CN114381386B - Culture medium for producing avermectin through fermentation - Google Patents

Culture medium for producing avermectin through fermentation Download PDF

Info

Publication number
CN114381386B
CN114381386B CN202111382342.0A CN202111382342A CN114381386B CN 114381386 B CN114381386 B CN 114381386B CN 202111382342 A CN202111382342 A CN 202111382342A CN 114381386 B CN114381386 B CN 114381386B
Authority
CN
China
Prior art keywords
steep liquor
fermentation
corn steep
yeast powder
starch
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.)
Active
Application number
CN202111382342.0A
Other languages
Chinese (zh)
Other versions
CN114381386A (en
Inventor
程曦
王学伟
耿中坤
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Hebei Xingbai Agricultural Technology Co ltd
Original Assignee
Hebei Xingbai Agricultural Technology Co ltd
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Hebei Xingbai Agricultural Technology Co ltd filed Critical Hebei Xingbai Agricultural Technology Co ltd
Priority to CN202111382342.0A priority Critical patent/CN114381386B/en
Publication of CN114381386A publication Critical patent/CN114381386A/en
Application granted granted Critical
Publication of CN114381386B publication Critical patent/CN114381386B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Classifications

    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12NMICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00Microorganisms, 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
    • C12N1/20Bacteria; Culture media therefor
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12PFERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00Preparation of compounds containing saccharide radicals
    • C12P19/44Preparation of O-glycosides, e.g. glucosides
    • C12P19/60Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin
    • C12P19/62Preparation of O-glycosides, e.g. glucosides having an oxygen of the saccharide radical directly bound to a non-saccharide heterocyclic ring or a condensed ring system containing a non-saccharide heterocyclic ring, e.g. coumermycin, novobiocin the hetero ring having eight or more ring members and only oxygen as ring hetero atoms, e.g. erythromycin, spiramycin, nystatin
    • C12P19/623Avermectin; Milbemycin; Ivermectin; C-076

Landscapes

  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Virology (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Fodder In General (AREA)

Abstract

The invention discloses a culture medium for producing avermectin by fermentation, which comprises 170-180g/L of corn starch, 15-25g/L of soybean meal, 5-15g/L of yeast powder, 2-10g/L of corn steep liquor, 1-2g/L of light calcium carbonate, 0.1-0.5g/L of solid alkali, 2-6g/L of defoamer and amylase: starch=0.0002-0.0003. The invention increases the dosage of corn steep liquor, reduces the dosage of yeast powder, means that quick-acting nitrogen sources are increased, is more beneficial to the rapid growth and propagation of thalli, shortens the logarithmic growth phase, relatively prolongs the productive phase, and is beneficial to the improvement of fermentation titer. Compared with yeast powder, the corn steep liquor also contains a certain amount of saccharides, the carbon-nitrogen ratio is also improved, more energy support can be provided for continuous synthesis of abamectin by the streptomyces avermitilis, the fermentation titer is more beneficial to increase, the corn steep liquor is turned into wealth, the application of the corn steep liquor is deeply developed, and the added value of the corn steep liquor is further improved.

