A process for producing pheromones and applications thereof

Abstract

The present disclosure relates to a process for producing moth pheromones or precursors thereof, comprising a) obtaining a immobilized and / or permeabilized first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; and contacting with a first medium comprising a first substrate to obtain a first reaction mixture comprising mono- or poly-unsaturated C6-C24 fatty acid; b) obtaining a immobilized and / or permeabilized second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme; and contacting with a second medium comprising the first reaction mixture or a second substrate to obtain a second reaction mixture comprising mono- or poly¬ unsaturated C6-C24 fatty alcohol; and c) mixing an aqueous solution of laccase enzyme, the second reaction mixture, a mediator, and an emulsifying agent under a continuous supply of a gas, to obtain the pheromones or precursors thereof.

Description

A PROCESS FOR PRODUCING PHEROMONES AND APPLICATIONS THEREOFFIELD OF INVENTION

[0001] The present disclosure relates to the field of pheromones. Particularly, the present disclosure relates to a process for producing pheromones or precursors thereof using permeabilized yeast cells.BACKGROUND OF INVENTION

[0002] Intensified pest control seems to be the best way to increase food production. During the past few decades, use of pesticide has increased agriculture outputs worldwide. Yet in percentage terms, crop losses have remained unchanged since 1950s. The ever-increasing use of conventional pesticides leads to pests resisting the same, severely altering natural ecology and damaging the environment. The number of insects and other species developing resistance to pesticides is growing steadily, forcing chemical companies to develop new pesticide formulations.

[0003] In response to the problems caused by the increased use of conventional pesticides, the concept of integrated pest management (IPM) was developed. IPM combines chemical, biological, and agrochemical approaches to achieve pest control at reasonable cost, while minimizing damage to the environment. The first step in IPM is effective monitoring using pheromones. Pheromones which are vectors of communication between individuals of the same species, are dispersed by one individual or by several, and their perception results in a change in the behaviour of the receiving individual. Pheromones can also be used for mass trapping or causing “confusion”, especially when the pest population is rather diffuse. The first sex pheromone, that of silkworm moth Bombyx mori was isolated in the late 1950s. Today more than 1000 pheromones of various species are known [Pimentel, D. Chem.Ber. (1991) 27, 646; Karlson, P.; Luscher, M. Nature (1959) 183, 55; Jutsum, A.R.; Gordon, R.F.S. Insect Pheremones in Plant Protection; Wiley and Sons; New York, 1989; Ridgway, R.L et al., M.N. Behavior-ModifyingChemicals for Insect management; Marcel-Dekker ; New York, 1990; Shani,S. CHEMTECH (1998) 30].

[0004] Structurally, major pheromone components are fatty acid related derivatives, such as long chain alcohols and aldehydes possessing double and triple bonds at various positions in the structural framework. Since production of such compounds through natural means is very difficult, most of these applications use synthetic copies of pheromones that mediate either attraction or aggregation.

[0005] The insect sex pheromones are being chemically synthesized at a commercial level, nevertheless, their chemical synthesis requires expensive substrates and catalysts and generates hazardous wastes (Herbert, M. B., Marx, V. M., Pederson, R. L., and Grubbs, R. H. (2013). Concise syntheses of insect pheromones using Z-selective cross metathesis. Angew. Chem. Int. Ed. Engl. 52, 310-314; Turczel, G., Kovacs, E., Merza, G., Coish, P., and Tuba, R. (2018). Synthesis of semiochemicals via olefin metathesis. ACS Sustain. Chem. Eng. 7, 33- 48). In contrast, the biocatalytic method using recombinant cells is operationally simple, and eco-friendly.

[0006] Great progress has also been made in the functional characterization of the candidate genes involved in the sex -pheromone biosynthetic pathway of insects. Different expression systems like, insect cell lines, and yeast were used to validate these candidate genes. Known processes based on recombinant cell technology produce pheromones by heterologous expression of insect and other genes and / or diverting the fatty acid synthesis pathway. These biocatalytic methods using recombinant cells as bio-factories to produce pheromones in the cell, are single use, and require maintaining sterile conditions. Further, the product is contaminated with other saturated and unsaturated fatty acids present in the cells and the purification of pheromones is challenging.

[0007] Therefore, there is a need to develop a sustainable, economical, and naturefriendly method of producing pheromones, which is commercially feasible and effective in controlling insect populations.SUMMARY OF THE INVENTION

[0008] In an aspect of the present disclosure, there is provided a process for producing pheromones, or precursors thereof, said process comprising: a) obtaining a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; contacting the first yeast cell with a first medium comprising a first substrate to obtain a first reaction mixture comprising mono- or polyunsaturated C6-C24 fatty acid; wherein the first yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell; wherein the first substrate is a saturated or unsaturated C6-C24 fatty acid or derivative thereof; b) obtaining a second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme; contacting the second yeast cell with a second medium comprising the first reaction mixture or a second substrate to obtain a second reaction mixture comprising mono- or poly-unsaturated C6-C24 fatty alcohol; wherein the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell; wherein the second substrate is a mono or poly unsaturated C6-C24 fatty acid or derivative thereof; and c) mixing an aqueous solution of laccase enzyme, the second reaction mixture, a mediator, and an emulsifying agent under a continuous supply of a gas, to obtain the pheromones or precursors thereof.

[0009] In an aspect of the present disclosure, there is provided a process for producing mono- or poly- unsaturated C6-C24 fatty acid or derivative thereof, said process comprising: obtaining a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; contacting the first yeast cell with a first medium comprising a first substrate to obtain mono- or poly-unsaturated C6- C24 fatty acid; wherein the first yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell; and wherein the first substrate is a saturated or unsaturated C6-C24 fatty acid or derivative thereof.

[0010] In an aspect of the present disclosure, there is provided a process for producing mono- or poly-unsaturated C6-C24 fatty alcohol, said process comprising: obtaining a second yeast cell comprising at least one heterologous geneencoding fatty acid reductase enzyme; contacting the second yeast cell with a second medium comprising a second substrate to obtain mono- or poly-unsaturated C6-C24 fatty alcohol; wherein the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell; and wherein the second substrate is a mono or poly unsaturated C6-C24 fatty acid or derivative thereof.

[0011] In an aspect of the present disclosure, there is provided a process for producing pheromones, or precursors thereof, said process comprising: mixing an aqueous solution of enzyme laccase and mono- or poly-unsaturated C6-C24 fatty alcohol followed by addition of a mediator and an emulsifying agent under a continuous supply of a gas, to obtain pheromones or precursors thereof.

[0012] In an aspect of the present disclosure, there is provided a composition comprising pheromones or precursors thereof, produced from the process as disclosed herein.

[0013] These and other features, aspects, and advantages of the present subject matter will be better understood with reference to the following description and appended claims. This summary is provided to introduce a selection of concepts in a simplified form. This summary is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used to limit the scope of the claimed subject matter.BRIEF DESCRIPTION OF DRAWINGS

[0014] The following drawings form a part of the present specification and are included to further illustrate aspects of the present disclosure. The disclosure may be better understood by reference to the drawings in combination with the detailed description of the specific embodiments presented herein.

[0015] Figure 1 is a schematic representation of the vector map of A) p YES -fatty acid desaturase (FAD) plasmid and B) pYES-FAR (fatty acid reductase) plasmid, in accordance with an embodiment of the present disclosure.

[0016] Figure 2 is a schematic representation of the vector map of A) pPICZaA- FAD plasmid and B) pPICZaA-FAR plasmid, in accordance with an embodiment of the present disclosure.

[0017] Figure 3 depicts A) GCMS chromatogram for the conversion of fatty acid to an unsaturated fatty acid with first yeast cells expressing insect FAD, and B) GCMS chromatogram for acid to alcohol conversion, with second yeast cells expressing insect FAR, in accordance with an embodiment of the present disclosure.

[0018] Figure 4 depicts the Gas Chromatography mass spectrometry (GC-MS) analysis of FAMEs from yeast expression system, A. SEQ ID NO. 6 desaturases supplemented with methyl ester of Z-9-tetradecenoic acid (Z9-C14: Me); B. SEQ ID NO. 8 desaturases supplemented with methyl ester of Z- 11 -hexadecenoic acid (Zl l-16: Me); C. SEQ ID NO. 14 reductase supplemented with Z9-C14; D. SEQ ID NO. 16 reductase supplemented with Z- 11 -hexadecenoic acid (Z11-C16), in accordance with an embodiment of the present disclosure.

[0019] Figure 5 depicts the mechanism of laccase catalyzed oxidation of alcohols in the presence of mediators, in accordance with an embodiment of the present disclosure.

[0020] Figure 6 depicts changes in the UV absorbance spectrum at 420 nm to analyse effect of pH on laccase activity at 30 °C in 0.05M citrate-phosphate buffer, in accordance with an embodiment of the present disclosure.

[0021] Figure 7 depicts the stability of laccase (a) solubilized in buffer and (b) immobilized on TiO2, in accordance with an embodiment of the present disclosure.

[0022] Figure 8 depicts the GC chromatogram illustrating the conversion of alcohol to aldehyde, in accordance with an embodiment of the present disclosure.DETAILED DESCRIPTION OF THE INVENTION

[0023] Those skilled in the art will be aware that the present disclosure is subject to variations and modifications other than those specifically described. It is to be understood that the present disclosure includes all such variations and modifications. The disclosure also includes all such steps, features, compositions, and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations of any or more of such steps or features.Definitions

[0024] For convenience, before further description of the present disclosure, certain terms employed in the specification, and examples are delineated here. These definitions should be read in light of the remainder of the disclosure and understood as by a person of skill in the art. The terms used herein have the meanings recognized and known to those of skill in the art, however, for convenience and completeness, particular terms and their meanings are set forth below.

[0025] The articles “a”, “an” and “the” are used to refer to one or more than one (i.e., to at least one) of the grammatical object of the article.

[0026] The terms “comprise” or “contain” and “comprising” or “containing” are used in the inclusive, open sense, meaning that additional elements may be included. It is not intended to be construed as “consists of only”.

[0027] The term "at least one" is used to mean one or more and thus includes individual components as well as mixtures / combinations.

[0028] Throughout this specification, unless the context requires otherwise the word “comprise”, and variations such as “comprises” and “comprising”, will be understood to imply the inclusion of a stated element or step or group of element or steps but not the exclusion of any other element or step or group of element or steps.

[0029] The term “including” is used to mean “including but not limited to”, “including” and “including but not limited to” are used interchangeably.

[0030] The term “pheromone” refers to a chemical substance that act as vectors of communication between individuals of the same species, especially a mammal or an insect, affecting the behaviour or physiology of others of its species. The word pheromone is derived from two greek words pherin, meaning to carry, and hormon, meaning to set in motion, in order to excite the pests located, and its diffusion in the field or orchard is determined.

[0031] The term “desaturated” and “unsaturated”, used interchangeably herein, refers to a compound comprising one or more double or triple carbon-carbon bonds.

[0032] The term “saturated”, as used herein, refers to a state of a chemical compound characterized by the absence of double or triple carbon-carbon bonds.

[0033] The term “fatty acid”, as used herein, refers to a carboxylic acid with an aliphatic carbon chain of length ranging from 6 to 24, such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, and 24. The fatty acid may be saturated or unsaturated.

[0034] The term “fatty acid desaturase”, as used herein, refers to enzymes that catalyze the insertion of at least one double bond, preferably at the delta position, of fatty acids. The terms “fatty acid desaturase”, “desaturase”, “FAD”, and “fatty acyl CoA desaturase” are used interchangeably in the present disclosure. The double bonds may be inserted at any position of the fatty acid. Further, the double bonds may be inserted at one or more positions. For example, a fatty acid desaturase that introduces a double bond at the 9th position is termed Delta 9-fatty acid desaturase. For example, a fatty acid desaturase that introduces a double bond at 11th position is termed Delta 11 -fatty acid desaturase. For example, a fatty acid desaturase that introduces a double bond at 12th position is termed Delta 12-fatty acid desaturase. For example, a fatty acid desaturase that introduces a double bond at the 14th position is termed Delta 14-fatty acid desaturase. For example, a bifunctional desaturase can insert double bonds in fatty acid substrates containing a preexisting double bond. Accordingly, a fatty acid desaturase that introduces double bond at the 11th position or at 12th position is termed Delta 11 and Delta 12 bifunctional desaturase.

[0035] The term “fatty acid reductase”, as used herein, refers to the enzymes that catalyze the reduction of a saturated or an unsaturated fatty acid to fatty alcohol. The terms “fatty acid reductase”, “reductase”, and “FAR” are used interchangeably in the present disclosure.

[0036] The term “laccase” refers to the nontoxic, highly stable, blue multi-copper oxidase enzyme which can be used as a catalyst in the fields of waste detoxification, textile dye transformation, biosensors, food industry, and pulp bleaching.

[0037] The term “mediator” refers to the electron transfer reagents which acts as a sort of ‘electron shuttle’; once it is oxidized by laccase, it diffuses away from the enzymatic pocket and in turn oxidizes any substrate that, due to its size, could not directly enter the enzymatic pocket. In an aspect of the present disclosure, themediators are selected from electron transfer reagents such as (2,2,6,6- tetramethylpiperidin-l-yl)oxyl (TEMPO), hydroxy benzotriazole (HOBT), 2,2'- azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) or combinations thereof.

[0038] The term “emulsifying agent” refers to the substances that provides stability to an emulsion and prevents the separation of the liquid phase from the solid phase. In an aspect of the present disclosure the emulsifying agent is polyethylene glycol (PEG) having molecular weight ranging from 200 to 2000, preferably having molecular weight of 200 - 600.

[0039] The term “wm” refers to the volume of air bubbled per unit volume of the medium per minute, which is calculated by dividing measured airflow rate (L / m) with the volume (L) of medium. In the present disclosure, continuous supply of a oxygen or air through the reaction mass is carried out at a rate in the range of 0.1 vvm to 0.5 vvm.

[0040] All percentages, parts and ratios are based upon the total weight of the composition of the present disclosure unless otherwise indicated. Ratios, concentrations, amounts, and other numerical data may be presented herein in a range format. It is to be understood that such range format is used merely for convenience and brevity and should be interpreted flexibly to include not only the numerical values explicitly recited as the limits of the range, but also to include all the individual numerical values or sub-ranges encompassed within that range as if each numerical value and sub-range is explicitly recited. For example, a weight percentage in the range of 0.1% to 5%, should be interpreted to include not only the explicitly recited limits of 0.1% to 5% but also to include sub-ranges, such as 1 to 5%, 0.9 to 3.5% and so forth, as well as individual amounts, within the specified ranges, such as 3.2%, and 0.5%.

[0041] Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this disclosure belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of thedisclosure, the preferred methods and materials are now described. All publications mentioned herein are incorporated herein by reference.

[0042] The present disclosure is not to be limited in scope by the specific embodiments described herein, which are intended for the purposes of exemplification only. Functionally equivalent products, compositions, and methods are clearly within the scope of the disclosure, as described herein.

