CN105506041A - Method for producing norvancomycin by fermentation - Google Patents

Method for producing norvancomycin by fermentation Download PDF

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Publication number
CN105506041A
CN105506041A CN201610097985.3A CN201610097985A CN105506041A CN 105506041 A CN105506041 A CN 105506041A CN 201610097985 A CN201610097985 A CN 201610097985A CN 105506041 A CN105506041 A CN 105506041A
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fermentation
norvancomycin
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microballoon
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张雪霞
任风芝
王耀耀
赵颖
李晓露
耿文飞
郑智慧
段宝玲
路新华
高任龙
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NCPC New Drug Research and Development Co Ltd
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Priority to CN201710092838.1A priority patent/CN106801078B/en
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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
    • C12P21/00—Preparation of peptides or proteins
    • C12P21/02—Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
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    • C07K9/00—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof
    • C07K9/006—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence being part of a ring structure
    • C07K9/008—Peptides having up to 20 amino acids, containing saccharide radicals and having a fully defined sequence; Derivatives thereof the peptide sequence being part of a ring structure directly attached to a hetero atom of the saccharide radical, e.g. actaplanin, avoparcin, ristomycin, vancomycin
    • C—CHEMISTRY; METALLURGY
    • 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
    • C12N1/20—Bacteria; Culture media therefor

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Abstract

The invention belongs to the field of fermentation engineering and particularly relates to a method for producing norvancomycin by fermentation. In this method, by adding super-paramagnetic microspheres in a norvancomycin synthesis phase to adsorb norvancomycin in a fermentation broth, a reduction is given to the feedback inhibition of a fermentation product upon generated bacteria and an improvement is given to the biological synthesis of norvancomycin. After fermentation, fermentation unit of the norvancomycin is up to 22593 ug/ml; compared with a method adding no super-paramagnetic microspheres, this method provides an increase of greater than 35% in norvancomycin yield, an improvement in productivity and equipment utilization and a reduction in wastewater and waste residue discharge.