Description

Culture medium for producing avermectin through fermentation
Technical Field
The invention relates to the technical field of fermentation, in particular to a culture medium for producing abamectin through fermentation.
Background
The avermectin is an insecticidal acaricide obtained by aeration, stirring and submerged fermentation of streptomyces avermitilis in a culture medium containing nutrient elements such as starch, soybean meal, yeast powder, corn steep liquor and the like. As a high-efficiency low-toxicity biological pesticide and a medical intermediate, the novel pesticide is gradually replaced by other pesticides in agriculture. After the industrialized production for nearly 20 years, the market is stable, the process is mature, and in the composition of the manufacturing cost, the raw materials account for about 50% of the cost. In addition to improving the production level, further reducing the cost; the use of inexpensive raw materials instead of expensive raw materials is another way of reducing production costs.
Disclosure of Invention
The invention aims to provide a culture medium for producing avermectin by fermentation aiming at the defects of the prior art.
In order to solve the problems, the invention adopts the following technical scheme:
the culture medium for producing avermectin through fermentation comprises 170-180g/L of corn starch, 15-25g/L of soybean meal, 5-15g/L of yeast powder, 2-10g/L of corn steep liquor, 1-2g/L of light calcium carbonate, 0.1-0.5g/L of solid alkali, 2-6g/L of defoaming agent and amylase: starch=0.0002-0.0003.
Further, the culture medium comprises 174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 5g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025.
Further, the corn steep liquor quality standard: the total nitrogen is more than 10%, the mass concentration of the reducing sugar is more than 2%, the mass fraction of the dissolved phosphorus is less than 1%, the dry matter is more than 45%, and the pH is 3.8-4.2.
Further, the yeast powder quality standard: total nitrogen is more than 6.7 percent, and water content is less than or equal to 8 percent.
Further, the soybean cake has the following quality standard: total nitrogen is more than 6.4% and water content is less than or equal to 8%.
Further, shaking culture conditions: 250mL shaking bottles are adopted, 40mL culture medium is filled, the rotation speed of a shaking table is 250r/min, and the culture temperature is 27-28 ℃.
Further, fermenter culture conditions: volume after elimination is 91-93 m 3 The inoculation amount is 8%, the temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, the stirring frequency is 35-42 Hz.
The beneficial effects of adopting above-mentioned technical scheme to produce lie in:
the main components of the microbial fermentation medium are a carbon source and a nitrogen source, and the nitrogen source refers to a nutrient substance for providing nitrogen for the growth and propagation of microorganisms and synthetic products. Functionally, the method is divided into a quick-acting nitrogen source and a slow-acting nitrogen source, wherein the quick-acting nitrogen source refers to substances with relatively small molecular weight and easy to be utilized by thalli, such as amino acid and ammonium sulfate; the slow-release nitrogen source refers to substances with relatively large molecular weight, such as soybean cake powder, peanut cake powder, cotton seed cake powder and the like. The quick-acting nitrogen source is mainly used for the growth and propagation of thalli in the earlier stage of fermentation, and has great influence on the concentration of the thalli; and the slow-acting nitrogen source mainly maintains the metabolism of the middle-late stage bacteria.
The avermectin is synthesized after the growth and propagation of streptomyces avermitilis reaches a certain stage, the corn steep liquor is a byproduct of preparing corn starch by a wet grinding method, is an inexpensive organic nitrogen source, is rich in amino acid, polypeptide and other nutrient elements, and is beneficial to increasing the dosage of corn steep liquor, reducing the dosage of yeast powder, increasing quick-acting nitrogen sources, being more beneficial to the rapid growth and propagation of thalli, shortening the logarithmic growth phase, relatively prolonging the productive phase and improving the fermentation titer. Compared with yeast powder, the corn steep liquor also contains a certain amount of saccharides, the carbon-nitrogen ratio is also improved, more energy support can be provided for continuous synthesis of abamectin by the streptomyces avermitilis, the fermentation titer is more beneficial to increase, the corn steep liquor is turned into wealth, the application of the corn steep liquor is deeply developed, the added value of the corn steep liquor is further improved, and the production cost is reduced.
Drawings
FIG. 1 is a graph showing the comparison of the titer of example 1 and the average monthly titer in the present invention;
FIG. 2 is a graph showing the comparison of the titer of example 2 with the monthly average titer in the present invention.
Detailed Description
Embodiments of the present invention are described in further detail below with reference to the accompanying drawings and examples. The following examples are illustrative of the invention but are not intended to limit the scope of the invention.
In the description of the present invention, it should be noted that the directions or positional relationships indicated by the terms "center", "upper", "lower", "left", "right", "vertical", "horizontal", "inner", "outer", etc. are based on the directions or positional relationships shown in the drawings, are merely for convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be configured and operated in a specific orientation, and thus should not be construed as limiting the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
The culture medium for producing avermectin through fermentation comprises 170-180g/L of corn starch, 15-25g/L of soybean meal, 5-15g/L of yeast powder, 2-10g/L of corn steep liquor, 1-2g/L of light calcium carbonate, 0.1-0.5g/L of solid alkali, 2-6g/L of defoaming agent and amylase: starch=0.0002-0.0003.
Further preferably, the culture medium comprises 174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 5g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025.