[0043] The present disclosure provides a process for producing of pheromones or precursors thereof. According to embodiments herein, pheromones refer to naturally occurring compounds that are secreted by organisms, such as insects. In one embodiment the insect is a moth. In some embodiments, the moth is a species selected from the group consisting of Argyrotaenia velutinana, Ostrinia nubilalis, Planotortrix octo, Argyrotaenia velutinana, Epiphyas postvittana, Helicoverpa assulta, Choristoneura rosaceana, Choristoneura parallela, Spodoptera littoralis, Thaumetopoea pityocampa, Lampronia capitella, Dendrolimus punctatus, Mamestra brassicae, Antheraea pernyi, Yponomeuta padella, Ctenopseustis herana, Ctenopseustis obliquana, Planotortrix excessana, Spodoptera exigua, Bicyclus anynana, Spodoptera litura, Heliothis virescens, Heliothis virescens, Heliothis subflexa, Agrotis segetum, Agrotis ipsilon, Cydia pomonella, Helicoverpa armigera, Ascotis selenaria cretacea, Arctia plantaginis, Pieris macdunnoughi, Parnassius apollo, Plutella xylostella, Danaus chrysippus, Diatraea saccharalis, Chrysodeixis includens, Brenthis ino, Iphiclides podalirius, Euphydryas editha, Manduca sexta, Eumeta japonica, Manduca sexta, Drosophila bipectinate, Drosophila birchii, Drosophila pseudoananassae, Phthorimaea operculella, Choristoneura fumiferana, Dendrolimus kikuchii, Tuta absoluta, Operophtera brumata, Bombyx mori, Amyelois transitella, Danaus plexippus plexippus, Chilo suppressalis, Streltzoviella insularis, Dioryctria abietella, Mythimna separata, Glyphodes pyloalis, Eeptidea sinapis, Anopheles gambiae str. PEST, Papilio polytes, Papilio Xuthus, Papilio machaon, Drosophila ananassae, Pieris rapae, Bicyclus anynana, Hyposmocoma kahamanoa, Ctenocephalides felis, Vanessa tameamea, Trichoplusia ni, Galleria mellonella, Bombyx mandarina, Ostrinia furnacalis, Maniola hyperantus, Spodoptera frugiperda, Glossina fuscipes,Hermetia illucens, Zerene cesonia, Pararge aegeria, Aricia agestis, Melitaea cinxia, Colias croceus, Pieris brassicae, Maniola jurtina, Vanessa cardui, Helicoverpa zea, Vanessa atalanta, Leguminivora glycinivorella, Pectinophora gossypiella, Nymphalis io, Yponomeuta evonymella, Yponomeuta rorrellus, Ostrinia latipennis, Ostrinia palustralis, Ostrinia scapulalis, Ostrinia nr. zaguliaevi JML-2013, Ostrinia zaguliaevi, Ostrinia zealis, Agrotis segetum, Sesamia inferens, Cotesia congregate, Astyanax mexicanus, Habropoda laboriosa, Eufriesea Mexicana, Cynoglossus semilaevis, Fopius arisanus, Pieris napi, Hypomesus transpacificus, Cotesia glomerata, Plectropomus leopardus, Anguilla anguilla, Megalopta genalis, Sphaeramia orbicularis, Myripristis murdjan, Mastacembelus armatus, Leptinotarsa decemlineata, Ephestia cautella, and Pseudomyrmex gracilis.

[0044] The pheromones disclosed herein are characterized by an unsaturated aliphatic carbon chain of length ranging from 6 to 24, such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, and 24, and terminally ending with an alcohol, aldehyde, or ester function group.

[0045] In one embodiment of the present disclosure, the pheromones are selected from an unsaturated C6 to C24 fatty alcohol, an unsaturated C6 to C24 fatty aldehyde, or an unsaturated C6 to C24 fatty acid ester.

[0046] The term “fatty alcohol”, as used herein, refers to an alcohol having an unsaturated aliphatic carbon chain of length ranging from 6 to 24, such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, and 24, and an alcohol functional group.

[0047] The term “fatty aldehyde”, as used herein, refers to an alcohol having an unsaturated aliphatic carbon chain of length ranging from 6 to 24, such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, and 24, and an aldehyde functional group.

[0048] The term “fatty acid ester”, as used herein, refers to an acid ester having an unsaturated aliphatic carbon chain of length ranging from 6 to 24, such as 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, and 24, and an ester functional group. In some embodiments, the ester group is an acetate, and the fatty acid esteris a fatty acyl acetate. The terms “fatty acid ester”, “fatty acyl acetate”, or “fatty acetate” are used interchangeably in the present disclosure.

[0049] In an embodiment of the present disclosure, the pheromones or precursors thereof is selected from the group consisting of (Z,Z,E)-3,6,8-dodecatrien-l-ol, (E,E)- 10, 14-hexadecadienal, (Z,Z)-9,2-tetradecadien- l-ol, (Z,Z)-9, 12- tetradecadienyl acetate, (Z,Z)-9,l l-tetradecadienylacetate, (Z,Z)-8,10- tetradecadienal, (Z,Z)-8,10-dodecadienyl acetate, (Z,Z)-l l,13-hexadecadienyl acetate, (Z,Z)-8,10-dodecadien-ol, (Z,Z)-l l,13-hexadecadien-l-ol, (Z,Z)-7,9- dodecadienyl acetate, (Z,Z)-10,12-hexadecadienal, (Z,Z)-7,9-dodecadien-l-ol, (Z,E)-10,12-hexadecadienal, (Z,Z)-7,l l-tridecadienyl acetate, (Z)- 11 -heptadecenyl acetate, (Z,Z)-5,9-tridecadienylacetate, (Z)-l l-heptadecen-l-ol, (Z,Z)-5,8- tetradecadienyl acetate, (Z,Z)-5,8-tetradecadien-l-ol, (Z,Z,Z)-9, 12,15- octadecatrienal, (Z,Z)-5,8-tetradecadienal, (Z,Z)-5,7-dodecadienal, (E,E)-10,12- hexadecadien-l-ol, (Z,Z)-4,7-tridecadienyl acetate, (Z)-8-heptadecen-l-ol, (Z,Z)- 4,7-tridecadien-l-ol, (E)-8-heptadecenylacetate, (Z,Z)-4, 7 -decadienyl acetate, (Z,Z)-9,11-pentadecadienal, (Z,Z)-4,7-decadien-l-ol, (E,Z)-9,11-pentadecadienal,(Z,Z)-3 ,8-dodecadien- 1 -ol, (Z,E)-7, 11-hexadecadienal, (Z,Z)-2,4-decadienal,(E,Z)-8, 10-pentadecadienyl acetate, (Z,Z)-9,l l-tetradecadienal, (Z,Z)-9,11- tetradecadien-l-ol, (Z,Z)-5,7-dodecadienyl acetate, (Z,Z)-9,1 1-hexadecadienal,(Z,E,E)-3,6,8-dodecatrien-l-ol, (Z,Z)-11,13-hexadecadienal, (Z,E)-9,11- tetradecadienyl acetate, (Z,E)-8,10-dodecadienyl acetate, (Z,E)-11,13- hexadecadienyl acetate, (Z,E)-7,9-dodecadienyl acetate (Z,E)-10,12- hexadecadienyl acetate, (Z,E)-7,9-dodecadien-l-ol, (E,Z)-10,2-hexadecadienal,(Z,E)-5,7-dodecadienyl acetate, (E,Z)-9,11-hexadecadienal, (Z,E)-5,7- dodecadienal, (Z,E)-9, 1-hexadecadienal, (Z,E)-3,5-tetradecadienyl acetate, (Z,Z)- 9,12-octadecadienal, (Z,E)-3,5-dodecadienyl acetate, (Z,E)-7, 11 -hexadecadienyl acetate, (Z,E)-3,5-decadienyl acetate, (Z,Z)- 8, 10-pentadecadienyl acetate, (Z,E)- 5,9-tridecadienyl acetate, (E)-lO-heptadecenyl acetate, (Z,E)-9,12-tetradecadienyl acetate, (Z,E)-9,12-tetradecadienal, (Z,E)-9,12-tetradecadien-l-ol, (Z,E)-9,11- tetradecadienal, (Z,E)-9, 11 -tetradecadien-l-ol, (Z,E)-8,10-tetradecadienyl acetate, (Z,E)-8 , 10-dodecadienal, (Z,E)- 1 , 13 -hexadecadienal, (Z,E)- 8 , 10-dodecadien- l-ol,(Z,E)- 11 , 13-hexadecadien- 1 -ol, (Z,E)-5,7 -dodecadien- 1 -ol, (E,Z)-9, 1 - hexadecadienyl acetate, (Z,E)-8,10-tetradecadien-l-ol, (Z)-S-tetradecenyl acetate, (Z)-13-octadecen-l-ol, (Z)-lO-tridecenyl acetate, (E,E,Z,Z)-4,6,11,13- hexadecatetraenal, (Z)-lO-tetradecenyl acetate, (Z,Z)-2,13-octadecadien-l-ol, (Z)- 10-dodecenyl acetate, (Z,Z)-7,10-hexadecadienyl acetate, (Z)-9-undecenyl acetate, (E)-6-hexadecenyl acetate, (Z)-9-tridecenyl acetate, (E, E,Z)-10, 12,14- hexadecatrienal, (Z)-9-tetradecenyl acetate, (E,Z)-2,13-octadecadienyl acetate, (Z)-9-tetradecenal, (E,Z)-2,13-qctadecadienal, (Z)-9-tetradecen-l-ol, (Z,E)-7,9- dodecadienyl acetate, (Z,E)-10,12-hexadecadienyl acetate, (Z,E)-7,9-dodecadien- l-ol, (Z,E)-5, 7 -dodecadienyl acetate, (Z,E)-3,5-tetradecadienyl acetate, (Z,E)-3,5- dodecadienyl acetate, (Z,E)-7,l l-hexadecadienyl acetate, (Z,E)-3,5-decadienyl acetate, (Z,Z)-8,10-pentadecadienyl acetate, (Z,E)-5,9-tridecadienyl acetate, (E)-10-heptadecenyl acetate, (Z,E)-9,12-tetradecadienyl acetate, (Z,E)-9,12- tetradecadien-l-ol, (Z,E)-9,11-tetradecadien-l-ol, (Z,E)-8,10-tetradecadienyl acetate, (Z,E)-l l,13-hexadecadienal, (Z,E)-8,10-dodecadien-l-ol, (Z,E)-11,13- hexadecadien-l-ol, (Z,E)-5,7-dodecadien-l-ol, (E,Z)-9,1 -hexadecadienyl acetate, (Z,E)-8,10-tetradecadien-l-ol, (Z)-S -tetradecenyl acetate, (Z)-13-octadecen-l-ol, (Z)-lO-tridecenyl acetate, (Z)-lO-tetradecenyl acetate, (Z,Z)-2,13-octadecadien-l- ol, (Z)-lO-dodecenylacetate, (Z,Z)-7,10-hexadecadienyl acetate, (Z)-9-undecenyl acetate, (E)-6-hexadecenyl acetate, (Z)-9-tridecenyl acetate, (Z)-9-tetradecenyl acetate, (E,Z)-2,13-octadecadienyi acetate, (E,Z)-2,13-octadecadienal, (Z)-9- tetradecen-l-ol, (E)-9-hexadecenal, (Z)-5-dodecen-l-ol, (E)-9-hexadecenol-ol, (Z)-5-decenyl acetate, (E)-12-pentadecenyl acetate, (Z)-5-decen-l-ol, (Z)-10- pentadecenal, (Z)-4-tridecenyl acetate, (E,Z,Z)-4,6,10-hexadecatrienyl acetate, (Z)- 4-tridecenal, (E,E,Z)-4,6,10-hexadecatrienyl acetate, (Z)-4-decenyl acetate, (Z)-8- pentadecenyl acetate, (Z)-4-decenal, (Z)-9-pentadecenyl acetate, (Z)-3-tetradecenyl acetate, (E)-2-octadecenal, (Z)-3-Tetradecen-l-ol, (E)-2-octadecenyl acetate, (Z)- 3-dodecenyl acetate, (Z)-7-hexadecen-l-ol, (Z)-3-dodecen-l-ol, (E)-7-hexadecenyl acetate, (Z)-2-tridecenyl acetate, (E,E,Z)-4,6,10-hexadecatrien-ol, (Z)-12- tetradecenyl acetate, (E,E)-5,9-octadecadien-l-ol, (Z)- 11 -tridecenyl acetate, (Z)-2- heptadecenal, (Z)- 11 -tetradecenyl acetate, (Z,E)-3,13-octadecadienyl acetate, (Z)-11-tetradecenal, (Z,Z)-3, 13 -octadecadienyl acetate, (Z)-lO-dodecen-l-ol, (Z,Z)- 7,10-hexadecadien-l-ol, (Z)-7-undecenyl acetate, (Z)-5-hexadecen-l-ol, (Z)-5- decenal, (Z)-12-pentadecenyl acetate, (Z)-l l-tetradecen-l-ol, (E,Z)-3,13- octadecadienal, (E,Z,Z)-4,7,10-tridecatrienyl acetate, (E,E)-4,8-heptadecadienyl acetate, (E,Z)-9, 11 -tetradecadienyl acetate, (E,Z)-8,10-tetradecadienyl acetate, (E,Z)-8,10-tetradecadienal, (E,Z)-8,10-dodecadienyl acetate, (E,Z)-11,13- hexadecadienyl acetate, (E,Z)-8,10-dodecadienal, (E,Z)-l l,3-hexadecadienal, (E,Z)-8,10-dodecadien-l-ol, (E,Z)-11,3-hexadecadien-l-ol, (E,Z)-7,9-dodecadienyl acetate, (E,Z)-10,12-hexadecadien-l-ol, (E,Z)-7,9-dodecadienal, (E,Z)-10,12- hexadecadienyl acetate, (E,Z)-5,9-tridecadienyl acetate, (Z)-9-heptadecenal, (E,Z)- 5,7-dodecadienyl acetate, (E,Z)-8,l l-hexadecadienal, (E,Z)-5,7-dodecadienal, (E,E)-9,l l-hexadecadienal, (E,Z)-4,9-tetradecadienyl acetate, (Z,Z)-13,15- octadecadienal, (E,Z)-4,9-tetradecadienal, (Z,Z,Z)-3,6,9-octadecatrienyl acetate, (E,Z)-4,7-tridecadienyl acetate, (E)-8-heptadecen-l-ol, (E,Z)-4,10-tetradecadienyl acetate, (E,E,E)-9,2,5-octadecatrien-l-ol, (E,Z)-3,8-tetradecadienyl acetate, (E,E)- 11,14-octadecadienal, (E,Z)-3,5-tetradecadienyl acetate, (Z,Z)-9,12-octadecadienyl acetate, (E,Z)-3,5-dodecadienyl acetate, (Z,E)-7,l l-hexadecadien-l-ol, (E,Z)-2,4- decadienal, (E,Z)-8,10-pentadecadien-l-ol, (E,Z)-7,9-dodecadien-l-ol, (E,E)- 10, 12-hexadecadienal, (E,Z)-5,7 -dodecadien- l-ol, (Z,Z)-8, 10-hexadecadienyl acetate, (E,Z)-3,7-tetradecadienyl acetate, (Z,Z)-l l,13-octadecadienal, (E,E,E)-10,12, 14-hexadecatrienyl, (E,E)-9, 11 -tetradecadienyl acetate, (E,E)-8, 10- dodecadienyl acetate, (E,E)-l,13-hexadecadienyl acetate, (E,E)-7,9- dodecadienylacetate, (E,E)-10,12-hexadecadienyl acetate, (E,E)-5,8- tetradecadienal, (Z,Z,Z)-9,12,15-octadecatrienyl acetate, (E,E)-5,7-dodecadienyl acetate, (Z,Z)-7,l l-hexadecadienal, (E,E)-5,7-dodecadien-l-ol, (Z,Z)-7,1- hexadecadienyl acetate, (E,E)-4,10-dodecadienyl acetate, (Z,Z)-7,l-hexadecadien- l-ol, (E,E)-3,5-tetradecadienyl acetate, (E,E)-9,12-octadecadien-l-ol, (E,E)-3,5- decadienylacetate, (Z,E)-8,10-pentadecadienyl acetate, (E, E,Z)-10, 12,14- hexadecatrienyl, (E,E)-9,2-tetradecadienyl acetate, (E,E)-8,10-tetradecadienyi acetate, (E,E)-8,10-tetradecadienal, (E,E)-8,10-dodecadien-l-ol, (E,E)-11,13- hexadecadien-l-ol, (E,E)-8,10-dodecadienal, (E,E)-l l,13-hexadecadienal, (E,E)-2,4-tetradecadienal, (E,E)-5,9-octadecadienyl acetate, (E,E)-2,4-decadienal, (E,E)-8.10-pentadecadienyl acetate, (E)-5-tridecenyl acetate, (E,E,Z)-4,6,10- hexadecatrienyl acetate, (E)-5-tetradecenal, (E)-9-octadecen-l-ol, (E)-5- tetradecen-l-ol, (Z)-2-octadecenyl acetate, (E)-5-dodecenyl acetate, (E)-5- hexadecenyl acetate, (Z)-lO-pentadecenyl acetate, (E)-lO-tetradecenyl acetate, (Z, E)-2,13-octadecadienyl acetate, (E)-lO-dodecenyl acetate, (E,Z)-6,11- hexadecadienyl acetate, (E)-lO-dodecenal, (E,Z)-6,l l-hexadecadienal, (E)-9- tridecenyl acetate, (E)-9-tetradecen-l-ol, (Z)-13-octadecenal, (E)-9-dodecenal, (Z)- 14-hexadecenyl acetate, (E)-9-dodecen-l-ol, (Z)-12-hexadecenal, (E)-9- tetradecenyl acetate, (E)-14-octadecenal, (E)-9-dodecenyl acetate, (E)-14- hexadecenal, (E)-8-tridecenyl acetate, (E)-8-tetradecenyl acetate, (E)-13- octadecenyl acetate, (E)-8-Dodecenyl acetate, (Z)-9-hexadecen-l-ol, (Z)-l l- hexadecen-l-ol, (E)-8-dodecenal, (Z)- 11 -hexadecenyl acetate, (E)-8-dodecen-l-ol, (E)-l l-hexadecenal, (E)-8-decen-l-ol, (Z,Z)-6,9-pentadecadienal, (E)-7- tetradecenyl acetate, (E)-l l-octadecenal, (E)-7-tetradecen-l-ol, (E)-l l-octadecen- l-ol, (E)-7 -dodecenyl acetate, (E)-lO-hexadecenal, (E)-7-dodecenal, (Z)-10- hexadecenyl acetate, (E)-7-dodecen-l-ol, (E)-lO-hexadecen-l-ol, (E)-7-decenyl acetate, (Z,Z)-8,9-pentadecadien-l-ol, (E)-6-tridecenyl acetate, (E,E,Z)-4,6,11- hexadecatrienyl acetate, (E)-6-tetradecenyl acetate, (Z)-9-octadecenyl acetate, (E)- 6-dodecenal, (Z)-9-hexadecenal, (E)-6-dodecen-l-ol, (Z)-9-hexadecen-l-ol, (E)-5- dodecen-l-ol, (Z)-7-hexadecenal, (E)-5-decen-l-ol, (E)-9-pentadecenyl acetate, (E)-5-tetradecenyl acetate, (Z)-2-octadecenal, (E)-4-tridecenyl acetate, (E,Z,Z)-4.6.10-hexadecatrien-l-ol, (E)-4-dodecenyl acetate, (Z)-7-hexadecenyi acetate, (E)-4-decenyl acetate, (Z)-8-pentadecen-l-ol, (E)-3-tetradecenyl acetate, (Z,Z)- 8, 11 -heptadecadienyl acetate, (E)-3-tetradecen-l-ol, (Z,Z)-8,10-heptadecadien-l- ol, (E)-3-dodecenyl acetate, (E)-7-hexadecenal, (E)-2-undecenyl acetate, (Z)-3- hexadecenyi acetate, (E)-2-undecenal, (E)-5-hexadecen-l-ol, (E)-2-tridecenyl acetate, (Z,E)-l,14-hexadecadienyl acetate, (E)-2-dodecenal, (E)-7-hexadecen-l- ol, (E)-12-tetradecenyl acetate, (Z,Z)-3,13-octadecadienal, (E)- 11 -tetradecenyl acetate, (E,E)-3, 13 -octadecadienyl acetate, (E)-lO-dodecen-l-ol, (E,Z)-4,6- hexadecadienal, (E)-l-tetradecen-l-ol, (Z,Z)-2,13-octadecadienyl acetate, (E)-l l-tridecenyl acetate, (E)-2-heptadecenal, (E)-l l-tetradecenal, (E,Z)-3, 13- octadecadienyl acetate, (E,E)-8,10-tetradecadien-l-ol, (Z)-9-tetradecenol, (E)-l l- hexadecenol, (Z)- 11 -hexadecenol, (E,E)-10,12-hexadecadienol, (Z)-l l- hexadecenal, (Z)-9-tetradecenyl acetate, (E,Z,Z)-3,8,l l-tetradecatrienyl acetate (TDTA), (Z,E)-9, 11 -tetradecadienyl acetate, (Z,E)-12,9-tetradecadienyl acetate, (E)-l 1 -hexadecenyl acetate, (Z)-l 1 -hexadecenyl acetate, (Z,E)-7,11 -hexadec adien- 1-yl acetate, (Z,Z)-7,l l-hexadecadien-l-yl acetate, (Z)- 13 -octadecenyl acetate, (Z)-7-dodecenyl acetate, (E,Z)-7,9-dodecadienyl acetate, (E,Z)-10,12- tetradecadienyl acetate, (Z,E)- 11,11 -tetradecadienyl acetate, (E)-2-decenal, (Z)-2- decenal, (Z)-5-dodecenal, (Z)-7-dodecenal, (Z)-9-dodecenal, (E)-lO-dodecenal, (Z)-4-tridecenal, (Z)-5-tetradecenal, (Z)-7-tetradecenal, (Z)-8-tetradecenal, (Z)-l l- tetradecenal, (E,E)-10,12-tetradecadienal, (Z)-lO-hexadecenal, (E)-7-hexadecenal, (Z,E)-9,11-hexadecadienal, (Z,Z)-9,11-hexadecadienal, (E,Z)-10,12- hexadecadienal, (Z,E)-10,12-hexadecadienal, (E,Z)-11,13-hexadecadienal, (E,E,Z)-4,6,11-hexadecatrienal, (E,E,E)-10,12,14-hexadecatrienal, (Z)-2- heptadecenal, (E)-9-octadecenal, (Z)-9-octadecenal, (Z)-l l-octadecenal, (E)-13- octadecenal, undecanal, (Z,Z)-9,11-hexadecadienal, (E,E,Z)-4,6,11- hexadecatrienal, (Z)-6,14-pentadecadienal, (Z)-9,13-tetradecadien-l l-ynal, (Z)-13- hexadecen-l l-ynal, (Z)-hexadec-9-enal, (Z)-hexadec-l l-enal, (11Z, 13Z)- hexadeca-l l,13-dienal, (10E, 12E)-hexadeca-10,12-dienal, (E)-hexadec-lO-enal, (Z)-octadec-13-enal, and (7Z, 11Z, 13E)-hexadeca-7,l l,13-trienal. In another embodiment of the present disclosure, the pheromone or precursors thereof is selected from (Z)-hexadec-9-enal, (Z)-hexadec-l l-enal, (11Z, 13Z)-hexadeca- 11,13-dienal, (10E, 12E)-hexadeca-10,112-dienal, (E)-hexadec-lO-enal, (Z)- octadec-13-enal, (7Z, 11Z, 13E)-hexadeca-7,l l,13-trienal, or combinations thereof.