Description

A kind of method of fermentative production Norvancomycin
Technical field
The invention belongs to field of fermentation engineering, be specifically related to a kind of method of fermentative production Norvancomycin.
Background technology
Norvancomycin hydrochloride is the unique independent development of China and is successfully applied to clinical glycopeptide antibiotics, is produced, be used for the treatment of gram positive bacteria infection by streptomycete fermentation.Norvancomycin have with vancomycin like chemical structure, pharmacological properties, anti-microbial activity and antimicrobial spectrum, its mechanism of action is the synthesis of anti-bacteria cell walls, is the choice drug of the severe infections that clinical treatment is caused by methicillin-resistant staphylococcus aureus.
Norvancomycin molecule is made up of two basic structures, and namely the center seven peptide core of glycosyl part α-o-vancosamine-β-o-glucosyl and peptidyl moiety, uses Norvancomycin hydrochloride clinically, and molecular formula is C 65h 73cl 2n 9o 24hCl, molecular weight is 1471.71.Norvancomycin hydrochloride is white or off-white color amorphous powder, and very easily water-soluble, dissolve in aqueous methanol, be insoluble to higher alcohols, acetone or ether, low urea can increase its water solubility.
Norvancomycin is the secondary metabolite of fermentable, be present in fermented liquid, Norvancomycin not only kills or restraining effect other biological, also kills or restraining effect producing strains self, and namely microbiotic also exists toxic action to producing strains self.Along with Norvancomycin concentration in fermented liquid constantly increases, its producing strains is subject to the feedback regulation effect of high density product, the potentiality of producing strains biomass cells and enzyme can not be given full play to, cause the productive rate of Norvancomycin to be difficult to improve, thus become the bottleneck that restriction improves Norvancomycin fermentation level.The producing strains of seed selection energy enduring high-concentration Norvancomycin no doubt can make productive capacity be improved, but can not tackle the problem at its root.
Summary of the invention
The object of the invention is to solve fermentable in prior art and prepare high density product that Norvancomycin exists to the feedback inhibition of producing strains and the low problem of fermentation level, there is provided a kind of by optimizing fermentative medium formula, fermentation condition, and add superparamagnetism microballoon during the fermentation, to adsorb Norvancomycin in fermented liquid, reduce the feedback inhibition to producing strains, promote the biosynthesizing of Norvancomycin, improve the method for Norvancomycin output.
Key ideas of the present invention is: adopt Hebei amycolatosis bacterial strain to be starting strain, produce in the process of Norvancomycin at fermentable, add superparamagnetism microballoon, optimize the condition such as joining day, add-on, pretreatment mode of superparamagnetism microballoon, Norvancomycin in absorption fermented liquid, reduce the feedback inhibition to producing strains, improve Norvancomycin output.
The method of a kind of fermentative production Norvancomycin provided by the present invention, comprises the following steps:
The seed liquor of a, preparation Hebei amycolatosis bacterial classification
The slant culture of Hebei amycolatosis bacterial classification or spore liquid are inoculated in seed culture medium, 26-30 DEG C, cultivate in shaking flask or fermentor tank 48-72h obtain seed liquor.
Wherein said seed culture medium obtains by the following method: W-Gum 2.0-4.0 gram, glucose 10.0-40.0 gram, hot moulding soybean cake powder 2.0-10.0 gram, yeast powder 5.0-10.0 gram, corn steep liquor 2.0-8.0 gram, sodium-chlor 0.5-2.0 gram, magnesium sulfate heptahydrate 0.5-2.0 gram, calcium carbonate 3.0-6.0 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min.
Bacterial classification of the present invention is that North China pharmacy gathers from positive definite vegetable garden for 1996, through being accredited as Hebei amycolatosis bacterial strain (Amycolatopsishebeiensis) NCPC1023 bacterial strain, this bacterial strain was preserved in China typical culture collection center in 2003, and preserving number is: CCTCCNO:203099).
The fermented liquid of b, preparation Hebei amycolatosis bacterial classification
Above-mentioned seed liquor is inoculated in fermention medium with the inoculum size of volume percent 6-10%, in shaking flask or fermentation cylinder for fermentation, ferments between 48-72 hour and add superparamagnetism microballoon, continue fermentation, total fermentation time 168-192h, leavening temperature 27-29 DEG C, obtains fermented liquid.
Wherein said fermention medium obtains by the following method: glucose 20.0-30.0 gram, Semen Maydis powder 20.0-40.0 gram, hot moulding soybean cake powder 0-20.0 gram, cottonseed flour 0-10.0 gram, yeast powder 0-10.0 gram, corn steep liquor 5.0-10.0 gram, potassium primary phosphate 3.0-5.0 gram, sodium-chlor 1.0-2.0 gram, magnesium sulfate heptahydrate 1.0-1.5 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min.
Preferably, the particle diameter of wherein said superparamagnetism microballoon is 5-30 μm, and size distribution is single dispersing, and the functional group on surface is hydroxy-acid group; Described superparamagnetism microballoon is the acid structure after HCl treatment.
Preferably, wherein the microballoon of superparamagnetism described in step b addition is that often liter of fermented liquid adds 5-15g.
Preferably, superparamagnetism microballoon addition is that often liter of fermented liquid adds 8-12g.
Preferably, the particle diameter of superparamagnetism microballoon is 10-20 μm.
Preferably, the pressure that ferments in described step b is 0.05 ± 0.01Mpa, ventilation ratio is 1:0.8-1.2, and stirring velocity is 300-400rpm.
Preferably, in wherein said step b, the interpolation time of superparamagnetism microballoon adds for fermenting between 54-64 hour.
Superparamagnetism microballoon pretreatment mode is the HCl that every gram of microballoon adds the 2moL/L of 3-5ml, after static immersing 2-4 hour filter, removing hydrochloric acid, then with deionized water wash to pH value 4.0-5.0.
After having fermented, superparamagnetism microballoon is with the Norvancomycin of ethanol sour water buffer solution elution absorption after externally-applied magnetic field reclaims, and acidic aqueous solution is phosphoric acid-phosphate buffered saline buffer, and pH value is 4.0, and ethanol volume percent is in the solution 15%.
Owing to have employed technique scheme, the technical progress that the present invention obtains is:
Superparamagnetism microballoon is a kind of novel magnetic particle that development in recent years is got up, and this magnetic-particle is with the Fe of magnetic 3o 4for core, take polymkeric substance as outer shell.Carboxy functional group is introduced on the surface of microballoon, and the present invention finds that it has strong adsorption to Norvancomycin.During producing strains synthesis Norvancomycin, in fermented liquid, add superparamagnetism microballoon, the Norvancomycin in absorption fermented liquid, reduce tunning to the feedback inhibition of producing strains, thus significantly improve the output of Norvancomycin.
Beneficial effect of the present invention:
Above-mentioned optimum condition, obtain Norvancomycin output bring up to 22593 μ g/ml from 11658 μ g/ml, the suitability for industrialized production for Norvancomycin provides a kind of new method.The present invention by optimization of fermentation conditions, ensure that the consistence of meta-bolites and fermentation level between different fermentations batch; By adding superparamagnetism microballoon during the fermentation, Norvancomycin in absorption fermented liquid, reduce the feedback inhibition of tunning to producing strains, remarkable promotion Norvancomycin generates, under the condition that other culture condition are identical, and do not add compared with superparamagnetism microballoon, Norvancomycin output increased more than 35%, improve production capacity and plant factor, reduce the discharge of waste water and waste residue.
Accompanying drawing explanation
Fig. 1 is the hypha form that strain fermentation is cultivated 48 hours.
Fig. 2 is the hypha form that strain fermentation is cultivated 72 hours.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited thereto.Experiment material used in following embodiment is industrial raw material if no special instructions; Superparamagnetism microballoon used is provided by Suzhou Nano-Micro Bio-technology Co., Ltd..Other reagent is commercially available prod.
Embodiment 1,500mL shake flask fermentation prepare Norvancomycin
The preparation of Hebei amycolatosis bacterial strain seed liquor
The slant culture of Hebei amycolatosis bacterial strain or spore liquid are inoculated in seed culture medium, 26 DEG C, 220rpm, cultivation 72h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: W-Gum 0.2 gram, glucose 4.0 grams, hot moulding soybean cake powder 0.2 gram, yeast powder 1.0 grams, corn steep liquor 0.8 gram, 0.05 gram, sodium-chlor, magnesium sulfate heptahydrate 0.05 gram, 0.3 gram, calcium carbonate, add tap water and be settled to 100mL, pH6.5,121 DEG C of sterilizing 30min.
The culture presevation number of Norvancomycin producing strains of the present invention is CCTCC203099.
Fermention medium forms:
Glucose 20.0 grams, Semen Maydis powder 40.0 grams, hot moulding soybean cake powder 20.0 grams, corn steep liquor 10.0 grams, potassium primary phosphate 3.0 grams, 1.0 grams, sodium-chlor, magnesium sulfate heptahydrate 1.0 grams, add tap water and be settled to 1000mL, pH6.5,121 DEG C of sterilizing 30min, 500mL shake flask fermentation is cultivated.
Fermentation condition: the triangular flask of 500ml loading amount 6 bottles, every bottled 50ml substratum, seed culture was inoculated after 72 hours, inoculum size is 10%, culture temperature is 28 DEG C, shaking speed 220rpm, after being cultured to 48 hours, (strain fermentation cultivate 48 hours hypha form see Fig. 1), every bottle adds the super suitable Nano microsphere medium 0.75g that particle diameter is 5 μm, and the hypha form that fermentation culture 168h(strain fermentation is cultivated 72 hours is shown in Fig. 2).
After fermentation ends, after being merged by 6 bottles of fermented liquids, magnetic resolution reclaims the superparamagnetism microballoon in fermented liquid.In superparamagnetism microballoon, add the ethanol-acidic aqueous buffer wash-out Norvancomycin of 23ml after recovery, measure the Norvancomycin unit in elutriant and fermented liquid respectively.Calculate Norvancomycin fermentation unit.
The results are shown in Table 1, under the fermentation condition optimized, adopt traditional soybean cake powder as organic nitrogen source, add superparamagnetism microballoon during the fermentation, the fermentation unit of Norvancomycin is 15738 μ g/ml.
Embodiment 2,5L Fermentation fermentation are for Norvancomycin
The preparation of Hebei amycolatosis bacterial strain seed liquor
The slant culture of Hebei amycolatosis bacterial strain or spore liquid are inoculated in seed culture medium, 28 DEG C, 220rpm, cultivation 64h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: W-Gum 3.0 grams, glucose 20.0 grams, hot moulding soybean cake powder 10.0 grams, yeast powder 5.0 grams, corn steep liquor 5.0 grams, 1.0 grams, sodium-chlor, magnesium sulfate heptahydrate 1.0 grams, 4.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH6.8,121 DEG C of sterilizing 30min.
The culture presevation number of Norvancomycin producing strains of the present invention is CCTCC203099.
Fermention medium forms:
Glucose 90.0 grams, Semen Maydis powder 90.0 grams, hot moulding soybean cake powder 45.0 grams, cottonseed flour 15.0 grams, corn steep liquor 15.0 grams, potassium primary phosphate 15.0 grams, 6.0 grams, sodium-chlor, magnesium sulfate heptahydrate 4.5 grams, add tap water and be settled to 3000mL, pH6.8,121 DEG C of sterilizing 30min, 5L Fermentation fermentation culture.
Fermentation condition: the pressure in tank is 0.05 ± 0.01Mpa, ventilation ratio 1:0.8vvm, and charge amount is 3L, and seed culture was inoculated after 64 hours, and inoculum size is 8%, and culture temperature is 29 DEG C, and rotating speed is 400r/min.After fermentation culture starts 56 hours, add the super suitable Nano microsphere medium 15g that particle diameter is 30 μm, fermentation culture 192 hours.
After fermentation ends, magnetic resolution reclaims the superparamagnetism microballoon in fermented liquid.In superparamagnetism microballoon, add the Norvancomycin of the ethanol-acidic aqueous buffer wash-out absorption of 75ml after recovery, measure the Norvancomycin unit in elutriant and fermented liquid respectively.The fermentation unit calculating Norvancomycin is 18703 μ g/ml.
The results are shown in Table 1, under the fermentation condition optimized, adopt the soybean cake powder and cottonseed flour optimized as organic nitrogen source, add superparamagnetism microballoon during the fermentation, the fermentation unit of Norvancomycin is 18703 μ g/ml.
Embodiment 3,10L Fermentation fermentation are for Norvancomycin
The preparation of Hebei amycolatosis bacterial strain seed liquor
The slant culture of Hebei amycolatosis bacterial strain or spore liquid are inoculated in seed culture medium, 30 DEG C, 220rpm, cultivation 48h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: W-Gum 4.0 grams, glucose 10.0 grams, hot moulding soybean cake powder 8.0 grams, yeast powder 8.0 grams, corn steep liquor 2.0 grams, 2.0 grams, sodium-chlor, magnesium sulfate heptahydrate 2.0 grams, 6.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min.
