CN103012514A - Membrane separation and extraction method for micronomicin sulfate - Google Patents
Membrane separation and extraction method for micronomicin sulfate Download PDFInfo
- Publication number
- CN103012514A CN103012514A CN2012105651584A CN201210565158A CN103012514A CN 103012514 A CN103012514 A CN 103012514A CN 2012105651584 A CN2012105651584 A CN 2012105651584A CN 201210565158 A CN201210565158 A CN 201210565158A CN 103012514 A CN103012514 A CN 103012514A
- Authority
- CN
- China
- Prior art keywords
- filtration unit
- group
- ultra filtration
- stage
- parts
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Images
Landscapes
- Separation Using Semi-Permeable Membranes (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention relates to a membrane separation and extraction method for micronomicin sulfate. The membrane separation and extraction method adopts the technical scheme that the method comprises the steps that a micronomicin sulfate analytic solution is decolorized by an ultrafiltration membrane system, and concentrated by a nanofiltration membrane system, wherein the ultrafiltration membrane system comprises six ultrafiltration membranes, every three ultrafiltration membranes are in series connection to form a group; the two groups are in parallel connection; and the nanofiltration membrane system is formed by connecting three nanofiltration membranes in series. The membrane separation and extraction method has the advantages that a technique is simple and easy to operate; the loss of effective ingredients is less; the product quality is stable; and the energy and material consumption is reduced.
Description
Technical field
The present invention relates to a kind of membrane sepn extracting method, be specifically related to the membrane sepn extracting method of Micronomicin Sulfate.
Background technology
Micronomicin Sulfate is porous solid or the powder of white or off-white color; Odorless, have draw moist.It is easily molten in water, and is almost insoluble in methyl alcohol, ethanol, acetone, ethyl acetate or trichloromethane.Be aminoglycoside antibiotics, its impede protein matter is synthetic, to the sensitive organism demonstration anti-microbial activity of gram-negative bacteria and gram positive organism.Be used for the various infection due to the gram negative bacilli, such as microbemia, respiratory system, bile duct, urinary tract infections etc.Local eye drop is treated responsive microbial blepharitis, dacryocystitis, conjunctivitis, keratitis etc.Existing technique Micronomicin Sulfate is by deep red micromonospora 201-9-8-53-84-37
#(Micromonospora purpura 201-9-8-53-84-37
#) generate.Deep red micromonospora 201-9-8-53-84-37
#(Micromonospora purpura 201-9-8-53-84-37
#), be actinomyces, Micromonosporaceae, in the center preservation of Chinese Typical Representative culture collection, preserving number is: CCTCC NO:M 2012471; Depositary institution address: Wuhan, China Wuhan University, preservation date: on November 21st, 2012.
Existing Micronomicin Sulfate extractive technique, with the Micronomicin Sulfate desorbed solution, through 711 type strongly basic anion exchange resin beds, behind the dynamic adsorption pigment again through the film under vacuum upgrading tower, use the steam heating evaporating solvent, get the concentrated solution of original volume 1/40, this concentrated solution adsorbs residual pigment with powdered active carbon at last, thereby reaches processing requirement.This technical process is complicated, the cycle is long, energy consumption is high, and effective constituent runs off more serious, and the product yield is low, and employed organic solvent, has not only increased extraction cost, and easily environment is caused serious pollution.As selecting traditional culture medium prescription and traditional technology in " producing 15 tons of Micronomicin Sulfate plant design per year ", the Micronomicin Sulfate desorbed solution concentrates, turns the techniques such as salt, decolouring and make product through pre-treatment, ion exchange resin absorption, steam, 20 tons of resins of annual about loss cause a large amount of wastes.Be optimization technological process, improve the quality of products and stability, save cost, seek new producing and manufacturing technique extremely urgent.
Background technology of the present invention is as follows:
Certain traditional technology first is as follows:
The preparation of A, micronomicin seed:
A1, female bottle slant culture: the traditional culture medium prescription of female bottle; Cultivated 6-7 days in 35 ℃;
A2, son bottle slant culture: the traditional substratum of son bottle; In 37 ℃ of cultivations 10-15 days, make spore, vacuum-drying saves backup;
A3, spore suspension: the spore of son bottle slant culture is made suspension;
The preparation of B, Micronomicin Sulfate fermented liquid:
B1, first class seed pot are cultivated: first order seed tradition substratum; In 35-37 ℃, ventilation 1:2V/V.m, stirring velocity 320-350rpm, PH nature, incubation time 40-45h;
B2, secondary seed tank are cultivated: secondary seed tradition substratum; In 33-35 ℃, ventilation 1:1-1.5V/V.m, stirring velocity 250-300rpm, PH nature, incubation time 13-15h;
B3, with the seed cultivated with 10% inoculum size access fermentation flask, cultivate in 32-34 ℃ and to get fermented liquid in 5-7 days, power of agitator 2-4KW/m3 wherein, rotating speed 150-180r/min, air flow quantity 50-70m3/m3.h, tank pressure 0.03-0.05Mpa;
The preparation of C, Micronomicin Sulfate desorbed solution:
C1, acidifying removal of impurities: 98% sulfuric acid acidation with 1 times of amount of fermented liquid quality filters to get acidizing fluid, removes miscellaneous bacteria and other impurity;
C2, neutralization: it is 7 that the NaOH that adds 3mol/L is neutralized to PH with acidizing fluid;
C3,732 resin absorption;
Through 711 type strongly basic anion exchange resin beds, again through the film under vacuum upgrading tower, use the steam heating evaporating solvent behind the dynamic adsorption pigment, get the concentrated solution of original volume 1/40, this concentrated solution adsorbs residual pigment with powdered active carbon at last, thereby reaches processing requirement.
Certain traditional technology second such as " producing 15 tons of Micronomicin Sulfate plant design per year ", has many similarities with certain traditional technology first.
For remedying the defective of above-mentioned prior art, the problem that needs most at present solution is: optimization technological process, reduce the effective constituent loss, and improve the product yield, stabilized product quality reduces the energy and supplies consumption, reduces pollutant emission, reduces production costs.
