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Hygromycin B, molecular formula C20H37N3O13, CAS 31282-04-9, is an antibiotic produced by Streptomyces hygroscopicus and belongs to the aminoglycoside antibiotic family. It usually exists in the form of a slightly yellowish brown powder, which may vary slightly due to differences in production process and purity. Its powder state is delicate, without obvious particle sensation, suitable for various biochemical experiments and pharmaceutical preparations. It is soluble in methanol, ethanol, and water, and the polarity of these solvents allows it to interact with polar groups in the molecule, thereby increasing its solubility. However, it is almost insoluble in solvents with lower polarity such as ether, chloroform, or benzene, reflecting the specific polarity characteristics of its molecular structure. As a weak base, it can react with many organic and inorganic acids to generate corresponding salts. This acid-base property makes it widely applicable in both in vivo and in vitro experiments, and its solubility and activity can be controlled by adjusting the pH value of the solution. In the laboratory, it is used to select and maintain prokaryotic and eukaryotic cells with hygromycin resistance genes. The hygromycin resistance gene encodes a kinase (hygromycin kinase) that is inactivated through phosphorylation. Since the discovery of the resistance gene for hygromycin, it has been used as a standard screening method in transgenic experiments. At present, this substance is commonly used for screening in plant tissue culture.
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Chemical Formula |
C20H37N3O13 |
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Exact Mass |
527 |
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Molecular Weight |
528 |
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m/z |
527 (100.0%), 528 (21.6%), 529 (2.5%), 529 (2.2%), 528 (1.1%) |
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Elemental Analysis |
C, 45.54; H, 7.07; N, 7.97; O, 39.43 |
Hygromycin B is an antibiotic produced by Streptomyces hygroscopicus and belongs to the aminoglycoside antibiotic family. Its wide range of functions and application areas make it an important tool in scientific research and the pharmaceutical industry.
1. Antibacterial and antifungal effects
Inhibition of protein synthesis:
The main mechanism of action is to kill bacteria, fungi, and some higher eukaryotic organisms by inhibiting protein biosynthesis in their cells. It stabilizes the tRNA binding site on the large subunit of the ribosome, preventing empty tRNA from detaching from the ribosome and thus inhibiting the continuation of translation. This inhibitory effect is effective for ribosomes in both prokaryotic and eukaryotic cells.
Broad spectrum antibacterial activity:
It has a wide range of antibacterial activities against various bacteria, fungi, and higher eukaryotic cells. This makes it useful as an antibacterial agent in agriculture, animal husbandry, and aquaculture for the prevention and treatment of diseases caused by these microorganisms.
Antiviral effect:
In addition to antibacterial and antifungal effects, it also has certain antiviral activity. It can selectively enter cells that become permeable due to viral infection, thereby inhibiting the translation process of the virus and suppressing its replication and spread.
2. Application In Transgenic Experiments
Standard screening method:
Since the discovery of the resistance gene for hygromycin, it has been widely used as a standard screening method in transgenic experiments. It allows researchers to select and maintain prokaryotic and eukaryotic cells that possess the hygromycin resistance gene. This screening method has the characteristics of high efficiency, reliability, and ease of operation.
Selection of genetically transformed cells:
In transgenic experiments, it is often used as a selector to screen cells that have successfully transformed and expressed the hygromycin resistance gene. These cells can grow and reproduce in a medium containing the substance, while untransformed cells will be killed.
Filter tags:
In genetic engineering and cell biology research, it is often used as a screening marker. By introducing plasmids carrying the hygromycin resistance gene into target cells, only cells that have successfully transformed and expressed the gene can survive in culture media containing the substance, thereby achieving the screening and purification of target cells.
Cell culture and screening:
It can also be used for cell screening and purification in cell culture experiments. By adjusting its concentration, stable cell lines expressing the target gene can be screened, providing a reliable source of cells for subsequent experimental research. This method is not only efficient and reliable, but also easy to operate, and has become one of the standard screening methods in the field of genetic engineering.
3. Drug Development And Application
Insect repellent drugs:
Have a wide range of applications in the veterinary field. Long term feeding can effectively control infections of pig roundworms, esophageal nematodes, and hairy head nematodes. This is because it is not only effective against adults and larvae, but also inhibits female oviposition, resulting in the loss of reproductive ability of the insect body. Therefore, adding this substance to the full feed of pregnant sows can protect piglets from roundworm infection during lactation.
