CN113980818A - Fungus dehydrogenation seed liquid culture method - Google Patents
Fungus dehydrogenation seed liquid culture method Download PDFInfo
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- CN113980818A CN113980818A CN202111069004.1A CN202111069004A CN113980818A CN 113980818 A CN113980818 A CN 113980818A CN 202111069004 A CN202111069004 A CN 202111069004A CN 113980818 A CN113980818 A CN 113980818A
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- 238000009630 liquid culture Methods 0.000 title claims abstract description 74
- 238000000034 method Methods 0.000 title claims abstract description 34
- 238000006356 dehydrogenation reaction Methods 0.000 title claims abstract description 26
- 241000233866 Fungi Species 0.000 title description 2
- 239000001963 growth medium Substances 0.000 claims abstract description 71
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 33
- 230000001954 sterilising effect Effects 0.000 claims abstract description 19
- 239000007788 liquid Substances 0.000 claims abstract description 18
- 241000894007 species Species 0.000 claims abstract description 13
- 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 abstract description 11
- 239000001888 Peptone Substances 0.000 claims abstract description 11
- 108010080698 Peptones Proteins 0.000 claims abstract description 11
- 240000008042 Zea mays Species 0.000 claims abstract description 11
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 claims abstract description 11
- 235000002017 Zea mays subsp mays Nutrition 0.000 claims abstract description 11
- 229940041514 candida albicans extract Drugs 0.000 claims abstract description 11
- 235000005822 corn Nutrition 0.000 claims abstract description 11
- 239000008103 glucose Substances 0.000 claims abstract description 11
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims abstract description 11
- 235000019796 monopotassium phosphate Nutrition 0.000 claims abstract description 11
- 235000019319 peptone Nutrition 0.000 claims abstract description 11
- 239000012138 yeast extract Substances 0.000 claims abstract description 11
- LYCAIKOWRPUZTN-UHFFFAOYSA-N Ethylene glycol Chemical compound OCCO LYCAIKOWRPUZTN-UHFFFAOYSA-N 0.000 claims abstract description 9
- 238000012258 culturing Methods 0.000 claims abstract description 8
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims abstract description 7
- 238000010438 heat treatment Methods 0.000 claims description 11
- 238000005070 sampling Methods 0.000 claims description 10
- 238000004659 sterilization and disinfection Methods 0.000 claims description 10
- 238000007599 discharging Methods 0.000 claims description 7
- 241000186063 Arthrobacter Species 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 5
- 238000009826 distribution Methods 0.000 claims description 4
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 claims description 4
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 claims description 4
- 238000011218 seed culture Methods 0.000 claims description 3
- -1 natural pond killer Substances 0.000 claims 1
- 238000002360 preparation method Methods 0.000 claims 1
- 239000002994 raw material Substances 0.000 abstract description 17
- 238000004519 manufacturing process Methods 0.000 abstract description 3
- 238000000855 fermentation Methods 0.000 description 14
- 230000004151 fermentation Effects 0.000 description 14
- 239000000243 solution Substances 0.000 description 8
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 6
- 238000005406 washing Methods 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 238000011081 inoculation Methods 0.000 description 4
- 238000001816 cooling Methods 0.000 description 3
- 229920000742 Cotton Polymers 0.000 description 2
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 239000006260 foam Substances 0.000 description 2
- 239000002054 inoculum Substances 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 230000002035 prolonged effect Effects 0.000 description 2
- 238000007789 sealing Methods 0.000 description 2
- 238000002791 soaking Methods 0.000 description 2
- LPXPTNMVRIOKMN-UHFFFAOYSA-M sodium nitrite Chemical compound [Na+].[O-]N=O LPXPTNMVRIOKMN-UHFFFAOYSA-M 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 210000001503 joint Anatomy 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000010288 sodium nitrite Nutrition 0.000 description 1
- 239000003270 steroid hormone Substances 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 238000001308 synthesis method Methods 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
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- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P1/00—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes
- C12P1/02—Preparation of compounds or compositions, not provided for in groups C12P3/00 - C12P39/00, by using microorganisms or enzymes by using fungi
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Abstract
The invention discloses a mould dehydrogenation species seed liquid culture method, which comprises the following steps: preparing a seed liquid culture medium; sterilizing the seed liquid culture medium; inoculating the seed liquid culture medium; and culturing the seed liquid culture medium to obtain the mould dehydrogenation seed liquid. Selecting glucose, corn steep liquor, peptone, ethylene glycol, water, yeast extract and potassium dihydrogen phosphate as raw materials of a seed liquid culture medium, and carrying out mass production, wherein the dosage of each raw material is adjusted, and the weight ratio is 32: 54-56: 20: 1.2: 4000: 2: and 2, the yield of the mould dehydrogenation seed liquid is highest, and the mould dehydrogenation seed liquid can be rapidly, safely and efficiently industrially produced in large scale.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to a mould dehydrogenation species seed liquid culture method.