Further, the corn steep liquor quality standard: the total nitrogen is more than 10%, the mass concentration of the reducing sugar is more than 2%, the mass fraction of the dissolved phosphorus is less than 1%, the dry matter is more than 45%, and the pH is 3.8-4.2; appearance: brown yellow, viscous, aromatic.
Further, the yeast powder quality standard: total nitrogen is more than 6.7 percent, and water content is less than or equal to 8 percent; appearance: pale yellow to yellowish orange powder.
Further, the soybean cake has the following quality standard: total nitrogen is more than 6.4 percent, and water content is less than or equal to 8 percent; appearance: light yellow, no moths and no mildewing.
Further, shaking culture conditions: 250mL shaking bottles are adopted, the filling amount of the culture medium is 40mL, the rotation speed of a shaking table is 250r/min, the culture temperature is 27-28 ℃, 3 shaking bottles are connected to each shaking bottle, and the fermentation titer is an average value of 3 bottles.
Further, fermenter culture conditions: volume after elimination is 91-93 m 3 The inoculation amount is 8%, the temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, the stirring frequency is 35-42 Hz.
The avermectin is a product of secondary fermentation of the streptomyces avermitilis, and is cooled for standby when hypha grows well in a seed tank, namely hypha is gathered into a bar-shaped ball, the bacterial concentration reaches 30% -40%, and the mixed bacteria pollution is avoided.
And calculating the feeding amount of the fermentation tank according to the proportion and the volume after elimination, and preparing materials.
Putting the prepared materials into a tank, metering the volume, cooling to 29-30 ℃ after sterilization, introducing seed liquid with the volume fraction of 8%, ventilating and stirring, sampling and microscopic examination every 8 hours, observing the conditions of sterility and hypha growth, starting to measure the initial titer for 50 hours, and testing every 24 hours until fermentation culture is finished.
The preparation method comprises mixing three nitrogen sources of soybean cake powder, corn steep liquor and yeast powder in a shaker under the condition of unchanged proportion of other raw materials to obtain L9 (3) 4 ) Performing an orthogonal test;
TABLE 1 factor level Table
Remarks: a, soybean cake powder; b corn steep liquor; c yeast powder
Table 2 L9 (3) 4 ) Results of the orthogonal test
The k value data shows that the optimal level combination is A3B2C2, namely 24g/L of soybean meal, 5g/L of corn steep liquor and 8g/L of yeast powder.
The effect of nitrogen source optimization on fermentation titers was verified by the following specific examples. Parallel experiments are carried out on industrialized mass production inconvenient, and average fermentation titers in the month are used as a control.
Example 1
(1) Fermentation cylinder (120 m) 3 ) Formulation of
174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 5g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025, post-consumer volume 91-93 m 3
(2) Inoculum size and formulation conditions
The inoculation amount is about 8m 3 The temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, stirring frequency is 35-42 Hz, and culturing time is 357h.
As shown in the data of FIG. 1, FIG. 1 is a graph showing the comparison of the titer and the average monthly titer in example 1, and the average fermentation titer of the soybean cake powder is improved by 7.8% compared with the average monthly level after the nitrogen source is optimized, namely, the soybean cake powder is prepared from 21g/L to 24g/L, the yeast powder is prepared from 10g/L to 8g/L, the corn steep liquor is prepared from 3g/L to 5 g/L. The corn steep liquor is used as a mild quick-acting organic nitrogen source, is more beneficial to the early-stage growth and propagation of the streptomyces avermitilis, is equivalent to shortening the growth and propagation period, prolonging the productive period, further improving the fermentation titer of the streptomyces avermitilis, and repeating the phenomenon after the nitrogen source in the formula is optimized, namely, the tank-placing titer of the streptomyces avermitilis is improved by about 8%.
Example 2
Example 1 shows that increasing the content of quick-acting organic nitrogen source corn steep liquor, reducing the content of soybean cake powder and yeast powder, and improving the pot-release titer by 7.8%; whether the corn steep liquor is added more or not is better, in order to further examine the influence of quick-acting organic nitrogen sources on the tank release titer of the industrialized production of the avermectin, the corn steep liquor is adjusted from 5g/L to 7g/L of the original corn steep liquor, other nitrogen sources and carbon sources are unchanged, and the test result is shown in figure 2
(1) Fermentation cylinder (120 m) 3 ) Formulation of
174g/L of corn starch, 24g/L of soybean meal, 8g/L of yeast powder, 7g/L of corn steep liquor, 1.6g/L of light calcium carbonate, 0.3g/L of solid alkali, 4.3g/L of defoamer and amylase: starch=0.00025, post-consumer volume 91-93 m 3
(2) Inoculum size and formulation conditions
The inoculation amount is about 8m 3 The temperature is 27-28 ℃, and the air quantity is 0.5-0.8 m 3 And/min, stirring frequency is 35-42 Hz, and culturing time is 357h.
As the data in FIG. 2 show, FIG. 2 is a graph of the titer of example 2 versus the average monthly titer, when the corn steep liquor mass concentration was adjusted from 5g/L to 7g/L, the discharge titer was 5% lower than the control. The method is characterized in that when the quick-acting nitrogen source in the avermectin fermentation formula is too much, mycelium is overgrown and high in fungus concentration are caused, rheological property of fermentation liquor is further influenced, energy and mass transfer in the fermentation liquor are influenced, the growth period is prolonged, the productive period is shortened, and the tank release titer is reduced.
Finally, it should be noted that: the above embodiments are only for illustrating the technical solution of the present invention, and are not limiting; although the invention has been described in detail with reference to the foregoing embodiments, it will be understood by those of ordinary skill in the art that: the technical scheme described in the foregoing embodiments can be modified or some of the technical features thereof can be replaced by equivalents; such modifications and substitutions do not depart from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims (1)

