Methods and compositions for making bacteriocins and antimicrobial peptides
Inventors
Assignees
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Publication Number
US-12195780-B2
Publication Date
2025-01-14
Expiration Date
Abstract
Methods and compositions for making bacteriocins are described in some embodiments herein. In some embodiments, pro-polypeptide comprising the bacteriocins in the desired ratios in cis, and separated by cleavage sited can be produced by a microbial cell comprising a nucleic acid encoding the pro-polypeptide. In some embodiments microfluidic devices and methods for making specified mixtures of antimicrobial peptides and/or bacteriocins are described.
Core Innovation
The invention provides a microfluidic device for producing a specified mixture of antimicrobial peptides and/or bacteriocins. The device includes discrete coding substrates, each encoding a different antimicrobial peptide and/or bacteriocin, where the coding substrates are comprised within separate chambers. An in vitro transcription/translation solution and a fluidic reservoir are included, and valves disposed on fluidic paths regulate flow between each discrete coding substrate and the fluidic reservoir.
The microfluidic device is configured to be placed in data communication with a processor. Based on the specified mixture, the processor configures the valves to place selected discrete coding substrates in fluidic communication with the fluidic reservoir at a stoichiometry defined by a user. Incubation of the in vitro transcription/translation solution with the discrete coding substrates produces the antimicrobial peptides and/or bacteriocins of the specified mixture, and the produced antimicrobial peptides and/or bacteriocins flow through the valves into the fluidic reservoir.
The invention further includes controlling flow of fluid in the fluidic reservoir so that movement of the antimicrobial peptides and/or bacteriocins in the fluidic reservoir produces the specified mixture. Additional embodiments include communicating the fluidic reservoir with tissue of a subject and mixing the produced mixture with a reservoir comprising chemical and/or phage antibiotics.
Claims Coverage
The independent claims are directed to a microfluidic device that uses discrete coding substrates in separate chambers and valve-controlled routing to produce a user-specified mixture of antimicrobial peptides and/or bacteriocins at a user-defined stoichiometry, with an in vitro transcription/translation solution and fluidic reservoir. Across the independent claims, five main inventive features are specified.
Processor-configured valve routing for user-defined stoichiometry
Configuring valves based on a specified mixture so that discrete coding substrates that encode the antimicrobial peptides and/or bacteriocins of the mixture are placed in fluidic communication with the fluidic reservoir at a stoichiometry defined by a user, but not other discrete coding substrates.
In vitro transcription/translation incubation with selected coding substrates
Permitting incubation of the in vitro transcription/translation solution with the discrete coding substrates that encode the antimicrobial peptides and/or bacteriocins of the specified mixture, whereby the antimicrobial peptides and/or bacteriocins of the specified mixture are produced.
Valve-controlled transfer into a fluidic reservoir to form the specified mixture
Permitting flow of the antimicrobial peptides and/or bacteriocins through the valves into the fluidic reservoir, and controlling flow of fluid in the fluidic reservoir such that movement of the antimicrobial peptides and/or bacteriocins in the fluidic reservoir produces the specified mixture of antimicrobial peptides and/or bacteriocins in the fluidic reservoir.
Discrete chambers encoding different peptide/bacteriocin mixtures at user-defined stoichiometry
Using discrete coding substrates comprised within separate chambers, where each discrete coding substrate encodes a different mixture of antimicrobial peptide(s) and/or bacteriocin(s) at a stoichiometry defined by a user.
Fluidic communication with an in vitro transcription/translation reservoir via valves
Using valves disposed on a fluidic path connected to a discrete coding substrate, each valve configured to regulate flow to or from the discrete coding substrate, wherein the device is configured to be placed in fluid communication with a fluidic reservoir of an in vitro transcription/translation solution.
The claim set covers microfluidic production of user-specified antimicrobial peptide and/or bacteriocin mixtures by valve-controlled routing of discrete coding substrates in separate chambers, using an in vitro transcription/translation solution and a fluidic reservoir, with stoichiometry defined by user input.
Stated Advantages
Documented Applications
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