Bacillus based delivery system and methods of use
AssigneeThe Curators of the University of Missouri
InventorsGeorge C. Stewart, Brian Matthew Thompson
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Abstract
Herein a Bacillus exosporium molecule delivery (BEMD) system that provides a means to deliver molecules of interest (MOIs) to an environment is disclosed. The system results in the display of MOIs on the exosporium surface of Bacillus family members such that they can be delivered to an environment in a stable and active form. In addition, methods of making and using the system are described.
Claims22
A method of reducing contaminants in an environment comprising distributing to a contaminated environment a recombinant Bacillus cereus family member expressing a fusion construct, wherein the fusion construct comprises at least one molecule of interest (MOI) and:
- a) a targeting sequence consisting of a 16 amino acid sequence having at least about 43% identity with amino acids 20-35 of SEQ ID NO. 7, wherein the identity with amino acids 25-35 is at least about 54%;
- b) a targeting sequence consisting of amino acids 1-35 of SEQ ID NO. 7;
- c) a targeting sequence consisting of amino acids 20-35 of SEQ ID NO. 7;
- d) a targeting sequence comprising SEQ ID NO. 13;
- e) a targeting sequence consisting of SEQ ID NO. 7;
- f) a targeting sequence comprising amino acids 1-27 of SEQ ID NO. 8;
- g) a targeting sequence comprising amino acids 12-27 of SEQ ID NO. 8;
- h) a targeting sequence comprising SEQ ID NO. 8;
- (i) an amino acid sequence comprising the amino acid sequence encoded by the entire open reading frame of BAS3290, wherein BAS3290 comprises an N-terminal domain comprising SEQ ID NO:
- 8;
- j) a targeting sequence comprising amino acids 1-38 of SEQ ID NO. 9;
- k) a targeting sequence comprising amino acids 23-38 of SEQ ID NO. 9;
- l) a targeting sequence comprising SEQ ID NO. 9;
- or
- m) a targeting sequence comprising a methionine residue covalently bound to an amino acid sequence consisting of 16 amino acids having at least 43% identity with amino acids 20-35 of SEQ ID NO.7, wherein the identity with amino acids 25-35 is at least 54%;
- wherein the MOI is physically attached to the exosporium of the recombinant Bacillus cereus family member.
A method of producing an energy source comprising distributing to a biomass material a recombinant Bacillus cereus family member expressing a fusion construct, wherein the fusion construct comprises at least one molecule of interest (MOI) and:
- a) a targeting sequence consisting of a 16 amino acid sequence having at least about 43% identity with amino acids 20-35 of SEQ ID NO. 7, wherein the identity with amino acids 25-35 is at least about 54%;
- b) a targeting sequence consisting of amino acids 1-35 of SEQ ID NO. 7;
- c) a targeting sequence consisting of amino acids 20-35 of SEQ ID NO. 7;
- d) a targeting sequence comprising SEQ ID NO. 13;
- e) a targeting sequence consisting of SEQ ID NO. 7;
- f) a targeting sequence comprising amino acids 1-27 of SEQ ID NO. 8;
- g) a targeting sequence comprising amino acids 12-27 of SEQ ID NO. 8;
- h) a targeting sequence comprising SEQ ID NO. 8;
- (i) an amino acid sequence comprising the amino acid sequence encoded by the entire open reading frame of BAS3290, wherein BAS3290 comprises an N-terminal domain comprising SEQ ID NO:
- 8;
- j) a targeting sequence comprising amino acids 1-38 of SEQ ID NO. 9;
- k) a targeting sequence comprising amino acids 23-38 of SEQ ID NO. 9;
- l) a targeting sequence comprising SEQ ID NO. 9;
- or
- m) a targeting sequence comprising a methionine residue covalently bound to an amino acid sequence consisting of 16 amino acids having at least 43% identity with amino acids 20-35 of SEQ ID NO.7, wherein the identity with amino acids 25-35 is at least 54%;
- wherein the MOI is physically attached to the exosporium of the recombinant Bacillus cereus family member.
Dependencies are inferred from the claim text.