CN104195203B - A kind of antifungal streptomycete fermentation produces the culture medium and feed process of enramycin - Google Patents
A kind of antifungal streptomycete fermentation produces the culture medium and feed process of enramycin Download PDFInfo
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- CN104195203B CN104195203B CN201410432903.7A CN201410432903A CN104195203B CN 104195203 B CN104195203 B CN 104195203B CN 201410432903 A CN201410432903 A CN 201410432903A CN 104195203 B CN104195203 B CN 104195203B
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Abstract
The present invention relates to the culture medium and feed process that a kind of antifungal streptomycete fermentation produces enramycin, the present invention produces the seed culture medium of enramycin and the compatibility of fermentation medium for strain fermentation by the antifungal streptomycete that is suitable for having been acknowledged, and using feed process as control method, the fermentation unit of enramycin is finally caused to reach more than 10000u/ml with reference to the enramycin fermentating controling process announced both at home and abroad at present, production cycle is no more than 240h, compared with domestic technique, fermentation unit improves 20%, and the production cycle reduces more than 20h.
Description
Technical field
The invention belongs to fermentation technical field, more particularly to a kind of antifungal streptomycete fermentation produces the training of enramycin
Support base and feed process.
Background technology
Enramycin (enramycin) is obtained by antifungal streptomycete fermentation, is unrighted acid and ten several ammonia
The polypeptide antibiotics that base acid is combined, key component has enramycin A and enramycin B, mainly with hydrochloride form application.
At present, the fermenting and producing of domestic enramycin use second order fermentation pattern, which exist subject matter have it is following some：
1 enramycin fermentation level is relatively low, and its fermentation unit is less than 8000u/ml.
Glucose is added in the culture medium of 2 domestic producing enramycin by fermentation, through high-temperature sterilization, part grape is easily caused
Sugar charcoal, reduces total sugar content, influences zymotic fluid quality.
Cost of raw needed for 3 enramycin fermenting and producings is higher, is commonly used particularly in enramycin culture medium
The organic nitrogen source market price it is higher, improve fermentation costs, reduce the market competitiveness.
The fermented and cultured cycle of 4 domestic enramycins is longer, typically in more than 260h, causes energy consumption higher.
The content of the invention
The purpose of the present invention is that the defect for overcoming above-mentioned prior art effectively improves fermentation unit there is provided one kind, drop
Low production cost, the antifungal streptomycete fermentation for realize that enramycin is stable, efficiently producing produces the culture medium of enramycin；
It is a further object of the present invention to provide the feed process that above-mentioned antifungal streptomycete fermentation produces enramycin.
The technical scheme taken to achieve the above object is：
A kind of antifungal streptomycete fermentation produces the culture medium of enramycin, including seed culture medium and fermentation medium,
It is characterized in that the seed culture medium composition is：15~20ml/L of corn oil, 5~10g/L of alpha-lactose, lactase 0.01~
0.03g/L, 20~25g/L of formula fish meal, the dry 15~20g/L of slag of brewer's yeast, 10~15g/L of Dried Corn Steep Liquor Powder, earthworm powder 5~
10g/L, 0.2~0.5g/L of ammonium sulfate, 1~5g/L of precipitated calcium carbonate；
The fermentation medium is constituted：20~30ml/L of corn oil, 20~25g/L of alpha-lactose, lactase 0.02~
0.05g/L, 30~40g/L of formula fish meal, the dry 15~20g/L of slag of brewer's yeast, 15~20g/L of Dried Corn Steep Liquor Powder, earthworm powder 10
~15g/L, 0.3~0.7g/L of ammonium sulfate, 1~5g/L of precipitated calcium carbonate, 0.03~0.06g/L of magnesium sulfate, zinc sulfate 0.02~
0.04g/L, 0.5~0.9g/L of potassium dihydrogen phosphate, 0.3~0.7g/L of sodium chloride, 0.2~0.5g/L of potassium chloride, surfactant
1.5~2.5g/L, 4~7g/L of the carrier of oxygen.