Summary of the invention
The membrane sepn extracting method that the purpose of this invention is to provide a kind of Micronomicin Sulfate.Main step comprises: the acquisition of Micronomicin Sulfate desorbed solution, the ultra-filtration membrane decoloration process of Micronomicin Sulfate desorbed solution, the nanofiltration membrane concentration technology of Micronomicin Sulfate desorbed solution.
Purpose one of the present invention is the 711 type strongly basic anion exchange resin bed decoloration process that substitute traditional technology with the ultra-filtration membrane decoloration process, substitute the steam concentration technology of traditional technology with the nanofiltration membrane concentration technology, reduce the waste of resin, and a large amount of losses of the energy.
Purpose two of the present invention is by improving culture medium prescription, and be combined with appropriate ultra-filtration membrane decoloration process, nanofiltration membrane concentration technology, such as aperture, the working pressure of the molecular weight cut-off of ultra-filtration membrane, nanofiltration membrane, see through the optimization of flow etc., not only can reduce the waste of resin, and a large amount of losses of energy consumption, the higher Micronomicin Sulfate of effective component content can also be prepared with, tiring and purity of product can be improved.Under the prerequisite that obtains more efficient valency product, the suitable medium prescription can effectively reduce the content of impurity, and then reduces the link of ultra-filtration membrane decoloration process, and for example the ultra-filtration membrane unit piece can be kept to six unit by ten unit.
Lay special stress on of the present invention: technical characteristics of the present invention is ultra-filtration membrane decoloration process, nanofiltration membrane concentration technology.The acquisition of Micronomicin Sulfate desorbed solution is not technical characteristics of the present invention, the Micronomicin Sulfate desorbed solution can obtain according to the technical clarification of " producing 15 tons of Micronomicin Sulfate plant design per year ", also can and separate acquisition according to other culture medium prescriptions cultivations.
Micronomicin Sulfate belongs to amino acid glycoside Broad spectrum antibiotics, and it can be by a strain deep red micromonospora 201-9-8-53-84-37
#(Micromonospora purpura 201-9-8-53-84-37
#) produce.Deep red micromonospora 201-9-8-53-84-37
#(Micromonospora purpura 201-9-8-53-84-37
#), be actinomyces, Micromonosporaceae, in the center preservation of Chinese Typical Representative culture collection, preserving number is: CCTCC NO:M 2012471; Depositary institution address: Wuhan, China Wuhan University, preservation date: on November 21st, 2012.
The principle of work of membrane separation technique is, sees through film as separating medium take selectivity, certain impellent in addition in the film both sides.Make feed side compositional selecting ground see through film, thereby reach the purpose of separating or purifying.According to the selection perviousness of film and varying in size of membrane pore size, can selectively with the material concentrating and separating of different-grain diameter, reclaim useful matter.The aperture of film is generally micron order, and the difference (or being called molecular weight cut-off) according to its aperture can be divided into film microfiltration membrane (MF), ultra-filtration membrane (UF), nanofiltration membrane (NF) and reverse osmosis membrane (RO) etc.Wherein, ultra-filtration membrane is that a kind of aperture specification is consistent, and specified pore diameter range is the micropore filtering film of 0.001-0.02 micron.And nanofiltration membrane is the semi-permeable membranes that allows solvent molecule or some low molecular weight solutes or low price ion to see through.
Do not undergo phase transition in the membrane sepn process, separation factor is larger, and is energy-conservation, efficient, non-secondary pollution, and at normal temperatures operate continuously is specially adapted to heat-sensitive substance.And this process need not to add other material from the external world, and is when avoiding effective constituent destroyed, also cost-saved like this, reduces pollutant emission.
Technical scheme of the present invention is as follows:
The preparation of A, micronomicin seed:
Get deep red micromonospora (Micromonospora purpura), the implant carrier bottle is cultivated;
A1, female bottle slant culture: substratum is made by the supplementary material of following weight proportioning, Zulkovsky starch 8-12 part, sodium-chlor 0.3-0.8 part, dipotassium hydrogen phosphate 0.2-0.5 part, saltpetre 0.8-1.2 part, asparagine 0.01-0.03 part, sal epsom 0.3-0.7 part, wheat bran 15-20 part, glycerine 0.3-0.8 part, agar 16-20 part; Cultivated 6-7 days in 35 ℃;
A2, son bottle slant culture: substratum is made by the supplementary material of following weight proportioning, Zulkovsky starch 10-15 part, sodium-chlor 0.3-0.8 part, dipotassium hydrogen phosphate 0.3-0.6 part, saltpetre 1.0-1.5 part, asparagine 0.02-0.04 part, sal epsom 0.3-0.7 part, wheat bran 16-22 part, glycerine 0.4-1.0 part, agar 18-22 part; In 37 ℃ of cultivations 10-15 days, make spore, vacuum-drying saves backup;
A3, spore suspension: the spore of son bottle slant culture is made suspension;
The preparation of B, Micronomicin Sulfate fermented liquid:
B1, first class seed pot are cultivated: substratum is by made soybean cake powder 12-18 part, glucose 0.8-1.5 part, Semen Maydis powder 18-24 part, fish meal 1-3 part, urea 0.8-1.2 part, calcium carbonate 0.8-1.2 part, starch 36-45 part by the supplementary material of following weight proportioning; In 35-37 ℃, ventilation 1:2V/V.m, stirring velocity 320-350rpm, PH nature, incubation time 40-45h;
B2, secondary seed tank are cultivated: substratum is by made groundnut meal 15-20 part, glucose 1.0-1.8 part, Semen Maydis powder 20-26 part, urea 0.8-1.2 part, saltpetre 0.8-1.2 part, calcium carbonate 0.8-1.2 part, starch 36-45 part by the supplementary material of following weight proportioning; In 33-35 ℃, ventilation 1:1-1.5V/V.m, stirring velocity 250-300rpm, PH nature, incubation time 13-15h;
B3, with the seed cultivated with 10% inoculum size access fermentation flask, cultivate in 32-34 ℃ and to get fermented liquid in 5-7 days, power of agitator 2-4KW/m3 wherein, rotating speed 150-180r/min, air flow quantity 50-70m3/m3.h, tank pressure 0.03-0.05Mpa;
The preparation of C, Micronomicin Sulfate desorbed solution:
C1, acidifying removal of impurities: 98% sulfuric acid acidation with 1 times of amount of fermented liquid quality filters to get acidizing fluid, removes miscellaneous bacteria and other impurity;
C2, neutralization: it is 7 that the NaOH that adds 3mol/L is neutralized to PH with acidizing fluid;
C3,732 resin absorption;
D, bleaching process: the Micronomicin Sulfate desorbed solution decolours through ultrafiltration membrane system;
Ultrafiltration membrane system wherein comprises first group of ultra filtration unit, second group of ultra filtration unit; First group of ultra filtration unit is comprised of first group of first step ultra filtration unit, first group of second stage ultra filtration unit, first group of third stage ultra filtration unit; Second group of ultra filtration unit is comprised of second group of first step ultra filtration unit, second group of second stage ultra filtration unit, second group of third stage ultra filtration unit; Wherein: first group of first step ultra filtration unit, first group of second stage ultra filtration unit and the group third stage ultra filtration unit of being connected connect successively, and second group of first step ultra filtration unit, second group of second stage ultra filtration unit and the group third stage ultra filtration unit of being connected connect successively; First group of ultra filtration unit and second group of ultra filtration unit are connected in parallel;
The decolouring step: the Micronomicin Sulfate desorbed solution is parallel through first group of ultra filtration unit and second group of ultra filtration unit decolouring, be that the Micronomicin Sulfate desorbed solution is abreast successively through first group of first step ultra filtration unit, first group of second stage ultra filtration unit, first group of third stage ultra filtration unit, with second group of first step ultra filtration unit, second group of second stage ultra filtration unit, second group of third stage ultra filtration unit, the Micronomicin Sulfate desorbed solution after the decolouring enters enrichment process;
The ultrafiltration membrane system working parameter is as follows: it is the ultra-filtration membrane of holding back the 1000-5000 molecular weight that ultra filtration unit is selected the aperture, and working pressure is controlled at 0.3-0.8Mpa, sees through flow is 2-5T/h;
E, enrichment process: the Micronomicin Sulfate desorbed solution after the decolouring is concentrated through the nanofiltration membrane system;
Nanofiltration membrane system wherein comprises first step nano-filtration unit, second stage nano-filtration unit, third stage nano-filtration unit; Wherein first step nano-filtration unit, second stage nano-filtration unit, third stage nano-filtration unit connect successively;
Enrichment step: the Micronomicin Sulfate desorbed solution after the decolouring concentrates through first step nano-filtration unit, second stage nano-filtration unit, third stage nano-filtration unit successively; Obtain Micronomicin Sulfate.
The nanofiltration membrane system operational parameters is as follows: it is the nanofiltration membrane of 1nm-50nm that nano-filtration unit is selected the aperture; Working pressure is controlled at 1.0-3.0Mpa, sees through flow is 2-5T/h.
The present invention has following advantage:
1, simplifies the technical process of traditional Micronomicin Sulfate separation and Extraction, shorten the production cycle;
2, avoid the loss of product effective ingredient in the course of processing or rotten, improve product yield;
3, stabilized product quality;
4, membrane sepn does not undergo phase transition, and carries out at normal temperatures, does not add other materials, reduces the energy and supplies consumption, reduces pollutant emission, reduces production costs.
Description of drawings
Fig. 1 is the structural representation of ultrafiltration membrane system of the present invention and nanofiltration membrane system.
Reference numeral: first group of first step ultra filtration unit 1, first group of second stage ultra filtration unit 2,3, the second groups of first step ultra filtration unit 4 of first group of third stage ultra filtration unit, second group of second stage ultra filtration unit 5, second group of third stage ultra filtration unit 6, first step nano-filtration unit 7, second stage nano-filtration unit 8, third stage nano-filtration unit 9.
Specific embodiment
The invention will be further described below in conjunction with specific embodiment.
Embodiment 1,
The membrane sepn extracting method of Micronomicin Sulfate (Micronomicin Sulfate) specifically obtains as follows:
The preparation of A, micronomicin seed:
Get deep red micromonospora (Micromonospora purpura), the implant carrier bottle is cultivated;
A1, female bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 10 parts of Zulkovsky starches, 0.5 part in sodium-chlor, 0.3 part of dipotassium hydrogen phosphate, 1.0 parts in saltpetre, 0.02 part of asparagine, 0.5 part in sal epsom, 18 parts in wheat bran, 0.6 part of glycerine, 18 parts in agar; Cultivated 7 days in 35 ℃;
A2, son bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 13 parts of Zulkovsky starches, 0.5 part in sodium-chlor, 0.4 part of dipotassium hydrogen phosphate, 1.3 parts in saltpetre, 0.03 part of asparagine, 0.5 part in sal epsom, 18 parts in wheat bran, 0.8 part of glycerine, 20 parts in agar; In 37 ℃ of cultivations 12 days, make spore, vacuum-drying saves backup;
A3, spore suspension: the spore of son bottle slant culture is made suspension;
The preparation of B, Micronomicin Sulfate fermented liquid:
B1, first class seed pot are cultivated: substratum is by made 15 parts of soybean cake powders, 1.2 parts of glucose, 22 parts of Semen Maydis powder, 2 parts of fish meal, 1.0 parts in urea, 1.0 parts in calcium carbonate, 40 parts of starch by the supplementary material of following weight proportioning; In 37 ℃, ventilation 1:2V/V.m, stirring velocity 350rpm, PH nature, incubation time 42h;
B2, secondary seed tank are cultivated: substratum is by made 18 parts of groundnut meals, 1.5 parts of glucose, 24 parts of Semen Maydis powder, 1.0 parts in urea, 1.0 parts in saltpetre, 0.8 part in calcium carbonate, 42 parts of starch by the supplementary material of following weight proportioning; In 35 ℃, ventilation 1:1.2V/V.m, stirring velocity 280rpm, PH nature, incubation time 15h;
B3, with the seed cultivated with 10% inoculum size access fermentation flask, cultivated 6 days to get fermented liquid in 34 ℃, power of agitator 3KW/m3 wherein, rotating speed 160r/min, air flow quantity 60m3/m3.h, tank pressure 0.04Mpa;
The preparation of C, Micronomicin Sulfate desorbed solution:
C1, acidifying removal of impurities: 98% sulfuric acid acidation with 1 times of amount of fermented liquid quality filters to get acidizing fluid, removes miscellaneous bacteria and other impurity;
C2, neutralization: it is 7 that the NaOH that adds 3mol/L is neutralized to PH with acidizing fluid;
C3, resin absorption removal of impurities;
D, bleaching process: the Micronomicin Sulfate desorbed solution decolours through ultrafiltration membrane system;
Ultrafiltration membrane system wherein comprises first group of ultra filtration unit, second group of ultra filtration unit; First group of ultra filtration unit is comprised of first group of first step ultra filtration unit 1, first group of second stage ultra filtration unit 2, first group of third stage ultra filtration unit 3; Second group of ultra filtration unit is comprised of second group of first step ultra filtration unit 4, second group of second stage ultra filtration unit 5, second group of third stage ultra filtration unit 6; Wherein: first group of first step ultra filtration unit 1, first group of second stage ultra filtration unit 2 and the group third stage ultra filtration unit 3 of being connected connect successively, and second group of first step ultra filtration unit 4, second group of second stage ultra filtration unit 5 and the group third stage ultra filtration unit 6 of being connected connect successively; First group of ultra filtration unit and second group of ultra filtration unit are connected in parallel;
The decolouring step: the Micronomicin Sulfate desorbed solution is parallel through first group of ultra filtration unit and second group of ultra filtration unit decolouring, be that the Micronomicin Sulfate desorbed solution is abreast successively through first group of first step ultra filtration unit 1, first group of second stage ultra filtration unit 2, first group of third stage ultra filtration unit 3, with second group of first step ultra filtration unit 4, second group of second stage ultra filtration unit 5, second group of third stage ultra filtration unit 6, the Micronomicin Sulfate desorbed solution after the decolouring enters enrichment process;
The ultrafiltration membrane system working parameter is as follows: it is the ultra-filtration membrane of holding back 5000 molecular weight that first group of first step ultra filtration unit 1, second group of first step ultra filtration unit 4 are selected the aperture, it is the ultra-filtration membrane of holding back 4000 molecular weight that first group of second stage ultra filtration unit 2, second group of second stage ultra filtration unit 5 are selected the aperture, it is the ultra-filtration membrane of holding back 3000 molecular weight that 3, second groups of third stage ultra filtration unit 6 of first group of third stage ultra filtration unit composition are selected the aperture, and working pressure all is controlled at 0.5Mpa, sees through flow is 3T/h;
E, enrichment process: the Micronomicin Sulfate desorbed solution after the decolouring is concentrated through the nanofiltration membrane system;
Nanofiltration membrane system wherein comprises first step nano-filtration unit 7, second stage nano-filtration unit 8, third stage nano-filtration unit 9; Wherein first step nano-filtration unit 7, second stage nano-filtration unit 8, third stage nano-filtration unit 9 connect successively;
Enrichment step: the Micronomicin Sulfate desorbed solution after the decolouring concentrates through first step nano-filtration unit 7, second stage nano-filtration unit 8, third stage nano-filtration unit 9 successively; Obtain Micronomicin Sulfate.
The nanofiltration membrane system operational parameters is as follows: it is the nanofiltration membrane of 35nm that first step nano-filtration unit 7 is selected the aperture, and it is the nanofiltration membrane of 20nm that second stage nano-filtration unit 8 is selected the aperture, and it is the nanofiltration membrane of 10nm that third stage nano-filtration unit 9 is selected the aperture; Working pressure all is controlled at 1.5Mpa, sees through flow is 3T/h.
The ultrafiltration membrane system working parameter is as follows: it is the ultra-filtration membrane of holding back 3000 molecular weight that first group of first step ultra filtration unit 1, second group of first step ultra filtration unit 4 are selected the aperture, it is the ultra-filtration membrane of holding back 2000 molecular weight that first group of second stage ultra filtration unit 2, second group of second stage ultra filtration unit 5 are selected the aperture, it is the ultra-filtration membrane of holding back 1000 molecular weight that 3, second groups of third stage ultra filtration unit 6 of first group of third stage ultra filtration unit composition are selected the aperture, and working pressure all is controlled at 0.3Mpa, sees through flow is 2T/h; All the other are with embodiment 1.
Embodiment 3,
The ultrafiltration membrane system working parameter is as follows: it is the ultra-filtration membrane of holding back 4000 molecular weight that first group of first step ultra filtration unit 1, second group of first step ultra filtration unit 4 are selected the aperture, it is the ultra-filtration membrane of holding back 3000 molecular weight that first group of second stage ultra filtration unit 2, second group of second stage ultra filtration unit 5 are selected the aperture, it is the ultra-filtration membrane of holding back 2000 molecular weight that 3, second groups of third stage ultra filtration unit 6 of first group of third stage ultra filtration unit composition are selected the aperture, and working pressure all is controlled at 0.8Mpa, sees through flow is 5T/h; All the other are with embodiment 1.
The nanofiltration membrane system operational parameters is as follows: it is the nanofiltration membrane of 20nm that first step nano-filtration unit 7 is selected the aperture, and it is the nanofiltration membrane of 10nm that second stage nano-filtration unit 8 is selected the aperture, and it is the nanofiltration membrane of 5nm that third stage nano-filtration unit 9 is selected the aperture; Working pressure all is controlled at 1.0Mpa, sees through flow is 2T/h; All the other are with embodiment 1.
The nanofiltration membrane system operational parameters is as follows: it is the nanofiltration membrane of 50nm that first step nano-filtration unit 7 is selected the aperture, and it is the nanofiltration membrane of 30nm that second stage nano-filtration unit 8 is selected the aperture, and it is the nanofiltration membrane of 20nm that third stage nano-filtration unit 9 is selected the aperture; Working pressure all is controlled at 3.0Mpa, sees through flow is 5T/h; All the other are with embodiment 1.