Background
The mould dehydrogenation product is an intermediate for preparing steroid hormones, is mainly prepared by a chemical synthesis method and a biological synthesis method, and has the problems of long process route, special reaction, complex process, low total yield and the like.
Disclosure of Invention
Aiming at the problems, the invention discloses a mould dehydrogenation species seed liquid culture method, which comprises the following steps:
preparing a seed liquid culture medium;
sterilizing the seed liquid culture medium;
inoculating the seed liquid culture medium;
and culturing the seed liquid culture medium to obtain the mould dehydrogenation seed liquid.
Furthermore, the raw materials of the seed liquid culture medium are glucose, corn steep liquor, peptone, sodium nitrite, water, yeast extract and potassium dihydrogen phosphate.
Further, the weight ratio of the glucose, the corn steep liquor, the peptone, the foam enemy, the water, the yeast extract and the monopotassium phosphate is 32: 54-56: 20: 1.2: 4000: 2: 2.
furthermore, the pH value of the prepared seed liquid culture medium is 8.5-9.
Further, the seed liquid medium is sterilized by steam.
Further, the steam enters the seed tank through a sampling tube, an air distribution tube and a bottom discharge tube.
Further, the specific sterilization steps of the seed liquid culture medium are as follows:
heating the seed liquid culture medium to 90 ℃;
introducing steam into a seed tank, heating the seed liquid culture medium to 100 ℃, wherein the pressure of the seed tank is 0.1-0.12 Mpa;
heating the seed liquid culture medium to 112-118 ℃, keeping the pressure of a seed tank at 0.1-0.12 MPa for 20 min;
and opening condensed water to cool the seed liquid culture medium to 29-31 ℃, discharging steam in the seed tank, and introducing sterile air.
Furthermore, the strain inoculated by the seed liquid culture medium is arthrobacter, and the inoculation number is 115-150 bottles.
Furthermore, the culture process of the seed liquid culture medium comprises the steps of 29-31 ℃ of temperature, 0.04-0.06 MPa of pressure of a seed tank, 110-150 r/min of stirring speed and 30-40 m of flow of sterile air3/h。
Furthermore, the first sampling detection is carried out after the seed liquid culture medium is inoculated with the strains for 8 hours, and the strains are planted into the fermentation tank when the pH value is 7.0-7.2.
Compared with the prior art, the invention has the beneficial effects that: selecting glucose, corn steep liquor, peptone, ethylene glycol, water, yeast extract and potassium dihydrogen phosphate as raw materials of a seed liquid culture medium, and carrying out mass production, wherein the dosage of each raw material is adjusted, and the weight ratio is 32: 54-56: 20: 1.2: 4000: 2: and 2, the yield of the mould dehydrogenation seed liquid is highest, and the mould dehydrogenation seed liquid can be rapidly, safely and efficiently industrially produced in large scale.
Additional features and advantages of the invention will be set forth in the description which follows, and in part will be obvious from the description, or may be learned by practice of the invention. The objectives and other advantages of the invention will be realized and attained by the process particularly pointed out in the written description and claims hereof as well as the appended drawings.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and those skilled in the art can also obtain other drawings according to the drawings without creative efforts.
FIG. 1 shows a flow diagram for the cultivation of an inoculum of a dehydrogenated species of mold according to an embodiment of the invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all, embodiments of the present invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
FIG. 1 shows a flow diagram for the cultivation of an inoculum of a dehydrogenated species of mold according to an embodiment of the invention. As shown in FIG. 1, the present invention provides a method for culturing mould dehydrogenation species in seed solution, comprising the following steps:
preparing a seed liquid culture medium; the seed liquid culture medium is prepared from glucose, corn steep liquor, peptone, sodium chloride, water, yeast extract and potassium dihydrogen phosphate in a weight ratio of 32: 54-56: 20: 1.2: 4000: 2: 2; the pH value of the prepared seed liquid culture medium is 8.5-9;
sterilizing the seed liquid culture medium; sterilizing the seed liquid culture medium by steam, and feeding the steam into a seed tank through a sampling pipe, an air distribution pipe and a bottom discharging pipe;
the specific sterilization steps of the seed liquid culture medium are as follows:
heating the seed liquid culture medium to 90 ℃;
introducing steam into a seed tank, heating the seed liquid culture medium to 100 ℃, wherein the pressure of the seed tank is 0.1-0.12 Mpa;
heating the seed liquid culture medium to 112-118 ℃, keeping the pressure of a seed tank at 0.1-0.12 MPa for 20 min;
opening condensed water to cool the seed liquid culture medium to 29-31 ℃, discharging steam in the seed tank, and introducing sterile air;
inoculating the seed liquid culture medium; wherein, the inoculation strain is arthrobacter, the inoculation quantity is 115-150 bottles, and the arthrobacter is purchased commercially and stored in a Kirschner flask.