The formula fish meal is according to weight ratio 7 by fish meal, bone meal and feather meal:3:1 is obtained by mixing.
The carrier of oxygen is Tween 80 or perfluocarbon.
The surfactant is polyxyethylated alkylolamides or glyceride or polyglycerol ester or polyethylene glycol stearic acid
Ester or octadecyl dihydroxy ethyl amine oxide.
Using the feed process of above-mentioned culture medium producing enramycin by fermentation, it is characterized in that：During the fermentation using stream
Addition carries out feed supplement, and feed supplement includes mending culture medium, moisturizing, mends ammonium sulfate, pH controls and mend amino acid, wherein
A mends culture medium：
Preceding 70h ferment without carrying out feed supplement,
Fermentation time mends carbon source and nitrogen source in 71~90h according to fermentative medium formula, and its amount of filling into is fermentation medium
The 3~5% of cumulative volume,
Fermentation time mends carbon source and nitrogen source in 110h~130h according to fermentative medium formula, and its amount of filling into is trained for fermentation
The 2~4% of base cumulative volume are supported,
Fermentation time mends carbon source and nitrogen source in 150h~180h according to fermentative medium formula, and its amount of filling into is trained for fermentation
Support the 1~2% of base cumulative volume；
B moisturizings：
Preceding 80h ferment without carrying out moisturizing,
Fermentation time, when cell concentration is higher than 55%, adds aqua sterilisa, it is desirable to control zymotic fluid thalline dense in 81~160h
Degree detects zymotic fluid cell concentration 50~55% every 6~8h,
Fermentation time, when cell concentration is higher than 50%, adds aqua sterilisa, it is desirable to control zymotic fluid in 161h~fermentation ends
Cell concentration detects zymotic fluid cell concentration 40~50% every 6~8h；
C fermentation process pH control techniques：
Fermentation before 80h, pH without control,
Fermentation time is in 81~100h, if pH ＜ 6.5, adds 10~20% sodium hydroxide, adjusts pH to 6.6~6.9；
If pH ＞ 7,20% ammonium sulfate is added, pH is to 6.6~6.9 for regulation,
Fermentation time is in 101~180h, if pH ＜ 6.2, adds 10~20% sodium hydroxide, and regulation pH controlled to 6.3~
6.6；If pH ＞ 6.6,30% ammonium sulfate is added, pH to 6.3~6.6 is adjusted；
D fills into amino acid：
80h before fermentation, without filling into amino acid,
Ferment and fill into the serine that concentration is 0.2~0.5mg/L, cysteine and smart ammonia respectively in 81h, 130h and 170h
Acid, its amount of filling into is the 0.1~0.3% of fermentating liquid volume（This process is to be separately added into these three, or adds mixing every time
Liquid）
E mends ammonium sulfate：
In 80h, 150h and 220h of fermentation, 30% ammonium sulfate is filled into respectively, and its consumption is controlled in fermentating liquid volume
0.05~0.1%.
Lactose is used in culture medium of the present invention, it is to avoid glucose carbonization phenomenon occur.
The present invention is trained by the antifungal streptomycete that is suitable for having been acknowledged for the seed that strain fermentation produces enramycin
The compatibility of base and fermentation medium is supported, and using feed process as control method, draws mould with reference to the grace announced both at home and abroad at present
Plain fermentating controling process finally causes the fermentation unit of enramycin to reach more than 10000u/ml, and the production cycle is no more than 240h,
Compared with domestic technique, fermentation unit improves 20%, and the production cycle reduces more than 20h.
The present invention is applied to scale producing enramycin by fermentation, disclosure satisfy that the production for producing 100 tons of enramycins per year is needed
Ask.
Specific implementation method
The present invention is explained with example, it should be understood that example is to be used to illustrate rather than to this below
The limitation of invention.The scope of the present invention is determined with core content according to claims.