Comparative Examples 1,
The membrane sepn extracting method of Micronomicin Sulfate specifically obtains as follows:
The preparation of A, micronomicin seed:
Micronomicin Sulfate belongs to amino acid glycoside Broad spectrum antibiotics, and it is to be produced by a strain deep red micromonospora (Micromonospora purpura).
A1, female bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 8 parts of Zulkovsky starches, 1 part in calcium carbonate, 0.5 part of dipotassium hydrogen phosphate, 1.0 parts of vitriolate of tartar, 0.02 part of asparagine, 0.5 part in sal epsom, 20 parts in wheat bran, 18 parts in agar; Cultivated 7 days in 35 ℃;
A2, son bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 10 parts of Zulkovsky starches, 1 part in calcium carbonate, 0.6 part of dipotassium hydrogen phosphate, 1.2 parts of vitriolate of tartar, 0.02 part of asparagine, 0.5 part in sal epsom, 22 parts in wheat bran, 16 parts in agar; In 37 ℃ of cultivations 12 days, make spore, vacuum-drying saves backup;
A3, spore suspension: the spore of son bottle slant culture is made suspension;
The preparation of B, Micronomicin Sulfate fermented liquid:
B1, first class seed pot are cultivated: substratum is by made 0.5 part of soybean cake powder, 4 parts of glucose, 0.5 part in calcium carbonate, 55 parts of starch, 0.05 part in ammonium sulfate, 0.05 part in saltpetre, 0.3 part of peptone by the supplementary material of following weight proportioning; In 37 ℃, ventilation 1:2V/V.m, stirring velocity 350rpm, PH nature, incubation time 42h;
B2, secondary seed tank are cultivated: substratum is by made 15 parts in Zulkovsky starch powder, 3 parts of glucose, 15 parts of Semen Maydis powder, 20 parts of analysis for soybean powder, 2 parts of peptones, 5 parts in calcium carbonate, 2 parts of yeast powders by the supplementary material of following weight proportioning; In 35 ℃, ventilation 1:1.2V/V.m, stirring velocity 280rpm, PH nature, incubation time 15h;
B3, with the seed cultivated with 10% inoculum size access fermentation flask, cultivated 6 days to get fermented liquid in 34 ℃, power of agitator 3KW/m3 wherein, rotating speed 160r/min, air flow quantity 60m3/m3.h, tank pressure 0.04Mpa;
The preparation of C, Micronomicin Sulfate desorbed solution:
C1, acidifying removal of impurities: 98% sulfuric acid acidation with 1 times of amount of fermented liquid quality filters to get acidizing fluid, removes miscellaneous bacteria and other impurity;
C2, neutralization: it is 7 that the NaOH that adds 3mol/L is neutralized to PH with acidizing fluid;
C3, resin absorption removal of impurities;
D, bleaching process: the Micronomicin Sulfate desorbed solution decolours through ultrafiltration membrane system;
Ultrafiltration membrane system wherein comprises first group of ultra filtration unit, second group of ultra filtration unit; First group of ultra filtration unit is comprised of first group of first step ultra filtration unit 1, first group of second stage ultra filtration unit 2, first group of third stage ultra filtration unit 3; Second group of ultra filtration unit is comprised of second group of first step ultra filtration unit 4, second group of second stage ultra filtration unit 5, second group of third stage ultra filtration unit 6; Wherein: first group of first step ultra filtration unit 1, first group of second stage ultra filtration unit 2 and the group third stage ultra filtration unit 3 of being connected connect successively, and second group of first step ultra filtration unit 4, second group of second stage ultra filtration unit 5 and the group third stage ultra filtration unit 6 of being connected connect successively; First group of ultra filtration unit and second group of ultra filtration unit are connected in parallel;
The decolouring step: the Micronomicin Sulfate desorbed solution is parallel through first group of ultra filtration unit and second group of ultra filtration unit decolouring, be that the Micronomicin Sulfate desorbed solution is abreast successively through first group of first step ultra filtration unit 1, first group of second stage ultra filtration unit 2, first group of third stage ultra filtration unit 3, with second group of first step ultra filtration unit 4, second group of second stage ultra filtration unit 5, second group of third stage ultra filtration unit 6, the Micronomicin Sulfate desorbed solution after the decolouring enters enrichment process;
The ultrafiltration membrane system working parameter is as follows: it is the ultra-filtration membrane of holding back 5000 molecular weight that first group of first step ultra filtration unit 1, second group of first step ultra filtration unit 4 are selected the aperture, it is the ultra-filtration membrane of holding back 4000 molecular weight that first group of second stage ultra filtration unit 2, second group of second stage ultra filtration unit 5 are selected the aperture, it is the ultra-filtration membrane of holding back 3000 molecular weight that 3, second groups of third stage ultra filtration unit 6 of first group of third stage ultra filtration unit composition are selected the aperture, and working pressure all is controlled at 0.5Mpa, sees through flow is 3T/h;
E, enrichment process: the Micronomicin Sulfate desorbed solution after the decolouring is concentrated through the nanofiltration membrane system;
Nanofiltration membrane system wherein comprises first step nano-filtration unit 7, second stage nano-filtration unit 8, third stage nano-filtration unit 9; Wherein first step nano-filtration unit 7, second stage nano-filtration unit 8, third stage nano-filtration unit 9 connect successively;
Enrichment step: the Micronomicin Sulfate desorbed solution after the decolouring concentrates through first step nano-filtration unit 7, second stage nano-filtration unit 8, third stage nano-filtration unit 9 successively; Obtain Micronomicin Sulfate.
The nanofiltration membrane system operational parameters is as follows: it is the nanofiltration membrane of 35nm that first step nano-filtration unit 7 is selected the aperture, and it is the nanofiltration membrane of 20nm that second stage nano-filtration unit 8 is selected the aperture, and it is the nanofiltration membrane of 10nm that third stage nano-filtration unit 9 is selected the aperture; Working pressure all is controlled at 1.5Mpa, sees through flow is 3T/h.