And culturing the seed liquid culture medium to obtain the mould dehydrogenation seed liquid. It is composed ofIn the method, the culture process of the seed liquid culture medium is carried out at the temperature of 29-31 ℃, and preferably at the temperature of 30 ℃; the pressure of the seed tank is 0.04-0.06 MPa, preferably 0.05 MPa; the stirring speed is 110-150 r/min, preferably 130 r/min; the flow rate of the sterile air is 30-40 m3H; the culture time is 14-15 h.
Sampling and detecting for the first time after the seed liquid culture medium is inoculated with the strains for 8 hours, and planting the strains to a fermentation tank when the pH value is 7.0-7.2.
Some of the names and specifications of the raw materials used in the following examples are shown in Table 1 below, and the raw materials used in the examples are commercially available.
Table 1 raw material names and specification
Example 1
1. Culture medium for preparing seed liquid
1.1, opening a seed tank cover, checking whether the seed tank is clean and has accumulated water, if so, discharging the seed tank from the bottom, and washing the seed tank with process water; wherein the process water is sterilized water.
1.2, checking whether each pipeline valve is intact and whether mechanical equipment is normal;
1.3, checking whether the raw materials are intact, whether the package is damaged, whether the weight is consistent with the weight of the batching list and the like;
1.4, weighing 32kg of glucose, 56kg of corn steep liquor, 20kg of peptone, 2kg of yeast extract and 2kg of monopotassium phosphate according to the proportion;
1.5, opening a water valve for process water, adding the process water to a required amount of mark according to the ratio, and taking the condensed water of the steam into consideration when the process water is added, wherein the total amount of the added water is 4000 kg;
1.6, pouring the weighed glucose, corn steep liquor, peptone, yeast extract and potassium dihydrogen phosphate into a seed tank, rechecking the raw materials by two persons when pouring the raw materials, taking out the fermentation sheet in the bag, checking the variety and the weight, and pouring the fermentation sheet and the material receiving sheet into the seed tank after checking the fermentation sheet and the material receiving sheet without errors;
1.7, washing the raw material powder on the opening of the seed tank and the inner wall of the tank by a small amount of process water, sealing the seed tank cover after washing, heating to 40 ℃, adjusting the pH value of the solution in the seed tank to 9.0 by using a 20% sodium hydroxide aqueous solution, adding 1.2kg of natural killer to obtain a seed liquid culture medium, tightening the seed tank cover, and preparing for sterilization;
2. sterilization
2.1, starting a stirrer to uniformly stir the raw materials;
2.2, opening a drain valve of the coil (jacket), opening a steam inlet valve of the coil (jacket), and finally opening a steam valve to drain condensed water in the coil (jacket), closing the drain valve of the coil (jacket) when a large amount of steam is discharged, keeping the coil (jacket) slightly opened to drain the condensed water, simultaneously preventing the waste of the steam, and controlling the pressure of the coil (jacket) to be less than 0.2 MPa;
2.3, opening each path of emptying valve at the top of the seed tank, and discharging cold air in the tank;
2.4, when the temperature of the seed liquid culture medium in the seed tank rises to above 90 ℃, closing a steam inlet valve of a coil pipe (a jacket), closing stirring, opening a drain valve on a steam pipe, draining condensed water until the steam is emitted, slowly opening a main steam valve, slowly raising the pressure, and not too fast to avoid damaging a steam filter, and when the pressure of the seed tank rises to 0.3MPa, opening a steam inlet valve of a sampling pipe, a steam inlet valve of an air distribution pipe and a steam inlet valve of a bottom discharging pipe of the seed tank, and keeping three paths of steam inlet for sterilization;