[0050] In an embodiment of the present disclosure, the mono- or poly-unsaturated C6-C24 fatty alcohols is selected from (E)-2-decenol, (Z)-2-decenol, (Z)-4-decenol, (Z)-5-decenol, (E,E)-2,4-decadienol, (E,Z)-2,4-decadienol, (Z,Z)-2,4-decadienol, (E)-2-undecenol, (E)-2-dodecenol, (Z)-5-dodecenol, (E)-6-dodecenol, (Z)-7- dodecenol, (E)-8-dodecenol, (E)-9-dodecenol, (Z)-9-dodecenol, (E)-lO-dodecenol,(E,Z)-5,7-dodecadienol, (Z,E)-5,7-dodecadienol, (Z,Z)-5,7-dodecadienol, (E,Z)-7.9-dodecadienol, (E,E)-8,10-dodecadienol, (E,Z)-8,10-dodecadienol, (Z,E)-8,10- dodecadienol, (Z) -4 -tridecenol, (E)-5-tetradecenol, (Z)-5-tetradecenol, (Z)-7- tetradecenol, (Z)- 8 -tetradecenol, (E)- 11 -tetradecenol, (Z)- 11 -tetradecenol, (E,E)- 2,4-tetradecadienol, (E,Z)-4,9-tetradecadienol, (E,E)-5,8-tetradecadienol, (Z,Z)- 5,8-tetradecadienol, (E,E)-8,10-tetradecadienol, (E,Z)-8,10-tetradecadienol, (Z,Z)-8.10-tetradecadienol, (Z,E)-9,11 -tetradecadienol, (Z,Z)-9,11 -tetradecadienol,(Z,E)-9,12-tetradecadienol, (E,E)-10,12-tetradecadienol, (Z)-lO-pentadecenol, (Z,Z)-6,9-pentadecadienol, (E,Z)-9,11 -pentadecadienol, (Z,Z)-9,11- pentadecadienol, (Z)-9-hexadecenol, (E)-lO-hexadecenol, (Z)-lO-hexadecenol, (E)- 11 -hexadecenol, (Z)- 11 -hexadecenol, (Z)-12-hexadecenol, (E)-14- hexadecenol, (E)-7-hexadecenol, (Z)-7-hexadecenol, (E)-9-hexadecenol, (E,Z)- 4,6-hexadecadienol, (E,Z)-6, 11 -hexadecadienol, (Z,E)-7, 11 -hexadecadienol, (Z,Z)-7,11 -hexadecadienol, (E,Z)-8,11 -hexadecadienol, (E,E)-9,11- hexadecadienol, (E,Z)-9, 11 -hexadecadienol, (Z,E)-9, 11 -hexadecadienol, (Z,Z)-9.11 -hexadecadienol, (E,E)-10,12-hexadecadienol, (E,Z)-10,12-hexadecadienol, (Z,E)-10,12-hexadecadienol, (Z,Z)-10,12-hexadecadienol, (E,E)-11,13- hexadecadienol, (E,Z)-11,13 -hexadecadienol, (Z,E)-11,13 -hexadecadienol, (Z,Z)-11.13 -hexadecadienol, (E,E)-10,14-hexadecadienol, (E,E,Z)-4,6,11- hexadecatrienol, (E,E,E)-10,12,14-hexadecatrienol, (E,E,Z)- 10, 12,14- hexadecatrienol, (E,E,Z,Z)-4,6,l l,13-hexadecatetraenol, (E)-2-heptadecenol, (Z)- 2-heptadecenol, (Z)-9-heptadecenol, (E)-2-octadecenol, (E)-9-octadecenol, (Z)-9- octadecenol, (E)- 11 -octadecenol, (Z)- 11 -octadecenol, (E)-13-octadecenol, (Z)-13- octadecenol, (E)-14-octadecenol, (E,Z)-2,13-octadecadienol, (E,Z)-3,13- octadecadienol, (Z,Z)-3,13-octadecadienol, (Z,Z)-9,12-octadecadienol, (Z,Z)-11.13 -octadecadienol, (E,E)-11,14-octadecadienol, (Z,Z)-13,15-octadecadienol,(Z,Z,Z)-9, 12, 15-octadecatrienol,(Z)-9-hexadecenol,(Z)-13-octadecenol, undecanol, (E)-7-dodecenol, (E)-l 1 -tetradecenol, (Z)-5-tetradecenol, (Z)-9-tetradecenol, (Z,Z,E)-7,11,13 -hexadecatrienol, (Z)-6,14-pentadecadienol, (Z)-9,13- tetradecadien-l l-ynol, (Z)-13-hexadecen-l l-ynol, or combinations thereof.

[0051] Embodiments herein include a process for producing pheromones or precursors thereof.

[0052] In an embodiment, the process comprising: a) obtaining a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; contacting the first yeast cell with a first medium comprising a first substrate to obtain a first reaction mixture comprising mono- or poly-unsaturated C6-C24 fatty acid; wherein the first yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell; wherein the first substrate is a saturated or unsaturated C6-C24 fatty acid or derivative thereof; b) obtaining a second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme; contacting the second yeast cell with a second medium comprising the first reaction mixture or a second substrate to obtain a second reaction mixture comprising mono- or poly-unsaturated C6-C24 fatty alcohol; wherein the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell; wherein the second substrate is a mono or poly unsaturated C6-C24 fatty acid or derivative thereof; and c) mixing an aqueous solution of laccase enzyme, the second reaction mixture, a mediator, and an emulsifying agent under a continuous supply of a gas, to obtain the pheromones or precursors thereof.

[0053] In an embodiment of the present disclosure, there is provided a process for producing mono- or poly- unsaturated C6-C24 fatty acid or derivative thereof, said process comprising: obtaining a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; contacting the first yeast cell with a first medium comprising a first substrate to obtain mono- or poly-unsaturated C6- C24 fatty acid; wherein the first yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell; and wherein the first substrate is a saturated or unsaturated C6-C24 fatty acid or derivative thereof.

[0054] In an embodiment of the present disclosure, there is provided a process for producing mono- or poly-unsaturated C6-C24 fatty alcohol, said process comprising: obtaining a second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme; contacting the second yeast cell with a second medium comprising a second substrate to obtain mono- or poly-unsaturatedC6-C24 fatty alcohol; wherein the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell; and wherein the second substrate is a mono or poly unsaturated C6-C24 fatty acid or derivative thereof.

[0055] In an embodiment of the present disclosure, wherein the yeast cell is selected from a group consisting of Hansenula polymorpha, Kluyveromyces lactis, Komagataella pastoris, Komagataella phaffii (Pichia pastoris), Komagataella pseudopastoris, Saccharomyces cerevisiae, Saccharomyces bayanus, Saccharomyces pastorianus, Saccharomyces uvarum, Saccharomyces cerevisiae (var. diastaticus), Saccharomyces kudravzevii, Saccharomyces mikatae, Saccharomyces paradoxus, Schizosacchromyces pombe, and Yarrowia lipolytica', and the yeast cells are present in an amount in a range of 20 wt% to 80 wt%.

[0056] In another embodiment of the present disclosure, the yeast cell is preferably selected from Saccharomyces cerevisiae or Pichia pastoris.

[0057] In an embodiment, the yeast cell is a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme or a a second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme.

[0058] The term “heterologous gene”, as used herein, refers to a gene encoding a polypeptide, such as a protein or an enzyme, which shall herein be understood to be a polypeptide, which is not naturally present in a wild type yeast cell.

[0059] In an embodiment of the present disclosure, wherein the amino acid sequence of the fatty acid desaturase is selected from the group consisting of SEQ ID NO.17, SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20, SEQ ID NO.21, SEQ ID NO.22, SEQ ID NO.23, and SEQ ID NO.24; and the amino acid sequence of the fatty acid reductase is selected from the group consisting of SEQ ID NO.25, SEQ ID NO.26, SEQ ID NO.27, SEQ ID NO.28, SEQ ID NO.29 SEQ ID NO.30, SEQ ID NO.31, and SEQ ID NO.32.

[0060] In an embodiment of the present disclosure, the gene encoding the fatty acid desaturase is selected from SEQ ID NO.l, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5, SEQ IN NO.6, SEQ ID NO.7, or SEQ ID NO.8; and the gene encoding said fatty acid reductase is selected from SEQ ID NO.9, SEQ ID NO.10,SEQ ID NO.11, SEQ ID NO.12, SEQ ID NO.13, SEQ ID NO.14, SEQ ID NO.15, or SEQ ID NO.16.

[0061] In some embodiments, the heterologous FAD enzyme is selected from the enzymes listed in Table 1.

[0062] Table 1

[0063] In some embodiments, the heterologous FAR enzyme is selected from the group consisting of enzymes listed in Table 2.

[0064] Table 2

[0065] The introduction of heterologous genes into yeast cells can be performed using various methods well-known in the art. In an embodiment, the heterologous gene is introduced into the yeast cell through a vector, such as a plasmid. In another embodiment, the vector is selected from pYES or pPICZaA. The gene encoding the heterologous desaturase and the heterologous FAR may further be codon optimized for the yeast cell using method generally known in the art. All such optimizations and improvements are understood to be included within the scope of the present invention.

