California Institute of Technology

Provivi, Inc.

Gevo, Inc.

California University of Technology

Peter Meinhold

Matthew W Peters

Frances H Arnold

Pedro Coelho

Keith Michael Wampler

Aristos Aristidou

Mike M Y Chen

Jun Urano

Rudi Fasan

Andrew Hawkins

Catherine Asleson Dundon

Vu Bui

Thomas Heel

Thomas Buelter

Effendi Leonard

Micah Sheppard

Reid M Renny Feldman

Uvini Gunawardena

Hasan Mehdi

Patrick R Gruber

Levente Ondi

William Andrew Evanko

David A Glassner

Christopher Smith

Stephanie Porter-Scheinman

Julie Kelly

Ruth Berry

Justas Jancauskas

Melissa Dey

James Wade

Doug Lies

Sabine Bastian

Marco Landwehr

Aharon Eyal

Kent Evans

Fudu Miao

Michael Cockrem

Lynne Albert

Toni M Lee

Catherine Mee-Hie Cho

Mike Ming Yu Chen

Eva Eckl

Renny Feldman

Mathew W Gevo Incl Peters

This invention relates to modified hydroxylases. The invention further relates to cells expressing such modified hydroxylases and methods of producing hydroxylated alkanes by contacting a suitable substrate with such cells.

Alkane oxidation by modified hydroxylases

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.

In one aspect, the invention provides a method for synthesizing a fatty olefin derivative. The method includes: a) contacting an olefin according to Formula I with a metathesis reaction partner according to Formula IIb in the presence of a metathesis catalyst under conditions sufficient to form a metathesis product according to Formula IIIb: andb) converting the metathesis product to the fatty olefin derivative. Each Ris independently selected from H, Calkyl, and Calkenyl; Ris Calkyl; subscript y is an integer ranging from 0 to 17; and subscript z is an integer ranging from 0 to 17. In certain embodiments, the metathesis catalyst is a tungsten catalyst or a molybdenum catalyst. In various embodiments, the fatty olefin derivative is a pheromone. Pheromone compositions and methods of using them are also described.

Production of fatty olefin derivatives via olefin metathesis

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.

In one aspect, the invention provides a method for synthesizing a fatty olefin derivative. The method includes: a) contacting an olefin according to Formula I with a metathesis reaction partner according to Formula IIb in the presence of a metathesis catalyst under conditions sufficient to form a metathesis product according to Formula IIIb: and

In certain aspects, the present invention provides methods for producing terminally hydroxylated alkenes and alkynes by contacting an unsaturated or saturated hydrocarbon substrate with a hydroxylase enzyme. Exemplary terminal hydroxylases useful for carrying out the methods of the invention exhibit strong selectivity towards one terminal carbon of a hydrocarbon substrate and include, but are not limited to, non-heme diiron alkane monooxygenases, cytochromes P450 (e.g., cytochromes P450 of the CYP52 and CYP153 family), as well as long chain alkane hydroxylases. In some embodiments, the terminally hydroxylated alkene or alkyne is further converted to a terminal alkenal. In certain embodiments, terminally hydroxylated alkenes and alkynes are useful as insect pheromones which modify insect behavior. In other embodiments, terminally hydroxylated alkenes and alkynes are useful intermediates for producing pheromones via acetylation or oxidation of the alcohol moiety.

Synthesis of olefinic alcohols via enzymatic terminal hydroxylation

The present disclosure provides pheromone compositions. In some aspects, the compositions taught herein comprise a pheromone chemically corresponding to the pheromone naturally produced by a given insect, along with at least one positional isomer of said pheromone. In various aspects, pheromone compositions of the present disclosure are able to modulate the response of the insect based on the ratio of natural pheromone to its positional isomer.

Agricultural pheromone compositions comprising positional isomers

The present invention relates, in part, to an improved process for oxidation of alcohols containing oxidatively sensitive functional groups, using inexpensive reagents under mild reaction conditions to provide high yields of carbonyl products such as aldehydes or ketones. In certain embodiments, an aldehyde product is obtained by contacting an oxidatively sensitive alcohol, such as an alkenol, with an oxidant and a TEMPO catalyst under conditions sufficient to convert the alkenol to the aldehyde.

Process for oxidation of fatty alkenols to alkenal pheromone products

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Pasadena, CA, United States of America

Peter Meinhold