2.5, when the temperature of the seed liquid culture medium is increased to be more than 100 ℃, the pressure of the seed tank is 0.1Mpa, a large amount of steam is emitted, each emptying valve is closed, and partial steam is kept to pass through, so that the pressure and the temperature of the seed tank are increased in a balanced manner;
2.6, when the temperature of the seed liquid culture medium is raised to 118 ℃ and the pressure of the seed tank is 0.12MPa, closing each steam inlet valve, closing each emptying valve, keeping the temperature and the pressure for 20min, and sterilizing the seed liquid culture medium;
2.7, during pressure maintaining, steam is fed into the butt joint pipe for sterilization, and during sterilization, the steam is required to pass through all parts of the pipeline, so that dead corners which are not reached by the steam are avoided;
2.8, after pressure maintaining, opening condensed water to cool the seed liquid culture medium, opening an exhaust valve to exhaust steam in the tank, introducing sterile air when the pressure of the seed tank is reduced to 0.06MPa, starting a stirrer at the rotating speed of 150r/min, and cooling the seed liquid culture medium to 31 ℃ for later use;
2.9, keeping the sterilized seed liquid culture medium at positive pressure all the time to prevent contamination, sterilizing the sampling tube, sampling the seed liquid culture medium by using a triangular flask, marking the batch number of the seed liquid culture medium, and sending the seed liquid culture medium into a sterile room for storage for later use;
3. inoculation of
3.1, tearing a small amount of cotton into strips or soaking gauze strips in alcohol, and placing the strips or gauze strips into gaps of seed pot covers;
3.2, taking out the hot air ring, and soaking the hot air ring in alcohol for later use;
3.3, preparing two sets of wrenches;
3.4, preparing a towel strip soaked by basin water or water for extinguishing the wind and fire circle;
3.5, igniting the cotton on the seed tank cover, and burning for 1min to disinfect the seed tank cover;
3.6, rapidly disassembling the screw of the seed tank cover by using a spanner, and closing the emptying valve when the seed tank cover is opened to discharge air from the opening of the seed tank so as to avoid bacterial contamination;
3.7, igniting the air-fire ring when the seed tank cover is completely disassembled, and placing the air-fire ring at the opening of the seed tank when the seed tank cover is opened;
3.8, pouring 150 bottles of prepared arthrobacter strains into a seed tank quickly through a wind-fire ring;
3.9, burning the seed tank cover on the wind-fire ring for about 20s, removing the wind-fire ring, putting the seed tank cover into water or putting the seed tank cover out by using a wet towel, quickly tightening the seed tank cover, opening the emptying valve, and adjusting the flow of the sterile air to be 40m3/h；
3.10, recording time, and starting seed liquid culture for 15 hours to obtain mould dehydrogenation seed liquid; wherein, the first sampling is carried out after the seed liquid culture medium is inoculated with the strain for 8 hours, when the microscopic examination does not pollute the mixed bacteria, the fermentation tank can be fed and sterilized for standby, when the pH value of the seed liquid culture medium is 7.2, the seed can be transplanted, the seed transplanting time can be properly prolonged, but the longest time of the first-level seed culture can not exceed 15 hours, and when the specified pH value is not reached, the mixed bacteria can be transplanted to the fermentation tank without pollution for the second-level fermentation;
4. transplanting seeds
4.1, sterilizing the culture medium of the fermentation tank (30t) before the seed transfer according to a sterilization operation method, and cooling for later use;
4.2, the seed transferring pipeline is also sterilized before seed transferring for standby;
4.3, transferring the seed liquid into a fermentation tank according to a seed transferring operation method after the mould dehydrogenation seed liquid is detected to be qualified, and performing fermentation culture;
4.4, after the seed liquid is transferred, the seed tank and the pipeline need to be washed clean by water and sterilized by steam for 30 min.