Antibacterial therapy:
Can also be used to treat infections caused by sensitive bacteria. Although its clinical application is relatively limited, it may serve as an alternative drug for the treatment of refractory infections in certain specific situations.
4. Laboratory Research Tools
Research on protein synthesis:
It has become an important tool for studying protein synthesis due to its unique mechanism of inhibiting protein synthesis. Researchers can use it to explore the regulatory mechanisms of protein synthesis, the structure and function of ribosomes, and various molecular events in the translation process.
Cell biology research:
It can also be used for cell screening and purification in cell biology research. By selectively killing untransformed or specific types of cells, researchers can obtain purified target cell populations for further research.
5. Cell Biology Research
Study cell apoptosis:
Widely used for studying cellular biological processes such as apoptosis, signal transduction, and cell cycle. Its unique mechanism of action makes it one of the important tools for studying these complex biological processes.
By observing and analyzing the cellular changes after treatment with hygromycin B, we can gain a deeper understanding of the molecular mechanisms and signaling pathways of cell apoptosis, providing new ideas and methods for disease treatment and drug development.
Antiviral research:
It also has antiviral effects and can selectively enter cells that become permeable due to viral infection, inhibiting the translation process of the virus and thus suppressing its replication and spread. This makes it have potential application value in antiviral research.
Insect repellent function:
It can also be mixed into animal feed to have deworming function, and has effective killing effect on parasites such as pig roundworms, esophageal nematodes, and hairy head nematodes.
6. Biological Applications
Combined use with other antibiotics:
Due to the differences in the mode of action of hygromycn B compared to other antibiotics such as G418, Zeocin, and Blasticidin S, it is often used for the selection of dual resistance positive cell lines. By combining with these antibiotics, the stability and accuracy of screening can be further improved.
Screening and maintaining prokaryotic or eukaryotic cells containing hygromycin resistance genes:
Due to its ability to inhibit protein synthesis, hygromycn B can be used as a selective marker for screening and maintaining cultured prokaryotic or eukaryotic cells successfully transfected with hygromycin resistance genes. These cells, carrying resistance genes to hygromycin (such as the hph gene), are capable of encoding hygromycn B phosphotransferase, which converts hygromycn B into biologically inactive phosphorylation products, thereby playing a detoxifying role.
When screening stable transfected cells, it is necessary to establish a killing curve to determine the minimum concentration of antibiotics that can kill untransfected host cells. This is usually achieved by selecting multiple concentration gradients and observing the survival of cells at different concentrations.
As an important antibiotic, the history of hygromycin can be traced back to the mid-20th century.
Discovery time:
It was first discovered in 1950 and is metabolized by Streptomyces hygroscopicus. This discovery marks the birth of another important member of the aminoglycoside antibiotic family.
Early use:
In its early stages of discovery, it was mainly used as an animal medicine and added to the feed of chickens and pigs, playing a role in deworming and insect resistance. This application demonstrates the killing effect of hygromycin on parasites in animals.
Discovery of resistance genes:
In the early 1980s, scientists discovered related resistance genes. This discovery lays the foundation for the application of hygromycin in genetic engineering and cell biology research.
Chaomycin can inhibit microbial growth by suppressing protein synthesis and has broad-spectrum antibacterial effects. It has a strong inhibitory effect on Gram positive bacteria, viral infections, Gram negative bacteria, and other microorganisms. As a broad-spectrum antibiotic, hygromycin is widely used in scientific research, medicine, veterinary medicine, pesticides and other fields. Chaomycin can prevent adult ovulation and prevent larval growth. Adding it to pig and poultry feed can effectively control infections such as pig roundworms and esophageal nematodes.
According to the Forecast Report on the Supply and Demand Status and Development Trend of China's Hygromycin Industry Market in 2023-2027 released by the New ThinkWorld Industry Research Center, the production and application of Hygromycin are mainly concentrated in China, India, Europe and the United States worldwide. China is a large livestock breeding country in the world, and the market demand for hygromycin is broad. But with increasingly strict regulation of antibiotics by governments around the world, the application of hygromycin will be subject to certain restrictions, and the market space may gradually shrink in the future.
