Methods, formulations, and kits for bacterial degradation

Abstract

Methodologies, formulations, and kits suitable for decontaminating environments containing bacterial spores by degrading the spores. Formulations contain papain, at least one germinant, and optionally one or more additional enzymes. Methods for killing a bacterial spore include contacting a bacterial spore with the formulation for a duration sufficient to kill the spore, or for a duration sufficient to render the spore susceptible to being killed by a reagent and then contacting the spore with the reagent for a duration sufficient to kill the spore.

1 . A formulation for killing a bacterial spore having a spore coat surrounding a spore cortex and spore contents and optionally an exosporium surrounding the spore coat, the formulation containing papain, at least one germinant, and optionally one or more additional enzymes. 
     
     
         2 . The formulation according to  claim 1 , wherein the at least one germinant comprises at least one of L-alanine and inosine. 
     
     
         3 . The formulation according to  claim 1 , wherein the formulation contains at least one of the additional enzymes. 
     
     
         4 . The formulation according to  claim 1 , wherein the at least one additional enzyme is pancreatin or pronase. 
     
     
         5 . The formulation according to  claim 1 , wherein the formulation does not contain an enzyme other than papain. 
     
     
         6 . A method of killing a bacterial spore by contacting the spore with the formulation of  claim 1 . 
     
     
         7 . The method of  claim 6 , wherein the bacterial spore is contacted by the formulation for a duration sufficient to kill the spore. 
     
     
         8 . The method of  claim 6 , wherein the contacting step comprises contacting the spore with the formulation for a duration sufficient to render the spore susceptible to being killed by a reagent, and the method further comprises contacting the spore with the reagent for a duration sufficient to kill the spore. 
     
     
         9 . The method according to  claim 6 , wherein the bacterial spore is a  Bacillus anthracis  spore. 
     
     
         10 . The method according to  claim 6 , wherein the bacterial spore is a  Clostridium difficile  spore. 
     
     
         11 . A kit comprising the formulation of  claim 1 . 
     
     
         12 . The kit according to  claim 11 , the kit further comprising at least one decontamination reagent chosen from the group consisting of ethanol and mild detergents. 
     
     
         13 . A method of killing a bacterial spore having a spore coat surrounding a spore cortex and spore contents and optionally an exosporium surrounding the spore coat, the method comprising:
 contacting the spore with a formulation containing papain, at least one germinant, and optionally one or more additional enzymes, the spore being contacted for a duration sufficient to kill the spore or for a duration sufficient to render the spore susceptible to being killed by a reagent and then contacting the spore with the reagent for a duration sufficient to kill the spore.   
     
     
         14 . The method according to  claim 13 , wherein the bacterial spore is killed by contacting the spore with the formulation and then contacting the spore with the reagent, wherein the formulation renders the spore susceptible to being killed by the reagent and the reagent would be unable to kill the spore without the spore first being rendered susceptible by the formulation. 
     
     
         15 . The method according to  claim 13 , wherein the reagent is chosen from the group consisting of ethanol and mild detergents. 
     
     
         16 . The method according to  claim 13 , wherein the bacterial spore is a  Bacillus anthracis  spore. 
     
     
         17 . The method according to  claim 13 , wherein the bacterial spore is a  Clostridium difficile  spore. 
     
     
         18 . The method according to  claim 13 , wherein the contacting step comprises breaching the spore coat of the bacterial spore with the papain. 
     
     
         19 . The method according to  claim 13 , wherein the contacting step comprises at least partially inducing germination of the bacterial spore with the at least one germinant. 
     
     
         20 . The method according to  claim 13 , wherein the formulation contains at least 10 milligrams of papain per milliliter of a solution containing up to 10 7  of the bacterial spores per milliliter of the solution. 
     
     
         21 . The method according to  claim 13 , wherein the formulation contains at least 50 mM of the at least one germinant to a solution containing up to 10 7  of the bacterial spores per milliliter of the solution. 
     
     
         22 . The method according to  claim 13 , wherein the bacterial spore is present in a facility and the contacting step comprises dispensing the formulation within the facility to kill a plurality of the bacterial spore. 
     
     
         23 . The method according to  claim 22 , wherein the facility is a hospital and the formulation is applied to surfaces within the hospital. 
     
     
         24 . The method according to  claim 22 , wherein the facility produces weaponized bacteria and the formulation is dispersed within the environment of the facility.