The strain of following embodiments selects antifungal streptomycete：The strain source of production and application is female bottle zymotic fluid.Female bottle
Zymotic fluid quality requirement is：PH6~8；Cell concentration 10~30%；Microscopy is without miscellaneous bacteria；2000~3000u/ml of fermentation unit.
Fermentation process in following examples uses known, disclosed with the life of antifungal streptomycete fermentation both at home and abroad at present
Production. art, such as
Seed culture technique：
Seed culture medium is sterilized first, cooled down, and uses filtrated air pressurize, then under flame protection, will have been trained
The antifungal strepto- starter bottle zymotic fluid access seeding tank supported is cultivated, and inoculum concentration is controlled the 5 of seed culture medium volume
~10%.Seed culture condition is：Tank presses 0.04~0.06MPa；28~30 DEG C of tank temperature；Air mass flow：0~12h, 30~
50m3/h；12h~culture transferring：50~70m3/h；80~100r/min of speed of agitator；PH6~8；30~35h of incubation time.Seed
Culture terminates, its cell concentration 20~30%；PH value 6~8；Without other living contaminantses.
Fermentating culturing process：
First fermentation medium is sterilized, cooled down, and uses filtrated air pressurize, seed liquor is all then moved into fermentation tank enters
Row culture.Fermentation culture conditions are：Fat control is 2~3% after medium sterilization；29~31 DEG C of cultivation temperature；Speed of agitator
100~120r/min；Tank presses 0.05~0.06MPa；230~240h of incubation time；PH controls are 6.0~7.5 in fermentation process；
Bacterium inspection is carried out in fermentation process, it is desirable to without other miscellaneous bacterias；0~120h：600~800m3/h；121h~fermentation ends：1000~
1500m3/h.Fermented and cultured stop condition is：Every 6~8h samplings the detection of fermentation titer, the fermentation unit difference of forward and backward detection exists
Within 500u/ml；Cell concentration 30~40%；Fermentation unit is in more than 10000u/mL.
Embodiment 1
10m3Seeding tank：Corn oil 150L, alpha-lactose 50kg, lactase 0.1kg, formula fish meal 200kg, brewer's yeast are done
Slag 150kg, Dried Corn Steep Liquor Powder 100kg, earthworm powder 50kg, ammonium sulfate 2kg, precipitated calcium carbonate 10kg.
Seed culture technology controlling and process：
Seed culture medium is sterilized first, cooled down, and uses filtrated air pressurize, then under flame protection, will have been trained
The antifungal strepto- starter bottle zymotic fluid access seeding tank supported is cultivated, and inoculum concentration is controlled in seed culture medium volume
5%.Seed culture condition is：Tank presses 0.04~0.06MPa；28~30 DEG C of tank temperature；Air mass flow：0~12h, 30m3/h；12h
~culture transferring：50m3/h；Speed of agitator 80r/min；PH6~8；Incubation time 30h.Seed culture terminates, its cell concentration 21%；
PH value 7.8；Without other living contaminantses.
60m3Fermentation tank：Corn oil 1200L, alpha-lactose 1200kg, lactase 1.2kg, formula fish meal 1800kg, beer ferment
Female slag does 900kg, Dried Corn Steep Liquor Powder 900g/L, earthworm powder 600kg, ammonium sulfate 18kg, precipitated calcium carbonate 60kg, magnesium sulfate
1.8kg, zinc sulfate 1.2kg, potassium dihydrogen phosphate 30kg, sodium chloride 18kg, potassium chloride 12kg, polyxyethylated alkylolamides
90kg, Tween 80 240kg.
