Bacterial strains, plasmids, method of producing bacterial strains capable of chemolithotrophic arsenites oxidation and uses thereof

Abstract

The invention provides novel strains Agrobacterium tumefaciens KKP 2039p and Paracoccus alcaliphilus KKP 2040p, the plasmid pSinA and its functional derivative, method for producing bacterial strains capable of chemolithotrophic arsenite oxidation and novel bacterial strains produced by this method. The invention also relates to the composition, comprising the novel bacterial strain or the plasmid pSinA and the use of these novel strains, as well as the method of bioaugmentation of an arsenic contaminated environment, particularly the method for the removal of arsenic from waters.

1 . An isolated or novel bacteria strain  Agrobacterium tumefaciens  deposited in the IAFB Collection of Industrial Microorganisms of Institute of Agricultural and Food Industry under the number KKP 2039p. 
     
     
         2 . An isolated or novel bacteria strain  Paracoccus alcaliphilus  deposited in the The IAFB Collection of Industrial Microorganisms of Institute of Agricultural and Food Industry under the number KKP 2040p. 
     
     
         3 . A novel or isolated or non-naturally occurring plasmid pSinA having a nucleotide sequence shown in SEQ ID NO: 1. 
     
     
         4 . A method for producing a bacterial strain capable of chemolithotrophic arsenite oxidation, comprising introducing the plasmid of  claim 3  into the bacterial strain. 
     
     
         5 . The method according to  claim 4 , wherein the introducing is carried out by a process comprising:
 (i) triparental mating with the use of a donor strain harbouring the plasmid, and a helper strain harbouring a helper plasmid, or,   (ii) biparental mating with the use of a donor strain harbouring the plasmid.   
     
     
         6 . The method according to  claim 5  wherein the donor strain is i Agrobacterium tumefaciens deposited under the number KKP 2039p or  Paracoccus alcaliphilus  deposited under the number KKP 2040p. 
     
     
         7 . The method according to  claim 4 , including introducing a gene encoding a selection marker into the bacterial strain. 
     
     
         8 . The method of  claim 7  wherein the selection marker comprises antibiotic resistance. 
     
     
         9 . The method according to  claim 7 , wherein the introducing is by a plasmid. 
     
     
         10 . The method of  claim 9  wherein the introducing is by triparental mating including a bacterial strain harbouring the plasmid containing the gene encoding the selection marker and a helper strain harbouring a helper plasmid. 
     
     
         11 . The method according to  claim 4 , wherein the bacterial strain is isolated from a natural environment. 
     
     
         12 . The method of  claim 4  wherein the bacterial strain is isolated from an arsenic contaminated environment. 
     
     
         13 . The method according to  claim 4 , wherein the bacterial strain is an  Alphaproteobacteria  or  Gammaproteobacteria  bacterial strain. 
     
     
         14 . An isolated, novel non-naturally occurring bacterial strain capable of chemolithotrophic arsenite oxidation, produced by the method according to  claim 4 . 
     
     
         15 . A composition comprising the novel bacterial strain according to  claim 1 . 
     
     
         16 . A composition comprising the novel bacterial strain according to  claim 2 . 
     
     
         17 . A composition comprising the isolated, novel, non-naturally occurring bacterial strain according to  claim 14 . 
     
     
         18 . A composition comprising a novel, non-naturally occurring, isolated bacteria containing the plasmid according to  claim 3 . 
     
     
         19 . A method of bioaugmentation of arsenic contaminated environment, comprising introducing the isolated novel strain according to  claim 1 , or the isolated novel strain according to  claim 2 , or the novel, non-naturally occurring isolated strain of  14 , or a bacteria harbouring the plasmid according to  claim 3 , or the composition according to  claim 15 , or the composition according to  claim 16 , or the composition according to  claim 17 , or the composition according to  claim 18 , or a combination thereof, into an arsenic contaminated environment. 
     
     
         20 . A method for the removal or recovery of arsenic comprising chemolithotrophic arsenite oxidation by the isolated novel strain according to  claim 1 , or the isolated novel strain according to  claim 2 , or the novel, non-naturally occurring isolated strain of  14 , or a bacteria harbouring the plasmid according to  claim 3 , or the composition according to  claim 15 , or the composition according to  claim 16 , or the composition according to  claim 17 , or the composition according to  claim 18 , or a combination thereof. 
     
     
         21 . The method according to  claim 20 , wherein the chemolithotrophic arsenite oxidation is followed by precipitation of the resulting arsenates and/or adsorption of arsenates. 
     
     
         22 . The method according to  claim 21  wherein the precipitation or adsorption is carried out using burnt lime (CaO), calcium hydroxide Ca(OH) 2 , bog iron ores or a combination thereof. 
     
     
         23 . An isolated or novel or non-naturally occurring plasmid comprising the nucleotide sequence corresponding to nucleotides 24376-34453 in SEQ ID NO: 1. 
     
     
         24 . An isolated or novel or non-naturally occurring bacterial strain comprising the plasmid of  claim 23  or the nucleotides 24376-34453 of SEQ ID NO: 1. 
     
     
         25 . Use of the plasmid defined in  claim 23  or the strain defined in  claim 24  for arsenite oxidation. 
     
     
         26 . An isolated or novel or non-naturally plasmid comprising the nucleotide sequence corresponding to nucleotides 43229-50772 in SEQ ID NO: 1. 
     
     
         27 . An isolated or novel or non-naturally bacterial strain comprising the plasmid of  claim 26 , or the nucleotides 43229-50772 of SEQ ID NO: 1. 
     
     
         28 . Use of the plasmid defined in  claim 26  or the nucleotide sequence comprising the 43229-50772 of SEQ ID NO: 1 for the production of a strain with increased resistance to arsenic. 
     
     
         29 . Use of the novel strain according to  claim 1  or  2 , the novel non-naturally occurring isolated strain bacterial strain according to  claim 14 , the plasmid according to  claim 3 , the composition according to  claim 15  or  16 , or combination thereof, in the processes of biological removal of arsenic characterized in that the removal of arsenic comprises bioaugmentation or biometallurgy of arsenic. 
     
     
         30 . Use of the isolated novel bacterial strain according to  claim 1  or  2 , the novel, non-naturally occurring bacterial strain according to  claim 14 , the plasmid according to  claim 3 , the composition according to  claim 15  or  16 , or combination thereof, for constructing bacterial strains capable of chemolithotrophic arsenite oxidation.