The culture presevation number of Norvancomycin producing strains of the present invention is CCTCC203099.
Fermention medium forms:
Glucose 150.0 grams, Semen Maydis powder 100.0 grams, cottonseed flour 50.0 grams, yeast powder 50.0 grams, corn steep liquor 40.0 grams, potassium primary phosphate 20.0 grams, 5.0 grams, sodium-chlor, magnesium sulfate heptahydrate 5.0 grams, add tap water and be settled to 5000mL, pH7.0,121 DEG C of sterilizing 30min, 10L Fermentation fermentation culture.
Fermentation condition: the pressure in tank is 0.05 ± 0.01Mpa, ventilation ratio 1:1.2vvm, and charge amount is 5L, and seed culture was inoculated after 48 hours, and inoculum size is 6%, and culture temperature is 27 DEG C, and rotating speed is 300r/min.Start 72 hours in fermentation culture, add the super suitable Nano microsphere medium 50g that particle diameter is 10 μm, fermentation culture 180 hours.The hypha form of different fermentations incubation time is shown in Fig. 1 and Fig. 2.
After fermentation ends, magnetic resolution reclaims the superparamagnetism microballoon in fermented liquid.In superparamagnetism microballoon, add the Norvancomycin of the ethanol-acidic aqueous buffer wash-out absorption of 250ml after recovery, measure the Norvancomycin unit in elutriant and fermented liquid respectively.
The results are shown in Table 1, under the fermentation condition optimized, adopt the cottonseed flour and yeast powder optimized as organic nitrogen source, add superparamagnetism microballoon during the fermentation, the fermentation unit of Norvancomycin is 22593 μ g/ml.
Comparative example 1,500mL shake flask fermentation prepare Norvancomycin
The preparation of Hebei amycolatosis bacterial strain seed liquor
The slant culture of Hebei amycolatosis bacterial strain or spore liquid are inoculated in seed culture medium, 26 DEG C, 220rpm, cultivation 72h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: W-Gum 0.2 gram, glucose 4.0 grams, hot moulding soybean cake powder 0.2 gram, yeast powder 1.0 grams, corn steep liquor 0.8 gram, 0.05 gram, sodium-chlor, magnesium sulfate heptahydrate 0.05 gram, 0.3 gram, calcium carbonate, add tap water and be settled to 100mL, pH6.5,121 DEG C of sterilizing 30min.
The culture presevation number of Norvancomycin producing strains of the present invention is CCTCC203099.
Fermention medium forms:
Glucose 20.0 grams, Semen Maydis powder 40.0 grams, hot moulding soybean cake powder 20.0 grams, corn steep liquor 10.0 grams, potassium primary phosphate 3.0 grams, 1.0 grams, sodium-chlor, magnesium sulfate heptahydrate 1.0 grams, add tap water and be settled to 1000mL, pH6.5,121 DEG C of sterilizing 30min, 500mL shake flask fermentation is cultivated.
Fermentation condition: the triangular flask of 500ml loading amount 6 bottles, every bottled 50ml substratum, seed culture was inoculated after 72 hours, and inoculum size is 10%, and culture temperature is 28 DEG C, rotating speed 220rpm, fermentation culture 168h.
The results are shown in Table 1, under the fermentation condition optimized, adopt traditional soybean cake powder as organic nitrogen source, when not adding superparamagnetism microballoon, after fermentation ends, the fermentation unit of Norvancomycin is 11658 μ g/ml.
Comparative example 2,5L Fermentation fermentation are for Norvancomycin
The preparation of Hebei amycolatosis bacterial strain seed liquor
The slant culture of Hebei amycolatosis bacterial strain or spore liquid are inoculated in seed culture medium, 28 DEG C, 220rpm, cultivation 64h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: W-Gum 3.0 grams, glucose 20.0 grams, hot moulding soybean cake powder 10.0 grams, yeast powder 5.0 grams, corn steep liquor 5.0 grams, 1.0 grams, sodium-chlor, magnesium sulfate heptahydrate 1.0 grams, 4.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH6.8,121 DEG C of sterilizing 30min.
The culture presevation number of Norvancomycin producing strains of the present invention is CCTCC203099.
Fermention medium forms:
Glucose 90.0 grams, Semen Maydis powder 90.0 grams, hot moulding soybean cake powder 45.0 grams, cottonseed flour 15.0 grams, corn steep liquor 15.0 grams, potassium primary phosphate 15.0 grams, 6.0 grams, sodium-chlor, magnesium sulfate heptahydrate 4.5 grams, add tap water and be settled to 3000mL, pH6.8,121 DEG C of sterilizing 30min, 5L Fermentation fermentation culture.
Fermentation condition: the pressure in tank is 0.05 ± 0.01Mpa, ventilation ratio 1:0.8, and charge amount is 3L, and seed culture was inoculated after 64 hours, and inoculum size is 8%, and culture temperature is 29 DEG C, and rotating speed is 400r/min, fermentation culture 192 hours.
The results are shown in Table 1, under the fermentation condition optimized, adopt the soybean cake powder and cottonseed flour optimized as organic nitrogen source, when not adding superparamagnetism microballoon, after fermentation ends, the fermentation unit of Norvancomycin is 13752 μ g/ml.
Comparative example 3,10L Fermentation fermentation are for Norvancomycin
The preparation of Hebei amycolatosis bacterial strain seed liquor
The slant culture of Hebei amycolatosis bacterial strain or spore liquid are inoculated in seed culture medium, 30 DEG C, 220rpm, cultivation 48h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: W-Gum 4.0 grams, glucose 10.0 grams, hot moulding soybean cake powder 8.0 grams, yeast powder 8.0 grams, corn steep liquor 2.0 grams, 2.0 grams, sodium-chlor, magnesium sulfate heptahydrate 2.0 grams, 6.0 grams, calcium carbonate, add tap water and be settled to 1000mL, pH7.0,121 DEG C of sterilizing 30min
The culture presevation number of Norvancomycin producing strains of the present invention is CCTCC203099.
Fermention medium forms:
Glucose 150.0 grams, Semen Maydis powder 100.0 grams, cottonseed flour 50.0 grams, yeast powder 50.0 grams, corn steep liquor 40.0 grams, potassium primary phosphate 20.0 grams, 5.0 grams, sodium-chlor, magnesium sulfate heptahydrate 5.0 grams, add tap water and be settled to 5000mL, pH7.0,121 DEG C of sterilizing 30min, 10L Fermentation fermentation culture.
Fermentation condition: the pressure in tank is 0.05 ± 0.01Mpa, ventilation ratio 1:1.2, and charge amount is 5L, and seed culture was inoculated after 48 hours, and inoculum size is 6%, and culture temperature is 27 DEG C, and rotating speed is 300r/min, fermentation culture 180 hours.
The results are shown in Table 1, under the fermentation condition optimized, adopt cottonseed flour and yeast powder as organic nitrogen source, when not adding superparamagnetism microballoon, after fermentation ends, the fermentation unit of Norvancomycin is 16254 μ g/ml.
Results contrast in table 1 is calculated known, add superparamagnetism microballoon compared with not adding, fermentation unit is all made to improve more than 35% under various formula, be respectively,: soybean cake powder: (15738-11658)/11658=35%, cottonseed flour and soybean cake powder: (18703-13752)/13752=36%, cottonseed flour and yeast powder: (22593-16254)/16254=39%.