Fermentating culturing process：
First fermentation medium is sterilized, cooled down, and uses filtrated air pressurize, seed liquor is all then moved into fermentation tank enters
Row culture.Fermentation culture conditions are：Fat control is 2.1% after medium sterilization；29~31 DEG C of cultivation temperature；Speed of agitator
100r/min；Tank presses 0.05~0.06MPa；PH controls are 6.0~7.5 in fermentation process.Bacterium inspection is carried out in fermentation process, it is desirable to
Without other miscellaneous bacterias；0~120h：600m3/h；121h~fermentation ends：1000m3/h.Fermented and cultured stops：Cell concentration is
31%, enramycin fermentation unit is 10276u/ml, production cycle 232h.
Feed supplement is carried out using stream addition during the fermentation, feed supplement includes mending culture medium, moisturizing, mends ammonium sulfate, pH controls
With mend amino acid, wherein
A mends culture medium technique control：
70h is without carrying out feed supplement before fermentation.
Fermentation time mends carbon source and nitrogen source in 71~90h according to fermentative medium formula, and its amount of filling into is fermentation medium
The 3~5% of cumulative volume.
Fermentation time mends carbon source and nitrogen source in 110h~130h according to fermentative medium formula, and its amount of filling into is trained for fermentation
Support the 2~4% of base cumulative volume.
Fermentation time mends carbon source and nitrogen source in 150h~180h according to fermentative medium formula, and its amount of filling into is trained for fermentation
Support the 1~2% of base cumulative volume.
B moisturizing technology controlling and process：
Preceding 80h ferment without carrying out moisturizing,
Fermentation time, when cell concentration is higher than 55%, adds aqua sterilisa, it is desirable to control zymotic fluid thalline dense in 81~160h
Degree detects zymotic fluid cell concentration 50~55% every 6~8h,
Fermentation time, when cell concentration is higher than 50%, adds aqua sterilisa, it is desirable to control zymotic fluid in 161h~fermentation ends
Cell concentration detects zymotic fluid cell concentration 40~50% every 6~8h；
C fermentation process pH control techniques：
Fermentation before 80h, pH without control,
Fermentation time is in 80~100h, if pH ＜ 6.5, adds 10~20% sodium hydroxide, adjusts pH to 6.6~6.9；
If pH ＞ 7,20% ammonium sulfate is added, pH to 6.6~6.9 is adjusted.
Fermentation time is in 101~180h, if pH ＜ 6.2, adds 10~20% sodium hydroxide, and regulation pH controlled to 6.3~
6.6；If pH ＞ 6.6,30% ammonium sulfate is added, pH to 6.3~6.6 is adjusted；
D fills into amino acid：
80h before fermentation, without filling into amino acid,
During enramycin fermented and cultured, the silk that concentration is 0.2~0.5mg/L is filled into 81h, 130h and 170h respectively
Propylhomoserin, cysteine and arginine, its amount of filling into are the 0.1~0.3% of fermentating liquid volume.
E mends ammonium sulfate
During enramycin fermented and cultured, 30% ammonium sulfate, its consumption are filled into 80h, 150h and 220h respectively
Control is the 0.05~0.1% of fermentating liquid volume.
Embodiment 2
10m3Seeding tank：Corn oil 160L, alpha-lactose 60kg, lactase 0.12kg, formula fish meal 210kg, brewer's yeast
Dry slag 160kg, Dried Corn Steep Liquor Powder 110kg, earthworm powder 60kg, ammonium sulfate 2.5kg, precipitated calcium carbonate 20kg.
Seed culture technique：
Seed culture medium is sterilized first, cooled down, and uses filtrated air pressurize, then under flame protection, will have been trained
The antifungal strepto- starter bottle zymotic fluid access seeding tank supported is cultivated, and inoculum concentration is controlled in seed culture medium volume
6%.Seed culture condition is：Tank presses 0.04~0.06MPa；28~30 DEG C of tank temperature；Air mass flow：0~12h, 35m3/h；12h
~culture transferring：55m3/h；Speed of agitator 85r/min；PH6~8；Incubation time 31h.Seed culture terminates, its cell concentration 22%；
PH value 7.4；Without other living contaminantses.
