CN106333059A - Mixed fermentation method of biological feed leavening agent - Google Patents
Mixed fermentation method of biological feed leavening agent Download PDFInfo
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- CN106333059A CN106333059A CN201610726612.8A CN201610726612A CN106333059A CN 106333059 A CN106333059 A CN 106333059A CN 201610726612 A CN201610726612 A CN 201610726612A CN 106333059 A CN106333059 A CN 106333059A
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- 238000000855 fermentation Methods 0.000 title claims abstract description 92
- 230000004151 fermentation Effects 0.000 title claims abstract description 92
- 238000000034 method Methods 0.000 title claims abstract description 15
- 235000010855 food raising agent Nutrition 0.000 title abstract 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 claims abstract description 26
- 239000001301 oxygen Substances 0.000 claims abstract description 26
- 229910052760 oxygen Inorganic materials 0.000 claims abstract description 26
- 241001148470 aerobic bacillus Species 0.000 claims abstract description 25
- 238000004519 manufacturing process Methods 0.000 claims abstract description 25
- 239000001963 growth medium Substances 0.000 claims abstract description 8
- 238000005070 sampling Methods 0.000 claims abstract description 7
- 230000000740 bleeding effect Effects 0.000 claims description 42
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 39
- 238000000386 microscopy Methods 0.000 claims description 18
- 239000000843 powder Substances 0.000 claims description 15
- 239000007787 solid Substances 0.000 claims description 13
- WCUXLLCKKVVCTQ-UHFFFAOYSA-M Potassium chloride Chemical compound [Cl-].[K+] WCUXLLCKKVVCTQ-UHFFFAOYSA-M 0.000 claims description 12
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 claims description 12
- 239000011575 calcium Substances 0.000 claims description 12
- 229910052791 calcium Inorganic materials 0.000 claims description 12
- 238000011081 inoculation Methods 0.000 claims description 12
- 235000013336 milk Nutrition 0.000 claims description 12
- 239000008267 milk Substances 0.000 claims description 12
- 210000004080 milk Anatomy 0.000 claims description 12
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 12
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 12
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 12
- 239000002609 medium Substances 0.000 claims description 10
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims description 10
- 229920000053 polysorbate 80 Polymers 0.000 claims description 10
- 239000013530 defoamer Substances 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 8
- 239000000463 material Substances 0.000 claims description 7
- 229920002261 Corn starch Polymers 0.000 claims description 6
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 6
- 239000001888 Peptone Substances 0.000 claims description 6
- 108010080698 Peptones Proteins 0.000 claims description 6
- 229940041514 candida albicans extract Drugs 0.000 claims description 6
- 238000004140 cleaning Methods 0.000 claims description 6
- 239000008120 corn starch Substances 0.000 claims description 6
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 claims description 6
- 238000007599 discharging Methods 0.000 claims description 6
- 230000008030 elimination Effects 0.000 claims description 6
- 238000003379 elimination reaction Methods 0.000 claims description 6
- 235000021050 feed intake Nutrition 0.000 claims description 6
- 239000011790 ferrous sulphate Substances 0.000 claims description 6
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 6
- 239000008103 glucose Substances 0.000 claims description 6
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 6
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical class CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 6
- 239000000203 mixture Substances 0.000 claims description 6
- 238000006213 oxygenation reaction Methods 0.000 claims description 6
- 235000019319 peptone Nutrition 0.000 claims description 6
- 239000001103 potassium chloride Substances 0.000 claims description 6
- 235000011164 potassium chloride Nutrition 0.000 claims description 6
- 239000011780 sodium chloride Substances 0.000 claims description 6
- 238000004659 sterilization and disinfection Methods 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 210000005239 tubule Anatomy 0.000 claims description 6
- 239000012138 yeast extract Substances 0.000 claims description 6
- 235000015097 nutrients Nutrition 0.000 claims description 5
- 241000193755 Bacillus cereus Species 0.000 claims description 4
- 241000194108 Bacillus licheniformis Species 0.000 claims description 4
- 244000199885 Lactobacillus bulgaricus Species 0.000 claims description 4
- 235000013960 Lactobacillus bulgaricus Nutrition 0.000 claims description 4
- 240000006024 Lactobacillus plantarum Species 0.000 claims description 4
- 235000013965 Lactobacillus plantarum Nutrition 0.000 claims description 4
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 4
- 229940004208 lactobacillus bulgaricus Drugs 0.000 claims description 4
- 229940072205 lactobacillus plantarum Drugs 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 241000194020 Streptococcus thermophilus Species 0.000 claims description 3
- DPJRMOMPQZCRJU-UHFFFAOYSA-M thiamine hydrochloride Chemical compound Cl.[Cl-].CC1=C(CCO)SC=[N+]1CC1=CN=C(C)N=C1N DPJRMOMPQZCRJU-UHFFFAOYSA-M 0.000 claims description 3
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 2
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 2
