Hygromycin B Explained

Hygromycin B is an antibiotic produced by the bacterium Streptomyces hygroscopicus. It is an aminoglycoside that kills bacteria, fungi and higher eukaryotic cells by inhibiting protein synthesis.[1]

History

Hygromycin B was originally developed in the 1950s for use with animals and is still added into swine and chicken feed as an anthelmintic or anti-worming agent (product name: Hygromix). Hygromycin B is produced by Streptomyces hygroscopicus, a bacterium isolated in 1953 from a soil sample. Resistance genes were discovered in the early 1980s.[2] [3]

Mechanism of action

Hygromycin B, along with aminoglycosides, inhibits protein synthesis by strengthening the interaction of tRNA binding in the ribosomal A-site. Hygromycin B also prevents mRNA and tRNA translocation by an unknown mechanism.[4]

Use in research

In the laboratory it is used for the selection and maintenance of prokaryotic and eukaryotic cells that contain the hygromycin resistance gene. The resistance gene is a kinase that inactivates hygromycin B through phosphorylation.[5] Since the discovery of hygromycin-resistance genes, hygromycin B has become a standard selection antibiotic in gene transfer experiments in many prokaryotic and eukaryotic cells. Based on impurity monitor method,[6] four different kinds of impurities are discovered in commercial hygromycin B from different suppliers and toxicities of different impurities to the cell lines are described in the following external links.

Use in plant research

Hygromycin resistance gene is frequently used as a selectable marker in research on plants. In rice Agrobacterium-mediated transformation system, hygromycin is used at about 30–75 mg L−1, with an average of 50 mg L−1. The use of hygromycin at 50 mg L−1 demonstrated highly toxic to non-transformed calli. Thus, it can be efficiently used to select transformants.[7]

Fungus Coniothyrium minitans was transformed with the hygromycin B resistance gene to improve the infection rates of Sclerotinia sclerotiorum, a fungal parasite of many crops.[8]

Notes and References

  1. Pittenger RC, Wolfe RN, Hoehn MM, Marks PN, Daily WA, McGUIRE JM . Hygromycin. I. Preliminary studies on the production and biologic activity of a new antibiotic . Antibiotics & Chemotherapy . 3 . 12 . 1268–1278 . December 1953 . 24542808 .
  2. Gritz L, Davies J . Plasmid-encoded hygromycin B resistance: the sequence of hygromycin B phosphotransferase gene and its expression in Escherichia coli and Saccharomyces cerevisiae . Gene . 25 . 2–3 . 179–188 . November 1983 . 6319235 . 10.1016/0378-1119(83)90223-8 .
  3. Kaster KR, Burgett SG, Rao RN, Ingolia TD . Analysis of a bacterial hygromycin B resistance gene by transcriptional and translational fusions and by DNA sequencing . Nucleic Acids Research . 11 . 19 . 6895–6911 . October 1983 . 6314265 . 326422 . 10.1093/nar/11.19.6895 .
  4. Web site: Hygromycin B . 2024-06-28 . TOKU-E . en.
  5. Rao RN, Allen NE, Hobbs JN, Alborn WE, Kirst HA, Paschal JW . Genetic and enzymatic basis of hygromycin B resistance in Escherichia coli . Antimicrobial Agents and Chemotherapy . 24 . 5 . 689–695 . November 1983 . 6318654 . 185926 . 10.1128/aac.24.5.689 .
  6. Kauffman JS. Analytical Strategies for Monitoring Residual Impurities Best methods to monitor product-related impurities throughout the production process. . BioPharm International . 23 . 1–3 . 2009 .
  7. Pazuki A, Asghari J, Sohani MM, Pessarakli M, Aflaki F . 2014 . Effects of Some Organic Nitrogen Sources and Antibiotics on Callus Growth of Indica Rice Cultivars . PDF . Journal of Plant Nutrition . 38 . 8 . 1231–1240 . 84495391 . November 17, 2014 . 10.1080/01904167.2014.983118 .
  8. Jones EE, Stewart A, Whipps JM . Use of Coniothyrium minitans transformed with the hygromycin B resistance gene to study survival and infection of Sclerotinia sclerotiorum sclerotia in soil . Mycological Research . 107 . Pt 3 . 267–276 . March 2003 . 12825495 . 10.1017/S0953756203007457 .