Comparative Examples 2,
The membrane sepn extracting method of Micronomicin Sulfate specifically obtains as follows:
The preparation of A, micronomicin seed:
Micronomicin Sulfate belongs to amino acid glycoside Broad spectrum antibiotics, and it is produced by a strain deep red micromonospora (Micromonospora purpura).
A1, female bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 8 parts of Zulkovsky starches, 1 part in calcium carbonate, 0.5 part of dipotassium hydrogen phosphate, 1.0 parts of vitriolate of tartar, 0.02 part of asparagine, 0.5 part in sal epsom, 20 parts in wheat bran, 18 parts in agar; Cultivated 7 days in 35 ℃;
A2, son bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 10 parts of Zulkovsky starches, 1 part in calcium carbonate, 0.6 part of dipotassium hydrogen phosphate, 1.2 parts of vitriolate of tartar, 0.02 part of asparagine, 0.5 part in sal epsom, 22 parts in wheat bran, 16 parts in agar; In 37 ℃ of cultivations 12 days, make spore, vacuum-drying saves backup;
A3, spore suspension: the spore of son bottle slant culture is made suspension;
The preparation of B, Micronomicin Sulfate fermented liquid:
B1, first class seed pot are cultivated: substratum is by made 0.5 part of soybean cake powder, 4 parts of glucose, 0.5 part in calcium carbonate, 55 parts of starch, 0.05 part in ammonium sulfate, 0.05 part in saltpetre, 0.3 part of peptone by the supplementary material of following weight proportioning; In 37 ℃, ventilation 1:2V/V.m, stirring velocity 350rpm, PH nature, incubation time 42h;
B2, secondary seed tank are cultivated: substratum is by made 15 parts in Zulkovsky starch powder, 3 parts of glucose, 15 parts of Semen Maydis powder, 20 parts of analysis for soybean powder, 2 parts of peptones, 5 parts in calcium carbonate, 2 parts of yeast powders by the supplementary material of following weight proportioning; In 35 ℃, ventilation 1:1.2V/V.m, stirring velocity 280rpm, PH nature, incubation time 15h;
B3, with the seed cultivated with 10% inoculum size access fermentation flask, cultivated 6 days to get fermented liquid in 34 ℃, power of agitator 3KW/m3 wherein, rotating speed 160r/min, air flow quantity 60m3/m3.h, tank pressure 0.04Mpa;
The preparation of C, Micronomicin Sulfate desorbed solution:
C1, acidifying removal of impurities: 98% sulfuric acid acidation with 1 times of amount of fermented liquid quality filters to get acidizing fluid, removes miscellaneous bacteria and other impurity;
C2, neutralization: it is 7 that the NaOH that adds 3mol/L is neutralized to PH with acidizing fluid;
C3, resin absorption removal of impurities;
D, bleaching process: the Micronomicin Sulfate desorbed solution decolours through ultrafiltration membrane system;
Ultrafiltration membrane system wherein comprises first group of ultra filtration unit, second group of ultra filtration unit; First group of ultra filtration unit is comprised of first group of first step ultra filtration unit, first group of second stage ultra filtration unit, first group of third stage ultra filtration unit, first group of fourth stage ultra filtration unit, first group of level V ultra filtration unit; Second group of ultra filtration unit is comprised of second group of first step ultra filtration unit, second group of second stage ultra filtration unit, second group of third stage ultra filtration unit, second group of fourth stage ultra filtration unit, second group of level V ultra filtration unit; Wherein: first group of first step ultra filtration unit, first group of second stage ultra filtration unit, first group of third stage ultra filtration unit, first group of fourth stage ultra filtration unit, first group of level V ultra filtration unit connect successively, and second group of first step ultra filtration unit, second group of second stage ultra filtration unit, second group of third stage ultra filtration unit, second group of fourth stage ultra filtration unit, second group of level V ultra filtration unit connect successively; First group of ultra filtration unit and second group of ultra filtration unit are connected in parallel;
The decolouring step: the Micronomicin Sulfate desorbed solution is parallel through first group of ultra filtration unit and second group of ultra filtration unit decolouring, be that the Micronomicin Sulfate desorbed solution is abreast successively through first group of first step ultra filtration unit, first group of second stage ultra filtration unit, first group of third stage ultra filtration unit, first group of fourth stage ultra filtration unit, first group of level V ultra filtration unit, with second group of first step ultra filtration unit, second group of second stage ultra filtration unit, second group of third stage ultra filtration unit, second group of fourth stage ultra filtration unit, second group of level V ultra filtration unit, the Micronomicin Sulfate desorbed solution after the decolouring enters enrichment process;
The ultrafiltration membrane system working parameter is as follows: first group of first step ultra filtration unit, it is the ultra-filtration membrane of holding back 5000 molecular weight that second group of first step ultra filtration unit selected the aperture, first group of second stage ultra filtration unit, it is the ultra-filtration membrane of holding back 4000 molecular weight that second group of second stage ultra filtration unit selected the aperture, first group of third stage ultra filtration unit forms, first group of fourth stage ultra filtration unit, first group of level V ultra filtration unit, second group of third stage ultra filtration unit, second group of fourth stage ultra filtration unit, it is the ultra-filtration membrane of holding back 3000 molecular weight that second group of level V ultra filtration unit selected the aperture, and working pressure all is controlled at 0.5Mpa, seeing through flow is 3T/h;
E, enrichment process: the Micronomicin Sulfate desorbed solution after the decolouring is concentrated through the nanofiltration membrane system;
Nanofiltration membrane system wherein comprises first step nano-filtration unit, second stage nano-filtration unit, third stage nano-filtration unit; Wherein first step nano-filtration unit, second stage nano-filtration unit, third stage nano-filtration unit connect successively;
Enrichment step: the Micronomicin Sulfate desorbed solution after the decolouring concentrates through first step nano-filtration unit, second stage nano-filtration unit, third stage nano-filtration unit successively; Obtain Micronomicin Sulfate.
The nanofiltration membrane system operational parameters is as follows: it is the nanofiltration membrane of 35nm that first step nano-filtration unit is selected the aperture, and it is the nanofiltration membrane of 20nm that second stage nano-filtration unit is selected the aperture, and it is the nanofiltration membrane of 10nm that third stage nano-filtration unit is selected the aperture; Working pressure all is controlled at 1.5Mpa, sees through flow is 3T/h.