[0066] The yeast cell comprising the heterologous gene is incubated under suitable conditions, for eg: in an appropriate medium and at an appropriate temperature. In an embodiment of the present disclosure, the first medium and the second medium comprises a carbon source selected from glucose, galactose, raffinose, glycerol, or a combination thereof.

[0067] In an embodiment of the present disclosure, the first medium and the second medium comprises a carbon source selected from glucose, galactose, raffinose, glycerol, or a combination thereof.

[0068] Suitable media comprising a carbon source that supports yeast growth and aids in the selection of those yeast cells comprising the heterologous gene are known in the art and may be used including, but not limited to, BMGY (Buffered Glycerol-complex Medium), BMMY (Buffered Methanol-complex Medium),YPDM (Yeast Extract Peptone Dextrose Medium), MGY (Minimal Glycerol Medium), RD (Regeneration Dextrose Medium), MD (Minimal Dextrose Medium), MM (Minimal Methanol Medium), BMG (Buffered Minimal Glycerol), or SD (synthetic minimal defined medium).

[0069] In an embodiment of the present disclosure, wherein the first medium or the second medium further comprises water or buffer.

[0070] Various buffer compositions are generally known in the art, any of which may be used in embodiments herein. In one embodiment, the buffer is selected from sodium citrate buffer, sodium phosphate buffer, potassium phosphate buffer, sodium acetate buffer, or combinations thereof.

[0071] In an embodiment of the present disclosure, the contacting is performed in a stirred tank reactor in batch mode. The term “contacting”, as used herein refers to culturing or reacting the yeast cell with the medium.

[0072] In an embodiment of the present disclosure, the contacting is carried out at a temperature ranging from 20 to 40°C for a duration ranging from 48 to 120 hours.

[0073] A person of skill in the art, in light of the disclosures herein, would be able to appropriately choose the desaturase and corresponding reductase, and various combinations thereof, for expressing in the yeast cell to produce the desired pheromones or precursors thereof.

[0074] In embodiment of the present disclosure, the first yeast cell or the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell or a permeabilized second yeast cell. In another embodiment, the first yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell. In yet another embodiment, the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell.

[0075] The term “permeabilized first yeast cell”, or “permeabilized second yeast cell”, as used herein, refers to a yeast cell with altered permeability of the cell membrane, which facilitates the diffusion of endogenous small molecules, substrates, products, enzymes, etc., in good yield into the medium, while also maintaining properties of the cells, such as their enzyme expression and enzymatic activity. The permeabilized yeast cells are spared from harsh chemical treatmentsconventionally carried out in the art for cell disruption processes, in order to release various substances into the medium.

[0076] In an embodiment of the present disclosure, the yeast cell and the permeabilizing agent in a weight ratio in the range of 0.1:1 to 0.01:8 is contacted for a duration in the range of 5 min to 48 hours. In another embodiment of the present disclosure, the yeast cell and the permeabilizing agent in a weight ratio in the range of 0.1:1 to 0.01:8 is contacted for a duration in the range of 8 min to 40 min.

[0077] In an embodiment of the present disclosure, the permeabilizing agent is selected from the group consisting of chloroform, cetyltrimethylammonium bromide (CTAB), Triton X-100, Tween 80, N-lauroylsarcosine, N-lauroylsarcosine sodium salt (sarkosyl / Sodium lauroyl sarcosinate), sorbitol, methanol, ethanol, 1- propanol, isopropyl alcohol, 2-propanol, 1-butanol, dimethyl sulfoxide (DMSO), toluene, benzene, benzene acetone, ethyl acetate, ethyl ether, acetone, benzalkonium chloride, and polyoxyethylene(23)lauryl ether; and the permeabilizing agent is in an amount in a range of 0.04 wt% to 80 wt%.

[0078] In an embodiment the permeabilizing agent is cetyltrimethylammonium bromide (CTAB) in an amount in the range of 0.04 wt % to 2 wt %.

[0079] In an embodiment, the permeabilizing agent is ethanol in an amount in the range of 20 wt% to 80 wt%. In another embodiment, the permeabilizing agent is ethanol in an amount in the range of 40 wt% to 60 wt%.

[0080] In embodiment of the present disclosure, the permeabilized first yeast cell or the permeabilized second yeast cell is contacted with a carrier to obtain an immobilized and permeabilized first yeast cell or an immobilized and permeabilized second yeast cell.

[0081] The term “immobilized yeast cells”, as used herein, refers to the physical confinement of yeast cells in or on a carrier for their stability and functional reuse. By employing this technique, yeast cells are made more efficient and cost-effective for reuse, while retaining the biological activity of the yeast cells. The yeast cells may be immobilized in or on a carrier by various methods known in the art, such as adsorption, aggregation, confinement, and entrapment.

[0082] In an embodiment of the present disclosure, the process disclosed herein further comprises contacting the permeabilized yeast cell with a carrier in a weight ratio in the range of 1 : 2 to 1 : 8 for a period in the range of 5 min to 48 h to obtain an immobilized and permeabilized yeast cell. In another embodiment of the present disclosure, the process disclosed herein further comprises contacting the permeabilized first yeast cell with a carrier in a weight ratio in the range of 1 : 2 to 1 : 8 for a period in the range of 5 min to 48 h to obtain an immobilized and permeabilized first yeast cell. In another embodiment of the present disclosure, the process disclosed herein further comprises contacting the permeabilized second yeast cell with a carrier in a weight ratio in the range of 1 : 2 to 1 : 8 for a period in the range of 5 min to 48 h to obtain an immobilized and permeabilized second yeast cell.

[0083] Examples of carrier that may be used in the present disclosure include but are not limited to from glutaraldehyde, silica sol-gel, sodium silicate, calcium alginate, agarose, polyethylene amine, ceramics, functional glass, activated carbon, diatomaceous earth, plastic, rubber, fiber, and foam. In an embodiment of the present disclosure, the carrier is selected from glutaraldehyde, silica sol-gel, or combinations thereof; and the carrier is in an amount in a range of 20 wt% to 80 wt%.

[0084] In an embodiment, the immobilizing agent is glutaraldehyde in an amount in a range of 20 wt% to 60 wt%.

[0085] In an embodiment, the immobilizing agent is silica-sol-gel in an amount in a range of 40 wt% to 80 wt%.

[0086] In an embodiment of the present disclosure, there is provided a process for producing mono- or poly- unsaturated C6-C24 fatty acid or derivative thereof, said process comprising: obtaining a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; contacting the first yeast cell with a first medium comprising a first substrate at a temperature ranging from 20 to 40°C for a period ranging from 48 to 120 hours to obtain mono- or poly-unsaturated C6- C24 fatty acid; wherein the first yeast cell is contacted with a permeabilizing agentto obtain a permeabilized first yeast cell; and wherein the first substrate is a saturated or unsaturated C6-C24 fatty acid or derivative thereof.

[0087] In an embodiment of the present disclosure, there is provided a process for producing mono- or poly-unsaturated C6-C24 fatty alcohol, said process comprising: obtaining a second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme; contacting the second yeast cell with a second medium comprising a second substrate at a temperature ranging from 20 to 40°C for a period ranging from 48 to 120 hours to obtain mono- or poly-unsaturated C6-C24 fatty alcohol; wherein the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell; and wherein the second substrate is a mono or poly unsaturated C6-C24 fatty acid or derivative thereof.

[0088] The term “substrate” refers to the saturated or unsaturated C6 to C24 fatty acid, including C6, C7, C8, C9, CIO, Cl l, C12, C13, C14, C15, C16, C17, C18, C19, C20, C21, C22, C23, and C24 fatty acid, or derivative thereof, converted to the desired pheromones, or precursors thereof, in the process disclosed herein.

[0089] According to embodiments herein, the substrate is a first substrate or a second substrate.

[0090] In an embodiment of the present disclosure, the first substrate is selected from the group consisting of dodecanoic acid, tetra decanoic acid, (E)-3- tetradecenoic acid, (Z)-8-tetradecenoic acid, (Z)- 11 -tetradecenoic acid, (E)-l l- tetradecenoic acid, (Z)-9-tetradecenoic acid, (E)-12- tetradecenoic acid, hexadecanoic acid, (Z) -7 -hexadecenoic acid, (E)-l l- hexadecenoic acid, (Z)-l l- hexadecenoic acid, (E)-lO-hexadecenoic acid, (E)-12-hexadecenoic acid, octadecanoic acid, (E)-9-octadecenoic acid, (Z)-7-octadecenoic acid, eicosanoic acid, (E)-7-eicosenoic acid, and (Z)-9-eicosenoic acid; and wherein the second substrate is selected from the group consisting of dodecanoic acid, tetra decanoic acid, hexadecanoic acid, octadecanoic acid, (E)-7-dodecenoic acid, (E)-8- dodecenoic acid, (Z)-9-dodecenoic acid, (E)-lO-dodecenoic acid, (E)-3- tetradecenoic acid, (Z)- 8 -tetradecenoic acid, (Z)-9-tetradecenoic acid, (E)-l l- tetradecenoic acid, (E)-12-tetradecenoic acid, (E,Z,Z)-3,8,11- tetradecatrienoicacid, (Z,E)-9, 11 -tetradecadienoic acid, (Z,E)-9,12- tetradecadienoic acid, (Z)-9- hexadecenoic acid, (E)-lO-hexadecenoic acid, (E)- 11 -hexadecenoic acid, (Z)ll- hexadecenoic acid, (Z,E)-7,11- hexadecadienoic acid, (Z,Z)-7,l l-hexadecadienoic acid, (E,E)-10, 12-15 hexadecadienoic acid, (Z)- 13 -hexadecenoic acid, and (Z)-13- eicosenoic acid.

[0091] In some embodiments, the substrate and the permeabilized yeast cell are reacted at a temperature ranging from 25 to 35 °C for a period ranging from 24 to 72 hours.

[0092] In one embodiment of the present disclosure, the first yeast cell in the process disclosed herein expresses a desaturase derived from E. cautella, wherein the desaturase comprises a polypeptide encoded by a nucleotide of sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity to SEQ ID NO 1. In an embodiment, the first yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 1 is used in the production of (Z)-9-tetradecenoic acid.

[0093] In one embodiment of the present disclosure, the first yeast cell in the process disclosed herein expresses a desaturase derived from E. cautella, wherein the desaturase comprises a polypeptide encoded by a nucleotide of sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% sequence identity SEQ ID NO 2. In another embodiment, the first yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 2 is used in the production of (Z)-9-tetradecenoic acid.

[0094] In an embodiment, the first and the second yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 3 and SEQ ID NO 9, respectively is used in the production of (E,Z)-l l,l l-tetradecadien-l-ol.

[0095] In an embodiment, the first and the second yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 3 and SEQ ID NO 10, respectively is used in the production of (E,Z)-l l,l l-tetradecadien-l-ol.

[0096] In an embodiment, the first and the second yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 4 and SEQ ID NO 8, respectively is used in the production of (Z)-7-dodecenol.

[0097] In one embodiment of the present disclosure, the first yeast cell in the process disclosed herein expresses a desaturase derived from S. exigua, wherein the desaturase comprises a polypeptide encoded by a nucleotide of sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity to SEQ ID NO 5. In another embodiment, the first yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 5 is used in the production of (E)-12-tetradecenoic acid.

[0098] In one embodiment of the present disclosure, the second yeast cell in the process disclosed herein expresses a reductase derived from H. armigera, wherein the reductase comprises a polypeptide encoded by a nucleotide of sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity to SEQ ID NO 12. In another embodiment, the first yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 12 is used in the production of (Z)-9-hexadecenol.

[0099] In one embodiment of the present disclosure, the first yeast cell in the process disclosed herein expresses a desaturase derived from Bombyx mori, wherein the desaturase comprises a polypeptide encoded by a nucleotide of sequence having at least 80%, at least 85%, at least 90%, at least 95%, at least 98%, at least 99%, or 100% identity to SEQ ID NO 13. In another embodiment, the first yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 13 is used in the production of (E, Z)-10,12-hexadecadienol.

[0100] In one embodiment of the present disclosure, the yeast cell disclosed herein expresses a desaturase derived from S. frugiperda, wherein the desaturase comprises a polypeptide encoded by a nucleotide of sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99%, or 100% sequence identity to SEQ ID NO 6. In an embodiment, the yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 6 is used in the production of (Z)-9-tetradecenoic acid. In another embodiment, the yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 6 is used in the production of (Z)-9-hexadecenoic acid.

[0101] In one embodiment of the present disclosure, the yeast cell disclosed herein expresses a desaturase derived from S. frugiperda, wherein the desaturase comprises a polypeptide encoded by a nucleotide of sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99%, or 100% sequence identity to SEQ ID NO 7. In another embodiment, the yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 7 is used in the production of (Z)- 11 -hexadecenoic acid.

[0102] In one embodiment of the present disclosure, the yeast cell disclosed herein expresses a desaturase derived from S. frugiperda, wherein the desaturase comprises a polypeptide encoded by a nucleotide of sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99%, or 100% sequence identity to SEQ ID NO 8. In another embodiment, the yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 8 is used in the production of (Z)- 11 -hexadecenoic acid.

[0103] In one embodiment of the present disclosure, the yeast cell disclosed herein expresses a reductase derived from S. frugiperda, wherein the reductase comprises a polypeptide encoded by a nucleotide of sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99%, or 100% sequence identity to SEQ ID NO 14. In another embodiment, the yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 14 is used in the production of tetradecadiene- 1 -ol.

[0104] In one embodiment of the present disclosure, the yeast cell disclosed herein expresses a reductase derived from S. frugiperda, wherein the reductase comprises a polypeptide encoded by a nucleotide of sequence having at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99%, or 100% sequence identity to SEQ ID NO 15. In another embodiment, the yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 15 is used in the production of dodecen-l-ol.

[0105] In one embodiment of the present disclosure, the yeast cell disclosed herein expresses a reductase derived from S. frugiperda, wherein the reductase comprises a polypeptide encoded by a nucleotide of sequence having at least 95%, at least96%, at least 97%, at least 98%, at least 99%, at least 99%, or 100% sequence identity to SEQ ID NO 16. In another embodiment, the yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 16 is used in the production of hexadecen-l-ol.

[0106] In an embodiment, the first yeast cell and the second yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 6 and SEQ ID NO 16, respectively is used in the production of (Z,E)-9,12-tetradecadien-l-ol. In yet another embodiment, the first yeast cell and the second yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 6 and SEQ ID NO 16, respectively is used in the production of (Z)-9-hexadecenal.

[0107] In an embodiment, the first yeast cell and the second yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 7 and SEQ ID NO 16, respectively is used in the production of (Z)- 11 -hexadecen-l-ol.

[0108] In an embodiment, the first yeast cell and the second yeast cell comprising the nucleotide of sequence as set forth in SEQ ID NO 6 and SEQ ID NO 16, respectively is used in the production of (Z)-l 1 -hexadecen-l-ol.

[0109] In an embodiment of the present disclosure, the process further comprises recovering the yeast cell for reuse. In another embodiment of the present disclosure, the process further comprises recovering the permeabilized and immobilized yeast cell for reuse. In an embodiment of the present disclosure, the yeast cell is recovered by filtration, or centrifugation, or a combination thereof.

[0110] In some embodiments of the present disclosure, the immobilized yeast cells are capable of being reused 3 to 10 times in the process disclosed herein for the production of pheromones or precursors thereof.

[0111] In some embodiments, the precursors of pheromones obtained using the disclosed process may be modified further; for example, desaturated fatty alcohols may be converted to the corresponding desaturated fatty aldehydes or fatty acid esters by chemical or biochemical processes well known in the art. In some embodiments the desaturated fatty alcohol is modified to fatty acetates biochemically using the fatty acetyl transferase enzymes.