Claims (6)

1. a method for fermentative production Norvancomycin, is characterized in that: comprise the following steps:
The seed liquor of a, preparation Hebei amycolatosis bacterial classification
The slant culture of Hebei amycolatosis bacterial classification or spore liquid are inoculated in seed culture medium, 26-30 DEG C, cultivate in shaking flask or fermentor tank 48-72h obtain seed liquor;
Wherein said seed culture medium obtains by the following method: W-Gum 2.0-4.0 gram, glucose 10.0-40.0 gram, hot moulding soybean cake powder 2.0-10.0 gram, yeast powder 5.0-10.0 gram, corn steep liquor 2.0-8.0 gram, sodium-chlor 0.5-2.0 gram, magnesium sulfate heptahydrate 0.5-2.0 gram, calcium carbonate 3.0-6.0 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min;
The fermented liquid of b, preparation Hebei amycolatosis bacterial classification
Above-mentioned seed liquor is inoculated in fermention medium with the inoculum size of volume percent 6-10%, in shaking flask or fermentation cylinder for fermentation, ferments between 48-72 hour and add superparamagnetism microballoon, continue fermentation, total fermentation time 168-192h, leavening temperature 27-29 DEG C, obtains fermented liquid;
Wherein said fermention medium obtains by the following method: glucose 20.0-30.0 gram, Semen Maydis powder 20.0-40.0 gram, hot moulding soybean cake powder 0-20.0 gram, cottonseed flour 0-10.0 gram, yeast powder 0-10.0 gram, corn steep liquor 5.0-10.0 gram, potassium primary phosphate 3.0-5.0 gram, sodium-chlor 1.0-2.0 gram, magnesium sulfate heptahydrate 1.0-1.5 gram, add water and be settled to 1000mL, pH6.5-7.0,121 DEG C of sterilizing 30min.
2. method according to claim 1, the particle diameter of the microballoon of superparamagnetism described in step b is 5-30 μm, and size distribution is single dispersing, and the functional group on surface is hydroxy-acid group.
3. method according to claim 1 and 2, the microballoon of superparamagnetism described in step b addition is that often liter of fermented liquid adds 5-15g.
4. method according to claim 3, superparamagnetism microballoon addition is that often liter of fermented liquid adds 8-12g.
5. method according to claim 2, the particle diameter of superparamagnetism microballoon is 10-20 μm.
6. method according to claim 1, the pressure that ferments in described step b is 0.05 ± 0.01Mpa, stirring velocity is 300-400rpm, and ventilation ratio is 1:0.8-1.2vvm.
CN201610097985.3A 2016-02-23 2016-02-23 Method for producing norvancomycin by fermentation Pending CN105506041A (en)

