The State University of New York

Suny Research Foundation

Christopher T Nomura

Benjamin R Lundgren

Ryan C Tappel

Qin Wang

Atahualpa Pinto

James P Nakas

Chengjun Zhu

Joseph A Perrotta

Methods and systems for producing prescribed unit size poly(3-hydroxyalkanoate) (PHA) polymers and copolymers are provided. The methods and systems can employ recombinant bacteria that are not native producers of PHA or lack enzymes to degrade PHA once synthesized, metabolize short to long chain fatty acids without induction, and express an (R)-specific enoyl-CoA hydratase and a PHA synthase, the (R)-specific enoyl-CoA hydratase and PHA synthase having wide substrate specificities. The recombinant bacteria are fed at least one fatty acid substrate that is equal in carbon length to the prescribed or desired unit size of the PHA polymer to be produced. The prescribed unit size PHA that is produced is then isolated and/or purified.

Engineered strain offor production of poly-R-3-hydroxyalkanoate polymers with defined monomer unit composition and methods based thereon

Methods and systems for producing prescribed unit size azido-poly(3-hydroxyalkanoate) (azido-PHA) polymers and copolymers are provided. The methods and systems can employ recombinant bacteria that are not native producers of PHA or lack enzymes to degrade PHA once synthesized, metabolize short to long chain fatty acids without induction, and express an (R)-specific enoyl-CoA hydratase and a PHA synthase, the (R)-specific enoyl-CoA hydratase and PHA synthase having wide substrate specificities. The recombinant bacteria are fed at least one &#x3c9;-azidofatty acid substrate that is equal in carbon length to the prescribed or desired unit size of an azido-PHA polymer to be produced. Azido-PHA polymers or azido-PHA co-polymers can be conjugated via copper-catalyzed alkyne-azide cycloaddition (CuAAC) or strain-promoted azide-alkyne cycloaddition (SPAAC) reactions. The prescribed unit size conjugated azido-PHA polymer or orthogonally conjugated azido-PHA co-polymer that is produced is then isolated and/or purified.

[object Object]

A synthetic peptide includes an amino acid sequence that: has 70-75%, 75-80%, 80-85%, 85-90%, 90-95% or 95-100% homology with an RpoN box of Region III of an RpoN protein; and binds specifically to a &#x2212;24, &#x2212;12, or &#x2212;24/&#x2212;12 site(s) of an RpoN promoter. The synthetic peptide is effective for repressing transcription and/or gene expression from a binding site of interest, and the binding site of interest is an RpoN binding site or a cryptic promoter upstream of an RpoN binding site.

Synthetic peptide for repressing transcription and/or gene expression from a binding site of interest

The present invention relates to, inter alia, a method for repressing transcription and/or gene expression from RpoN binding sites (or promoters) or cryptic promoters upstream of RpoN binding sites. The method comprises providing an agent that specifically and selectively binds to RpoN promoter sequences to inhibit or repress the expression of genes downstream of that promoter; and contacting the RpoN promoter with the agent. Agents for repressing transcription and/or gene expression from RpoN promoters are also provided. The agent can be a composition that binds specifically to the &#x2212;24, &#x2212;12, or &#x2212;24/&#x2212;12 site(s) for RpoN promoter interference. Synthetic peptides, vectors, and host cells are also provided.

Molecular roadblocks for RpoN binding sites

Methods are provided for producing biodegradable polyhydroxyalkanoates (PHAs) with desired geometry, molecular mass, mechanical and/or physical-chemical properties from glycerol, an inexpensive carbon source and byproduct of the biodiesel industry. Microorganisms capable of converting carbon to PHA can be used to convert biodiesel-glycerol to poly-3-hydroxybutyrate (PHB) or other monomer or copolymer PHAs via fermentation. The microorganisms are cultured in a medium comprising glycerol as a primary carbon source and one or more low molecular mass organic acids as a secondary carbon source. Biomass can be harvested from the culture medium and crude PHA extracted and purified, thereby recovering purified PHA with the desired property. After PHA isolation, a nucleating agent can be added to improve certain physical-chemical properties of the PHA, e.g., crystallization temperature, to enhance performance of the PHA during injection molding.

Growing community of inventors

Syracuse, NY, United States of America

Christopher T Nomura