Test example
Tiring of test example 1, Nephelometric Determination Micronomicin Sulfate
Get respectively the Micronomicin Sulfate of embodiment 1-5 and Comparative Examples 1 and Comparative Examples 2 preparations, precision weighing, make the solution of 1000U/mg with aqua sterilisa, accurate this liquid of absorption is an amount of respectively, with ditalimfos phthalate buffer (PH=7.0) dilution make 10, the high low dosage solution of 20U/mg concentration, ratio is 2:1 between the height dosage facility, precision measures each 1.0ml of mentioned solution respectively, put in the sterilization colorimetric cylinder, each 6 pipe, add respectively 3% escherichia coli liquid culture medium 9.0ml, shake up immediately, in turbidimetry, cultivate 3.5h.Use microorganism than turbid instrument, constant temperature culture, regularly vibration, calculation result.Negative control: PH=7.0 phosphate buffer 1 .0ml adds the not substratum of inoculation experiments bacterium of 9.0ml.The results are shown in Table 1.
The result that tires of table 1 Nephelometric Determination Micronomicin Sulfate compares
| Embodiment | (μ g/mg) tires | Micronomicin Sulfate C2b percentage composition (%) |
| Embodiment 1 | 630 | 96.1 |
| | 582 | 85.9 |
| Embodiment 3 | 618 | 92.7 |
| | 594 | 86.2 |
| | 604 | 93.5 |
| Comparative Examples 1 | 533 | 75.4 |
| Comparative Examples 2 | 605 | 89.3 |
Conclusion: by above data as can be known, change membrane pore size, working pressure in the film separating system, see through flow, larger on the product concentration impact.Micronomicin Sulfate content is all greater than 85% among the embodiment, and the Micronomicin Sulfate of embodiment 1 preparation is tired and Micronomicin Sulfate C2b percentage composition is all optimum, and visible embodiment 1 selected parameter is optimum.Comparative Examples 1 each parameter is all not as embodiment 1-5, and the desorbed solution preparation method after visible the improvement is better than traditional technology, but the Effective Raise quality product.Though the Micronomicin Sulfate C2b percentage composition of Comparative Examples 2 has used ten ultra-filtration membrane unit greater than 85% in its decoloration process, increased production cost and production time.
Claims (3)
1. the membrane sepn extracting method of Micronomicin Sulfate, it is characterized in that: the Micronomicin Sulfate desorbed solution decolours through ultrafiltration membrane system, and the Micronomicin Sulfate desorbed solution after the decolouring is concentrated through the nanofiltration membrane system again;
Ultrafiltration membrane system wherein comprises first group of ultra filtration unit, second group of ultra filtration unit; First group of ultra filtration unit is comprised of first group of first step ultra filtration unit (1), first group of second stage ultra filtration unit (2), first group of third stage ultra filtration unit (3); Second group of ultra filtration unit is comprised of second group of first step ultra filtration unit (4), second group of second stage ultra filtration unit (5), second group of third stage ultra filtration unit (6); Wherein: first group of first step ultra filtration unit (1), first group of second stage ultra filtration unit (2) and the group third stage ultra filtration unit (3) of being connected connect successively, and second group of first step ultra filtration unit (4), second group of second stage ultra filtration unit (5) and the group third stage ultra filtration unit (6) of being connected connect successively; First group of ultra filtration unit and second group of ultra filtration unit are connected in parallel;
The decolouring step: the Micronomicin Sulfate desorbed solution is parallel through first group of ultra filtration unit and second group of ultra filtration unit decolouring, be that the Micronomicin Sulfate desorbed solution is abreast successively through first group of first step ultra filtration unit (1), first group of second stage ultra filtration unit (2), first group of third stage ultra filtration unit (3), with second group of first step ultra filtration unit (4), second group of second stage ultra filtration unit (5), second group of third stage ultra filtration unit (6), the Micronomicin Sulfate desorbed solution after the decolouring enters enrichment process;
The ultrafiltration membrane system working parameter is as follows: it is the ultra-filtration membrane of holding back 5000 molecular weight that first group of first step ultra filtration unit (1), second group of first step ultra filtration unit (4) are selected the aperture, it is the ultra-filtration membrane of holding back 4000 molecular weight that first group of second stage ultra filtration unit (2), second group of second stage ultra filtration unit (5) are selected the aperture, and it is the ultra-filtration membrane of holding back 3000 molecular weight that first group of third stage ultra filtration unit 3, second groups of third stage ultra filtration unit of composition (6) are selected the aperture;
The nanofiltration membrane system comprises first step nano-filtration unit (7), second stage nano-filtration unit (8), third stage nano-filtration unit (9); Wherein first step nano-filtration unit (7), second stage nano-filtration unit (8), third stage nano-filtration unit (9) connect successively;
Enrichment step: the Micronomicin Sulfate desorbed solution after the decolouring passes through successively first step nano-filtration unit (7), second stage nano-filtration unit (8), third stage nano-filtration unit (9) and concentrates; Obtain Micronomicin Sulfate;
The nanofiltration membrane system operational parameters is as follows: it is the nanofiltration membrane of 35nm that first step nano-filtration unit (7) is selected the aperture, it is the nanofiltration membrane of 20nm that second stage nano-filtration unit (8) is selected the aperture, and it is the nanofiltration membrane of 10nm that third stage nano-filtration unit (9) is selected the aperture.
2. the membrane sepn extracting method of Micronomicin Sulfate as claimed in claim 1 is characterized in that: the working pressure in the ultrafiltration membrane system working parameter all is controlled at 0.5Mpa, sees through flow is 3T/h; Working pressure in the nanofiltration membrane system operational parameters all is controlled at 1.5Mpa, sees through flow is 3T/h.