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Cited By (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105755077A (en) * 2016-05-19 2016-07-13 宁夏泰瑞制药股份有限公司 Fermentation culture medium for producing vancomycin through fermentation of streptomyces orientalis and material supplementing method
CN109880863A (en) * 2019-01-25 2019-06-14 南开大学 The method for improving yield of streptomycete antibiotic as additive using nano material
CN115404175A (en) * 2022-10-12 2022-11-29 北京雷力海洋生物新产业股份有限公司 Fermentation medium of trichoderma viride, trichoderma agent and preparation method of trichoderma agent

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Publication number Priority date Publication date Assignee Title
CN105755077A (en) * 2016-05-19 2016-07-13 宁夏泰瑞制药股份有限公司 Fermentation culture medium for producing vancomycin through fermentation of streptomyces orientalis and material supplementing method
CN109880863A (en) * 2019-01-25 2019-06-14 南开大学 The method for improving yield of streptomycete antibiotic as additive using nano material
CN109880863B (en) * 2019-01-25 2022-07-22 南开大学 Method for improving yield of streptomycete antibiotics by using nano material as additive
CN115404175A (en) * 2022-10-12 2022-11-29 北京雷力海洋生物新产业股份有限公司 Fermentation medium of trichoderma viride, trichoderma agent and preparation method of trichoderma agent

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