60m3 fermentation tanks：Corn oil 1320L, alpha-lactose 1260kg, lactase 1.8kg, formula fish meal 1980kg, beer ferment
Female slag does 960kg, Dried Corn Steep Liquor Powder 960g/L, earthworm powder 660kg, ammonium sulfate 24kg, precipitated calcium carbonate 120kg, magnesium sulfate
2.4kg, zinc sulfate 1.5kg, potassium dihydrogen phosphate 36kg, sodium chloride 24kg, potassium chloride 18kg, glyceride 102kg, perfluocarbon
300kg。
Fermentating culturing process：
First fermentation medium is sterilized, cooled down, and uses filtrated air pressurize, seed liquor is all then moved into fermentation tank enters
Row culture.Fermentation culture conditions are：Fat control is 2.3% after medium sterilization；29~31 DEG C of cultivation temperature；Speed of agitator
105r/min；Tank presses 0.05~0.06MPa；PH controls are 6.0~7.5 in fermentation process；Bacterium inspection is carried out in fermentation process, it is desirable to
Without other miscellaneous bacterias；0~120h：650m3/h；121h~fermentation ends：1100m3/h.Fermented and cultured terminates, and cell concentration 30~
40%；Enramycin fermentation unit is 10723u/ml, production cycle 234h.
Feed supplement is carried out using stream addition during the fermentation, specific feed profile is referring to embodiment 1.
Embodiment 3
10m3Seeding tank：Corn oil 175L, alpha-lactose 75kg, lactase 0.2kg, formula fish meal 225kg, brewer's yeast are done
Slag 175kg, Dried Corn Steep Liquor Powder 125kg, earthworm powder 75kg, ammonium sulfate 3.5kg, precipitated calcium carbonate 30kg.
Seed culture technique：
Seed culture medium is sterilized first, cooled down, and uses filtrated air pressurize, then under flame protection, will have been trained
The antifungal strepto- starter bottle zymotic fluid access seeding tank supported is cultivated, and inoculum concentration is controlled in seed culture medium volume
7%.Seed culture condition is：Tank presses 0.04~0.06MPa；28~30 DEG C of tank temperature；Air mass flow：0~12h, 40m3/h；12h
~culture transferring：60m3/h；Speed of agitator 90r/min；PH6~8；Incubation time 33h.Seed culture terminates, its cell concentration 25%；
PH value 7.1；Without other living contaminantses.
60m3 fermentation tanks：Corn oil 1500L, alpha-lactose 1350kg, lactase 2.1kg, formula fish meal 2100kg, beer ferment
Female slag does 1050kg, Dried Corn Steep Liquor Powder 1050g/L, earthworm powder 750kg, ammonium sulfate 30kg, precipitated calcium carbonate 180kg, magnesium sulfate
2.7kg, zinc sulfate 1.8kg, potassium dihydrogen phosphate 42kg, sodium chloride 30kg, potassium chloride 21g, polyglycerol ester 120kg, perfluocarbon
330kg。
Zymotechnique is controlled：
First fermentation medium is sterilized, cooled down, and uses filtrated air pressurize, seed liquor is all then moved into fermentation tank enters
Row culture.Fermentation culture conditions are：Fat control is 2.5% after medium sterilization；29~31 DEG C of cultivation temperature；Speed of agitator
110r/min；Tank presses 0.05~0.06MPa；PH controls are 6.0~7.5 in fermentation process；Bacterium inspection is carried out in fermentation process, it is desirable to
Without other miscellaneous bacterias；0~120h：700m3/h；121h~fermentation ends：1200m3/h.Fermented and cultured terminates, cell concentration 35%；
Enramycin fermentation unit is 11386u/ml, production cycle 236h.
Feed supplement is carried out using stream addition during the fermentation, specific feed profile is referring to embodiment 1.