[0112] The existing processes for the conversion of alcohol to aldehyde form of pheromones results in aldehydes contaminated with other side products and requires tedious purification. Thus, there is still a need for an improved, cost- effective and environment friendly method for the preparation of aldehydes by oxidation of alcohol catalyzed by enzyme Laccase which obviates the drawbacks as discussed before.

[0113] In an embodiment of the present disclosure, there is provided a process for producing pheromones, or precursors thereof, said process comprising: mixing an aqueous solution of enzyme laccase and mono- or poly-unsaturated C6-C24 fatty alcohol followed by addition of a mediator and an emulsifying agent under a continuous supply of a gas, to obtain pheromones or precursors thereof.

[0114] In an embodiment of the present disclosure, the laccase enzyme and the mono- or poly-unsaturated C6-C24 fatty alcohol is in a weight ratio in a range of 0.1 : 1 to 1 :0.1. In another embodiment of the present disclosure, the laccase enzyme and the mono- or poly-unsaturated C6-C24 fatty alcohol is in a weight ratio in a range of 0.2:1 to 0.4:1

[0115] The enzyme laccase (benzenediol : oxygen oxidoreductase, EC 1.10.3.2), a blue multi-copper oxidase, has drawn considerable attention due to its nontoxic nature, high stability, and lack of substrate inhibition, and hence are ideal candidates for the conversion.

[0116] In an embodiment of the present disclosure, the laccase enzyme is sourced from Trametes versicolor, Trametes pubescens, Rhus vernicifera, Pyricularia oryzae, Agaricus bisporus, Mycelopthora termophila, or combinations thereof; and the enzyme is either in a soluble form or immobilized on a solid support.

[0117] In an embodiment of the present disclosure, the aqueous solution of laccase enzyme is obtained by dissolving 0.5 to 5g of laccase powder in 100-500 mL distilled water. In another embodiment of the present disclosure, wherein the aqueous solution of laccase enzyme is obtained by dissolving preferably 1 to 2 g of laccase powder in 200 mL of distilled water.

[0118] In an embodiment of the present disclosure, the aqueous solution of laccase enzyme and alcohol is obtained by dissolving 0.5 to 5g of laccase powder in 100-500 mL distilled water followed by mixing with 3 to 30g of alcohol and stirring at a temperature in the range of 10 °C - 60°C. In another embodiment of the present disclosure, stirring is carried out at a temperature in the range of 20-50°C. In yet another embodiment of the present disclosure, stirring is carried out at a temperature of 40°C.

[0119] In an embodiment of present disclosure, the laccases enzyme has an optimal temperature in a range of temperature from 25 to 40 °C, preferably in a range of 30- 35 °C and in the presence of air.

[0120] Laccase may be immobilized in several ways as known to a person skilled in the art. According to embodiments herein, laccase may be immobilized on several support materials, such as alginate beads, silica gel, zeolites, magnetic chitosan nanoparticles, and activated carbons using adsorption, covalent binding, entrapment, or cross-linking approaches. Immobilization of enzymes on solid structures can offer several benefits that soluble enzymes lack like high operational stability, product inhibition, easy workup, and recyclability of enzyme. Choosing suitable carrier materials is an essential factor for achieving successful immobilization technology. The most used materials are known to be divided into the inorganic, synthetic polymers, and biomass-derived materials. Inorganic materials include ceramics, functional glass, activated carbon, and diatomaceous earth. Synthetic polymer materials have the advantage that their pore size, specific surface area, and mass transfer performance can be controlled artificially, hence have been widely used as carriers to immobilize enzymes.

[0121] Combinations of TiCL-ZrCL (with TiCL: ZrCL molar ratio of 8:2) and TiCL- ZrCL -SiCL (with TiCL: ZrCL :SiO2 molar ratio of 8:1:1) oxide materials have been synthesized and used as supports for laccase immobilization. A combination of laccase and TiCh was used as a combined bio- and photo-catalysis materials. In most of the cases reported earlier, purified laccase was used and TiO2 was specially synthesised in the laboratory. However, increased purification steps and high cost are a few limitations of using purified laccase. On the other hand, the utilization of commercial TiO2 and semi-crude laccases for immobilization is economical. In present disclosure, commercially available crude laccase was immobilized on TiO2.Besides, the applications were mostly for dye removal which is usually present in ppm level. In the present case the substrates were used in milligram / L levels. Accordingly, in an embodiment of the present disclosure, the enzyme laccase has been immobilized on TiCh.

[0122] For the reactions where the substrate to be oxidized has a redox potential higher than laccase, or the substrate is too large to penetrate into the enzyme active site, the presence of a low-molecular weight chemical mediator is required to facilitate oxidative reactions. A mediator acts as a sort of ‘electron shuttle’, once it is oxidized by laccase, it diffuses away from the enzymatic pocket and in turn oxidizes any substrate that, due to its size, could not directly enter the enzymatic pocket, [(a) Galli, Cet.al J. Phys. Org. Chem.(2004) 17,973. (b) Morozova, O. V., et.al. Appl. Biochem. Microbiol. (2007) 43, 523. (c) Wells, Aet.al. Biochem. Soc. Trans. (2006), 34, 304. (d) Baiocco, P., et.al.,. Org.Biomol.Chem. (2003) 1, 191. (e). Li, K, et.al., Appl. Environ. Microbiol. (1999) 65,2654. (f) d’Acunzo, F., et.al, J. Biochem. (2002) 269, 5330.] The structures and abbreviated names of representative artificial laccase mediators are given below.

[0123] Structures of representative synthetic mediators:

[0124] Table 3

[0125] By using these ‘chemical mediators’, the redox potential of laccases can be extended which allows the oxidation of a wide range of non-phenolic substrates such as sugars, ethers, alkenes, amides, aromatic methyl groups, polycyclic aromatic hydrocarbons, lipids, and alcohols.

[0126] Various mediators are available [Chenjie Zhu, et al. Green Chem., 2014, 16, 1131-1138], however such efficient mediators are far too expensive to prepare the target aldehyde on a commercial scale. An environmentally compatible and sustainable approach leads toward aerobic oxidations with transition-metal catalysts (based on Pd, Ru, Fe, Cu, Pt, Au, Ir, Rh, etc.,) and dioxygen or hydrogen peroxide as oxidants. The use of molecular oxygen as a stoichiometric re-oxidant in combination with a catalytic metal has practical advantages due to the favorable economics associated with O2 and the formation of environmentally benign byproducts (water and hydrogen peroxide).

[0127] In an embodiment of the present disclosure, the mediator is an electron transfer reagent selected from 2,2,6,6-Tetramethylpiperidin-l-yl)oxyl (TEMPO) hydroxybenzotriazole (HOBT), 2,2’-azinobis-(3-ethylbenzothiazoline-6-sulfonate (ABTS), or combinations thereof.

[0128] In an embodiment of the present disclosure, the mediator and the alcohol is in a mole ratio in a range of 0.50: 1 to 0.10: 1. In another embodiment of the present disclosure, the mediator and the alcohol is in a mole ratio in a range of 0.50:1 to 0.10:10.15:0.30

[0129] Accordingly, a scalable, catalytic copper / TEMPO based oxidation process is developed using air as the terminal oxidant which selectively yields aldehyde as the product preventing over-oxidation. However, the cost for conversion of theintermediate alcohol to the final aldehyde can be high due to non- stoichiometric yield and expensive catalyst. An alternative methodology is hence desirable.

[0130] Accordingly, the present disclosure provides an efficient and environment friendly process for the preparation of aldehydes involving the following steps: mixing an aqueous solution of enzyme laccase and alcohol at select ratio, adding mediator and emulsifying agent followed by stirring under a continuous supply of oxygen or air till the conversion of alcohol to aldehyde is complete and extracting and purifying the crude aldehyde to obtain high purity aldehyde, wherein the process is carried out in an aqueous buffer in pH range of 2.5 to 6.5 and at temperatures between 10 and 65°C.

[0131] According to embodiments herein, the process for the preparation of aldehydes involves the following steps: mixing an aqueous solution of enzyme laccase immobilized on a solid support and alcohol at select ratio followed by addition of mediator and emulsifying agent under stirring with a continuous supply of oxygen or air till the conversion of alcohol to aldehyde is complete and extracting and purifying the crude aldehyde to obtain high purity aldehyde, wherein the process is carried out in an aqueous buffer in pH range of 2.5 to 6.5 and at temperatures between 10 and 65°C.

[0132] Reaction Scheme for preparing aldehydes:LaccaseR-CH2OH - ► R-CHOMediator1Scheme 1 wherein R = straight chained or branched Cf> to C20 alkyl, preferably n-hexyl, n- octyl, iso-octyl and the alkyl chain may possess double and / or triple bonds at various positions.

[0133] One of the embodiments of the present disclosure describes an efficient and environment friendly method for obtaining the following aldehydes (3-9) from their precursor alcohols via enzyme catalyzed oxidation.4: (Z)-hexadec-ll-enal9 : (7Z,1 lZ,13Zi)-hexadeca-7,l 1 ,13-trienal

[0137] In an embodiment of the present disclosure, the emulsifying agent is polyethylene glycol (PEG) having molecular weight ranging from 200 to 2000; and the emulsifying agent is in an amount in a range of 0.1% to 5%.

[0138] In an embodiment of the present disclosure, the gas is air or oxygen.

[0139] In an embodiment of the present disclosure, wherein the continuous supply of air or oxygen is at a rate of 0.1 vvm to 0.5 vvm

[0140] In an embodiment of the present disclosure, the mixing is performed in a stirred tank reactor or in a vessel under oxygen pressure. In another embodiment of the present disclosure, the mixing is performed in a vessel under oxygen pressure ranging from Ikgf to 3 kgf.

[0141] In an embodiment of the present disclosure, the mixing in step (c) of the disclosed process is carried out in the presence of an aqueous buffer with a pH in a range of 2.5 to 6.5 and at temperature in a range of 10 and 65 °C.

[0142] In an embodiment of the present disclosure, the process disclosed herein further comprises extracting the pheromones or precursors thereof using a solvent.

[0143] In an embodiment of the present disclosure, the solvent is selected from hexane, chloroform, methanol, heptane, methyl tert-butyl ether, ethyl acetate, or combinations thereof.

[0144] In an embodiment of the present disclosure, the process further comprises purifying the pheromones or precursors thereof by formation of a bisulfite adduct. In an embodiment of the present disclosure, wherein the extracted pheromones or precursors thereof is purified by preparing its bisulfite adduct and washing out the impurities from the precipitated adduct with an organic solvent selected from ethanol, ethyl acetate, hexane, or combinations thereof.

[0145] In an embodiment of the present disclosure, the isolated bisulfite adduct is treated with aqueous solution of formaldehyde to liberate the aldehyde which is extracted with an organic solvent such as ethyl acetate or hexane. The product is recovered from the extract by evaporation of the organic solvent.

[0146] Embodiments herein include a composition comprising pheromones or precursors thereof, produced from the process disclosed herein. Various methods are known in the art to formulate the composition. A person skilled in the art will be aware of choosing appropriate additives based on the efficient dispersion of the composition. For example, appropriate additives include vegetable oils, refined mineral oils or fractions thereof, rubbers, plastics, silica, diatomaceous earth, wax, or cellulose.

[0147] In an embodiment of the present disclosure, there is provided a composition comprising pheromones or precursors thereof, by the process as disclosed herein.

[0148] Embodiments herein include a use of the pheromones or precursors thereof produced by the process as disclosed herein or the composition as disclosed herein. In another embodiment of the present disclosure, wherein the pheromones or precursors thereof or the composition are used to control insects or pests.

[0149] Although the present disclosure has been described in considerable detail with reference to certain embodiments and implementations thereof, other embodiments are possible to cover the modifications and variations of the present disclosure.

[0150] Sequences used in the present disclosure:Examples

[0151] The disclosure will now be illustrated with working examples, which is intended to illustrate the working of disclosure and not intended to take restrictively to imply any limitations on the scope of the present disclosure. Unless defined otherwise, all technical and scientific terms used herein have the same meaning ascommonly understood to one of ordinary skill in the art to which this disclosure belongs. Although methods and materials similar or equivalent to those described herein can be used in the practice of the disclosed methods and compositions, the exemplary methods, devices, and materials are described herein. It is to be understood that this disclosure is not limited to particular methods, and experimental conditions described, as such methods and conditions may apply.Materials

[0152] For the purpose of the present disclosure, following raw materials with the specified grades / brands were used. Laccases supplied by M / s Aumgene Biosciences, and Americos Chemicals Pvt Ltd, India were used. 11-Z-hexadecanol was obtained from ATGC Biotech Pvt Ltd, India. TEMPO [(2, 2, 6, 6- Tetramethylpiperidin-l-yl) oxyl], Emulsifying agent polyethylene glycol (PEG- 400), solid support titanium dioxide, sodium bicarbonate, 2-methyl tetrahydrofuran, hexane, and anhydrous Magnesium sulfate were obtained from SRL (Sisco Research Laboratories Pvt. Ltd), India. Sodium bisulfite was obtained from FINAR Chemicals, India. The chemicals, such as cetyl trimethyl ammonium bromide (CT AB), isopropanol, sorbitol, and glutaraldehyde were procured from SRL, and ethanol was procured from CS -China.

[0153] The plasmids used in the present disclosure were procured from Invitrogen.

[0154] The yeast and pichia cells were procured from Invitrogen.

[0155] The culture media components were procured from Hi-media labs.EXAMPLE 1Preparation of yeast cells

[0156] Functionally validated insect FADs (SEQ ID NO. 1 to SEQ ID NO. 8) were cloned into Saccharomyces yeast (pYES) (Figure 1A) and Pichia (pPICZaA) expression vectors (Figure 2A) and transferred respectively into INVScl strain of S. cerevisiae (Invitrogen) and X33 strain of Pichia pastoris (Invitrogen) to obtain first yeast cells. Functionally validated insect FARs (SEQ ID NO. 9 to SEQ IDNO.16) were cloned into Saccharomyces yeast (pYES) (Figure IB) and Pichia (pPICZaA) expression vectors (Figure 2B) and transferred respectively into INVScl strain of S. cerevisiae (Invitrogen) and X33 strain of Pichia pastoris (Invitrogen) to obtain second yeast cell.

[0157] Recombinant saccharomyces yeast clones were selected on selection media. Selected yeast clones were grown in synthetic media lacking uracil for 48 h at 28 °C and 250rpm in a shaker incubator. After 48 h, the cell biomass was collected, transferred to induction media, and continued to grow for another 48 h in induction media containing galactose. After 48 h induction, biomass (recombinant yeast cells) was harvested. A similar method was followed for Pichia, except using Pichia- specific media, such as BMGY (Buffered Glycerol-complex Medium) for growing cells and BMMY (Buffered Methanol-complex Medium) for induction.