1. A method for producing avermectin by fermentation is characterized in that the specific fermentation conditions are as follows:
shake cultivation conditions: shaking a bottle by adopting a 250mL bottle, filling 40mL of fermentation medium, rotating a shaking table at the speed of 250r/min, and culturing at the temperature of 27-28 ℃;
fermentation tank culture conditions: 91-93 m percent of volume after elimination, 8 percent of inoculation amount, 27-28 ℃ of temperature and 0.5-0.8 m of air quantity 3 Stirring frequency is 35-42 Hz per minute;
the fermentation medium comprises corn starch 174g/L, soybean meal 24g/L, yeast powder 8g/L, corn steep liquor 5g/L, light calcium carbonate 1.6g/L, solid alkali 0.3g/L, defoamer 4.3g/L and amylase: starch=0.00025;
the corn steep liquor quality standard: the total nitrogen is more than 10%, the mass concentration of the reducing sugar is more than 2%, the mass fraction of the dissolved phosphorus is less than 1%, the dry matter is more than 45%, and the pH is 3.8-4.2;
the yeast powder quality standard: total nitrogen is more than 6.7 percent, and water content is less than or equal to 8 percent;
the quality standard of the soybean cake is as follows: total nitrogen is more than 6.4% and water content is less than or equal to 8%.
CN202111382342.0A 2021-11-21 2021-11-21 Culture medium for producing avermectin through fermentation Active CN114381386B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN202111382342.0A CN114381386B (en) 2021-11-21 2021-11-21 Culture medium for producing avermectin through fermentation

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN202111382342.0A CN114381386B (en) 2021-11-21 2021-11-21 Culture medium for producing avermectin through fermentation

Publications (2)

Publication Number Publication Date
CN114381386A CN114381386A (en) 2022-04-22
CN114381386B true CN114381386B (en) 2023-12-15

Family

ID=81195243

Family Applications (1)

Application Number Title Priority Date Filing Date
CN202111382342.0A Active CN114381386B (en) 2021-11-21 2021-11-21 Culture medium for producing avermectin through fermentation

Country Status (1)

Country Link
CN (1) CN114381386B (en)

Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101407775A (en) * 2008-11-27 2009-04-15 中国科学院微生物研究所 Method for preparing avermectin and special bacterial strain thereof
CN102634471A (en) * 2012-04-18 2012-08-15 南京工业大学 Avermectin B1a high-yielding strain and application thereof
CN103882080A (en) * 2014-03-17 2014-06-25 中国科学院成都生物研究所 Effective method for preparing avermectin
CN106191177A (en) * 2016-07-25 2016-12-07 宁夏泰益欣生物科技有限公司 A kind of culture medium utilizing streptomyces griseus fermenting and producing avilamycin
CN108060193A (en) * 2018-01-31 2018-05-22 天俱时工程科技集团有限公司 The production technology of avermectin
CN112391432A (en) * 2019-08-19 2021-02-23 上海医药工业研究院 Fermentation medium and fermentation method for producing pingyangmycin