Embodiment 4
10m3Seeding tank：Corn oil 180L, alpha-lactose 90kg, lactase 0.25kg, formula fish meal 240kg, brewer's yeast
Dry slag 190kg, Dried Corn Steep Liquor Powder 140kg, earthworm powder 90kg, ammonium sulfate 4kg, precipitated calcium carbonate 40kg.
Seed culture technique
Seed culture medium is sterilized first, cooled down, and uses filtrated air pressurize, then under flame protection, will have been trained
The antifungal strepto- starter bottle zymotic fluid access seeding tank supported is cultivated, and inoculum concentration is controlled in seed culture medium volume
8%.Seed culture condition is：Tank presses 0.04~0.06MPa；28~30 DEG C of tank temperature；Air mass flow：0~12h, 45m3/h；12h
~culture transferring：65m3/h；Speed of agitator 95r/min；PH6~8；Incubation time 34h.Seed culture terminates, its cell concentration 27%；
PH value 6.7；Without other living contaminantses.
60m3 fermentation tanks：Corn oil 1620L, alpha-lactose 1350kg, lactase 2.4kg, formula fish meal 2220kg, beer ferment
Female slag does 1080kg, Dried Corn Steep Liquor Powder 1140g/L, earthworm powder 840kg, ammonium sulfate 36kg, precipitated calcium carbonate 240kg, magnesium sulfate
3kg, zinc sulfate 2.1kg, potassium dihydrogen phosphate 48kg, sodium chloride 36kg, potassium chloride 24g, polyethylene glycol（100）Stearate
132kg, Tween 80 360kg.
Fermentating culturing process：
First fermentation medium is sterilized, cooled down, and uses filtrated air pressurize, seed liquor is all then moved into fermentation tank enters
Row culture.Fermentation culture conditions are：Fat control is 2.7% after medium sterilization；29~31 DEG C of cultivation temperature；Speed of agitator
115r/min；Tank presses 0.05~0.06MPa；PH controls are 6.0~7.5 in fermentation process；Bacterium inspection is carried out in fermentation process, it is desirable to
Without other miscellaneous bacterias；0~120h：750m3/h；121h~fermentation ends：1350m3/h.Fermented and cultured terminates, cell concentration 37%；
Enramycin fermentation unit is 11167u/ml, production cycle 238h.
Feed supplement is carried out using stream addition during the fermentation, specific feed profile is referring to embodiment 1.
Embodiment 5
10m3 seeding tanks：Corn oil 200L, alpha-lactose 100kg, lactase 0.3kg, formula fish meal 250kg, brewer's yeast
Dry slag 200kg, Dried Corn Steep Liquor Powder 150kg, earthworm powder 90kg, ammonium sulfate 5kg, precipitated calcium carbonate 50kg.
Seed culture technique
Seed culture medium is sterilized first, cooled down, and uses filtrated air pressurize, then under flame protection, will have been trained
The antifungal strepto- starter bottle zymotic fluid access seeding tank supported is cultivated, and inoculum concentration is controlled in seed culture medium volume
10%.Seed culture condition is：Tank presses 0.04~0.06MPa；28~30 DEG C of tank temperature；Air mass flow：0~12h, 50m3/h；12h
~culture transferring：70m3/h；Speed of agitator 100r/min；PH6~8；Incubation time 35h.Seed culture terminates, its cell concentration 30%；
PH value 6.2；Without other living contaminantses.
60m3 fermentation tanks：Corn oil 1800L, alpha-lactose 1500kg, lactase 3kg, formula fish meal 2400kg, brewer's yeast
Slag does 1200kg, Dried Corn Steep Liquor Powder 1200g/L, earthworm powder 900kg, ammonium sulfate 42kg, precipitated calcium carbonate 300kg, magnesium sulfate
3.6kg, zinc sulfate 2.4kg, potassium dihydrogen phosphate 54kg, sodium chloride 42kg, potassium chloride 30g, octadecyl dihydroxy ethyl amine oxide
150kg, Tween 80 420kg.