[0158] Harvested biomass of first yeast cell and second yeast cell were further subjected to permeabilization and / or immobilization.EXAMPLE 2Preparation of permeabilized yeast cells

[0159] In the present disclosure, the yeast cells have been permeabilized in different ways and used for the process of producing pheromones or precursors thereof.1. Permeabilization with cetyl trimethylammonium bromide (CT AB)

[0160] The yeast cells obtained from Example 2 were harvested from 50 ml of broth by centrifugation (5000 rpm) for 15 min at 4 °C and washed twice with phosphate buffer (0.1 M, pH 7.0). CTAB (permeabilizing agent), (50 mL, 0.04 to 2.0 %, w / v), the permeabilizing agent, was added to the yeast biomass, and the contents were mixed on a vortex mixer and incubated for 10 min under shaking conditions. After this, the cells were re-centrifuged and washed twice with the same buffer. The permeabilized cells were used for producing pheromones or precursors thereof.2. Permeabilization with ethanol

[0161] The yeast cell pellet (1 g, wet) obtained using Example 2 was stirred at 15 °C with 20 to 80% ethanol-water (5 mL) (permeabilizing agent) for 40 min. Thecell pellet was collected by centrifugation at 5000 rpm for 15 min, washed with 5 mL 0.1M citrate phosphate buffer (pH 6.3). The permeabilized first and second yeast cells were used for producing pheromones or precursors thereof.3. Cell permeabilization with sorbitol

[0162] The yeast cell pellet (1g, wet) obtained using Example 2 was stirred at 30 °C with 5 to 25% sorbitol solution (permeabilizing agent) in water (5 mL) for 40 min. The cell pellet was collected by centrifugation at 5000 rpm for 15 min, washed with 5 mL 0.1M citrate phosphate buffer (pH 6.3). The permeabilized first and second yeast cells were used for producing pheromones or precursors thereof.EXAMPLE 3Preparation of permeabilized and immobilized yeast cells

[0163] In the present disclosure, permeabilized yeast cells obtained from Example 2 were either cross-linked with glutaraldehyde (carrier) or entrapped with silica solgel (carrier) for immobilization.1. Immobilization by glutaraldehyde crosslinking

[0164] Yeast or Pichia cells, after 72 h of growth in 100 mL culture medium, were collected, washed with distilled water, and then stirred with 100 mL buffer solution (0.1 M phosphate, pH 6.8). 6 mL of 25% glutaraldehyde solution (carrier) was added and the suspension was shaken for 4 h at room temperature. It was then centrifuged, and the cells were washed several times with phosphate buffer to remove excess glutaraldehyde. The pellet was stored at -20 °C for further use2. Immobilization in silica-sol-gel

[0165] The cell pellet after 48 h of growth in 20 mL culture medium was suspended in 2 mL of 0.1 to 2.0 M citric acid solution and mixed with 5 mL Ludox (40% SiCL). The pH of the mixture was about 2.5. To this mixture, 1 mL of sodium silicate (26.5%) (carrier) was added with good mixing. A further 1 mL of sodium silicate solution was added. A gel containing entrapped cells was formed within 5 min. The gel was kept in the refrigerator overnight and then washed with buffer (0.1M Phosphate buffer pH6.8, 2 x 25 mL). The washed gel was then used for biotransformation, as described in Example 5.EXAMPLE 4A process for producing mono- or poly- unsaturated C6-C24 fatty acid

[0166] The saturated fatty acids (first substrate) were added to the water (medium) at the concentration of 0.5mM to 20mM and permeabilized and immobilized first yeast cells obtained from Example 3 were added to the medium. The biotransformation reaction was carried out at 25 to 35 °C with shaking at 250 to 300 rpm for 48 to 72h. After completion of the reaction, the first reaction mixture was extracted with solvents like hexane and analyzed in GC / GC-MS to obtain mono- or poly-unsaturated C6-C24 fatty acid. The used yeast cells were recovered from the process by filtration(size) or centrifugation at 2000 to 3000 rpm for 3 to 5 min. After recovery, cells were suspended in water and centrifuged / filtered to recover the cell. These immobilized cells were reused 3 to 10 times for the pheromones production. Results

[0167] In Figure 3A, 4A and 4B the GCMS Chromatogram shows the saturated fatty acid to unsaturated fatty acid conversion with first yeast cells expressing FAD.

[0168] Figure 3A is a GCMS chromatogram for the conversion of fatty acid to an unsaturated fatty acid with first yeast cells expressing insect FAD, and Figure 4A shows SEQ ID NO. 6 desaturases supplemented with methyl ester of Z-9- tetradecenoic acid (Z9-C14: Me); Figure 4B shows SEQ ID NO. 8 desaturases supplemented with methyl ester of Z-l 1 -hexadecenoic acid (Z11-16: Me).EXAMPLE 5A process for producing mono- or poly- unsaturated C6-C24 fatty alcohol

[0169] The unsaturated fatty acids (second substrate) were added to the water (medium) at the concentration of 0.5mM to 20mM and permeabilized and immobilized first yeast cells obtained from Example 3 were added to the medium. The biotransformation reaction was carried out at 25 to 35 °C with shaking at 250 to 300 rpm for 48 to 72h. After completion of the reaction, the second reaction mixture was extracted with solvents like hexane and analyzed in GC / GC-MS toobtain mono- or poly-unsaturated C6-C24 fatty alcohol. The used yeast cells were recovered from the process by filtration(size) or centrifugation at 2000 to 3000 rpm for 3 to 5 min. After recovery, cells were suspended in water and centrifuged / filtered to recover the cell. These immobilized cells were reused 3 to 10 times for the pheromones production.Results

[0170] In Figure 3B, and 4C, the GCMS Chromatogram shows unsaturated fatty acid to fatty alcohol conversion, with second yeast cells expressing insect FAR. Figure 4C depicts the Gas Chromatography mass spectrometry (GC-MS) analysis of FAMEs from yeast expression system for SEQ ID NO. 14 reductase supplemented with Z9-C14; D., and SEQ ID NO. 16 reductase supplemented with Z- 11 -hexadecenoic acid (Z 11 -Cl 6)

[0171] After permeabilization, conversion (product formation) was increased by 3 to 7 folds compared to untreated cells, i.e., which were not permeabilized prior to the biotransformation reaction. Among the different permeabilizing agents used, the highest conversion (6.37-fold over untreated control, 45.88%) was observed when ethanol was used as the permeabilizing agent as shown in Table 4.

[0172] Table 4EXAMPLE 6A process for producing pheromones (fatty aldehydes)1. Preparation of 11-Z-hexadecanal with free enzyme

[0173] 1 g of commercially procured laccase powder was dissolved in 200 ml of distilled water and the alcohol 11-Z-hexadecanol (3-20 g, mono- or polyunsaturated C6-C24 fatty alcohol) was added to the enzyme solution and stirred at 40°C while passing air through the reaction mass. The solution of TEMPO (0.5 to 1 g, mediator) in 20 ml of distilled water was added and the reaction mass was stirred for 10-24 h. The product was extracted with hexane (2 x 100 mL, solvent) and recovered after evaporation of hexane. The GC analysis of the reaction mass showed 86% conversion of the fatty alcohol to fatty aldehyde and the conversion process is depicted in Figure 5.2. Preparation of 11-Z-hexadecanal with laccase immobilized on TiCPurification of commercial HO2

[0174] Commercial sample of titanium dioxide (100 g) was stirred with 1 L of IN nitric acid for 1 h and then separated by centrifugation. The process was repeated till the supernatant was free of chloride ions as tested with silver nitrate. The residue was washed with distilled water and then with 1% citric acid solution. The pH of supernatant was 3.5. The residue was finally washed with distilled water and dried in oven at 200°C to obtain purified TiCh.3. Immobilization of laccase on HO 2

[0175] A solution of 2 g laccase powder in 20 mL distilled water was stirred with 12 g of purified titanium dioxide powder for 1 h at room temperature. The binding of laccase to TiCL was followed by measuring laccase activity in the supernatant. When laccase activity in the supernatant was almost undetectable, the reaction mass was centrifuged and the residue with immobilized laccase was washed once with distilled water and stored in refrigerator.4. Determination of laccase activity

[0176] Laccase activity was determined by following the kinetics of the oxidation of 2,2'-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) ABTS to ABTS radical by spectroscopic analysis. Briefly, ABTS (1 mM, 2 mL in 0.05 M citrate-phosphate buffer pH 3.0-6.0) was taken in a cuvette and 10 pL of laccase solution or 10 mg of immobilized laccase were added. Increase in absorbance was monitored at 420 nm for 5 min reaction for laccase solution. In case of immobilized laccase, the slopes(absorbance / min) were calculated by recording the absorbance at 0 min and after 5 min. One unit laccase activity is the amount of enzyme that oxidized 1 pmol ABTS / min under these conditions. (AE = 36,000 M-1cm-1).

[0177] The ABTS assay exhibited linear absorption with an increase over time. Best activity was observed at pH 3.0 as shown in Figure 6. There was no observable shift in pH-activity profile on immobilization on TiO2. Further studies were performed in 0.1 M acetate buffer, pH 3.0. The figure indicated the effect of pH on laccase activity at 30°C in 0.05 M citrate -phosphate buffer.5. Preparation of fatty aldehyde using immobilized enzyme

[0178] The immobilized laccase obtained by the process explained above was suspended in 50 mL of 0.1 M acetate buffer of pH 3.0. 500 mg of PEG-400 (emulsifying agent) was added, and the contents were aerated for 30 mins while stirring at 100 rpm. Solution was 0.32g TEMPO in 2 mL ethyl acetate and 2.4 g alcohol in 2.5 mL ethyl acetate were added portion wise (0.5 mL every 30 min) while passing air through the reaction mass. The reaction mass was stirred till alcohol spot was not observed in TLC analysis (20 h). The reaction mass was centrifuged, and the supernatant was extracted with hexane. TiO2-Laccase mass was also extracted with hexane to collect the aldehyde. The obtained product was extracted with hexane (2 x 100 mL) and recovered after evaporation of hexane. The laccase immobilized on TiCL was reused for another cycle.Results

[0179] Figure 7 shows the stability of laccase (A) solubilized in buffer as compared to that (B) immobilized on TiCh . It was observed that the immobilized laccase was stable for 24 h in 0.1 M acetate buffer (pH 3.0) at 30 °C and slowly lost its activity.

[0180] The GC analysis of the reaction mass showed 86% conversion of the alcohol to aldehyde.EXAMPLE 7A process for producing pheromones

[0181] In a stirred tank reactor or a vessel under pressure the following steps were carried out to produce pheromones in batch mode of operation.1. Production of mono- or poly- unsaturated C6-C24 fatty acid

[0182] The first yeast cell was provided in a first medium comprising a first substrate into the stirred tank reactor and allowed to react to obtain a first reaction mixture comprising mono- or poly-unsaturated C6-C24 fatty acid. The first yeast cell may be recovered from the reactor for reuse.2. Production of mono- or poly- unsaturated C6-C24 fatty alcohol

[0183] After the production of mono- or poly-unsaturated C6-C24 fatty acid, the second yeast cell was provided either in a second medium comprising a second substrate or provided directly into the reactor comprising first reaction mixture and allowed to react to obtain a second reaction mixture comprising mono- or polyunsaturated C6-C24 fatty alcohol. The second yeast cell may be recovered from the reactor for reuse.3. Production of mono- or poly- unsaturated C6-C24 fatty alcohol

[0184] An aqueous solution of laccase enzyme was added to the second reaction mixture in the vessel under oxygen pressure along with, a mediator, and an emulsifying agent under a continuous supply of a gas and mixed to obtain fatty aldehydes, the pheromones. The pheromones were further extracted using hexane as described in Example 6.EXAMPLE 8Purification of pheromones (fatty aldehyde)

[0185] The fatty aldehyde obtained in example 6 or 7 after removal of the solvent (10 g) was added to a solution of 30% sodium bisulfite dissolved in ethanol (100 mL) and which was added dropwise to a solution of sodium bisulfite (16 g) in water (30 mL) at room temperature. The resulting suspension was stirred for 6 h, cooled in ice and the precipitate was filtered through a Buchner funnel. The residue was washed with ethanol, followed by washing with isopropanol and hexane to remove unreacted alcohol. The washings were pooled, and the organic solvent was removed on rotavapor. The residual aqueous solution with unreacted alcohol was extracted with hexane (3 x 50 mL). Evaporation of hexane provided the unreacted alcohol (6 g).The washed bisulfite adduct was suspended in water (50 mL) and cooled in icebath. 6N HC1 was added dropwise to the suspension with stirring till all the solid decomposed to give clear aqueous layer with an organic layer of aldehyde. The aldehyde was extracted with hexane, washed with water, and dried over anhydrous magnesium sulfate. Removal of the solvent on rotavapor resulted in the aldehyde 11 -hexadecanal with >99% purity (3.8 g), as determined from the elution peak at 5.31 min in the GC chromatogram as provided in Table 5 and Figure 8.

[0186] Table 5

[0187] In another example, the fatty aldehyde obtained in example 7 or 8 after removal of the solvent was added to a solution of sodium bisulfite (16 g) in water(30 mL) at room temperature. pH of the solution was adjusted to 6.0 with sodium bicarbonate and the resulting suspension was stirred for 6 h, cooled in ice and the precipitate was filtered through a Buchner funnel. The residue was washed with 2- methyl tetrahydrofuran, followed by washing with hexane to remove unwanted impurities and unreacted alcohol. Recovery of the aldehyde was performed in the following manner. A solution comprising 5-20% formaldehyde (preferably 5-10%) containing 10-20% of magnesium sulfate (preferably 15-20% ) was mixed with the 2-5 g of bisulfite adduct and stirred till all the solid decomposed, to result in clear aqueous layer and an organic layer of aldehyde. The aldehyde was extracted with hexane, washed with water, and dried over anhydrous magnesium sulfate. Removal of the solvent on rotavapor resulted in the aldehyde with >99% purity.Advantages of the present disclosure

[0188] The present disclosure provides a process of producing pheromones or precursors thereof, which exhibits the following advantages. a) Permeabilization of the yeast cells results in improved product formation and therefore, product recovery and production of commercially relevant titers of the pheromone in yeast for pest control.b) Immobilization of the yeast cells used allows to overcome most of the process restrictions, improves recovery of the yeast cells for reuse, offers better stability, activity, and selectivity of the molecules, higher resistance against inhibition, helps the elimination of unnecessary separation and purification steps, and consequently produces the desired pheromones with more efficiency. c) The combined approach of immobilizing and permeabilizing the yeast cells makes the production process of pheromones more cost-effective. d) The present disclosure provides an efficient and environment-friendly process to obtain a high purity aldehyde from of pheromones (98 to 99.8%) by oxidation of mono or poly unsaturated alcohol catalyzed by enzyme laccase. Hence, the disclosed process does not generate harmful waste and is an environmentally sustainable method of producing pheromones.

Claims

I / We Claim:

1. A process for producing pheromones, or precursors thereof, said process comprising: a) obtaining a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; contacting the first yeast cell with a first medium comprising a first substrate to obtain a first reaction mixture comprising mono- or poly-unsaturated C6-C24 fatty acid; wherein the first yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell; wherein the first substrate is a saturated or unsaturated C6- C24 fatty acid or derivative thereof; b) obtaining a second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme; contacting the second yeast cell with a second medium comprising the first reaction mixture or a second substrate to obtain a second reaction mixture comprising mono- or poly-unsaturated C6-C24 fatty alcohol; wherein the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell; wherein the second substrate is a mono or poly unsaturated C6-C24 fatty acid or derivative thereof; and c) mixing an aqueous solution of laccase enzyme, the second reaction mixture, a mediator, and an emulsifying agent under a continuous supply of a gas, to obtain the pheromones or precursors thereof.

2. A process for producing mono- or poly- unsaturated C6-C24 fatty acid or derivative thereof, said process comprising: obtaining a first yeast cell comprising at least one heterologous gene encoding fatty acid desaturase enzyme; contacting the first yeast cell with a first medium comprising a first substrate to obtain mono- or polyunsaturated C6-C24 fatty acid; wherein the first yeast cell is contacted with a permeabilizing agent to obtain a permeabilized first yeast cell; and wherein the first substrate is a saturated or unsaturated C6-C24 fatty acid or derivative thereof.

3. A process for producing mono- or poly-unsaturated C6-C24 fatty alcohol, said process comprising: obtaining a second yeast cell comprising at least one heterologous gene encoding fatty acid reductase enzyme; contacting the second yeast cell with a second medium comprising a second substrate to obtain mono- or polyunsaturated C6-C24 fatty alcohol; wherein the second yeast cell is contacted with a permeabilizing agent to obtain a permeabilized second yeast cell; and wherein the second substrate is a mono or poly unsaturated C6-C24 fatty acid or derivative thereof.

4. A process for producing pheromones, or precursors thereof, said process comprising: mixing an aqueous solution of enzyme laccase and mono- or polyunsaturated C6-C24 fatty alcohol followed by addition of a mediator and an emulsifying agent under a continuous supply of a gas, to obtain pheromones or precursors thereof.

