tank;

4-1, canned production

When the liquid culture bacteria in the fermentation tank reaches fermentation termination, after qualified Upon Strict Inspection, be transported to by aseptic pipeline in the separating box of sterile vacuum filling production lines, by the sterile vacuum filling mechanism liquid spawn under vacuum state, can and seals the bung of PET polyester bucket under vacuum state with the disposable plastic intelligent lid in PET polyester bucket, then, it is pre-chilled to below 5 ℃, in 1-5 ℃ refrigerated room, preserves and sell by the form of chain direct selling; Its storage life is the longest to reach 6 months, and its product quality, performance are constant.

2. a kind of edible mushroom liquid culture bacteria commercialization production equipment of a kind of commercial production method for edible mushroom liquid culture bacteria special use as claimed in claim 1

The sterile vacuum filling production lines is by connecting gear, and vacuum filling mechanism manages and covers cover supplier, vavuum pump, motor, speed changer, clutch composition;

It is characterized in that:

Connecting gear comprises chain scraper conveyor, thumb wheel, and chain scraper conveyor also is listed in the annular rotary platform outside in the vacuum filling machine structure, vacuum filling mechanism below;

In sterile vacuum filling mechanism casing, the lower right is square double acting hydraulic cylinder, on the carrier bar on the annular rotary platform beaming roller below its piston is fixed on, grinding tooth wheel in the carrier bar right-hand member is fixed with, interior grinding tooth wheel is meshed with spur gear, and spur gear is connected with motor, speed changer, the clutch of below by power transmission shaft, angular wheel again; The piston left side of square hydraulic cylinder is provided with impact switch, hydraulic cylinder body then is arranged in the cabinet, its top be fuel tank with the three-position electromagnetic reversal valve that links to each other of fluid cylinder cylinder body and pressure switch, pipeline, the left side is the device that declutches, the device that declutches is connected with semicircle sealing tile fragment, be fixed with second half circular seal tile fragment on the cabinet wall of left side on the other side, be provided with impact switch in this tile fragment; The fuel tank left side is an electric oil pump, its export pipeline place is provided with pressure switch, the oil pump top is two three-position electromagnetic reversal valves, be connected with the hydraulic cylinder that is vertically fixed on cabinet left side cavity inner top with the hydraulic cylinder of crosswise fixed respectively, be connected with the pushing cover sealing baffle on the piston of top, right side hydraulic cylinder at top, cabinet right side; Be connected with on the piston of left side top hydraulic cylinder and cover groove, lid groove right openings, built-in jump ring, with on the lid channel opening corresponding cavity onesize opening is arranged also, open lower end, horizontal direction are provided with the bung slide plate, the slide plate top has the bung inlet on the cabinet casing; Be provided with the slide plate chamber in the sealing link stopper; Arranged outside has near switch below the device left side casing that declutches;

Declutch above the device, in the cavity that can seal in oil pump left side, the right side is provided with linear electric motors, and its mover top is connected with flow controller and limit switch and perfusing hole;

Flow controller is connected by the outer catheter of bellows and cavity, and the catheter other end links to each other with separating box by metal tube; Vacuum tube and catheter also are listed in cavity outside upper center, and opening is to communicate with cavity at cavity wall, the low-vacuum load-tripping device of the other end and upper end, casing left side, magnetic valve is connected, magnetic valve links to each other with gettering container by metal catheter again, separating box is fixed on gettering container upper end, and the gettering container lower end is fixed on the rolling bearing on the support; Air intake duct links to each other with vavuum pump by the rotatory sealing loose joint; The separating box top is connected with the sealing rotary guide pipe and links to each other with receiver;

Send and cover capping machine and remove head and be provided with near switch.

3. the automatic sterilized inoculator of the supporting special use of a kind of commercial production method for edible mushroom liquid culture bacteria as claimed in claim 1;

By the vaccination mechanism of frame inner and upper and below indexing transfer mechanism and motor form; It is characterized in that: in vaccination mechanism, anion purifier is arranged on the lower end and links to each other with body, slide-bar is the left and right sides above it, and the lower end is fixed on the frame beam, and the upper end links to each other with screw rod, the screw rod upper end is fixed on the frame top, be connected with support and slide plate by slide block on the slide-bar, continuous syringe is horizontally arranged in the support, and its cylindrical shell is fixed on the rack beam, the pipette of lower end links to each other with the bellows of hopper mechanism, and its piston then is fixed on the slide plate below;

Inoculation syringe needle upper end links to each other with the syringe lower end, the lower end is arranged in anion purifier utmost point grid gap, the cylinder stent top is connected with the chain bar at grade with the slide plate top, and cylinder stent lifting chain bar and power transmission shaft branch are listed in the left and right sides of the rotten and power transmission shaft of slide plate lifting chain; Power transmission shaft all is fixed on the screw rod and can regulates on the pedestal of shift position with nut up and down according to rule, syringe rack lifting chain bar and power transmission shaft are fixed on the pedestal below, piston slide plate lifting chain bar and power transmission shaft are fixed on the top, they use synchronous cog belt, toothed belt wheel is connected, slide plate lifting chain bar power transmission shaft passes through synchronous cog belt again, toothed belt wheel and the upper fixed final drive shaft on housiung separator, regulating wheel is connected, the final drive shaft rear end is a toothed belt wheel, by the speed change power transmission shaft of synchronous cog belt and below, synchronous cog belt, toothed wheel and motor are linked;

In the indexing transfer mechanism, directed beaming roller is positioned at two ends, the inboard left and right sides of frame, the axle of its rear and front end is fixed in the rolling bearing on the frame, the back shaft roll row is listed in the frame top, its upper end-face edge and directed beaming roller upper end-face edge are at grade, attached moving axis roller is positioned at and supports the beaming roller below, and its lower edge and directed beaming roller lower edge are at grade; The conveyer belt locating wheel is between the attached moving axis roller and directed beaming roller on right side, front and back two rows, and be fixed on the frame by axle bed, conveyor belt loop is around directed beaming roller, support beaming roller, the outside of attached moving axis roller and locating wheel, the conveyer belt surface is provided with movable tooth grid, conveyer belt is fixed with jump ring on the edge, the right side is sliding sleeve and slide block below conveyer belt, and sliding sleeve is fixed on the housiung separator, and slide block is in sliding sleeve, the slide block left end links to each other with support, two groups of front and back are connected with crossbeam, and chain bar one end is fixed on the crossbeam middle part, the other end is connected on the speed change power transmission shaft, links to each other with motor by synchronous cog belt and belt wheel;

Hopper mechanism is arranged on vaccination mechanism frame upper right side, links to each other with frame by support; The support upper end is antibacterial smart seat, built-in anion purifier, antibacterial smart seat is connected with following fluid storage tank by the loose joint ball valve, and fluid storage tank is provided with diffluence pass, terminate on the bellows on the diffluence pass of fluid storage tank, the lower end is connected on the continuous syringe pipette in the vaccination mechanism; Motor and anion purifier switch are fixed on the frame of middle part.

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