- 241000193830 Bacillus <bacterium> Species 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 4
- 241001148471 unidentified anaerobic bacterium Species 0.000 abstract 3
- 230000001580 bacterial effect Effects 0.000 abstract 2
- 238000011109 contamination Methods 0.000 abstract 2
- 230000000249 desinfective effect Effects 0.000 abstract 1
- 241000894006 Bacteria Species 0.000 description 9
- 238000009423 ventilation Methods 0.000 description 6
- FYYHWMGAXLPEAU-UHFFFAOYSA-N Magnesium Chemical compound [Mg] FYYHWMGAXLPEAU-UHFFFAOYSA-N 0.000 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
- 239000011777 magnesium Substances 0.000 description 4
- 229910052749 magnesium Inorganic materials 0.000 description 4
- 239000001117 sulphuric acid Substances 0.000 description 4
- 235000011149 sulphuric acid Nutrition 0.000 description 4
- 244000063299 Bacillus subtilis Species 0.000 description 3
- 235000014469 Bacillus subtilis Nutrition 0.000 description 3
- 244000046052 Phaseolus vulgaris Species 0.000 description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 3
- 150000002333 glycines Chemical class 0.000 description 3
- 238000011169 microbiological contamination Methods 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 150000003722 vitamin derivatives Chemical class 0.000 description 3
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 241001660259 Cereus <cactus> Species 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 230000000529 probiotic effect Effects 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 239000010902 straw Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/123—Bulgaricus
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/21—Streptococcus, lactococcus
- A23V2400/249—Thermophilus
Landscapes
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a method for preparing a feed leavening agent through a mixed fermentation technology, and mainly aims at solving the problems that due to the fact that at preset, aerobic bacteria and anaerobic bacteria are cultured separately, more production links are needed, and the bacterial contamination probability is increased. The method comprises the steps that a seed solution A, a seed solution B and a synthetic culture medium are prepared, and a fermentation tank is disinfected; feeding, disinfecting and inoculating are conducted in sequence, standing is conducted for two hours after inoculating is conducted, an exhaust valve arranged on the fermentation tank is switched on, oxygen is introduced for aerobic culture, the oxygen is introduced for 8 hours, sampling is conducted for a microscopic examination, the growth rates of the aerobic bacteria and the anaerobic bacteria are observed, when the aerobic bacteria reach 75%, oxygen adding is stopped, an air valve is switched off for anaerobic culture for about 8 hours, then a microscopic examination is conducted, when the anaerobic bacteria account for 70%, the air valve arranged on the fermentation tank is switched on for aerobic culture for 8 hours, then a microscopic examination is conducted, when the aerobic bacteria account for 80% or above, oxygen adding is stopped, anaerobic culture is conducted completely, and culture is finished after 48 hours. The method has the advantages of reducing the production links and the bacterial contamination probability.
Description
Technical field:
The present invention relates to a kind of method preparing feed fermentation agent using mixed fermentation technology.
Background technology:
The fermentation technology of standard biologic feed fermentation agent is aerobic bacteria and anaerobe is cultivated respectively, and similar, cultural characters are identical
Or similar strain can be in same fermentation tank mixed fermentation.It is disadvantageous in that aerobic bacteria and anaerobe cultivate production respectively
Link is many, increases microbiological contamination chance, and production cost is high.
Content of the invention:
The technical problem to be solved is to provide a kind of strain mixed fermentation by different aerobic-types, thus reducing production
Link, reduces microbiological contamination chance, reduces the mixed fermentation method of the biological feedstuff leaven of production cost.
Above-mentioned purpose is achieved in that
One, the preparation of seed liquor:
Make a seed liquor: weigh 1 gram of lactic acid class solid mycopowder and be inoculated in 100 milliliters of triangles equipped with 100 milliliters of synthetic mediums
In bottle, inoculate 5 bottles；32 DEG C of quiescent culture 72 hours, use plastic wraps bottleneck, Anaerobic culturel；Obtain a seed liquor；
Make b seed liquor: weigh 1 gram of spore class solid mycopowder and be inoculated in 100 milliliters of triangles equipped with 100 milliliters of synthetic mediums
In bottle, inoculate 5 bottles；Place in constant-temperature table, 32 DEG C, cultivate 24 hours under the conditions of 150 revs/min, obtain b seed liquor；
The manufacture method of synthetic medium is as follows: takes 20 grams of glucose, 10 grams of corn starch, 1 gram of dipotassium hydrogen phosphate, magnesium sulfate
0.5 gram, 1 gram of sodium chloride, 1 gram of potassium chloride, 0.01 gram of ferrous sulfate, 3 grams of yeast extract, 5 grams of peptone, vitamin b1 10 (every
10 milligrams of piece) it is added in 1000 milliliters of water, mix homogeneously, ph is natural；In each 100 milliliters of triangular flask, 100 milliliters of subpackage is comprehensive
Close culture medium；Autoclaving 15 minutes at a temperature of 121 DEG C.