3. the membrane sepn extracting method of Micronomicin Sulfate as claimed in claim 1 or 2, it is characterized in that: the Micronomicin Sulfate desorbed solution obtains as follows,
The preparation of A, micronomicin seed:
Get deep red micromonospora, the implant carrier bottle is cultivated;
A1, female bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 10 parts of Zulkovsky starches, 0.5 part in sodium-chlor, 0.3 part of dipotassium hydrogen phosphate, 1.0 parts in saltpetre, 0.02 part of asparagine, 0.5 part in sal epsom, 18 parts in wheat bran, 0.6 part of glycerine, 18 parts in agar; Cultivated 7 days in 35 ℃;
A2, son bottle slant culture: substratum is made by the supplementary material of following weight proportioning, 13 parts of Zulkovsky starches, 0.5 part in sodium-chlor, 0.4 part of dipotassium hydrogen phosphate, 1.3 parts in saltpetre, 0.03 part of asparagine, 0.5 part in sal epsom, 18 parts in wheat bran, 0.8 part of glycerine, 20 parts in agar; In 37 ℃ of cultivations 12 days, make spore, vacuum-drying saves backup;
A3, spore suspension: the spore of son bottle slant culture is made suspension;
The preparation of B, Micronomicin Sulfate fermented liquid:
B1, first class seed pot are cultivated: substratum is by made 15 parts of soybean cake powders, 1.2 parts of glucose, 22 parts of Semen Maydis powder, 2 parts of fish meal, 1.0 parts in urea, 1.0 parts in calcium carbonate, 40 parts of starch by the supplementary material of following weight proportioning; In 37 ℃, ventilation 1:2V/V.m, stirring velocity 350rpm, PH nature, incubation time 42h;
B2, secondary seed tank are cultivated: substratum is by made 18 parts of groundnut meals, 1.5 parts of glucose, 24 parts of Semen Maydis powder, 1.0 parts in urea, 1.0 parts in saltpetre, 0.8 part in calcium carbonate, 42 parts of starch by the supplementary material of following weight proportioning; In 35 ℃, ventilation 1:1.2V/V.m, stirring velocity 280rpm, PH nature, incubation time 15h;
B3, with the seed cultivated with 10% inoculum size access fermentation flask, cultivated 6 days to get fermented liquid in 34 ℃, power of agitator 3KW/m3 wherein, rotating speed 160r/min, air flow quantity 60m3/m3.h, tank pressure 0.04Mpa;
The preparation of C, Micronomicin Sulfate desorbed solution:
C1, acidifying removal of impurities: 98% sulfuric acid acidation with 1 times of amount of fermented liquid quality filters to get acidizing fluid, removes miscellaneous bacteria and other impurity;
C2, neutralization: it is 7 that the NaOH that adds 3mol/L is neutralized to PH with acidizing fluid;
C3, resin absorption removal of impurities.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210565158.4A CN103012514B (en) | 2012-12-24 | 2012-12-24 | Membrane separation and extraction method for micronomicin sulfate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210565158.4A CN103012514B (en) | 2012-12-24 | 2012-12-24 | Membrane separation and extraction method for micronomicin sulfate |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103012514A true CN103012514A (en) | 2013-04-03 |
| CN103012514B CN103012514B (en) | 2015-05-20 |
Family
ID=47961663
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210565158.4A Active CN103012514B (en) | 2012-12-24 | 2012-12-24 | Membrane separation and extraction method for micronomicin sulfate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103012514B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107673958A (en) * | 2017-10-10 | 2018-02-09 | 浙江新和成股份有限公司 | A kind of method that Co-Q10 isolates and purifies |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1974585A (en) * | 2006-12-20 | 2007-06-06 | 福州大学 | Membrane separating and purifying process for aminoglycoside antibiotics |
-
2012
- 2012-12-24 CN CN201210565158.4A patent/CN103012514B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1974585A (en) * | 2006-12-20 | 2007-06-06 | 福州大学 | Membrane separating and purifying process for aminoglycoside antibiotics |
Non-Patent Citations (1)
| Title |
|---|
| 刘均忠等: "小诺霉素生产菌株选育及发酵工艺改进的研究", 《山东医药工业》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107673958A (en) * | 2017-10-10 | 2018-02-09 | 浙江新和成股份有限公司 | A kind of method that Co-Q10 isolates and purifies |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103012514B (en) | 2015-05-20 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN103320362B (en) | One strain is produced the bacterial strain of L-Glutamic decarboxylase and is produced the method for γ-aminobutyric acid with it | |
| CN103695489B (en) | A kind of arginine process for refining | |
| CN103695487B (en) | A kind of fermentable produces arginine technique | |
| CN105418694A (en) | Preparation method of trehalose | |
| CN103709235B (en) | A kind of method reducing the extraction high-purity enramycin that solvent uses | |
| CN103087126A (en) | Preparation method of zhongshengmycin bulk drug | |
| CN103667381B (en) | A kind of method improving yield of arginine | |
| CN108949850B (en) | Online separation and purification method of rhamnolipid fermentation liquor | |
| CN104402976B (en) | A kind of method preparing enramycin fine powder | |
| CN103012514B (en) | Membrane separation and extraction method for micronomicin sulfate | |
| CN103224960B (en) | Method for producing erythritol by continuously fermenting and extracting Torula.sp strain | |
| CN102838624A (en) | Method for purifying clavulanic acid from fermentation liquor | |
| CN113321580A (en) | Method for producing malic acid | |
| CN103695490A (en) | High-purity arginine production process | |
| CN102812050A (en) | Purification Method For Hyaluronic Acid And/or Salts Thereof | |
| CN107858174B (en) | Special embryo culture mineral oil for in vitro fertilization and preparation method thereof | |
| Camelini et al. | Nanofiltration of polysaccharides from Agaricus subrufescens | |
| CN103695488B (en) | A kind of arginine preparation method | |
| CN106518700A (en) | Glutamicacid membrane method production process | |
| CN105111285A (en) | Daptomycin extraction method | |
| CN112409426B (en) | Preparation method of sisomicin sulfate | |
| CN104694614A (en) | Novel extraction process of L-tryptophan | |
| CN109136313A (en) | Utilize the method for Michigan's Klebsiella synthesis 2'-deoxyadenosine | |
| CN111100823B (en) | Polymyxin B sulfate production strain, preparation method and application of polymyxin B sulfate | |
| CN107873734A (en) | A kind of biological mildew inhibitor and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant |