(12) INTERNATIONAL APPLICATION PUBLISHED UNDER THE PATENT COOPERATION TREATY (PCT) (19) World Intellectual Property Organization International Bureau (10) International Publication Number (43) International Publication Date WO 2018/213554 Al 22 November 2018 (22.11.2018) W ! P O PCT (51) International Patent Classification: (72) Inventors: SHEPPARD, Micah; c/o Provivi, Inc., 1701 C12P 7/04 (2006 .0 1) C12N 9/02 (2006 .01) Colorado Avenue, Santa Monica, California 90404 (US). C12P 7/24 (2006 .0 1) C12N 9/10 (2006 .01) HEEL, Thomas; c/o Provivi, Inc., 1701 Colorado Avenue, C12P 7/64 (2006.01) Santa Monica, California 90404 (US). MEINHOLD, Pe¬ ter; c/o Provivi, Inc., 1701 Colorado Avenue, Santa Mon (21) International Application Number: ica, California 90404 (US). WAMPLER, Keith; c/o Pro PCT/US20 18/033 15 1 vivi, Inc., 1701 Colorado Avenue, Santa Monica, Califor (22) International Filing Date: nia 90404 (US). COELHO, Pedro; c/o Provivi, Inc., 1701 17 May 2018 (17.05.2018) Colorado Avenue, Santa Monica, California 90404 (US). LEONARD, Effendi; c/o Provivi, Inc., 1701 Colorado Av (25) Filing Language: English enue, Santa Monica, California 90404 (US). (26) Publication Language: English (74) Agent: HOLLY, David Christopher et al; Cooley LLP, (30) Priority Data: 1299 Pennsylvania Avenue, N.W., Suite 700, Washington, 62/507,654 17 May 2017 (17.05.2017) US District of Columbia 20004-2400 (US). (71) Applicant: PROVIVI, INC. [US/US]; 1701 Colorado Av (81) Designated States (unless otherwise indicated, for every enue, Santa Monica, California 90404 (US). kind of national protection available): AE, AG, AL, AM, AO, AT, AU, AZ, BA, BB, BG, BH, BN, BR, BW, BY, BZ, (54) Title: MICROORGANISMS FOR THE PRODUCTION OF INSECT PHEROMONES AND RELATED COMPOUNDS r -A kan Fatty ecid Triglyceride 1 Fatty alcohol Fatty aldehyde Fatty acid c -Hy xyf a tt y acid Fatty acyl-CoA Cr ) D s t ra se (1, 2, 3... ) ci-Hydrcxyfatty aldehyde r .;. - En y -C A Mono a n p o y- s t rat d fatty acyl-CoA Reductase j , 2, 3„N| Dica rb x Ei d p-Kydroxyacyl-CoA i oxidant o r oxidase - t acy l-C A acetyjchlorideorac. transferase Fatty acyl-CoA {Crt-2 5 A y -C A FIGURE 1 (57) Abstract: The present application relates to recombinant microorganisms useful in the biosynthesis of unsaturated C6-C24 fatty alcohols, aldehydes, and acetates which may be useful as insect pheromones, fragrances, flavors, and polymer intermediates. The re combinant 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 C6-C24 fatty alcohols, aldehydes, and acetates. The C6-C24 fatty 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 -C24 fatty 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 o alcohols, aldehydes, or acetates. o [Continued on nextpage] WO 2018/213554 Al llll II II 11III II I I II III I I I I Hill II I II CA, CH, CL, CN, CO, CR, CU, CZ, DE, DJ, DK, DM, DO, DZ, EC, EE, EG, ES, FI, GB, GD, GE, GH, GM, GT, HN, HR, HU, ID, IL, IN, IR, IS, JO, JP, KE, KG, KH, KN, KP, KR, KW, KZ, LA, LC, LK, LR, LS, LU, LY, MA, MD, ME, MG, MK, MN, MW, MX, MY, MZ, NA, NG, NI, NO, NZ, OM, PA, PE, PG, PH, PL, PT, QA, RO, RS, RU, RW, SA, SC, SD, SE, SG, SK, SL, SM, ST, SV, SY, TH, TJ, TM, TN, TR, TT, TZ, UA, UG, US, UZ, VC, VN, ZA, ZM, ZW. (84) Designated States (unless otherwise indicated, for every kind of regional protection available): ARIPO (BW, GH, GM, KE, LR, LS, MW, MZ, NA, RW, SD, SL, ST, SZ, TZ, UG, ZM, ZW), Eurasian (AM, AZ, BY, KG, KZ, RU, TJ, TM), European (AL, AT, BE, BG, CH, CY, CZ, DE, DK, EE, ES, FI, FR, GB, GR, HR, HU, IE, IS, IT, LT, LU, LV, MC, MK, MT, NL, NO, PL, PT, RO, RS, SE, SI, SK, SM, TR), OAPI (BF, BJ, CF, CG, CI, CM, GA, GN, GQ, GW, KM, ML, MR, NE, SN, TD, TG). Declarations under Rule 4.17: — as to applicant's entitlement to apply for and be granted a patent (Rule 4.1 7(H)) Published: — with international search report (Art. 21(3)) — before the expiration of the time limit for amending the claims and to be republished in the event of receipt of amendments (Rule 48.2(h)) — with sequence listing part of description (Rule 5.2(a)) MICROORGANISMS FOR THE PRODUCTION OF INSECT PHEROMONES AND RELATED COMPOUNDS CROSS REFERENCE T O RELATED APPLICATIONS [0001] This application claims priority to U.S. Provisional Application Serial No. 62/507,654, filed on May 7, 2017, which is hereby incorporated by reference in its entirety. STATEMENT REGARDING THE SEQUENCE LISTING [0002] The Sequence Listing associated with this application is provided in text format in lieu of a paper copy, and is hereby incorporated by reference into the specification. The name of the text file containing the Sequence Listing is PRVI 020 0 WO SeqList ST25.txt. The text file is about 240 KB, was created on May 17, 2 , and is being submitted electronically viaEFS-Web. TECHNICAL FIELD [0003] This application relates to recombinant microorganisms useful in the biosynthesis of unsaturated C -C2 fatty alcohols, aldehydes, and acetates which may be useful as insect pheromones, fragrances, flavors, and polymer intermediates. The application further relates to methods of producing unsaturated C&-C24 fatty alcohols, aldehydes, and acetates using the recombinant microorganisms, as well as compositions comprising one or more of these compounds and/or the recombinant microorganisms. BACKGROUND [0004] As the global demand for food grows, there is an increasing need for effective pest control. Conventional insecticides are among the most popular chemical control agents because they are readily available, rapid acting, and highly reliable. However, the overuse, misuse, and abuse of these chemicals have led to resistant pests, alteration of the natural ecology, and in some cases, environmental damage. [0005] The use of insect pheromones to control pest populations has gained increasing popularity as a viable, safe, and environmentally friendly alternative to conventional insecticides. Since their discovery in the late 1950s, these molecules have shown efficacy in reducing insect populations through a variety of methods, including mass trappings, attract and kill, and mating disruption. The latter method in particular represents a non-toxic means of pest control and utilizes the ability of synthetic pheromones to mask naturally occurring pheromones, thereby causing confusion and mating disruption. [0006] Although pheromones have significant potential in agricultural insect control, the cost of synthesizing pheromones using currently available techniques is very high, which prohibits widespread use of this sustainable technology beyond high- value crops. Thus, there is an existing need to develop novel technologies for the cost-efficient production of insect pheromones and related fragrances, flavors, and polymer intermediates. The present inventors address tins need with the development of recombinant microorganisms capable of producing a wide-range of unsaturated CVC24 fatty alcohols, aldehydes, and acetates including synthetic insect pheromones from low-cost feedstocks. SUMMARY OF THE DISCLOSURE [0007] The present application relates to recombinant microorganisms having a biosynthesis pathway for the production of one or more compounds selected from unsaturated C6-C24 fatty- alcohols, aldehydes, and acetates. The recombinant microorganisms described herein may be used for the production of at least one compound, such as an insect pheromone, a fragrance, or a flavoring agent, selected from unsaturated C6-C24 fatty alcohols, aldehydes, and acetates. [0008] In one embodiment, the recombinant microorganism comprises a biosynthesis pathway for the production of an unsaturated C6-C24 fatty aldehyde or fatty alcohol . Accordingly, in a first aspect, the application relates to a recombinant microorganism capable of producing an unsaturated C6-C24 fatty aldehyde or fatty alcohol from an endogenous or exogenous source of saturated C6-C24 fatty acyl-CoA, wherein the recombinant microorganism expresses (a): at least one exogenous nucleic acid molecule encoding a fatty- acyl desaturase that catalyzes the conversion of a saturated C6-C24 fatty acyl-CoA to a corresponding mono- or poly-unsaturated C6-C24 fatty acyl-CoA; and (b): at least one exogenous nucleic acid molecule encoding a fatty aldehyde forming fatty-acyl reductase that catalyzes the conversion of the mono- or poly-unsaturated C6-C24 fatty acyl-CoA from into the corresponding mono- or poly-unsaturated C6-C24 fatty aldehyde. In some embodiments, the mono- or poly-unsaturated C6-C24 fatty aldehyde is an insect pheromone. In some embodiments, the mono- or poly-unsaturated C6-C24 fatty aldehyde is a fragrance or flavoring agent. In some embodiments, the recombinant microorganism further comprises at least one endogenous or exogenous nucieic acid molecule encoding an acetyl transferase capable of catalyzing the conversion of the mono- or poly-unsaturated C6-C24 fatty alcohol from (b) into a corresponding mono- or poly-unsaturated C6-C24 fatty acetate (c) at least one exogenous nucleic acid molecule encoding a fatty alcohol forming fatty-acyl reductase that catalyzes the conversion of the mono- or poly-unsaturated C6-C24 fatty acyl-CoA from (a) into the corresponding mono- or poly-unsaturated C6-C24 fatty alcohol. In some embodiments, the mono- or poly-unsaturated C6-C24 fatty alcohol is an insect pheromone. In some embodiments, the mono- or poly-unsaturated C6-C24 fatty alcohol is a fragrance or flavoring agent.