5. The process as claimed in claim 1 or claim 2 or claim 3, wherein the yeast cell and the permeabilizing agent in a weight ratio in the range of 0.1:1 to 0.01:8 is contacted for a duration in the range of 5 min to 48 hours.

6. The process as claimed in claim 5, wherein the permeabilizing agent is selected from the group consisting of chloroform, cetyltrimethylammonium bromide (CT AB), Triton X-100, Tween 80, N-lauroylsarcosine, N- lauroylsarcosine sodium salt (sarkosyl / Sodium lauroyl sarcosinate), sorbitol, methanol, ethanol, 1 -propanol, isopropyl alcohol, 2-propanol, 1- butanol, dimethyl sulfoxide (DMSO), toluene, benzene, benzene acetone, ethyl acetate, ethyl ether, acetone, benzalkonium chloride, and polyoxyethylene(23)lauryl ether; and the permeabilizing agent is in an amount in a range of 0.04 wt% to 80 wt%.

7. The process as claimed in claim 1 or claim 2 or claim 3, wherein the process further comprises contacting the permeabilized yeast cell with a carrier in a weight ratio in the range of 1 : 2 to 1 : 8 for a period in the range of 5 min to 48 h to obtain an immobilized and permeabilized yeast cell.

8. The process as claimed in claim 7, wherein the carrier is selected from glutaraldehyde, silica sol-gel, or combinations thereof; and the carrier is in an amount in a range of 20 wt% to 80 wt%.

9. The process as claimed in claim 1 or claim 2 or claim 3, wherein the yeast cell is selected from a group consisting of Hansenula polymorpha, Kluyveromyces lactis, Komagataella pastoris, Komagataella phaffii ( Pichia pastoris), Komagataella pseudopastoris, Saccharomyces cerevisiae, Saccharomyces bayanus, Saccharomyces pastorianus, Saccharomyces uvarum, Saccharomyces cerevisiae (var. diastaticus), Saccharomyces kudravzevii, Saccharomyces mikatae, Saccharomyces paradoxus, Schizosacchromyces pombe, and Yarrowia lipolytica', and the yeast cells are present in an amount in a range of 20 wt% to 80 wt%.

10. The process as claimed in claim 1 or claim 2 or claim 3, wherein the amino acid sequence of the fatty acid desaturase is selected from the group consisting of SEQ ID NO.17, SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20, SEQ ID NO.21, SEQ IN NO.22, SEQ ID NO.23, and SEQ ID NO.24; and the amino acid sequence of the fatty acid reductase is selected from the group consisting of SEQ ID NO.25, SEQ ID NO.26, SEQ ID NO.27, SEQ ID NO.28, SEQ ID NO.29 SEQ ID NO.30, SEQ ID NO.31, and SEQ ID NO.32.

11. The process as claimed in claim 1 or claim 2 or claim 3, wherein the gene encoding the fatty acid desaturase is selected from SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4, SEQ ID NO.5, SEQ IN NO.6, SEQ ID NO.7, or SEQ ID NO.8; and the gene encoding said fatty acid reductase is selected from SEQ ID NO.9, SEQ ID NO.10, SEQ ID NO.11, SEQ ID NO.12, SEQ ID NO.13, SEQ ID NO.14, SEQ ID NO.15, or SEQ ID NO.16.

12. The process as claimed in claim 1 or claim 4, wherein the pheromones or precursors thereof is selected from the group consisting of (Z,Z,E)-3,6,8- dodecatrien-l-ol, (E,E)-10,14-hexadecadienal, (Z,Z)-9,2-tetradecadien-l- ol, (Z,Z)-9,12-tetradecadienyl acetate, (Z,Z)-9,l l-tetradecadienylacetate, (Z,Z)-8,10-tetradecadienal, (Z,Z)-8,10-dodecadienyl acetate, (Z,Z)-11,13-hexadecadienyl acetate, (Z,Z)-8,10-dodecadien-ol, (Z,Z)-11,13- hexadecadien-l-ol, (Z,Z)-7,9-dodecadienyl acetate, (Z,Z)- 10,12- hexadecadienal, (Z,Z)-7,9-dodecadien-l-ol, (Z,E)-10,12-hexadecadienal, (Z,Z)-7,l l-tridecadienyl acetate, (Z)- 11 -heptadecenyl acetate, (Z,Z)-5,9- tridecadienylacetate, (Z)-l 1-heptadecen-l-ol, (Z,Z)-5,8-tetradecadienyl acetate, (Z,Z)-5,8-tetradecadien- l-ol, (Z,Z,Z)-9,12, 15-octadecatrienal, (Z,Z)-5,8-tetradecadienal, (Z,Z)-5,7-dodecadienal, (E,E)-10,12- hexadecadien-l-ol, (Z,Z)-4,7-tridecadienyl acetate, (Z)-8-heptadecen-l-ol, (Z,Z)-4,7-tridecadien-l-ol, (E)-8-heptadecenylacetate, (Z,Z)-4,7- decadienyl acetate, (Z,Z)-9,l l-pentadecadienal, (Z,Z)-4,7-decadien-l-ol, (E,Z)-9,11-pentadecadienal, (Z,Z)-3,8-dodecadien-l-ol, (Z,E)-7,11- hexadecadienal, (Z,Z)-2,4-decadienal, (E,Z)-8,10-pentadecadienyl acetate, (Z,Z)-9, 11-tetradecadienal, (Z,Z)-9, 11-tetradecadien- l-ol, (Z,Z)-5,7- dodecadienyl acetate, (Z,Z)-9,1 1-hexadecadienal, (Z,E,E)-3,6,8- dodecatrien-l-ol, (Z,Z)-11,13-hexadecadienal, (Z,E)-9,11 -tetradecadienyl acetate, (Z,E)-8,10-dodecadienyl acetate, (Z,E)-l l,13-hexadecadienyl acetate, (Z,E)-7,9-dodecadienyl acetate (Z,E)-10,12-hexadecadienyl acetate, (Z,E)-7,9-dodecadien-l-ol, (E,Z)-10,2-hexadecadienal, (Z,E)-5,7- dodecadienyl acetate, (E,Z)-9,11-hexadecadienal, (Z,E)-5,7-dodecadienal, (Z,E)-9, 1-hexadecadienal, (Z,E)-3,5-tetradecadienyl acetate, (Z,Z)-9,12- octadecadienal, (Z,E)-3,5-dodecadienyl acetate, (Z,E)-7,l l-hexadecadienyl acetate, (Z,E)-3,5-decadienyl acetate, (Z,Z)-8,10-pentadecadienyl acetate, (Z,E)-5,9-tridecadienyl acetate, (E)-lO-heptadecenyl acetate, (Z,E)-9,12- tetradecadienyl acetate, (Z,E)-9,12-tetradecadienal, (Z,E)-9,12- tetradecadien-l-ol, (Z,E)-9,11-tetradecadienal, (Z,E)-9,11-tetradecadien-l- ol, (Z,E)-8,10-tetradecadienyl acetate, (Z,E)-8,10-dodecadienal, (Z,E)- 1,13-hexadecadienal, (Z,E)-8,10-dodecadien-l-ol, (Z,E)-11,13- hexadecadien- 1 -ol, (Z,E)-5,7 -dodecadien- 1 -ol, (E,Z)-9, 1 -hexadecadienyl acetate, (Z,E)-8,10-tetradecadien-l-ol, (Z)-S-tetradecenyl acetate, (Z)-13- octadecen-l-ol, (Z)-lO-tridecenyl acetate, (E,E,Z,Z)-4,6,11,13- hexadecatetraenal, (Z)-lO-tetradecenyl acetate, (Z,Z)-2,13-octadecadien-l-ol, (Z)-lO-dodecenyl acetate, (Z,Z)-7,10-hexadecadienyl acetate, (Z)-9- undecenyl acetate, (E)-6-hexadecenyl acetate, (Z)-9-tridecenyl acetate, (E,E,Z)-10,12,14-hexadecatrienal, (Z)-9-tetradecenyl acetate, (E,Z)-2,13- octadecadienyl acetate, (Z)-9-tetradecenal, (E,Z)-2,13-qctadecadienal, (Z)-9-tetradecen-l-ol, (Z,E)-7,9-dodecadienyl acetate, (Z,E)- 10,12- hexadecadienyl acetate, (Z,E)-7,9-dodecadien-l-ol, (Z,E)-5,7-dodecadienyl acetate, (Z,E)-3,5-tetradecadienyl acetate, (Z,E)-3,5-dodecadienyl acetate, (Z,E)-7,l l-hexadecadienyl acetate, (Z,E)-3,5-decadienyl acetate, (Z,Z)- 8,10-pentadecadienyl acetate, (Z,E)-5,9-tridecadienyl acetate, (E)-10- heptadecenyl acetate, (Z,E)-9,12-tetradecadienyl acetate, (Z,E)-9,12- tetradecadien-l-ol, (Z,E)-9,l l-tetradecadien-l-ol, (Z,E)-8,10- tetradecadienyl acetate, (Z,E)-l l,13-hexadecadienal, (Z,E)-8,10- dodecadien-l-ol, (Z,E)-11,13-hexadecadien-l-ol, (Z,E)-5,7-dodecadien-l- ol, (E,Z)-9,1 -hexadecadienyl acetate, (Z,E)-8,10-tetradecadien-l-ol, (Z)-S- tetradecenyl acetate, (Z)-13-octadecen-l-ol, (Z)-lO-tridecenyl acetate, (Z)-10-tetradecenyl acetate, (Z,Z)-2,13-octadecadien-l-ol, (Z)-10- dodecenylacetate, (Z,Z)-7,10-hexadecadienyl acetate, (Z)-9-undecenyl acetate, (E)-6-hexadecenyl acetate, (Z)-9-tridecenyl acetate, (Z)-9- tetradecenyl acetate, (E,Z)-2,13-octadecadienyi acetate, (E,Z)-2,13- octadecadienal, (Z)-9-tetradecen-l-ol, (E)-9-hexadecenal, (Z)-5-dodecen-l- ol, (E)-9-hexadecenol-ol, (Z)-5-decenyl acetate, (E)-12-pentadecenyl acetate, (Z)-5-decen-l-ol, (Z)-lO-pentadecenal, (Z)-4-tridecenyl acetate, (E,Z,Z)-4,6,10-hexadecatrienyl acetate, (Z)-4-tridecenal, (E,E,Z)-4,6,10- hexadecatrienyl acetate, (Z)-4-decenyl acetate, (Z)-8-pentadecenyl acetate, (Z)-4-decenal, (Z)-9-pentadecenyl acetate, (Z)-3-tetradecenyl acetate, (E)- 2-octadecenal, (Z)-3-Tetradecen-l-ol, (E)-2-octadecenyl acetate, (Z)-3- dodecenyl acetate, (Z)-7-hexadecen-l-ol, (Z)-3-dodecen-l-ol, (E)-7- hexadecenyl acetate, (Z)-2-tridecenyl acetate, (E,E,Z)-4,6,10- hexadecatrien-ol, (Z)-12-tetradecenyl acetate, (E,E)-5,9-octadecadien-l-ol, (Z)- 11 -tridecenyl acetate, (Z)-2-heptadecenal, (Z)- 11 -tetradecenyl acetate, (Z,E)-3,13-octadecadienyl acetate, (Z)-l l-tetradecenal, (Z,Z)-3, 13-octadecadienyl acetate, (Z)-lO-dodecen-l-ol, (Z,Z)-7,10-hexadecadien-l- ol, (Z)-7-undecenyl acetate, (Z)-5-hexadecen-l-ol, (Z)-5-decenal, (Z)-12- pentadecenyl acetate, (Z)-l l-tetradecen-l-ol, (E,Z)-3,13-octadecadienal, (E,Z,Z)-4,7,10-tridecatrienyl acetate, (E,E)-4,8-heptadecadienyl acetate, (E,Z)-9, 11 -tetradecadienyl acetate, (E,Z)-8,10-tetradecadienyl acetate, (E,Z)-8,10-tetradecadienal, (E,Z)-8,10-dodecadienyl acetate, (E,Z)-11,13- hexadecadienyl acetate, (E,Z)-8,10-dodecadienal, (E,Z)-11,3- hexadecadienal, (E,Z)-8,10-dodecadien-l-ol, (E,Z)-11,3-hexadecadien-l-ol, (E,Z)-7,9-dodecadienyl acetate, (E,Z)-10,12-hexadecadien-l-ol, (E,Z)-7,9- dodecadienal, (E,Z)-10,12-hexadecadienyl acetate, (E,Z)-5,9-tridecadienyl acetate, (Z)-9-heptadecenal, (E,Z)-5,7-dodecadienyl acetate, (E,Z)-8,11- hexadecadienal, (E,Z)-5,7-dodecadienal, (E,E)-9,l l-hexadecadienal, (E,Z)-4,9-tetradecadienyl acetate, (Z,Z)-13,15-octadecadienal, (E,Z)-4,9- tetradecadienal, (Z,Z,Z)-3,6,9-octadecatrienyl acetate, (E,Z)-4,7- tridecadienyl acetate, (E)-8-heptadecen-l-ol, (E,Z)-4,10-tetradecadienyl acetate, (E,E,E)-9,2,5-octadecatrien-l-ol, (E,Z)-3,8-tetradecadienyl acetate, (E,E)-l l,14-octadecadienal, (E,Z)-3,5-tetradecadienyl acetate, (Z,Z)-9,12- octadecadienyl acetate, (E,Z)-3,5-dodecadienyl acetate, (Z,E)-7,11- hexadecadien-l-ol, (E,Z)-2,4-decadienal, (E,Z)-8,10-pentadecadien-l-ol, (E,Z)-7,9-dodecadien-l-ol, (E,E)-10,12-hexadecadienal, (E,Z)-5,7- dodecadien-l-ol, (Z,Z)-8,10-hexadecadienyl acetate, (E,Z)-3,7- tetradecadienyl acetate, (Z,Z)-l l,13-octadecadienal, (E, E,E)-10, 12,14- hexadecatrienyl, (E,E)-9, 11 -tetradecadienyl acetate, (E,E)-8,10- dodecadienyl acetate, (E,E)-l,13-hexadecadienyl acetate, (E,E)-7,9- dodecadienylacetate, (E,E)-10,12-hexadecadienyl acetate, (E,E)-5,8- tetradecadienal, (Z,Z,Z)-9,12,15-octadecatrienyl acetate, (E,E)-5,7- dodecadienyl acetate, (Z,Z)-7,l l-hexadecadienal, (E,E)-5,7-dodecadien-l- ol, (Z,Z)-7,l-hexadecadienyl acetate, (E,E)-4,10-dodecadienyl acetate, (Z,Z)-7,l-hexadecadien-l-ol, (E,E)-3,5-tetradecadienyl acetate, (E,E)-9,12- octadecadien-l-ol, (E,E)-3,5-decadienylacetate, (Z,E)-8,10- pentadecadienyl acetate, (E,E,Z)-10,12,14-hexadecatrienyl, (E,E)-9,2-tetradecadienyl acetate, (E,E)-8,10-tetradecadienyi acetate, (E,E)-8,10- tetradecadienal, (E,E)-8,10-dodecadien-l-ol, (E,E)-11,13-hexadecadien-l- ol, (E,E)-8,10-dodecadienal, (E,E)-l l,13-hexadecadienal, (E,E)-2,4- tetradecadienal, (E,E)-5,9-octadecadienyl acetate, (E,E)-2,4-decadienal, (E,E)-8,10-pentadecadienyl acetate, (E)-5-tridecenyl acetate, (E,E,Z)- 4,6,10-hexadecatrienyl acetate, (E)-5-tetradecenal, (E)-9-octadecen-l-ol, (E)-5-tetradecen-l-ol, (Z)-2-octadecenyl acetate, (E)-5-dodecenyl acetate, (E)-5-hexadecenyl acetate, (Z)-lO-pentadecenyl acetate, (E)-1O- tetradecenyl acetate, (Z, E)-2,13-octadecadienyl acetate, (E)-lO-dodecenyl acetate, (E,Z)-6, 11 -hexadecadienyl acetate, (E)-lO-dodecenal, (E,Z)-6,11- hexadecadienal, (E)-9-tridecenyl acetate, (E)-9-tetradecen-l-ol, (Z)-13- octadecenal, (E)-9-dodecenal, (Z)-14-hexadecenyl acetate, (E)-9-dodecen-I-ol, (Z)-12-hexadecenal, (E)-9-tetradecenyl acetate, (E)-14-octadecenal, (E)-9-dodecenyl acetate, (E)-14-hexadecenal, (E)-8-tridecenyl acetate, (E)- 8-tetradecenyl acetate, (E)-13-octadecenyl acetate, (E)-8-Dodecenyl acetate, (Z)-9-hexadecen-l-ol, (Z)-l l-hexadecen-l-ol, (E)-8-dodecenal, (Z)- 11 -hexadecenyl acetate, (E)-8-dodecen-l-ol, (E)-l l-hexadecenal, (E)- 8-decen-l-ol, (Z,Z)-6,9-pentadecadienal, (E) -7 -tetradecenyl acetate, (E)-I I-octadecenal, (E)-7-tetradecen-l-ol, (E)-l l-octadecen-l-ol, (E)-7- dodecenyl acetate, (E)-lO-hexadecenal, (E)-7-dodecenal, (Z)-10- hexadecenyl acetate, (E)-7-dodecen-l-ol, (E)-lO-hexadecen-l-ol, (E)-7- decenyl acetate, (Z,Z)-8,9-pentadecadien-l-ol, (E)-6-tridecenyl acetate, (E,E,Z)-4,6, 11 -hexadecatrienyl acetate, (E)-6-tetradecenyl acetate, (Z)-9- octadecenyl acetate, (E)-6-dodecenal, (Z)-9-hexadecenal, (E)-6-dodecen-l- ol, (Z)-9-hexadecen-l-ol, (E)-5-dodecen-l-ol, (Z)-7-hexadecenal, (E)-5- decen-l-ol, (E)-9-pentadecenyl acetate, (E)-5-tetradecenyl acetate, (Z)-2- octadecenal, (E)-4-tridecenyl acetate, (E,Z,Z)-4,6,10-hexadecatrien-l-ol, (E)-4-dodecenyl acetate, (Z)-7-hexadecenyi acetate, (E)-4-decenyl acetate, (Z)-8-pentadecen-l-ol, (E)-3-tetradecenyl acetate, (Z,Z)-8,11- heptadecadienyl acetate, (E)-3-tetradecen-l-ol, (Z,Z)-8,10-heptadecadien- l-ol, (E)-3-dodecenyl acetate, (E)-7-hexadecenal, (E)-2-undecenyl acetate,(Z)-3-hexadecenyi acetate, (E)-2-undecenal, (E)-5-hexadecen-l-ol, (E)-2- tridecenyl acetate, (Z,E)-l,14-hexadecadienyl acetate, (E)-2-dodecenal, (E)-7-hexadecen-l-ol, (E)-12-tetradecenyl acetate, (Z,Z)-3,13- octadecadienal, (E)- 11 -tetradecenyl acetate, (E,E)-3, 13 -octadecadienyl acetate, (E)-lO-dodecen-l-ol, (E,Z)-4,6-hexadecadienal, (E)-l-tetradecen-I-ol, (Z,Z)-2,13-octadecadienyl acetate, (E)- 11 -tridecenyl acetate, (E)-2- heptadecenal, (E)-l l-tetradecenal, (E,Z)-3, 13 -octadecadienyl acetate, (E,E)-8,10-tetradecadien-l-ol, (Z)-9-tetradecenol, (E)- 11 -hexadecenol, (Z)-I I -hexadecenol, (E,E)-10,12-hexadecadienol, (Z)-l l-hexadecenal, (Z)-9- tetradecenyl acetate, (E,Z,Z)-3,8,11 -tetradecatrienyl acetate (TDTA), (Z,E)-9.11 -tetradecadienyl acetate, (Z,E)-12,9-tetradecadienyl acetate, (E)-l l- hexadecenyl acetate, (Z)-l 1 -hexadecenyl acetate, (Z,E)-7,11-hexadecadien- 1-yl acetate, (Z,Z)-7,l l-hexadecadien-l-yl acetate, (Z)-13-octadecenyl acetate, (Z)-7 -dodecenyl acetate, (E,Z)-7,9-dodecadienyl acetate, (E,Z)- 10,12-tetradecadienyl acetate, (Z,E)-l l,l l-tetradecadienyl acetate, (E)-2- decenal, (Z)-2-decenal, (Z)-5-dodecenal, (Z)-7-dodecenal, (Z)-9-dodecenal, (E)-lO-dodecenal, (Z)-4-tridecenal, (Z)-5-tetradecenal, (Z)-7-tetradecenal, (Z)-8-tetradecenal, (Z)-l 1-tetradecenal, (E,E)-10,12-tetradecadienal, (Z)- 10-hexadecenal, (E)-7-hexadecenal, (Z,E)-9,l l-hexadecadienal, (Z,Z)-9.11-hexadecadienal, (E,Z)-10,12-hexadecadienal, (Z,E)-10,12- hexadecadienal, (E,Z)-l l,13-hexadecadienal, (E,E,Z)-4,6,11- hexadecatrienal, (E,E,E)- 10, 12, 14-hexadecatrienal, (Z)-2-heptadecenal, (E)-9-octadecenal, (Z)-9-octadecenal, (Z)-l l-octadecenal, (E)-13- octadecenal, undecanal, (Z,Z)-9,l l-hexadecadienal, (E,E,Z)-4,6,11- hexadecatrienal, (Z)-6,14-pentadecadienal, (Z)-9,13-tetradecadien-l 1-ynal, (Z)-13-hexadecen-l l-ynal, (Z)-hexadec-9-enal, (Z)-hexadec-l l-enal, (11Z, 13Z)-hexadeca-l l,13-dienal, (10E, 12E)-hexadeca-10,12-dienal, (E)- hexadec-10-enal, (Z)-octadec-13-enal, and (7Z, 11Z, 13E)-hexadeca- 7,11,13-trienal.

