Sulfolobal phosphotriesterase-like (pll) lactonases activity having enhanced properties and the uses thereof

Abstract

Mutated hyperthermophilic PTE having a lactonase activity derived from a hyperthermophilic phosphotriesterase corresponding to the consensus sequence of SEQ ID NO: 1, the mutated PTE including the at least one mutation chosen amongst 53 putative positions and the mutated PTE having enhanced properties. Also provided are compositions including the mutated hyperthermophilic PTE and the uses thereof, notably as bioscavenger of organophosphate compounds or as quorum quencher of the bacteria using lactones to communicate.

1 . An antibacterial composition comprising as active ingredient at least one mutated hyperthermophilic phosphotriesterase,
 said mutated hyperthermophilic phosphotrieterase has an increased lactonase catalytic activity in comparison of the lactonase activity of a non-mutated hyperthermophilic phosphotriesterase, wherein the non-mutated hyperthermophilic phosphotriesterase is a wild-type phosphotriesterase corresponding to the consensus sequence of SEQ ID NO:1, wherein the amino acid W in position 265 is substituted by an amino acid selected from the group consisting of isoleucine I, valine V, threonine T and alanine A within the mutated hyperthermophilic phosphotriesterase.   
     
     
         2 . The antibacterial composition according to  claim 1 , wherein hydrolysis of 3-oxo-C12 AHL by said mutated hyperthermophilic phosphotriesterase is increased by at least 2 times, in comparison to hydrolysis of 3-oxo-C12 AHL by said non-mutated hyperthermophilic phosphotriesterase. 
     
     
         3 . The antibacterial composition according to  claim 1 , wherein said mutated hyperthermophilic phosphotriesterase has a thermostability, which is substantially similar to the thermostability of said non-mutated hyperthermophilic phosphotriesterase. 
     
     
         4 . The antibacterial composition according to  claim 1 , wherein the amino acid in position 2 in SEQ ID NO: 1 of said non-mutated hyperthermophilic phosphotriesterase is missing. 
     
     
         5 . The antibacterial composition according to  claim 1 , wherein said non-mutated hyperthermophilic phosphotriesterase is selected from the group consisting of SEQ ID NO: 3 from  Sulfolobus solfataricus , SEQ ID NO: 5 from  Sulfolobus acidocalaricus , and from SEQ ID NO: 7  Sulfolobus islandicus,    wherein said sequences SEQ ID NO: 3, SEQ ID NO: 5 and SEQ ID NO: 7 belong to the consensus SEQ ID NO: 1, and for said mutated hyperthermophilic phosphotriesterase the amino acid in position 2 in SEQ ID NO: 1 being missing from SEQ ID NO: 5 and the amino acids in position 2 and 3 in SEQ ID NO: 1 being missing from SEQ ID NO: 3 and SEQ ID NO: 7.   
     
     
         6 . The antibacterial composition according to  claim 1 , wherein said amino acid W in position 265 is substituted by an amino acid Isoleucine I within said mutated hyperthermophilic phosphotriesterase. 
     
     
         7 . The antibacterial composition according to  claim 1 , said mutated hyperthermophilic PTE being chosen in the group consisting of: SEQ ID NO: 13, SEQ ID NO: 15, SEQ ID NO: 17, SEQ ID NO: 19, SEQ ID NO: 57, SEQ ID NO: 59, SEQ ID NO: 61, SEQ ID NO: 63, SEQ ID NO: 101, SEQ ID NO: 103, SEQ ID NO: 105 and SEQ ID NO: 107. 
     
     
         8 . A phytosanitary composition comprising the antibacterial composition according to  claim 1 . 
     
     
         9 . A pharmaceutical composition comprising the antibacterial composition according to  claim 1 , said antibacterial composition further comprising a pharmaceutically acceptable vehicle. 
     
     
         10 . The pharmaceutical composition according to  claim 9 , further comprising at least one antibiotic selected from the group consisting of gentamycine, ciprofloxacin, ceftazidime, imipenem, and tobramycine. 
     
     
         11 . A method of treating a bacterial infection, comprising administering to a patient in need thereof the antibacterial composition according to  claim 1 . 
     
     
         12 . The method of treating bacterial infections according to  claim 11 , wherein the bacterial infection is pneumonia or nosocomial diseases, caused by bacteria using homoserin lactone substrates to communicate, in particular in the blood, wounds, burn, skin, biomaterial-body contact area. 
     
     
         13 . A method of disrupting quorum-sensing in bacteria comprising administering the antibacterial composition according to  claim 1 . 
     
     
         14 . The method according to  claim 13 , wherein the antibacterial composition is administered to boats or other sea equipment and limits the formation of biofilms in boats or other sea equipment. 
     
     
         15 . The method according to  claim 13 , wherein the antibacterial composition is administered to plants or vegetables and inhibits fire blight in plants or rotting of vegetables.