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101407775A (en) * 2008-11-27 2009-04-15 中国科学院微生物研究所 Method for preparing avermectin and special bacterial strain thereof
CN102634471A (en) * 2012-04-18 2012-08-15 南京工业大学 Avermectin B1a high-yielding strain and application thereof
CN103882080A (en) * 2014-03-17 2014-06-25 中国科学院成都生物研究所 Effective method for preparing avermectin
CN106191177A (en) * 2016-07-25 2016-12-07 宁夏泰益欣生物科技有限公司 A kind of culture medium utilizing streptomyces griseus fermenting and producing avilamycin
CN108060193A (en) * 2018-01-31 2018-05-22 天俱时工程科技集团有限公司 The production technology of avermectin
CN112391432A (en) * 2019-08-19 2021-02-23 上海医药工业研究院 Fermentation medium and fermentation method for producing pingyangmycin

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
消泡剂对阿维菌素发酵过程生产收率的影响;郭鸿雁等;价值工程(第25期);289-290 *

Also Published As

Publication number Publication date
CN114381386A (en) 2022-04-22

Similar Documents

Publication Publication Date Title
CN100390272C (en) Fluorescent pseudomonads and its fermenting culture process and application
CN106222118B (en) One streptomycete category actinomyces and application thereof
CN102899377A (en) Preparation method of chlortetracycline premix
CN102660461A (en) Microbial preparation for shortening tobacco fermentation period and application of microbial preparation
CN110330370A (en) While a kind of turf biological organic fertilizer and its preparation method and application containing trichoderma and bacillus amyloliquefaciens
CN102154168A (en) Abamectin producing bacterium and preparation method thereof
CN104705347B (en) One cultivates peanut Slow_growing rhizobia agent, preparation method and use
CN107628894A (en) Composite bacteria agent increase soil fertility and its preparation method and application
CN114381386B (en) Culture medium for producing avermectin through fermentation
US11149293B2 (en) Fermentation method for producing gellan gum
KR100204983B1 (en) The method of culturing liquid starter of basidomycetis and the apparatus for culturing it and the starter
CN1498865A (en) Micro ecological agent for supporting water in fishing use and its prepn. method
CN110387343B (en) Solid culture medium for fermentation of streptomyces noursensis and preparation method and application thereof
CN102701831A (en) Microbial fertilizer and preparation method thereof
CN101967461B (en) Fermentation method for azospirillum, fermentation liquor prepared by method and application of fermentation liquor
CN1888045A (en) Composite microbial prepn for treating solid waste and its prepn process
CN113896786A (en) Artificially synthesized material with biological activity and agricultural application thereof
CN1163617C (en) Process for preparing evericin by fermentation method
CN110183260A (en) A kind of plant nutrition liquid and its application using fermented liquid for biologically producing hydrogen preparation
CN102154130B (en) Gossypol degrading strain and application thereof
CN110656131A (en) Preparation method of antibacterial secondary metabolite of actinomycetes
CN108841750A (en) A kind of effective biological agent and preparation method thereof for decomposing rice straw
CN114946531B (en) Lepista sordida cultivation method
CN113831167B (en) Microorganism rooting agent prepared from biogas slurry and preparation method thereof
CN1916162B (en) Legume inoculation agent prepared by using municipal sludge, and preparation method

Legal Events

Date Code Title Description
PB01 Publication
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB02 Change of applicant information

Address after: 051530 Hebei South Shijiazhuang City, Zhaoxian County South Park biological industrial park

Applicant after: Hebei Xingbai Agricultural Technology Co.,Ltd.

Address before: 051530 Hebei South Shijiazhuang City, Zhaoxian County South Park biological industrial park

Applicant before: HEBEI XINGBAI AGRICULTURAL SCIENCE AND TECHNOLOGY CO.,LTD.

CB02 Change of applicant information
GR01 Patent grant
GR01 Patent grant