Two, production process:
(1), 0.6 ton of fermentation tank is carried out disinfection:
First tubule on the air cock of fermentation tank is transferred on air inlet valve, opens lower section steam inlet valve, open top
Air bleeding valve, opens the condensed water in underlying discharging opening valve emptying fermentation tank, simultaneously while fermentation tank temperature rise
Note the condensed water elimination of air cleaning tank being connected with fermentation tank, after steam is discharged from check-valves, start the clock, high
Sterilize 20 minutes under warm (121 DEG C), high pressure (0.15 MPa) state；
(2), make production culture medium:
Weigh 6 kilograms of brown sugar, 0.35 kilogram of potassium dihydrogen phosphate, 1 kilogram of defat high-calcium milk powder, 200 milliliters of tween 80, defoamer
100 milliliters of (Oleum Glycines), dissolve in water brown sugar, potassium dihydrogen phosphate, defat high-calcium milk powder being put into 15 20 kilograms, then will
Tween 80, defoamer (Oleum Glycines) are added in solution, stir, and ph is natural；In supply fermentation tank, growth of microorganism supports
Point.
(3), feed intake:
The above-mentioned material having dissolved is added in fermentation tank, then adds water, treat that liquid reaches and stop adding water between two visors.Greatly
About plus 0.5 ton of water.
(4), sterilize:
Close dog-house, top air bleeding valve remains on, and then opens tank bottoms inlet valve and heats, and makes tank temperature reach 100 degree,
Close top air bleeding valve, when gauge hand reaches 0.05 MPa, opening air bleeding valve makes its pressure be zeroed, and turns off air bleeding valve
Gauge hand is made to reach 0.15 MPa, pressurize closed inlet valve after 30 minutes.
(5), inoculate
Treat fermentation tank temperature drop to 35 degree about, the described a seed liquor of flame protection inoculation and b seed liquor are in fermentation tank.
After inoculation, stand two hours, open the air bleeding valve on fermentation tank, logical oxygen carries out aerobic culture, logical oxygen 8 hours, leads to
Cross crack air bleeding valve to control tank pressure at 0.05 MPa.Sampling microscopy, observes the growth ratio of aerobic bacteria and anaerobe, is a good
Oxygen bacterium is high, when ratio reaches 75%, stops oxygenation (if ratio is not enough, ventilation carries out aerobic culture), closes air cock and carry out
Anaerobic culturel, carries out microscopy in 8 hours about again, when anaerobe accounts for 70%, opens air cock on fermentation tank (if ratio is not
Foot, then proceed Anaerobic culturel), carry out aerobic culture, microscopy after 8 hours, stop when aerobic bacteria accounts for more than 80% supplying
Support, carry out Anaerobic culturel completely, cultivate after 48 hours and terminate, now anaerobe accounts for 90% about.
Tank temperature controls always at 30 ± 2 DEG C.
Described lactic acid class solid mycopowder is Lactobacillus plantarum, streptococcus thermophiluss, Lactobacillus bulgaricus；Described spore
Class solid mycopowder is bacillus subtilises, bacillus licheniformis, Bacillus cereuss.
The invention has the advantage that mixed fermentation is by oxygen different demands, but do not have to be mutually exclusive, the not bacterium of mutually suppression
Kind it is blended in a fermentation tank, by the condition being suitable for its growth, ventilation are created to different strain to the control of ventilation
When, in a dormant state, oxygen is not lethal factor to it to anaerobe, aerobic bacteria amount reproduction；During quiescent culture, at aerobic bacteria
In dormancy, anaerobe is bred.
After strain accesses fermentation tank, 8 hours of first ventilating, aerobic bacteria amount reproduction, then quiescent culture, aerobic bacteria continues
Using the oxygen of remaining in fermentation liquid, make the anaerobic state of fermentation tank more preferably, be suitable for the growth of anaerobe, with the consumption of oxygen
To the greatest extent, aerobic bacillus cereuss adapt to adverse circumstances and form hypopuss-spore.Anaerobe starts amount reproduction, and final production goes out anaerobism
Mixed fermentation liquid based on bacterium, supplemented by aerobic bacteria.The advantage of mixed fermentation: the strain mixed fermentation of different aerobic-types, reduces life
Produce link, reduce microbiological contamination chance, reduce production cost.
In fermentation termination product, anaerobe can account for 90% about, and fermented product, as the Inoculant of the feedstuff substrate such as straw, passes through
Solid fermentation to feedstuff substrate, produce be conducive in a large number growth of animal, efficient containing abundant nutrition material and probiotic bacteria
High-quality biological feedstuff.
Specific embodiment:
One, the preparation of seed liquor:
Make a seed liquor: weigh 1 gram of lactic acid class solid mycopowder and be inoculated in 100 milliliters of triangles equipped with 100 milliliters of synthetic mediums
In bottle, inoculate 5 bottles；32 DEG C of quiescent culture 72 hours, use plastic wraps bottleneck, Anaerobic culturel；Obtain a seed liquor.
Make b seed liquor: weigh 1 gram of spore class solid mycopowder and be inoculated in 100 milliliters equipped with 100 milliliters of synthetic mediums
In triangular flask, inoculate 5 bottles；Place in constant-temperature table, 32 DEG C, cultivate 24 hours under the conditions of 150 revs/min, obtain b seed liquor.
The manufacture method of synthetic medium is as follows: takes 20 grams of glucose, 10 grams of corn starch, 1 gram of dipotassium hydrogen phosphate, sulphuric acid
0.5 gram of magnesium, 1 gram of sodium chloride, 1 gram of potassium chloride, 0.01 gram of ferrous sulfate, 3 grams of yeast extract, 5 grams of peptone, vitamin b110 piece
(every 10 milligrams) are added in 1000 milliliters of water, mix homogeneously, and ph is natural；100 milliliters of subpackage in each 100 milliliters of triangular flask
Synthetic medium；Autoclaving 15 minutes at a temperature of 121 DEG C.
Two, production process:
1,0.6 ton of fermentation tank is carried out disinfection:
First tubule on the air cock of fermentation tank is transferred on air inlet valve, opens lower section steam inlet valve, open top
Air bleeding valve, opens the condensed water in underlying discharging opening valve emptying fermentation tank, simultaneously while fermentation tank temperature rise
Note the condensed water elimination of air cleaning tank being connected with fermentation tank, after steam is discharged from check-valves, start the clock, high
Sterilize 20 minutes under warm (121 DEG C), high pressure (0.15 MPa) state.
2nd, make production culture medium:
Weigh 6 kilograms of brown sugar, 0.35 kilogram of potassium dihydrogen phosphate, 1 kilogram of defat high-calcium milk powder, 200 milliliters of tween 80, defoamer
100 milliliters, dissolve in water brown sugar, potassium dihydrogen phosphate, defat high-calcium milk powder being put into 15 20 kilograms, then by tween-
80th, defoamer (Oleum Glycines) is added in solution, stirs, and ph is natural.The nutrient of growth of microorganism in supply fermentation tank.
3rd, feed intake:
The above-mentioned material having dissolved is added in fermentation tank, then adds water, treat that liquid reaches and stop adding water between two visors.Greatly
About plus 0.5 ton of water.
4th, sterilize:
Close dog-house, top air bleeding valve remains on, and then opens tank bottoms inlet valve and heats, and makes tank temperature reach 100 degree,
Close top air bleeding valve, when gauge hand reaches 0.05 MPa, opening air bleeding valve makes its pressure be zeroed, and turns off air bleeding valve
Gauge hand is made to reach 0.15 MPa, pressurize closed inlet valve after 30 minutes.
5th, inoculate
Treat fermentation tank temperature drop to 35 degree about, the described a seed liquor of flame protection inoculation and b seed liquor are in fermentation tank.
After inoculation, stand two hours, open the air bleeding valve on fermentation tank, logical oxygen carries out aerobic culture, logical oxygen 8 hours, leads to
Cross crack air bleeding valve to control tank pressure at 0.05 MPa.Sampling microscopy, observes the growth ratio of aerobic bacteria and anaerobe, is a good
Oxygen bacterium is high, when ratio reaches 75%, stops oxygenation (if ratio is not enough, ventilation carries out aerobic culture), closes air cock and carry out
Anaerobic culturel, carries out microscopy in 8 hours about again, when anaerobe accounts for 70%, opens air cock on fermentation tank (if ratio is not
Foot, then proceed Anaerobic culturel), carry out aerobic culture, microscopy after 8 hours, stop when aerobic bacteria accounts for more than 80% supplying
Support, carry out Anaerobic culturel completely, cultivate after 48 hours and terminate, now anaerobe accounts for 90% about.
Tank temperature controls always at 30 ± 2 DEG C.
Described lactic acid class solid mycopowder is Lactobacillus plantarum, streptococcus thermophiluss, Lactobacillus bulgaricus；Described spore
Class solid mycopowder is bacillus subtilises, bacillus licheniformis, Bacillus cereuss.
Embodiment 1,
Make a seed liquor: weigh 1 gram of Lactobacillus plantarum and be inoculated in 100 milliliters of triangular flasks equipped with 100 milliliters of synthetic mediums
In, inoculate 5 bottles；32 DEG C of quiescent culture 72 hours, use plastic wraps bottleneck, Anaerobic culturel；Obtain a seed liquor.
Make b seed liquor: weigh 1 gram of bacillus subtilis, be inoculated in 100 milliliters equipped with 100 milliliters of synthetic mediums
In triangular flask, inoculate 5 bottles；Place in constant-temperature table, 32 DEG C, cultivate 24 hours under the conditions of 150 revs/min, obtain b seed liquor.
The manufacture method of synthetic medium is as follows: by 20 grams of glucose, 10 grams of corn starch, 1 gram of dipotassium hydrogen phosphate, sulphuric acid
0.5 gram of magnesium, 1 gram of sodium chloride, 1 gram of potassium chloride, 0.01 gram of ferrous sulfate, 3 grams of yeast extract, 5 grams of peptone, vitamin b1 10
(every 10 milligrams) are added in 1000 milliliters of water, mix homogeneously, and ph is natural；100 milliliters of subpackage in each 100 milliliters of triangular flask
Synthetic medium；Autoclaving 15 minutes at a temperature of 121 DEG C.
(1), 0.6 ton of fermentation tank is carried out disinfection:
First tubule on the air cock of fermentation tank is transferred on air inlet valve, opens lower section steam inlet valve, open top
Air bleeding valve, opens the condensed water in underlying discharging opening valve emptying fermentation tank, simultaneously while fermentation tank temperature rise
Note the condensed water elimination of air cleaning tank being connected with fermentation tank, after steam is discharged from check-valves, start the clock, high
Sterilize 20 minutes under warm (121 DEG C), high pressure (0.15 MPa) state.
(2), make production culture medium:
Weigh 6 kilograms of brown sugar, 0.35 kilogram of potassium dihydrogen phosphate, 1 kilogram of defat high-calcium milk powder, 200 milliliters of tween 80, defoamer
100 milliliters, dissolve in water brown sugar, potassium dihydrogen phosphate, defat high-calcium milk powder being put into 15 kilograms, then by tween 80, bean
Oil is added in solution, stirs, and ph is natural.The nutrient of growth of microorganism in supply fermentation tank.
(3), feed intake:
Then plus about 0.5 ton of water the above-mentioned material having dissolved is added in fermentation tank, treats that liquid reaches and stop between two visors
Only add water.
(4), sterilize:
Close dog-house, top air bleeding valve remains on, and then opens tank bottoms inlet valve and heats, and makes tank temperature reach 100 degree,
Close top air bleeding valve, when gauge hand reaches 0.05 MPa, opening air bleeding valve makes its pressure be zeroed, and turns off air bleeding valve
Gauge hand is made to reach 0.15 MPa, pressurize closed inlet valve after 30 minutes.
(5), inoculate:
Treat fermentation tank temperature drop to 35 degree about, the described a seed liquor of flame protection inoculation and b seed liquor are in fermentation tank.
After inoculation, stand two hours, open the air bleeding valve on fermentation tank, logical oxygen carries out aerobic culture, logical oxygen 8 hours, leads to
Cross crack air bleeding valve to control tank pressure at 0.05 MPa.Sampling microscopy, observes the growth ratio of aerobic bacteria and anaerobe, is a good
When oxygen bacterium reaches 75% at high proportion, stop oxygenation, close air cock and carry out Anaerobic culturel, carry out microscopy again within 8 hours about, when detesting
When oxygen bacterium accounts for 70%, open the air cock on fermentation tank, carry out aerobic culture, microscopy after 8 hours, when aerobic bacteria accounts for more than 80%
When stop support, carry out Anaerobic culturel completely, cultivate after 48 hours and terminate, now anaerobe accounts for 90% about.Tank temperature is controlled always
System is at 30 ± 2 DEG C.
Embodiment 2,
One, the preparation of seed liquor:
Make a seed liquor: weigh 1 gram of thermophilus mycopowder, be inoculated in 100 milliliters of triangles equipped with 100 milliliters of synthetic mediums
In bottle, inoculate 5 bottles；32 DEG C of quiescent culture 72 hours, use plastic wraps bottleneck, Anaerobic culturel；Obtain a seed liquor.
Make b seed liquor: weigh 1 gram of bacillus licheniformis powder, be inoculated in 100 millis equipped with 100 milliliters of synthetic mediums
Rise in triangular flask, inoculate 5 bottles；Place in constant-temperature table, 32 DEG C, cultivate 24 hours under the conditions of 150 revs/min, obtain b seed liquor.
The manufacture method of synthetic medium is as follows: by 20 grams of glucose, 10 grams of corn starch, 1 gram of dipotassium hydrogen phosphate, sulphuric acid
0.5 gram of magnesium, 1 gram of sodium chloride, 1 gram of potassium chloride, 0.01 gram of ferrous sulfate, 3 grams of yeast extract, 5 grams of peptone, vitamin b110 piece
(every 10 milligrams) are added in 1000 milliliters of water, mix homogeneously, and ph is natural；100 milliliters of subpackage in each 100 milliliters of triangular flask
Synthetic medium；Autoclaving 15 minutes at a temperature of 121 DEG C.
Two, production process:
(1), 0.6 ton of fermentation tank is carried out disinfection:
First tubule on the air cock of fermentation tank is transferred on air inlet valve, opens lower section steam inlet valve, open top
Air bleeding valve, opens the condensed water in underlying discharging opening valve emptying fermentation tank, simultaneously while fermentation tank temperature rise
Note the condensed water elimination of air cleaning tank being connected with fermentation tank, after steam is discharged from check-valves, start the clock,
121 DEG C, sterilize 20 minutes under 0.15 MPa of state of high pressure.
(2), make production culture medium:
Weigh 6 kilograms of brown sugar, 0.35 kilogram of potassium dihydrogen phosphate, 1 kilogram of defat high-calcium milk powder, 200 milliliters of tween 80, defoamer
100 milliliters, dissolve in water brown sugar, potassium dihydrogen phosphate, defat high-calcium milk powder being put into 18 kilograms, then by tween 80, bean
Oil is added in solution, stirs, and ph is natural.The nutrient of growth of microorganism in supply fermentation tank.
(3), feed intake:
Then plus about 0.5 ton of water the above-mentioned material having dissolved is added in fermentation tank, treats that liquid reaches and stop between two visors
Add water.
(4), sterilize:
Close dog-house, top air bleeding valve remains on, and then opens tank bottoms inlet valve and heats, and makes tank temperature reach 100 degree,
Close top air bleeding valve, when gauge hand reaches 0.05 MPa, opening air bleeding valve makes its pressure be zeroed, and turns off air bleeding valve
Gauge hand is made to reach 0.15 MPa, pressurize closed inlet valve after 30 minutes.
(5), inoculate
Treat fermentation tank temperature drop to 35 degree about, the described a seed liquor of flame protection inoculation and b seed liquor are in fermentation tank.
After inoculation, stand two hours, open the air bleeding valve on fermentation tank, logical oxygen carries out aerobic culture, logical oxygen 8 hours, leads to
Cross crack air bleeding valve to control tank pressure at 0.05 MPa.Sampling microscopy, observes the growth ratio of aerobic bacteria and anaerobe, is a good
When oxygen bacterium reaches 75%, stop oxygenation (if ratio is not enough, ventilation carries out aerobic culture), close air cock and carry out anaerobism training
Support, carry out microscopy again within 8 hours about, when anaerobe accounts for 70%, the air cock opened on fermentation tank (if ratio is not enough, continues
Continue and carry out Anaerobic culturel), carry out aerobic culture, microscopy after 8 hours, stop when aerobic bacteria accounts for more than 80% supporting, enter completely
Row Anaerobic culturel, cultivates after 48 hours and terminates, now anaerobe accounts for 90% about.
Tank temperature controls always at 30 ± 2 DEG C.
Embodiment 3,
Make a seed liquor: weigh 1 gram of Lactobacillus bulgaricus powder and be inoculated in 100 milliliter three equipped with 100 milliliters of synthetic mediums
In the bottle of angle, inoculate 5 bottles；32 DEG C of quiescent culture 72 hours, use plastic wraps bottleneck, Anaerobic culturel；Obtain a seed liquor.
Make b seed liquor: weigh 1 gram of Bacillus cereuss powder and be inoculated in 100 milliliters equipped with 100 milliliters of synthetic mediums
In triangular flask, inoculate 5 bottles；Place in constant-temperature table, 32 DEG C, cultivate 24 hours under the conditions of 150 revs/min, obtain b seed liquor.
The manufacture method of synthetic medium is as follows: takes 20 grams of glucose, 10 grams of corn starch, 1 gram of dipotassium hydrogen phosphate, sulphuric acid
0.5 gram of magnesium, 1 gram of sodium chloride, 1 gram of potassium chloride, 0.01 gram of ferrous sulfate, 3 grams of yeast extract, 5 grams of peptone, vitamin b110 piece
(every 10 milligrams) are added in 1000 milliliters of water, mix homogeneously, and ph is natural；100 milliliters of subpackage in each 100 milliliters of triangular flask
Synthetic medium；121 DEG C of autoclaving, 15 minutes.
Two, production process:
(1), 0.6 ton of fermentation tank is carried out disinfection:
First tubule on the air cock of fermentation tank is transferred on air inlet valve, opens lower section steam inlet valve, open top
Air bleeding valve, opens the condensed water in underlying discharging opening valve emptying fermentation tank, simultaneously while fermentation tank temperature rise
Note the condensed water elimination of air cleaning tank being connected with fermentation tank, after steam is discharged from check-valves, start the clock, high
Sterilize 20 minutes under warm (121 DEG C), high pressure (0.15 MPa) state.
(2), make production culture medium:
Weigh 6 kilograms of brown sugar, 0.35 kilogram of potassium dihydrogen phosphate, 1 kilogram of defat high-calcium milk powder, 200 milliliters of tween 80, defoamer
100 milliliters, dissolve in water brown sugar, potassium dihydrogen phosphate, defat high-calcium milk powder being put into 20 kilograms, then by tween 80, bean
Oil is added in solution, stirs, and ph is natural.The nutrient of growth of microorganism in supply fermentation tank.
(3), feed intake:
The above-mentioned material having dissolved is added in fermentation tank, then adds water, treat that liquid reaches and stop adding water between two visors.Greatly
About plus 0.5 ton of water.
(4), sterilize:
Close dog-house, top air bleeding valve remains on, and then opens tank bottoms inlet valve and heats, and makes tank temperature reach 100 degree,
Close top air bleeding valve, when gauge hand reaches 0.05 MPa, opening air bleeding valve makes its pressure be zeroed, and turns off air bleeding valve
Gauge hand is made to reach 0.15 MPa, pressurize closed inlet valve after 30 minutes.
(5), inoculate
Treat fermentation tank temperature drop to 35 degree about, the described a seed liquor of flame protection inoculation and b seed liquor are in fermentation tank.
After inoculation, stand two hours, open the air bleeding valve on fermentation tank, logical oxygen carries out aerobic culture, logical oxygen 8 hours, leads to
Cross crack air bleeding valve to control tank pressure at 0.05 MPa.Sampling microscopy, observes the growth ratio of aerobic bacteria and anaerobe, is a good
Oxygen bacterium is high, when ratio reaches 75%, stops oxygenation (if ratio is not enough, ventilation carries out aerobic culture), closes air cock and carry out
Anaerobic culturel, carries out microscopy in 8 hours about again, when anaerobe accounts for 70%, opens air cock on fermentation tank (if ratio is not
Foot, then proceed Anaerobic culturel), carry out aerobic culture, microscopy after 8 hours, stop when aerobic bacteria accounts for more than 80% supplying
Support, carry out Anaerobic culturel completely, cultivate after 48 hours and terminate, now anaerobe accounts for 90% about.
Tank temperature controls always at 30 ± 2 DEG C.
Claims (2)
1. a kind of mixed fermentation method of biological feedstuff leaven, is characterized in that:
One, the preparation of seed liquor:
Make a seed liquor: weigh 1 gram of lactic acid class solid mycopowder and be inoculated in 100 milliliters of triangles equipped with 100 milliliters of synthetic mediums
In bottle, inoculate 5 bottles；32 DEG C of quiescent culture 72 hours, use plastic wraps bottleneck, Anaerobic culturel；Obtain a seed liquor；
Make b seed liquor: weigh 1 gram of spore class solid mycopowder and be inoculated in 100 milliliters of triangles equipped with 100 milliliters of synthetic mediums
In bottle, inoculate 5 bottles；Place in constant-temperature table, 32 DEG C, cultivate 24 hours under the conditions of 150 revs/min, obtain b seed liquor；
The manufacture method of synthetic medium is as follows: takes 20 grams of glucose, 10 grams of corn starch, 1 gram of dipotassium hydrogen phosphate, magnesium sulfate
0.5 gram, 1 gram of sodium chloride, 1 gram of potassium chloride, 0.01 gram of ferrous sulfate, 3 grams of yeast extract, 5 grams of peptone, vitamin b1 10 plus
Enter in 1000 milliliters of water, mix homogeneously, ph is natural；100 milliliters of synthetic mediums of subpackage in each 100 milliliters of triangular flask；121
Autoclaving 15 minutes at a temperature of DEG C；
Two, production process:
(1), 0.6 ton of fermentation tank is carried out disinfection:
First tubule on the air cock of fermentation tank is transferred on air inlet valve, opens lower section steam inlet valve, open top
Air bleeding valve, opens the condensed water in underlying discharging opening valve emptying fermentation tank, simultaneously while fermentation tank temperature rise
