Part:BBa_K4607003
SH3B30 Domain
Description
The biobrick consists of the SH3 domain from the bacteriophage B30, which is capable of recognizing and binding to the cell wall of Streptococcus agalactiae, Streptococcus uberis, and Staphylococcus aureus. For this to be possible, the SH3B30 domain recognizes and binds to the highly specific glycine of the pentaglycine cross-bridge glycosidic bond in the heteropolymer of the S. aureus, S. agalactiae and S. uberis peptidoglycan, activating the catalytic domain. The enzyme has a length of 62 amino acids and a molecular weight of 7.162 kDa. It keeps its stability at a range of 4 to 37° C and pH from 7 to 8 [1] [2].
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Usage and Biology
As a brief contextualization, bovine mastitis is the result of the infection of the bovine mammary glands caused by pathogenic microorganisms, mainly gram-positive and negative bacteria. This disease reduces milk quality production to a great extent and produces painful damage to the bovine. The main treatment for mastitis is the use of diverse antibiotics, therefore the overuse and misuse of them has caused a real problem in the development of multidrug-resistant pathogens [3]. Our team has conducted an extensive investigation to find an alternative treatment for bovine mastitis without risking the environment.
To design our proposal of a novel non-antibiotic treatment against the losses of milk and bovine, capable of decreasing the effects on the milk industry and their consequences for the nutrition of the Mexican population, we took into account the SH3 domain from the endolysin of the bacteriophage B30. The domain is capable of recognizing a wide spectrum of the pathogenic bacteria that cause bovine mastitis, including Streptococcus agalactiae, Streptococcus uberis, and Staphylococcus aureus. This characteristic results beneficial for the development of the treatment considering that the intramammary infection caused by Streptococcus agalactiae often leads to subclinical mastitis, which can result in clinical mastitis [4].
A bacteriophage is a virus that targets a specific bacterial host and lyses its surface in order to degrade it at the end of its reproductive cycle. Endolysins have at least two domains: the N-terminal enzymatic activity domain for the lysis of the cell wall and the cell-wall binding domain (CBD) to attach to the cell wall of a specific host. If the CBD is fused with other domains of endolysins it can be used to attack particular bacteria in species or strain level [5] [6].
We intend to use the SH3 domain from bacteriophage B30 to increase the sensitivity of the enzyme for pathogenic bacteria, specifically Streptococcus uberis, Staphylococcus aureus, and Streptococcus agalactiae, which is completely safe for the host [1]. The use of enzybiotics represents an alternative to the misuse of antibiotics without loss of efficiency; it is a novel and environmentally friendly process. It supplies antibacterial protection to pathogenic bacteria but shows no toxic effects on mammalian cells.
References
[1] Jarábková, V., Tišáková, L., Benešík, M., & Godány, A. (2021). SH3 binding domains from phage endolysins: how to use them for detection of gram-positive pathogens. Journal of Microbiology, Biotechnology and Food Sciences, 2021, 1215-1220. https://doi.org/10.15414/jmbfs.2020.9.6.1215-1220
[2] Lade, H., & Kim, J.-S. (2021). Bacterial Targets of Antibiotics in Methicillin-Resistant Staphylococcus aureus. Antibiotics, 10(4), 398. https://doi.org/10.3390/antibiotics10040398
[3] World Health Organization. (2021, November 17). Antimicrobial resistance. Who.int; World Health Organization: WHO. https://www.who.int/news-room/fact-sheets/detail/antimicrobial-resistance
[4] Tong, J., Zhang, H., Zhang, Y., Xiong, B., & Jiang, L. (2019). Microbiome and metabolome analyses of milk from dairy cows with subclinical streptococcus agalactiae mastitis—potential biomarkers. Frontiers in microbiology, 10, 2547. https://doi.org/10.3389/fmicb.2019.02547
[5] Broendum, S. S., Buckle, A. M., & McGowan, S. (2018). Catalytic diversity and cell wall binding repeats in the phage‐encoded endolysins. Molecular microbiology, 110(6), 879-896. https://doi.org/10.1111/mmi.14134
[6] Cho, J. H., Kwon, J. G., O'Sullivan, D. J., Ryu, S., & Lee, J. H. (2021). Development of an endolysin enzyme and its cell wall–binding domain protein and their applications for biocontrol and rapid detection of Clostridium perfringens in food. Food Chemistry, 345, 128562. https://doi.org/10.1016/j.foodchem.2020.128562| None |