Provivi, Inc.

Calysta, Inc.

Effendi Leonard

Micah Sheppard

Thomas Heel

Joshua A Silverman

Keith Michael Wampler

Peter Meinhold

Pedro Coelho

Renee M Saville

Jon Edward Ness

Jeremy S Minshull

Mark D Welch

Jana Stumpe

Kathryn Christina Wu

Vu Bui

Konrad B Otte

Tom Purcell

The present application relates to methods to improve biomass or lipid production in a microorganism from one or more fatty acid and one or more simple carbon co-substrates. Produced lipids may include unsaturated C-Cfatty acids, alcohols, aldehydes, and acetates which may be useful as final products or precursors to insect pheromones, fragrances, flavors, and polymer intermediates. The application further relates to recombinant microorganisms modified for improved production of biomass or lipid, or improved lipid selectivity. Also provided are methods of producing one or more lipid using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally one or more of the product lipid.

Multi-substrate metabolism for improving biomass and lipid production

The present application relates to recombinant microorganisms useful in the biosynthesis of unsaturated C-Cfatty alcohols, aldehydes, and acetates which may be useful as insect pheromones, fragrances, flavors, and polymer intermediates. The recombinant microorganisms may express enzymes or enzyme variants useful for production of and/or may be modified to downregulate pathways to shunt production toward unsaturated C-Cfatty alcohols, aldehydes, and acetates. The C-Cfatty alcohols, aldehydes, and acetates described herein may be used as substrates for metathesis reactions to expand the repertoire of target compounds and pheromones. The application further relates to recombinant microorganisms co-expressing a pheromone pathway and a pathway for the production of a toxic protein, peptide, oligonucleotide, or small molecule suitable for use in an attract-and-kill pest control approach. The application further relates to microorganisms modified to express or downregulate enzymes useful for production of unsaturated short chain fatty alcohols, aldehydes, and acetates which may be useful as insect pheromones, fragrances, flavors, and polymer intermediates. Also provided are methods of producing unsaturated C-Cfatty alcohols, aldehydes, and acetates using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally one or more of the product alcohols, aldehydes, or acetates.

Microorganisms for the production of insect pheromones and related compounds

The present application relates to recombinant microorganisms useful in the biosynthesis of unsaturated C-Cfatty alcohols, aldehydes, and acetates which may be useful as insect pheromones, fragrances, flavors, and polymer intermediates. The C-Cfatty alcohols, aldehydes, and acetates described herein may be used as substrates for metathesis reactions to expand the repertoire of target compounds and pheromones. The application further relates to recombinant microorganisms co-expressing a pheromone pathway and a pathway for the production of a toxic protein, peptide, oligonucleotide, or small molecule suitable for use in an attract-and-kill pest control approach. Also provided are methods of producing unsaturated C-Cfatty alcohols, aldehydes, and acetates using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally one or more of the product alcohols, aldehydes, or acetates.

The present application relates to methods of producing one or more fatty alcohols and/or one or more fatty aldehydes from one or more unsaturated lipid moieties by combining the obtainment or production of the one or more unsaturated lipid moieties from a biological source with conversion by non-biological means of the one or more unsaturated lipid moieties to one or more fatty alcohols and/or one or more fatty aldehydes. The present application also relates to recombinant microorganisms having a biosynthesis pathway for the production of one or more unsaturated lipid moieties. The one or more fatty alcohols can further be chemically converted to one or more corresponding fatty acetates. The one or more fatty alcohols, one or more fatty aldehydes and/or one or more fatty acetates produced by the methods described herein may be one or more insect pheromones, one or more fragrances, one or more flavoring agents, or one or more polymer intermediates.

Semi-biosynthetic production of fatty alcohols and fatty aldehydes

The present application relates to recombinant microorganisms useful in the biosynthesis of unsaturated C-Cfatty alcohols, aldehydes, and acetates which may be useful as insect pheromones, fragrances, flavors, and polymer intermediates. The recombinant microorganisms may express enzymes or enzyme variants useful for production of and/or may be modified to down regulate pathways to shunt production toward unsaturated C-Cfatty alcohols, aldehydes, and acetates. The C-Cfatty alcohols, aldehydes, and acetates described herein may be used as substrates for metathesis reactions to expand the repertoire of target compounds and pheromones. Also provided are methods of producing unsaturated C-Cfatty alcohols, aldehydes, and acetates using the recombinant microorganisms, as well as compositions comprising the recombinant microorganisms and/or optionally one or more of the product alcohols, aldehydes, or acetates.

The present disclosure provides nucleic acids and vectors for use with methanotrophic bacteria. Related host cells and methods for using such nucleic acids and vectors for expressing polypeptides or other genetic manipulation of methanotrophic bacteria are also provided.

Nucleic acids and vectors for regulated target gene expression in methanotrophic bacteria

The present disclosure provides nucleic acids and vectors for use with methanotrophic bacteria. Related host cells and methods for using such nucleic acids and vectors for expressing polypeptides or other genetic manipulation of methanotrophic bacteria are also provided. In one aspect, the present disclosure is directed to a non-naturally occurring nucleic acid molecule, comprising (1) a promoter that is functional in a methanotrophic bacterium, and (2) a native or altered methanol dehydrogenase (MDH) ribosomal binding sequence, provided that when the promoter is an MDH gene promoter, the nucleic acid comprises an altered MDH ribosomal binding sequence.

Nucleic acids and vectors for use with methanotrophic bacteria

The present disclosure relates to genetically engineered methanotrophic bacteria with the capability of growing on a multi-carbon substrate (e.g., glucose) as a primary or sole carbon source and methods for growing methanotrophic bacteria on the multi-carbon substrate.

Engineering of multi-carbon substrate utilization pathways in methanotrophic bacteria

The present disclosure relates to genetically engineered methanotrophic bacteria with the capability of growing on a multi-carbon substrate (e.g., glycerol) as a primary or sole carbon source and methods for growing methanotrophic bacteria on the multi-carbon substrate.

The present disclosure relates to genetically engineered methanotrophic bacteria with the capability of growing on a multi-carbon substrate as a primary or sole carbon source and methods for growing methanotrophic bacteria on a multi-carbon substrate.

Provided herein is an alkane-metabolizing cell that is unable to convert propionyl-CoA into methylmalonyl-CoA or 2-metylcitrate synthase. Depending on which enzymes are present in the cell, the cell can produce acrylate or a precursor for the same (e.g., propionate, 3-hydroxypropionyl-CoA, 3-hydroxypropionate, acrylyl-CoA) that can be readily converted to acrylate enzymatically (e.g., in the cell) or by chemical treatment. In one embodiment, the cell may contain a cytochrome P450 or alkane oxidase enzyme that allows the production of 3-hydroxypropionyl-CoA, which can be readily converted to 3-hydroxypropionate. In order to make such compounds, the cell may be grown in the presence of an odd-numbered chain alkane (e.g., pentane or heptane), although another odd-numbered chain alkane may be used. In another embodiment, the cell may contain acyl-CoA oxidase, enoyl-CoA hydratase, and hydrolase.

Growing community of inventors

Anaheim, CA, United States of America

Effendi Leonard