Note the condensed water elimination of air cleaning tank being connected with fermentation tank, after steam is discharged from check-valves, start the clock, high
Sterilize 20 minutes under temperature, high pressure conditions；
(2), make production culture medium:
Weigh 6 kilograms of brown sugar, 0.35 kilogram of potassium dihydrogen phosphate, 1 kilogram of defat high-calcium milk powder, 200 milliliters of tween 80, defoamer
100 milliliters, dissolve in water brown sugar, potassium dihydrogen phosphate, defat high-calcium milk powder being put into 15 20 kilograms, then by tween-
80th, defoamer is added in solution, stirs, and ph is natural；The nutrient of growth of microorganism in supply fermentation tank；
(3), feed intake:
The above-mentioned material having dissolved is added in fermentation tank, then adds water, treat that liquid reaches and stop adding water between two visors；
(4), sterilize:
Close dog-house, top air bleeding valve remains on, and then opens tank bottoms inlet valve and heats, and makes tank temperature reach 100 degree,
Close top air bleeding valve, when gauge hand reaches 0.05 MPa, opening air bleeding valve makes its pressure be zeroed, and turns off air bleeding valve
Gauge hand is made to reach 0.15 MPa, pressurize closed inlet valve after 30 minutes；
(5), inoculate
Treat fermentation tank temperature drop to 35 degree about, the described a seed liquor of flame protection inoculation and b seed liquor are in fermentation tank；
After inoculation, stand two hours, open the air bleeding valve on fermentation tank, logical oxygen carries out aerobic culture, and logical oxygen 8 hours, by micro-
The air bleeding valve opened controls tank pressure at 0.05 MPa；Sampling microscopy, observes the growth ratio of aerobic bacteria and anaerobe, works as aerobic bacteria
Height, when ratio reaches 75%, stops oxygenation, closes air cock and carries out Anaerobic culturel, carries out microscopy again within 8 hours about, work as anaerobe
When accounting for 70%, open the air cock on fermentation tank, carry out aerobic culture, microscopy after 8 hours, stop when aerobic bacteria accounts for more than 80%
Only support, carry out Anaerobic culturel completely, cultivate after 48 hours and terminate, now anaerobe accounts for 90% about.
2., according to a kind of mixed fermentation method of the biological feedstuff leaven described in claim 1, it is characterized in that: described lactic acid
Class solid mycopowder is Lactobacillus plantarum, streptococcus thermophiluss, Lactobacillus bulgaricus；Described spore class solid mycopowder is hay bud
Spore bacillus, bacillus licheniformis, Bacillus cereuss.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108497158A (en) * | 2018-06-11 | 2018-09-07 | 厦门市科环海洋生物科技有限公司 | A kind of the biological and ecological methods to prevent plant disease, pests, and erosion fermented feed and its production method and production equipment of prevention vibriosis |
| CN114752512A (en) * | 2022-04-15 | 2022-07-15 | 成都香阁里科技有限公司 | Microbial flora pairing technology in fecal sewage biodegradation |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732451A (en) * | 2012-05-23 | 2012-10-17 | 绍兴虞林生物科技有限公司 | Composite strain mixing fermentation process |
| CN105685472A (en) * | 2016-01-25 | 2016-06-22 | 黑龙江省黑钻极光新能源科技有限公司 | Microbial fermentation additive and preparation method as well as application thereof |
| CN105767506A (en) * | 2016-03-17 | 2016-07-20 | 武汉绿富农业生物工程技术有限公司 | Special fermenting agent for aquatic products as well as preparation method and application of fermenting agent |
-
2016
- 2016-08-26 CN CN201610726612.8A patent/CN106333059A/en not_active Withdrawn
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102732451A (en) * | 2012-05-23 | 2012-10-17 | 绍兴虞林生物科技有限公司 | Composite strain mixing fermentation process |
| CN105685472A (en) * | 2016-01-25 | 2016-06-22 | 黑龙江省黑钻极光新能源科技有限公司 | Microbial fermentation additive and preparation method as well as application thereof |
| CN105767506A (en) * | 2016-03-17 | 2016-07-20 | 武汉绿富农业生物工程技术有限公司 | Special fermenting agent for aquatic products as well as preparation method and application of fermenting agent |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108497158A (en) * | 2018-06-11 | 2018-09-07 | 厦门市科环海洋生物科技有限公司 | A kind of the biological and ecological methods to prevent plant disease, pests, and erosion fermented feed and its production method and production equipment of prevention vibriosis |
| CN114752512A (en) * | 2022-04-15 | 2022-07-15 | 成都香阁里科技有限公司 | Microbial flora pairing technology in fecal sewage biodegradation |
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