Fermentating culturing process：
First fermentation medium is sterilized, cooled down, and uses filtrated air pressurize, seed liquor is all then moved into fermentation tank enters
Row culture.Fermentation culture conditions are：Fat control is 2.9% after medium sterilization；29~31 DEG C of cultivation temperature；Speed of agitator
120r/min；Tank presses 0.05~0.06MPa；PH controls are 6.0~7.5 in fermentation process；Bacterium inspection is carried out in fermentation process, it is desirable to
Without other miscellaneous bacterias；0~120h：800m3/h；121h~fermentation ends：1500m3/h.Fermented and cultured terminates, cell concentration 39%；
Enramycin fermentation unit is 10823u/ml, production cycle 240h.
Feed supplement is carried out using stream addition during the fermentation, specific feed profile is referring to embodiment 1.
Comparative example
10m3 seeding tanks：Soya-bean oil 175L, glucose 75kg, corn protein powder 225kg, groundnut meal 175kg, corn steep liquor
125kg, yeast extract 75kg, ammonium sulfate 3.5kg, precipitated calcium carbonate 30kg.
Seed culture technique
Seed culture medium is sterilized first, cooled down, and uses filtrated air pressurize, then under flame protection, will have been trained
The antifungal strepto- starter bottle zymotic fluid access seeding tank supported is cultivated, and inoculum concentration is controlled in seed culture medium volume
7%.Seed culture condition is：Tank presses 0.04~0.06MPa；28~30 DEG C of tank temperature；Air mass flow：0~12h, 40m3/h；12h
~culture transferring：60m3/h；Speed of agitator 90r/min；PH6~8；Incubation time 32h.Seed culture terminates, its cell concentration 18%；
PH value 6.8；Without other living contaminantses.
60m3 fermentation tanks：Soya-bean oil 1500L, glucose 1350kg, corn protein powder 2100kg, groundnut meal 1050kg, jade
Rice ＆ peanut milk 1050g/L, yeast extract 750kg, ammonium sulfate 30kg, precipitated calcium carbonate 180kg, magnesium sulfate 2.7kg, zinc sulfate 1.8kg, phosphorus
Acid dihydride K42 kg, sodium chloride 30kg, potassium chloride 21g, defoamer organic silicone oil 120L.
Fermentating culturing process
First fermentation medium is sterilized, cooled down, and uses filtrated air pressurize, seed liquor is all then moved into fermentation tank enters
Row culture.Fermentation culture conditions are：Fat control is 1.7% after medium sterilization；29~31 DEG C of cultivation temperature；Speed of agitator
110r/min；Tank presses 0.05~0.06MPa；PH controls are 6.0~7.5 in fermentation process；Bacterium inspection is carried out in fermentation process, it is desirable to
Without other miscellaneous bacterias；0~120h：700m3/h；121h~fermentation ends：1100m3/h.Fermented and cultured terminates, cell concentration 32%；
Enramycin fermentation unit is 8143u/ml, production cycle 271h.