13. The process as claimed in claim 1 or claim 2 or claim 3, wherein the first substrate is selected from the group consisting of dodecanoic acid, tetradecanoic acid, (E)-3-tetradecenoic acid, (Z)-8-tetradecenoic acid, (Z)-l l- tetradecenoic acid, (E)- 11 -tetradecenoic acid, (Z)-9-tetradecenoic acid, (E)- 12- tetradecenoic acid, hexadecanoic acid, (Z) -7 -hexadecenoic acid, (E)-l 1- hexadecenoic acid, (Z)- 11 -hexadecenoic acid, (E)-lO-hexadecenoic acid, (E)-12-hexadecenoic acid, octadecanoic acid, (E)-9-octadecenoic acid, (Z)- 7-octadecenoic acid, eicosanoic acid, (E)-7-eicosenoic acid, and (Z)-9- eicosenoic acid; and wherein the second substrate is selected from the group consisting of dodecanoic acid, tetra decanoic acid, hexadecanoic acid, octadecanoic acid, (E)-7-dodecenoic acid, (E)-8-dodecenoic acid, (Z)-9- dodecenoic acid, (E)-lO-dodecenoic acid, (E)- 3 -tetradecenoic acid, (Z)-8- tetradecenoic acid, (Z)-9-tetradecenoic acid, (E)-l 1 -tetradecenoic acid, (E)- 12-tetradecenoic acid, (E,Z,Z)-3,8,11- tetradecatrienoic acid, (Z,E)-9,11- tetradecadienoic acid, (Z,E)-9,12- tetradecadienoic acid, (Z)-9- hexadecenoic acid, (E)-lO-hexadecenoic acid, (E)- 11 -hexadecenoic acid, (Z) 11 -hexadecenoic acid, (Z,E)-7,11- hexadecadienoic acid, (Z,Z)-7,11- hexadecadienoic acid, (E,E)-10, 12-15 hexadecadienoic acid, (Z)-13- hexadecenoic acid, and (Z)-13-eicosenoic acid.

14. The process as claimed in claim 1 or claim 4, wherein the mixing in step (c) is carried out in the presence of an aqueous buffer with a pH in a range of 2.5 to 6.5 and at temperature in a range of 10 and 65 °C.

15. The process as claimed in claim 1 or claim 4, wherein the laccase enzyme and the alcohol is in a weight ratio in a range of 0.1 : 1 to 1:0.1.

16. The process as claimed in claim 1 or claim 4, wherein the laccase enzyme is sourced from Trametes versicolor, Trametes pubescens, Rhus vernicifera, Pyricularia oryzae, Agaricus bisporus, Mycelopthora termophila, or combinations thereof; and the enzyme is either in a soluble form or immobilized on a solid support.

17. The process as claimed in claim 1 or claim 4, wherein the mediator is an electron transfer reagent selected from 2,2,6,6-Tetramethylpiperidin-l- yl)oxyl (TEMPO) hydroxybenzotriazole (HOBT), 2,2’-azinobis-(3- ethylbenzothiazoline-6-sulfonate(ABTS), or combinations thereof.

18. The process as claimed in claim 1 or claim 4, wherein the mediator and the alcohol is in a mole ratio in a range of 0.50:1 to 0.10:1.

19. The process as claimed in claim 1 or claim 4, wherein the emulsifying agent is polyethylene glycol (PEG) having molecular weight ranging from 200 to 2000; and the emulsifying agent is in an amount in a range of 0.1% to 5%.

20. The process as claimed in claims 1 or claim 3, wherein the mono or poly unsaturated C6-C24 alcohol is selected from (E)-2-decenol, (Z)-2-decenol, (Z)-4-decenol, (Z)-5-decenol, (E,E)-2,4-decadienol, (E,Z)-2,4-decadienol, (Z,Z)-2,4-decadienol, (E)-2-undecenol, (E)-2-dodecenol, (Z)-5-dodecenol, (E)-6-dodecenol, (Z)-7-dodecenol, (E)-8-dodecenol, (E)-9-dodecenol, (Z)-9-dodecenol, (E)-lO-dodecenol, (E,Z)-5,7-dodecadienol, (Z,E)-5,7- dodecadienol, (Z,Z)-5,7-dodecadienol, (E,Z)-7,9-dodecadienol, (E,E)-8,10- dodecadienol, (E,Z)-8,10-dodecadienol, (Z,E)-8,10-dodecadienol, (Z)-4- tridecenol, (E)-5-tetradecenol, (Z)-5-tetradecenol, (Z)-7-tetradecenol, (Z)- 8-tetradecenol, (E)- 11 -tetradecenol, (Z)- 11 -tetradecenol, (E,E)-2,4- tetradecadienol, (E,Z)-4,9-tetradecadienol, (E,E)-5,8-tetradecadienol, (Z,Z)-5 , 8-tetradecadienol, (E,E)- 8 , 10-tetradecadienol, (E,Z)- 8 , 10- tetradecadienol, (Z,Z)-8,10-tetradecadienol, (Z,E)-9,11 -tetradecadienol, (Z,Z)-9,11 -tetradecadienol, (Z,E)-9,12-tetradecadienol, (E,E)-10,12- tetradecadienol, (Z)-lO-pentadecenol, (Z,Z)-6,9-pentadecadienol, (E,Z)-9.11 -pentadecadienol, (Z,Z)-9, 11 -pentadecadienol, (Z)-9-hexadecenol, (E)-10-hexadecenol, (Z)-lO-hexadecenol, (E)- 11 -hexadecenol, (Z)-l l- hexadecenol, (Z)-12-hexadecenol, (E)-14-hexadecenol, (E)-7-hexadecenol, (Z) -7 -hexadecenol, (E)-9-hexadecenol, (E,Z)-4,6-hexadecadienol, (E,Z)-6.11 -hexadecadienol, (Z,E)-7,l l-hexadecadienol, (Z,Z)-7,11- hexadecadienol, (E,Z)-8,11 -hexadecadienol, (E,E)-9,11 -hexadecadienol, (E,Z)-9,11 -hexadecadienol, (Z,E)-9,11 -hexadecadienol, (Z,Z)-9,11- hexadecadienol, (E,E)- 10, 12-hexadecadienol, (E,Z)- 10, 12-hexadecadienol, (Z,E)- 10, 12-hexadecadienol, (Z,Z)- 10, 12-hexadecadienol, (E,E)- 11 , 13 - hexadecadienol, (E,Z)-11,13 -hexadecadienol, (Z,E)-11,13-hexadecadienol, (Z,Z)-11,13 -hexadecadienol, (E,E)-10,14-hexadecadienol, (E,E,Z)-4,6,11-hexadecatrienol, (E,E,E)-10,12,14-hexadecatrienol, (E,E,Z)-10,12,14- hexadecatrienol, (E,E,Z,Z)-4,6,l l,13-hexadecatetraenol, (E)-2- heptadecenol, (Z)-2-heptadecenol, (Z)-9-heptadecenol, (E)-2-octadecenol, (E)-9-octadecenol, (Z)-9-octadecenol, (E)- 11 -octadecenol, (Z)-l l- octadecenol, (E)-13-octadecenol, (Z)- 13 -octadecenol, (E)-14-octadecenol, (E,Z)-2, 13 -octadecadienol, (E,Z)-3 , 13 -octadecadienol, (Z,Z)-3 , 13- octadecadienol, (Z,Z)-9,12-octadecadienol, (Z,Z)-11,13-octadecadienol, (E,E)-l l,14-octadecadienol, (Z,Z)-13,15-octadecadienol, (Z,Z,Z)-9, 12,15- octadecatrienol, (Z)-9-hexadecenol, ( (Z)-13-octadecenol, undecanol, (E)-7- dodecenol, (E)- 11 -tetradecenol, (Z)-5-tetradecenol, (Z)-9-tetradecenol, (Z,Z,E)-7,11,13 -hexadecatrienol, (Z)-6,14-pentadecadienol, (Z)-9,13- tetradecadien-l l-ynol, (Z)-13-hexadecen-ll-ynol, or combinations thereof.

21. The process as claimed in claim 1 or claim 4, wherein the gas is air or oxygen.

22. The process as claimed in claim 1 or claim 2 or claim 3, wherein the first medium and the second medium comprises a carbon source selected from glucose, galactose, raffinose, glycerol, or a combination thereof.

23. The process as claimed in claim 1 or claim 2 or claim 3, wherein the first medium or the second medium further comprises water or buffer.

24. The process as claimed in claim 1 or claim 4, wherein the process further comprises extracting the pheromones or precursors thereof using a solvent.

25. The process as claimed in claim 24, wherein the solvent is selected from hexane, chloroform, methanol, heptane, methyl tert-butyl ether, ethyl acetate, or combinations thereof.

26. The process as claimed in claim 24, wherein the process further comprises purifying the pheromones or precursors thereof by formation of a bisulfite adduct.

27. The process as claimed in any one of claims 1 to 3, wherein contacting the first yeast cell with a first medium or contacting the second yeast cell with a second medium is performed in a stirred tank reactor in batch mode.

28. The process as claimed in claim 1 or claim 4, wherein the mixing is performed in a stirred tank reactor or in a vessel under oxygen pressure.

29. The process as claimed in any one of claims 1 to 4, wherein the contacting is carried out at a temperature ranging from 20 to 40°C for a duration ranging from 48 to 120 hours.

30. The process as claimed in any one of claims 1 to 4, wherein the process further comprises: recovering the yeast cell for reuse.

31. The process as claimed in claim 30, wherein the yeast cell is recovered by filtration, or centrifugation, or a combination thereof.

32. A composition comprising pheromones or precursors thereof, produced from the process as claimed in any one of claims 1 to 31.

33. Use of the pheromones or precursors thereof produced by the process as claimed in any one of claims 1 to 31 or the composition as claimed in claim 32.

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