Example 2
The other steps in this example are the same as example 1, except that:
1.4, weighing 32kg of glucose, 54kg of corn steep liquor, 20kg of peptone, 2kg of yeast extract and 2kg of monopotassium phosphate according to the proportion;
1.7, washing the raw material powder on the opening of the seed tank and the inner wall of the tank by a small amount of process water, sealing the seed tank cover after washing, heating to 40 ℃, adjusting the pH value of the solution in the seed tank to 8.5 by using a 20% sodium hydroxide aqueous solution, adding 1.2kg of natural killer to obtain a seed liquid culture medium, tightening the seed tank cover, and preparing for sterilization;
2.6, when the temperature of the seed liquid culture medium is raised to 112 ℃ and the pressure of the seed tank is 0.1MPa, closing each steam inlet valve, closing each emptying valve, keeping the temperature and the pressure for 20min, and sterilizing the seed liquid culture medium;
2.8, after pressure maintaining, opening condensed water to cool the seed liquid culture medium, opening an exhaust valve to exhaust steam in the seed tank, introducing sterile air when the pressure of the seed tank is reduced to 0.04MPa, starting a stirrer at the rotating speed of 110r/min, and cooling the seed liquid culture medium to 29 ℃ for later use;
3.8, pouring the prepared 115 bottles of arthrobacter strains into a seed tank quickly through a wind-fire ring;
3.9, the seed pot cover is burned on the wind-fire ring for about 20s, the wind-fire ring is removed, and the seed pot is put into water or is swabbed by a wet towelKilling, quickly fastening the cover of seed pot, opening the emptying valve, and regulating the flow of sterile air to 30m3/h；
3.10, recording time, and starting seed liquid culture for 14 hours to obtain mould dehydrogenation seed liquid; wherein, the first sampling is carried out after the seed liquid culture medium is inoculated with the strain for 8 hours, when the microscopic examination does not pollute the mixed bacteria, the fermentation tank can be charged and sterilized for standby, when the pH value of the seed liquid culture medium is 7.0, the seed can be transplanted, the seed transplanting time can be properly prolonged, but the longest time of the first-level seed culture can not exceed 15 hours, and if the specified pH value can not be reached, the mixed bacteria can be transplanted to the fermentation tank without pollution, and the second-level fermentation is carried out.
The mould dehydrogenation species seed liquid culture method provided by the invention selects glucose, corn steep liquor, peptone, foam, water, yeast extract and potassium dihydrogen phosphate as raw materials of a seed liquid culture medium, and the raw materials are subjected to mass production, and the dosage of each raw material is adjusted, wherein the weight ratio is 32: 54-56: 20: 1.2: 4000: 2: and 2, the yield of the mould dehydrogenation seed liquid is highest, and the mould dehydrogenation seed liquid can be rapidly, safely and efficiently industrially produced on a large scale and used for producing mould dehydrogenation.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. A mould dehydrogenation species seed liquid culture method is characterized by comprising the following steps:
preparing a seed liquid culture medium;
sterilizing the seed liquid culture medium;
inoculating the seed liquid culture medium;
and culturing the seed liquid culture medium to obtain the mould dehydrogenation seed liquid.
2. The method of claim 1, wherein the seed culture medium is selected from the group consisting of glucose, corn steep liquor, peptone, ethylene glycol, water, yeast extract, and potassium dihydrogen phosphate.
3. The method of claim 2, wherein the glucose, corn steep liquor, peptone, natural pond killer, water, yeast extract, and monopotassium phosphate are present in a weight ratio of 32: 54-56: 20: 1.2: 4000: 2: 2.
4. the method for culturing the seed solution of the mold dehydrogenation species according to claim 1, wherein the pH value of the seed solution culture medium after the preparation is completed is 8.5-9.
5. The method of claim 1, wherein the seed broth is steam sterilized.
6. A process for the cultivation of mould dehydrogenated species in seed broth according to claim 5, characterised in that the steam enters the seed tank through sampling pipes, air distribution pipes and bottom blow pipes.
7. A method of cultivation of mould dehydrogenated species in seed solution according to claim 1 or 5, wherein the specific sterilization steps of the seed solution culture medium are as follows:
heating the seed liquid culture medium to 90 ℃;
introducing steam into a seed tank, heating the seed liquid culture medium to 100 ℃, wherein the pressure of the seed tank is 0.1-0.12 Mpa;
heating the seed liquid culture medium to 112-118 ℃, keeping the pressure of a seed tank at 0.1-0.12 MPa for 20 min;
and opening condensed water to cool the seed liquid culture medium to 29-31 ℃, discharging steam in the seed tank, and introducing sterile air.
8. The method for culturing mould dehydrogenation species seed solution according to claim 1, wherein the seed solution culture medium is inoculated with arthrobacter in an amount of 115-150 bottles.
9. The method for culturing seed liquid of mold dehydrogenation species according to claim 1, wherein the culture process of the seed liquid culture medium comprises the temperature of 29-31 ℃, the pressure of a seed tank of 0.04-0.06 MPa, the stirring speed of 110-150 r/min, and the flow of sterile air of 30-40 m3/h。
10. The method of claim 1, wherein the seed liquid culture medium is inoculated with the strain for 8 hours, sampled for the first time, and inoculated to the fermenter when the pH is 7.0-7.2.
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