Claims (2)
1. a kind of antifungal streptomycete fermentation produces the culture medium of enramycin, including seed culture medium and fermentation medium, its
It is characterised by that the seed culture medium composition is：15~20ml/L of corn oil, 5~10g/L of alpha-lactose, lactase 0.01~
0.03g/L, 20~25g/L of formula fish meal, the dry 15~20g/L of slag of brewer's yeast, 10~15g/L of Dried Corn Steep Liquor Powder, earthworm powder 5~
10g/L, 0.2~0.5g/L of ammonium sulfate, 1~5g/L of precipitated calcium carbonate；
The fermentation medium is constituted：20~30ml/L of corn oil, 20~25g/L of alpha-lactose, 0.02~0.05g/ of lactase
L, 30~40g/L of formula fish meal, the dry 15~20g/L of slag of brewer's yeast, 15~20g/L of Dried Corn Steep Liquor Powder, 10~15g/ of earthworm powder
L, 0.3~0.7g/L of ammonium sulfate, 1~5g/L of precipitated calcium carbonate, 0.03~0.06g/L of magnesium sulfate, 0.02~0.04g/ of zinc sulfate
L, 0.5~0.9g/L of potassium dihydrogen phosphate, 0.3~0.7g/L of sodium chloride, 0.2~0.5g/L of potassium chloride, surfactant 1.5~
2.5g/L, 4~7g/L of the carrier of oxygen；
The formula fish meal is according to weight ratio 7 by fish meal, bone meal and feather meal:3:1 is obtained by mixing；
The carrier of oxygen is Tween 80 or perfluocarbon；
The surfactant be polyxyethylated alkylolamides or glyceride or polyglycerol ester or polyethylene glycol stearate or
Octadecyl dihydroxy ethyl amine oxide.
2. a kind of feed process of the culture medium producing enramycin by fermentation described in utilization claim 1, it is characterized in that：In fermentation
During using stream addition carry out feed supplement, feed supplement include mend culture medium, moisturizing, mend ammonium sulfate, pH control and mend amino acid, wherein
A mends culture medium：
Preceding 70h ferment without carrying out feed supplement,
Fermentation time mends carbon source and nitrogen source in 71~90h according to fermentative medium formula, and its amount of filling into is that fermentation medium is overall
Long-pending 3~5%,
Fermentation time mends carbon source and nitrogen source in 110h~130h according to fermentative medium formula, and its amount of filling into is fermentation medium
The 2~4% of cumulative volume,
Fermentation time mends carbon source and nitrogen source in 150h~180h according to fermentative medium formula, and its amount of filling into is fermentation medium
The 1~2% of cumulative volume；
B moisturizings：
Preceding 80h ferment without carrying out moisturizing,
Fermentation time, when cell concentration is higher than 55%, adds aqua sterilisa, it is desirable to control zymotic fluid cell concentration to exist in 81~160h
50~55%, zymotic fluid cell concentration is detected every 6~8h,
Fermentation time, when cell concentration is higher than 50%, adds aqua sterilisa, it is desirable to control zymotic fluid thalline in 161h~fermentation ends
Concentration detects zymotic fluid cell concentration 40~50% every 6~8h；
C fermentation process pH control techniques：
Fermentation before 80h, pH without control,
Fermentation time is in 81~100h, if pH ＜ 6.5, adds 10~20% sodium hydroxide, adjusts pH to 6.6~6.9；If pH
＞ 7, the ammonium sulfate of addition 20%, pH is to 6.6~6.9 for regulation,
Fermentation time is in 101~180h, if pH ＜ 6.2, adds 10~20% sodium hydroxide, and regulation pH is controlled to 6.3~6.6；If
PH ＞ 6.6, the ammonium sulfate of addition 30% adjusts pH to 6.3~6.6；
D fills into amino acid：
80h before fermentation, without filling into amino acid,
Ferment and fill into serine, cysteine and the arginine that concentration is 0.2~0.5mg/L respectively in 81h, 130h and 170h,
Its amount of filling into is the 0.1~0.3% of fermentating liquid volume；
E mends ammonium sulfate：
In 80h, 150h and 220h of fermentation, 30% ammonium sulfate is filled into respectively, and its consumption is controlled in fermentating liquid volume
0.05~0.1%.
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| CN106148460A (en) * | 2015-04-27 | 2016-11-23 | 牡丹江佰佳信生物科技有限公司 | A kind of fermentation medium improving enramycin B component yield and fermentation process |
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| CN110157757B (en) * | 2019-02-15 | 2022-11-25 | 河北圣雪大成制药有限责任公司 | Yield improving method of enramycin monocomponent, culture medium and separation method |
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