A kind of enhanced microbial fermentation to prepare gentamicin sulfate and application method thereof

A technology of gentamicin and microbial strains, applied in microorganism-based methods, biochemical equipment and methods, microorganisms, etc., can solve the problems of pollution, interference, high material consumption, etc., to meet industrialization requirements, shorten production Cycle time, the effect of improving production capacity

Active Publication Date: 2021-12-28
青岛安惠仕生物制药有限公司
View PDF5 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The above two epimers have similar chemical structures and similar physical and chemical properties. It is very difficult to separate single-component gentamicin C2 from them, resulting in a shortage of gentamicin C2, high production costs and long chromatographic separation cycles. , low yield, high material consumption, serious pollution, complex extraction and refining process, and unstable product quality, and the by-product gentamicin C2a, which is similar in structure to gentamicin C2, interferes at any time, causing great problems for product quality control. Large uncertainties seriously affect the stability, safety and effectiveness of drugs, and are urgent scientific problems to be solved

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • A kind of enhanced microbial fermentation to prepare gentamicin sulfate and application method thereof
  • A kind of enhanced microbial fermentation to prepare gentamicin sulfate and application method thereof
  • A kind of enhanced microbial fermentation to prepare gentamicin sulfate and application method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Example 1 Atmospheric room temperature plasma mutagenesis technology screening

[0017] 1) With the gentamicin production strain Micromonospora rubrum SD2018 in the applicant's strain bank as the starting strain, use a sterilized 1ml straw to draw 0.1ml and inoculate it on the sterilized slant medium for slanting Spore culture, culture temperature is 35~37 ℃, stand still, culture time is 4 days, obtain the slant spore of SD2020 strain;

[0018] 2) Take 1.0cm 2 Step 1) Add the slanted spores prepared into a sterilized Erlenmeyer flask containing 5ml of saline with a mass concentration of 0.9%, and shake for 20 minutes to obtain a spore suspension;

[0019] 3) Take 500 μl of the spore suspension prepared in step 2), add 250 μl of 20% lithium chloride aqueous solution, 250 μl of 0.9% normal saline, shake and mix for 5 minutes, and obtain spores containing 5% lithium chloride Suspension; After shaking and mixing, draw 20 μl of spore suspension containing 5% lithium chlori...

Embodiment 2

[0021] The efficient fermentation of embodiment 2 gentamicin C2

[0022]The Plackett-Burman design method is a nearly saturated two-level experimental design method developed in the middle and late 20th century. Based on the principle of incomplete balance blocks, it can estimate the main effect of factors with the least number of trials, and quickly and effectively screen out the most important factors from many factors for further research. This method is simple in data processing. Compared with the currently commonly used partial factor test and random balance test, this method is the most efficient and accurate in screening important factors. Therefore, it has been widely used in the optimization of microbial culture conditions and achieved good results. In the embodiment, it is planned to investigate and evaluate many factors in the fermentation medium formula of gentamycin C2 through the Plackett-Burman experimental design of DesignExpert7.0, and screen out several impor...

Embodiment 3

[0036] The refining of embodiment 3 gentamicin C2

[0037] Utilize the above-mentioned fermentation method to carry out expanded cultivation, add 10% tap water to dilute the fermented liquid after expanded fermentation, add hydrochloric acid or sulfuric acid, acidify to pH1.0-3.0, fully stir 1-4 hour; pH5.0-7.0, put in 732NH 4+ Resin static adsorption for 6-8 hours. The amount of resin input is calculated at about 50,000 u / mL. Collect the adsorbed saturated resin and rinse it with tap water (until there is no floating mycelium). Pack the saturated resin into a column, pickle the saturated resin with 0.5M HCl solution twice the volume of the saturated resin, then wash it with ion-free water until neutral, and then use 0.01% ammonia water for alkaline washing. When the effluent reaches pH 9.0 or above, Connect in series to an equal volume of 711 resin column, use 4% ammonia water for elution, the amount of ammonia water is 8-10 times the volume of saturated resin, the elution...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention claims to protect an enhanced microbial fermentation preparation of gentamicin sulfate and its application method. The enhanced microbial engineering bacteria is a strain of Micromonospora purpurea C2 1591 strain. The engineering bacteria with directional high-yield gentamycin C2 screened by the present invention can directional high-yield gentamicin C2, and the component ratio of gentamicin C2 is increased from about 15% to more than 85% at present, realizing the optimization The highest fermentation potency of the final formula is 1184U / mL, which is 42% higher than that of the original formula. Compared with the starting strain, the yield of gentamicin C2 is increased by 3 times, and basically does not contain other isomers or impurities, which saves a lot of cost and meets the requirements of industrialization.

Description

technical field [0001] The invention belongs to the field of antibiotic pharmacy, and relates to an enhanced microbial fermentation to prepare gentamycin sulfate and its application method. Specifically, the invention relates to an engineering bacterium with a directional high-yield gentamicin C2 and its application. Antibiotic manufacturing. Background technique [0002] Gentamicin was first discovered by Weinstein in 1963. It is a multi-component aminoglycoside antibiotic produced by Micromonospora, including C1, C2, C1a, C2a and C2b components. The main components of gentamicin, C1, C2 and C1a, are widely used clinically, among which C1a has the highest antibacterial activity, which is the precursor of etimicin synthesis, followed by C2b, also known as sagamycin . These antibiotics can bind to 16SrRNA on the 30S subunit of bacterial ribosomes, causing misreading of the genetic code, thereby blocking the synthesis of bacterial proteins, so they are mainly used to treat b...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12N1/02C12P19/48C12R1/31
CPCC12N1/20C12N1/02C12P19/485
Inventor 王云松史京波张梵杨翠霞袁帅妮
Owner 青岛安惠仕生物制药有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products