The market price of hygromycin is influenced by various factors, including raw material costs, production process complexity, market supply and demand relationships, market competition level, policies and regulations, etc. Suppliers related to hygromycin include Thermo Fisher Scientific, Merck, Roche, Aibixin Biotechnology, Hunan Huibaishi Biotechnology, Huazhong Haiwei (Beijing) Gene Technology, etc. The market competition is quite fierce. Domestic enterprises can reduce production costs through economies of scale and transportation cost control, so compared with imported hygromycin, the market price of domestic hygromycin is generally lower.
The reaction conditions of hygromycin are harsh and the synthesis process is complex. The synthesis process of hygromycin involves steps such as strain extraction, fermentation culture, isolation, purification, and drying. The production cost of hygromycin is relatively high, and related enterprises still need to continuously improve and perfect their production processes to enhance the efficiency of hygromycin production and reduce production costs.
Technological innovation is one of the important factors driving the development of the hygromycin market. With the deepening of research and the advancement of technology, the production process of hygromycin is constantly being improved and perfected. Sun Yuhui's team from the School of Pharmacy at Wuhan University has developed a relatively complete biosynthetic pathway for hygromycin based on base editing technology, providing technical support for industrial production of hygromycin B. Industry analysts from Xinsijie stated that hygromycin has broad-spectrum antibacterial properties and has broad application value in fields such as scientific research, animal husbandry, agriculture, and medicine. However, the complex synthesis process of hygromycin and excessive use can easily lead to drug resistance, which limits its application in certain situations. The competition in the hygromycin market is fierce. With the improvement of synthesis technology, hygromycin enterprises will adopt more environmentally friendly, efficient, and economical production methods to improve product quality and their own competitiveness.
adverse reaction
Hygromycin B is an aminoglycoside antibiotic derived from Streptomyces hygroscopicus. Due to its broad-spectrum antibacterial activity and cytotoxicity against bacteria, fungi, and mammalian cells, it is widely used in genetic engineering, molecular biology research, and animal husbandry. In genetic engineering, Hygromycin B is often used as a screening marker to select prokaryotic or eukaryotic cells that have been successfully transfected with resistance genes (such as hyg or hph); In animal husbandry, its antibacterial properties are used to prevent and treat infections. However, the toxic effects of Hygromycin B cannot be ignored, as its residues and misuse may pose a risk of drug resistance and potential threats to human health and environmental safety.
Adverse reactions in scientific research applications
Cytotoxicity
In genetic engineering experiments, Hygromycin B is commonly used to screen cells transfected with resistance genes. However, even if cells express resistance genes (such as Hygromycin B phosphotransferase, which phosphorylates and inactivates Hygromycin B), high concentrations or prolonged exposure to Hygromycin B may still be toxic to cells. Research has shown that Hygromycin B can induce cell apoptosis or necrosis, manifested by changes in cell morphology (such as cell shrinkage, membrane vesicle formation), DNA fragmentation, and activation of caspase enzyme activity. This toxic effect may affect the accuracy of experimental results, such as leading to false negative results (resistant cells dying due to Hygromycin B toxicity) or false positive results (non resistant cells dying due to Hygromycin B toxicity being mistaken for resistant cells).
Risk of genetic mutations
The induction of incorrect translation of Hygromycin B may increase the risk of gene mutations. During protein synthesis, translation errors caused by Hygromycin B may result in abnormal proteins that may be toxic or trigger cellular stress responses, leading to DNA damage and gene mutations. Although there is currently no direct evidence to suggest that Hygromycin B significantly increases gene mutation rates in scientific applications, its potential mutagenicity still needs attention, especially in long-term or high-dose use.
Operational security risks
Hygromycin B has a certain toxic effect on the human body, and safety regulations must be strictly followed during operation. Direct contact with Hygromycin B powder or solution may cause skin or eye irritation, and inhalation of its dust or aerosols may cause damage to the respiratory tract. In addition, Hygromycin B can be absorbed or inhaled into the human body through the skin, and long-term or repeated exposure may cause systemic toxicity, such as liver and kidney function damage, neurological abnormalities, etc. Therefore, researchers should wear lab coats, gloves, and goggles when using Hygromycin B, and operate in a fume hood to avoid direct contact and inhalation.
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