Team:YAU-China/PelA PslG
PelA&PslG
Biofilms are complex communities of bacteria embedded in an extracellular matrix composed of proteins, eDNA and exopolysaccharides. Among them, extracellular polysaccharide plays a key role in the supporting skeleton of the biofilm structure, so we destroy the biofilm for it. In the biological genome, a hydrolase that decomposes the polysaccharide is often present in a gene cluster encoding an exopolysaccharide. Pseudomonas aeruginosa PAO1 is divided into two types according to the genetic ability of its biofilm extracellular polysaccharide: Psl, Pel.
After investigation, we obtained the glycoside hydrolase PslG and PelA to prevent and rapidly destroy Pseudomonas aeruginosa to produce extracellular polysaccharides in vitro, thereby destroying the formation of biofilm.
In order to prove the effect, we constructed the expression plasmids of these two kinds of extracellular polysaccharide hydrolase, transferred them into E. coli expression strains, induced the cells, collected the cells and obtained the crude extracts of the two enzymes, and acts on the biofilm formed by Pseudomonas aeruginosa PAO1.
It can be seen that the lysate containing the two extracellular polysaccharide hydrolyzing enzymes of PelA and PslG destroyed the Pseudomonas aeruginosa biofilm by 50%. Therefore, we are convinced that the experimental design is effective.
References:
[1] Baker P, Hill P J, Snarr B D, Alnabelseya N, Pestrak M J, Lee M J, Jennings L K, Tam J, Melnyk R A, Parsek M R, Sheppard D C, Wozniak D J, Howell P L. Exopolysaccharide biosynthetic glycoside hydrolases can be utilized to disrupt and prevent Pseudomonas aeruginosa biofilms[J]. Science Advances, 2016, 2(5): e1501632-e1501632.
[2] Colvin K M, Alnabelseya N, Baker P, Whitney J C, Howell P L, Parsekac M R. PelA deacetylase activity is required for Pel polysaccharide synthesis in Pseudomonas aeruginosa[J]. Journal of Bacteriology, 2013, 195(10): 2329-2339.
[3] Jackson K D, Starkey M, Kremer S, Parsek M R, Wozniak D J. Identification of psl, a locus encoding a potential exopolysaccharide that is essential for Pseudomonas aeruginosa PAO1 biofilm formation[J]. Journal of Bacteriology, 2004, 186(14): 4466-4475.