WO2017053574A1 - Cannabinoid glycoside prodrugs and methods of synthesis - Google Patents

Cannabinoid glycoside prodrugs and methods of synthesis Download PDF

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WO2017053574A1
WO2017053574A1 PCT/US2016/053122 US2016053122W WO2017053574A1 WO 2017053574 A1 WO2017053574 A1 WO 2017053574A1 US 2016053122 W US2016053122 W US 2016053122W WO 2017053574 A1 WO2017053574 A1 WO 2017053574A1
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cannabinoid
method
ugt76g1
glycoside
udp
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Brandon J. ZIPP
Janee' M. HARDMAN
Robert T. BROOKE
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Vitality Biopharma, Inc.
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Application filed by Vitality Biopharma, Inc. filed Critical Vitality Biopharma, Inc.
Publication of WO2017053574A1 publication Critical patent/WO2017053574A1/en

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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K47/00Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549Sugars, nucleosides, nucleotides or nucleic acids
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/335Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
    • A61K31/35Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
    • A61K31/352Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. cannabinols, methantheline
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7012Compounds having a free or esterified carboxyl group attached, directly or through a carbon chain, to a carbon atom of the saccharide radical, e.g. glucuronic acid, neuraminic acid
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    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7016Disaccharides, e.g. lactose, lactulose
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/70Carbohydrates; Sugars; Derivatives thereof
    • A61K31/7028Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages
    • A61K31/7034Compounds having saccharide radicals attached to non-saccharide compounds by glycosidic linkages attached to a carbocyclic compound, e.g. phloridzin
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P1/00Drugs for disorders of the alimentary tract or the digestive system
    • A61P1/10Laxatives
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P25/00Drugs for disorders of the nervous system
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00Non-central analgesic, antipyretic or anti-inflammatory agents, e.g antirheumatic agents; Non-steroidal anti-inflammatory drugs (NSAIDs)
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07HSUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/02Acyclic radicals, not substituted by cyclic structures
    • C07H15/04Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical
    • C07H15/10Acyclic radicals, not substituted by cyclic structures attached to an oxygen atom of the saccharide radical containing unsaturated carbon-to-carbon bonds
    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07HSUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
    • C07H15/00Compounds containing hydrocarbon or substituted hydrocarbon radicals directly attached to hetero atoms of saccharide radicals
    • C07H15/20Carbocyclic rings
    • C07H15/203Monocyclic carbocyclic rings other than cyclohexane rings; Bicyclic carbocyclic ring systems
    • AHUMAN NECESSITIES
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    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/0014Skin, i.e. galenical aspects of topical compositions
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/0019Injectable compositions; Intramuscular, intravenous, arterial, subcutaneous administration; Compositions to be administered through the skin in an invasive manner
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL, OR TOILET PURPOSES
    • A61K9/00Medicinal preparations characterised by special physical form
    • A61K9/0012Galenical forms characterised by the site of application
    • A61K9/0053Mouth and digestive tract, i.e. intraoral and peroral administration

Abstract

The present invention relates to cannabinoid glycoside prodrugs suitable for site- and tissue-specific delivery of cannabinoid molecules. The present invention also relates to methods of forming the cannabinoid glycoside prodrugs through glycosyltransferase mediated glycosylation of cannabinoid molecules.

Description

CANNABINOID GLYCOSIDE PRODRUGS AND METHODS OF SYNTHESIS

FIELD OF THE INVENTION

[001 ] The present invention pertains to the field of drug development and in particular to novel cannabinoid glycoside prodrugs and methods for their production by enzyme-mediated carbohydrate transfer.

BACKGROUND

[002] Phytocannabinoids from Cannabis sativa have long been used for altering mental states, but recent findings have illuminated the potential of specific cannabinoid compounds for treatment and maintenance of various diseases and conditions. Of particular importance is the non-psychotropic molecule cannabidiol (CBD) which has potential therapeutic application as an anti-psychotic, a neuroprotectant, and has potential for treatment of numerous other maladies (Zuardi 2012, luvone 2009, for review Mechoulam 2002, respectively). One shortcoming of CBD is that it is easily oxidized to THC and CBN derivatives by light, heat, and acidic or basic conditions, and another detrimental attribute to CBD is that its extremely hydrophobic nature makes it difficult for formulation and delivery. Additionally, current pharmaceutical compositions of CBD and THC have unpleasant organoleptic properties, and their hydrophobic nature results in a lingering on the palate.

[003] Cannabinoids are extremely hydrophobic in nature, complicating their use in drug formulations. Non-covalent methods have been found to improve the solubility of cannabinoids by utilizing carrier carbohydrates such as cyclized maltodextrins (Jarho 1998). Covalent chemical manipulations have produced novel CBD prodrugs with improved solubility (WO2009018389, WO 201201 1 1 12). Even fluorine substituted CBD compounds have been created through synthetic chemical manipulations in an effort to functionalize CBD (WO2014108899). The aforementioned strategies were somewhat successful in improving the solubility of CBD, but they create unnatural compositions which alter the composition and will release the unnatural prodrug moieties upon hydrolysis.

[004] A growing body of evidence shows that glycosides are capable of acting as prodrugs and also to have direct therapeutic effects. Glycoside prodrugs may enable improved drug bioavailability or improved drug pharmacokinetics including more site-specific or tissue- specific drug delivery, more consistent levels of drug in the plasma, and sustained or delayed release of the drug. Site-specific delivery of steroid glycosides to the colon has previously been demonstrated (Friend 1985, Friend 1984), and could enable treatment of local disorders such as inflammatory bowel disease. Glycosylation of steroids enabled survival of stable bioactive molecules in the acidic stomach environment and delivery into the large intestine, where the aglycones were liberated by glycosidases produced by colonic bacteria, and then absorbed into the systemic circulation. Glycosidases are also present universally in different tissues (Conchie 1959), so delivery of glycosides by methods that bypass the digestive tract and colon, such as intravenous delivery, will enable targeted delivery to other cells and tissues that have increased expression of glycosidases. In addition, the distribution of alpha-glycosidase and beta- glycosidase enzymes differ throughout the intestinal tract and other tissues, and different forms of glycosides may therefore provide unique pharmacokinetic profiles, including formulations that target delivery of specific diseased areas, or targeted release at locations that can promote or restrict systemic absorption of the cannabinoids and other compounds described herein. Many biologically active compounds are glycosides, including members of classes of compounds such as hormones, antibiotics, sweeteners, alkaloids, and flavonoids. While it is generally accepted that glycosides will be more water-soluble than the aglycones, literature reviews have analyzed structure-activity relationships and determined that it is nearly impossible to define a general pattern for the biological activities of glycosides across different classes of compounds (Kren 2008).

[005] As with synthetic chemistry, in vivo detoxification strategies serve as another model for improving the solubility of cannabinoids. CBD is glucuronidated in humans by the liver glucosyltransferases, but to date only minor activity has been demonstrated with UGT1 A9 and UGT2B7 in in vitro assays (US Patent No. 8,410,064). In vitro assays showed that cannabinol (CBN) is efficiently glucuronidated by the Human UGT1 A10 ( US Patent No. 8,410,064). The glucuronidation of CBD is one mechanism to increase CBD solubility and facilitate removal and excretion through the kidneys. Searching for glucosyltransferase activity towards cannabinoids, cannabinol was found to be glycosylated when incubated with in vitro cell culture of Pinellia ternata (Tanaka 1993). Similarly, cannabidiol was shown to be glycosylated when incubated with tissue cultures from Pinellia ternata and Datura inoxia, yielding CBD-6'-0^-D- glucopyranoside and CBD-(2',6')-0^-D-diglucopyranoside (Tanaka 1996). These biotransformation studies demonstrate the potential for limited glycosylation of these two compounds to occur by unknown plant glucosyltransferases, and for them to be produced in minute quantities, but to date, no specific plant glucosyltransferase proteins capable of glycosylation of cannabinoids have been identified, no cannabinoid glycosides been produced in large, purified quantities, and the biological activity or pharmaceutical properties of cannabinoid glycosides have never been characterized.

[006] Cannabinoids contain a hydroxylated hydrophobic backbone, similar to the steviol backbone of steviol glycosides found in the Stevia rebaudiana plant. UGT76G1 is a glucosyltransferase from Stevia that is capable of transferring a secondary glucose to the 3C- hydroxyl of the primary glycosylation on both C13-OH and C19-COOH position of the steviol glycoside, and thus its substrates include steviolmonoside, stevioside, rubusoside, RebA, RebD, RebG, RebE, etc. (Richman et al. 2005, Stevia First Corp unpublished work). The substrate recognition site of UGT76G1 is capable of binding and glycosylating multiple steviol glycosides, but it was previously not known to have glycosylation activity towards any other glycosides, and there previously was no established activity of UGT76G1 towards any aglycone compounds at all. As UGT76G1 is capable of glycosylating steviol glycosides on the primary sugar located on both C13 hydroxyl group and the C19 carboxyl group it demonstrates bi- functional glycosylation. Cyclodextrin glucanotransferase (CGTase, Toruzyme 3.0L, Novozymes Inc.) is a member of the amylase family of enzymes and is best known for its ability to cyclize maltodextrin chains. A lesser known activity of CGTase is disproportionation of linear maltodextrin chains and transfer to an acceptor sugar molecule (Li 2012).

[007] There are no known cannabinoid glycosides available as cannabinoid prodrugs. Nor is there a known method for the efficient regioselective production of cannabinoid glycosides, which is necessary in order to produce large, purified quantities of individual glycosides and to assess their pharmaceutical properties, including evaluation of in vivo drug pharmacokinetics and pharmacodynamics. To solve the aforementioned problem, screening of glucosyltransferase enzymes from various organisms has been conducted to identify candidates for the glycosylation of cannabinoids, and to identify cannabinoid glycosides as potential prodrugs of cannabinoids, and as novel cannabinoid compositions with novel properties and functions.

[008] This background information is provided to reveal information believed by the applicant to be of possible relevance to the present invention. No admission is necessarily intended, nor should be construed, that any of the preceding information constitutes prior art against the present invention. SUMMARY OF THE INVENTION

[009] The present invention relates to novel cannabinoid glycoside prodrugs and methods for their production by enzyme-mediated carbohydrate transfer.

[0010] An object of the present invention is to provide a cannabinoid glycoside prodrug. In accordance with an aspect of the present invention, there is provided a cannabinoid glycoside prodrug compound having formula (I

Figure imgf000005_0001

wherein R is H, β-D-glucopyranosyl, or 3-0^-D-glucopyranosyl^-D-glucopyranosyl; R' is H or β-D-glucopyranosyl, or 3-0- β -D-glucopyranosyl^-D-glucopyranosyl; and A is an aglycone moiety formed through reaction of a hydroxyl group on a cannabinoid compound, an endocannabinoid compound, or a vanilloid compound, or a pharmaceutically compatible salt thereof.

[0011] In accordance with another aspect of the present invention, there is provided a method for the site-specific delivery of a cannabinoid drug to a subject, comprising the step of administering a cannabinoid glycoside prodrug in accordance with the present invention to a subject in need thereof.

[0012] In accordance with another aspect of the present invention, there is provided a method of producing a cannabinoid glycoside, comprising incubating a cannabinoid aglycone with one or more sugar donors in the presence of one or more glycosyltransferases.

[0013] Further aspects of the technology described herein will be brought out in the following portions of the specification, wherein the detailed description is for the purpose of fully disclosing preferred embodiments of the technology without placing limitations thereon. BRIEF DESCRIPTION OF THE FIGURES

[0014] Figure 1 A illustrates aglycones employed in the glycosylation methods of the present invention. Figure 1 B illustrates the possible points of glycosylation on the aglycones.

[0015] Figure 2 illustrates possible products of the glycosylation of cannabidiol (CBD).

[0016] Figure 3 illustrates possible products of the glycosylation of cannabidivarin (CBDV).

[0017] Figure 4 illustrates possible rotational products of the glycosylation of cannabidiol (CBD).

[0018] Figure 5 illustrates possible rotational products of the glycosylation of cannabidivarin (CBDV).

[0019] Figure 6 illustrates the proposed superpositioning of the substrate cannabidiol (CBD) in the catalytic site of UGT76G1 .

[0020] Figure 7 illustrates possible products of the glycosylation of tetrahydrocannabinol (Δ9- THC).

[0021] Figure 8 illustrates possible products of the glycosylation of cannabinol (CBN).

[0022] Figure 9 illustrates possible products of the glycosylation of arachidonoyi ethanolamide

(AEA).

[0023] Figure 10 illustrates possible products of the glycosylation of 2-arachidonoyl ethanolamide (2-AG).

[0024] Figure 1 1 illustrates possible products of the glycosylation of 1 -arachidonoyi ethanolamide (1 -AG).

[0025] Figure 12 illustrates possible products of the glycosylation of N- docosahexaenoylethanolamine (DHEA).

[0026] Figure 13 illustrates possible products of the glycosylation of capsaicin.

[0027] Figure 14 illustrates possible products of the glycosylation of vanillin.

[0028] Figures 15A and 15B illustrate possible products of the glycosylation of curcumin.

[0029] Figure 16 is an HPLC linetrace of the reaction products of the glycosylation of CBD.

[0030] Figure 17 is an HPLC linetrace of the reaction products of the glycosylation of CBDV.

[0031] Figure 18 is an HPLC linetrace of the reaction products of the glycosylation of A9-THC.

[0032] Figure 19 is an HPLC linetrace of the reaction products of the glycosylation of CBN.

[0033] Figure 20 is an HPLC linetrace of the reaction products of the glycosylation of 1 -AG and 2-AG.

[0034] Figure 21 is an HPLC linetrace of the reaction products of the glycosylation of synaptamide (DHEA). [0035] Figure 22 is an HPLC linetrace of the reaction products of the glycosylation of AEA.

[0036] Figure 23 is an HPLC linetrace of the reaction products of the glycosylation of vanillin.

[0037] Figure 24 is an HPLC linetrace of the reaction products of the glycosylation of capsaicin.

[0038] Figure 25 is an HPLC linetrace of the reaction products of the glycosylation of CBDgl (VB104) with the glycosyltransf erase UGT76G1 .

[0039] Figure 26 is an HPLC linetrace of the reaction products of the glycosylation of CBDgl (VB104) with the glycosyltransf erase Os03g0702000

[0040] Figure 27 is a 1NMR spectrum of an isolated product, VB104, of the glycosylation of CBD.

[0041] Figure 28 is a 1NMR spectrum of an isolated product, VB1 10 of the glycosylation of CBD.

[0042] Figure 29 is a plot of C18 retention times vs cLogP values for selected cannabinoids and cannabinoid glycosides.

[0043] Figure 30A is a graphical presentation of the results of the analysis of the small intestine extracts of a bioavailability assay.

[0044] Figure 30B is a graphical presentation of the results of the analysis of the large intestine extracts of a bioavailability assay

DETAILED DESCRIPTION OF THE INVENTION

[0045] The following abbreviations are used throughout:

CB Cannabinoid

CBD Cannabidiol.

CBDV Cannabidivarin

CBG Cannabigerol

A9-THC or THC Tetrahydrocannabinol

CBN Cannabinol

CBNV Cannabinavarin

CBDA Cannabidiolic acid

THCV Tetrahydrocannabivarin

UGT UDPG-dependent glucosyltransferase

UDPG Uridine diphosphoglucose

UDP Uridine diphosphate

AEA Arachidonoyl ethanolamide (aka, anandamide) 2-AG 2-Arachidonoyl ethanolamide.

1 -AG 1 -Arachidonoyl ethanolamide.,

DHEA N-Docosahexaenoylethanolamine (aka, synaptamide)

SUS Sucrose synthase.

[0046] The term "glucopyranoside" is used for naming molecules and is shorthand for a β- D-glucose attached through the hydroxyl at the 1 -position (the anomeric carbon) of the glucose to the aglycone.

[0047] The term "aglycone" is used in the present application to refer to the non-glycosidic portion of a glycoside compound.

[0048] The term "prodrug" refers to a compound that, upon administration, must undergo a chemical conversion by metabolic processes before becoming an active pharmacological agent.

[0049] The term "cannabinoid glycoside prodrug" refers generally to the glycosides of cannabinoid compounds, endocannabinoid compounds and vanilloid compounds. The cannabinoid glycoside prodrug undergoes hydrolysis of the glycosidic bond, typically by action of a glycosidase, to release the active cannabinoid, endocannabinoid or vanilloid compounds to a desired site in the body of the subject. The cannabinoid glycoside prodrug of the present invention may also be referred to using the term "cannaboside".

[0050] The term "cannabinoid" is used in the present application to refer generally to compounds found in cannabis and which act on cannabinoid receptors. "Cannabinoid" compounds include, but are not limited to, cannabidiol (CBD), cannabidivarin (CBDV), cannabigerol (CBG), tetrahydrocannabinol (A9-THC or THC), cannabinol (CBN), cannabidiolic acid (CBDA), and tetrahydrocannabivarin (THCV). Particularly preferred cannabinoids compounds are CBD, CBDV, THC and CBN.

[0051] The term "endocannabinoid" is used in the present application to refer to compounds including arachidonoyl ethanolamide (anandamide, AEA), 2-arachidonoyl ethanolamide (2-AG), 1 -arachidonoyl ethanolamide (1 -AG), and docosahexaenoyl ethanolamide (DHEA, synaptamide), oleoyl ethanolamide (OEA), eicsapentaenoyl ethanolamide, prostaglandin ethanolamide, docosahexaenoyl ethanolamide, linolenoyl ethanolamide, 5(Z),8(Z),1 1 (Z)- eicosatrienoic acid ethanolamide (mead acid ethanolamide), heptadecanoul ethanolamide, stearoyl ethanolamide, docosaenoyl ethanolamide, nervonoyl ethanolamide, tricosanoyl ethanolamide, lignoceroyl ethanolamide, myristoyl ethanolamide, pentadecanoyl ethanolamide, palmitoleoyl ethanolamide, docosahexaenoic acid (DHA). Particularly preferred endocannabinoids are AEA, 2-AG, 1 -AG, and DHEA.

[0052] The term "vanilloid" is used in the present application to refer to compounds comprising a vanillyl group and which act on vanilloid receptors like TRPV1 . "Vanilloid" compounds include, but are not limited to, vanillin, capsaicin and curcumin.

[0053] As used herein, the term "about" refers to a +/-10% variation from the nominal value. It is to be understood that such a variation is always included in a given value provided herein, whether or not it is specifically referred to.

[0054] The term "subject" or "patient" as used herein refers to an animal in need of treatment. In one embodiment, the animal is a human.

[0055] 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 invention belongs.

[0056] In accordance with the present invention, cannabinoids, endocannabinoids and vanilloids are employed as substrates for glucosyltransferases to which one or more sugar molecules are attached to create novel cannabinoid glycoside prodrugs. The resulting cannabinoid glycoside prodrugs demonstrate site-specific or tissue-specific delivery, improved aqueous solubility for improved pharmacological delivery, and/or sustained or delayed release of the cannabinoid, endocannabinoid and vanilloid drug molecules.

[0057] Also in accordance with the present invention, the cannabinoid glycoside prodrugs are converted upon hydrolysis of the glycosidic bond to provide the active cannabinoid, endocannabinoid and vanilloid drug. Accordingly, the present invention has demonstrated that glycosides with a hydrophobic aglycone moiety undergo glucose hydrolysis in the gastrointestinal tract or in tissues having increased expression of glycosidases, yielding the hydrophobic cannabinoid compound in the targeted tissue or organ. [0058] The glucose residues of glycosides are commonly acid-hydrolyzed in the stomach or cleaved by glycosidase enzymes in the intestinal tract, including by alpha-glycosidases and beta-glycosidases, which are expressed by intestinal microflora across different regions of the intestine. Accordingly, glycosides are hydrolyzed upon ingestion to release the desired compound into the intestines or target tissues.

[0059] In one embodiment, glycosylation of cannabinoid drugs provides cannabinoid glycoside prodrugs capable of persisting in the acidic stomach environment upon oral administration, thereby allowing delivery of the prodrug into the large intestine, where the cannabinoid aglycones can be liberated by glycosidases produced by colonic bacteria.

[0060] In one embodiment, glycosylation of cannabinoid drugs provides cannabinoid glycoside prodrugs suitable for targeted delivery to tissues having increased expression of glycosidases. Upon parenteral administration of the cannabinoid glycoside prodrug formulation to the subject, the cannabinoid aglycones are liberated by the glycosidases in the target tissues.

[0061] It is also within the scope of the present invention that the cannabinoid glycoside prodrug are also useful as pharmaceutical agents without glucose cleavage, where they exhibit novel pharmacodynamic properties compared to the parent compound alone. The increased aqueous solubility of the cannabinoid glycoside prodrugs of the present invention also enables new formulations for delivery in transdermal or aqueous formulations that would not have been achievable if formulating hydrophobic cannabinoid, endocannabinoid and vanilloid molecules.

[0062] In one embodiment of the present invention, there are provided cannabinoid glycoside prodrug compounds having formula (I):

Figure imgf000010_0001
or a pharmaceutically compatible salt thereof, wherein R is H, β-D-glucopyranosyl, or 3-0-β-ϋ- glucopyranosyl^-D-glucopyranosyl; R' is H or β-D-glucopyranosyl, or 3-0^-D-glucopyranosyl- β-D-glucopyranosyl; and A is an aglycone moiety formed through reaction of a hydroxyl group on a cannabinoid compound, an endocannabinoid compound, or a vanilloid compound.

[0063] In accordance with one embodiment of the present invention, A is A', A" or A'";

wher ' is:

Figure imgf000011_0001

wherein A" is:

Figure imgf000011_0002
Figure imgf000012_0001
and wherein A'" is:

Figure imgf000012_0002
wherein G is H, β-D-glucopyranosyl, 3-0^-D-glucopyranosyl^-D-glucopyranosyl, or β-D- glucopyranosyl-(1→3)^-D-glucopyranosyl-(1→3)-D-glucopyranosyl; or a pharmaceutically compatible salt thereof.

[0064] In accordance with one embodiment of the present invention, the cannabinoid glycoside prodrug is a glycoside of a cannabinoid, wherein the prodrug has the formula (Γ):

Figure imgf000013_0001
wherein R is H, β-D-glucopyranosyl, or 3-0- β-D-glucopyranosyl- β-D-glucopyranosyl;

R' is H, β-D-glucopyranosyl, or 3-0- β-D-glucopyranosyl- β-D-glucopyranosyl; and

wherein A' is:

Figure imgf000013_0002
wherein G is β-D-glucopyranosyl, 3-0^-D-glucopyranosyl^-D-glucopyranosyl, or β-D- glucopyranosyl-(1 -3)-β-D-glucopyranosyl-(1 -3)-D-glucopyranosyl.

[0065] Compounds of Formula (Γ) include the compounds listed in Tables 1 to 4.

[0066] Exemplary cannabidiol (CBD)-glycosides falling within the scope of Formula (Γ), produced by the glycosylation of CBD (VB101 ) in accordance with the present invention, include:

Figure imgf000014_0001



Figure imgf000015_0001

[0067] Exemplary cannabidivarin (CBDV)-glycosides falling within the scope of Formula (Γ), produced by the glycosylation of CBDV (VB201 ) in accordance with the present invention, include:

Figure imgf000015_0002

Figure imgf000016_0001

[0068] Exemplary tetrahydrocannabinol (A9-THC)-glycosides falling within the scope of Formula (Γ), produced by the glycosylation of Δ9-ΤΗΟ (VB301 ) in accordance with the present invention, include:

Figure imgf000017_0001

VB305 and

[0069] Exemplary cannabinol (CBN)-glycosides falling within the scope of Formula (Γ), produced by the glycosylation of CBN (VB401 ) in accordance with the present invention, include:

Figure imgf000017_0002
Figure imgf000018_0001

VB405 , and

[0070] In accordance with one embodiment of the present invention, the cannabinoid glycoside prodrug is a glycoside of an endocannabinoid, the prodrug having the formula (I"):

Figure imgf000018_0002
(I") wherein

R is H, β-D-glucopyranosyl, or 3-0- β-D-glucopyranosyl- β-D-glucopyranosyl;

R' is H, β-D-glucopyranosyl, or 3-0- β-D-glucopyranosyl- β-D-glucopyranosyl; and

wherein A" is:

Figure imgf000018_0003
Figure imgf000019_0001

[0071] Compounds of Formula (I") include the compounds listed in Tables 5 to 8.

[0072] Exemplary arachidonoyi ethanolamide (AEA)-glycosides falling within the scope of Formula (I"), produced by the glycosylation of AEA (VB501 ) in accordance with the present invention, include:

Figure imgf000019_0002

VB502 V8503

Figure imgf000020_0001

VB504 VB505 and

Figure imgf000020_0002

VB508

[0073] Exemplary 2-arachidonoyl ethanolamide (2-AG)-glycosides falling within the scope of Formula (I"), produced by the glycosylation of 2-AG (VB601 ) in accordance with the present invention, include:

Figure imgf000020_0003
Figure imgf000021_0001

Figure imgf000021_0002
, and
Figure imgf000022_0001

[0074] Exemplary 1 -arachidonoyi ethanolamide (l -AG)-glycosides falling within the scope of Formula (I"), produced by the glycosylation of 1 -AG (VB701 ) in accordance with the present invention, include:

Figure imgf000022_0002
Figure imgf000023_0001

Figure imgf000024_0001

[0075] Exemplary N-docosahexaenoylethanolamine (DHEA)-glycosides falling within the scope of Formula (I"), produced by the glycosylation of DHEA (VB801 ) in accordance with the present invention, include:

Figure imgf000024_0002

Figure imgf000024_0003

VB803

Figure imgf000025_0001

Figure imgf000025_0002

, and

Figure imgf000025_0003

[0076] In accordance with one embodiment of the present invention, the cannabinoid glycoside prodrug is a glycoside of a vanilloid, the prodrug having the formula (!'"):

Figure imgf000026_0001
wherein

R is H, β-D-glucopyranosyl, or 3-0- β-D-glucopyranosyl- β-D-glucopyranosyl;

R' is H or β-D-glucopyranosyl, or 3-0- β-D-glucopyranosyl- β-D-glucopyranosyl; and, wherein A'" is:

Figure imgf000026_0002

[0077] Compounds of Formula (H) include the compounds listed in Tables 9 to 1 1 .

[0078] Exemplary capsaicin-glycosides falling within the scope of Formula (H), produced by the glycosylation of capsaicin (VB901 ) in accordance with the present invention, include:

Figure imgf000027_0001

[0079] Exemplary vanillin-glycosides falling within the scope of Formula (H), produced by the glycosylation of vanillin (VB1001 ) in accordance with the present invention, include:

Figure imgf000028_0001
Figure imgf000028_0002

[0080] Exemplary curcumin-glycosides falling within the scope of Formula (H), produced by the glycosylation of curcumin (VB1 101 ) in accordance with the present invention, include:

Figure imgf000028_0003
Figure imgf000029_0001
Figure imgf000029_0002
Figure imgf000029_0003
Figure imgf000029_0004
Figure imgf000030_0001
Figure imgf000030_0002
Figure imgf000030_0003

Figure imgf000031_0001

Figure imgf000031_0002
Figure imgf000032_0001
Figure imgf000032_0002
Figure imgf000032_0003

Figure imgf000033_0001

[0081] In one embodiment, there is provided a method for the site-specific delivery of a cannabinoid drug to a subject, comprising the step of administering to a subject in need thereof one or more cannabinoid glycoside prodrugs in accordance with the present invention. In one embodiment, the site of delivery is the large intestine. In one embodiment, the site of delivery is the rectum. In one embodiment, the site of delivery is the liver. In one embodiment, the site of delivery is the skin.

[0082] In one embodiment, there is provided a method for facilitating the transport of a cannabinoid drug to the brain through intranasal, stereotactic, or intrathecal delivery, or delivery across the blood brain barrier of a subject comprising administering a cannabinoid glycoside prodrug in accordance with the present invention to a subject in need thereof.

[0083] In accordance with the present invention, the cannabinoid glycoside prodrugs are useful in the treatment of conditions that benefit from or can be ameliorated with the administration of a cannabinoid drug. Conditions that can be treated or ameliorated through the administration of cannabinoid glycoside prodrugs of the present invention, include but are not limited to, inflammatory bowel disease including induction of remission from Crohn's disease, and colitis and induction of remission from ulcerative colitis. Among the benefits that can be achieved through the administration of cannabinoid glycoside prodrugs of the present invention are decreased inflammation of the intestines and rectum, decreased pain in the intestines, rectum, as well as decrease in neuropathic pain and abdominal pain, and inhibition of proliferation or cytotoxicity against colorectal cancer. Additional treatment indications, effects, or applications for cannabinoids or cannabinoid glycosides may include but are not limited to anorexia, nausea, emesis, pain, wasting syndrome, HIV-wasting, chemotherapy induced nausea and vomiting, epilepsy, schizophrenia, irritable bowel syndrome, cramping, spasticity, seizure disorders, alcohol use disorders, substance abuse disorders, addiction, cancer, amyotrophic lateral sclerosis, glioblastoma multiforme, glioma, increased intraocular pressure, glaucoma, cannabis use disorders, Tourette's syndrome, dystonia, multiple sclerosis, white matter disorders, demyelinating disorders, chronic traumatic encephalopathy, leukoencephalopathies, Guillain- Barre syndrome, inflammatory bowel disorders, gastrointestinal disorders, bacterial infections, MRSA, sepsis, septic shock, viral infections, arthritis, dermatitis, Rheumatoid arthritis, systemic lupus erythematosus, anti-inflammatory, anti-convulsant, anti-psychotic, anti-oxidant, neuroprotective, anti-cancer, immunomodulatory effects, neuropathic pain, neuropathic pain associated with post-herpetic neuralgia, diabetic neuropathy, shingles, burns, actinic keratosis, oral cavity sores and ulcers, post-episiotomy pain, psoriasis, pruritis, gout, chondrocalcinosis, joint pain, fibromyalgia, musculoskeletal pain, neuropathic-postoperative complications.

[0084] In one embodiment, the cannabinoid glycoside prodrug is administered in a pharmaceutical composition further comprising a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant. In one embodiment, the pharmaceutical compositions comprise one or more cannabinoid glycoside prodrugs and one or more pharmaceutically acceptable carriers, diluents, excipients and/or adjuvants. For administration to a subject, the pharmaceutical compositions can be formulated for administration by a variety of routes including but not limited to oral, topical, rectal, parenteral, and intranasal administration.

[0085] The pharmaceutical compositions may comprise from about 1 % to about 95% of a cannabinoid glycoside prodrug of the invention. Compositions formulated for administration in a single dose form may comprise, for example, about 20% to about 90% of the cannabinoid glycoside prodrug of the invention, whereas compositions that are not in a single dose form may comprise, for example, from about 5% to about 20% of the cannabinoid glycoside prodrug of the invention. Non-limiting examples of unit dose forms include tablets, ampoules, dragees, suppositories, and capsules.

[0086] In a preferred embodiment, the pharmaceutical compositions are formulated for oral administration. Pharmaceutical compositions for oral administration can be formulated, for example, as tablets, troches, lozenges, aqueous or oily suspensions, dispersible powders or granules, emulsion hard or soft capsules, or syrups or elixirs. Such compositions can be prepared according to standard methods known in the art for the manufacture of pharmaceutical compositions and may contain one or more agents selected from the group of sweetening agents, flavouring agents, colouring agents and preserving agents in order to provide pharmaceutically elegant and palatable preparations. Tablets contain the active ingredient in admixture with suitable non-toxic pharmaceutically acceptable excipients including, for example, inert diluents, such as calcium carbonate, sodium carbonate, lactose, calcium phosphate or sodium phosphate; granulating and disintegrating agents, such as corn starch, or alginic acid; binding agents, such as starch, gelatine or acacia, and lubricating agents, such as magnesium stearate, stearic acid or talc. The tablets can be uncoated, or they may be coated by known techniques in order to delay disintegration and absorption in the gastrointestinal tract and thereby provide a sustained action over a longer period. For example, a time delay material such as glyceryl monosterate or glyceryl distearate may be employed to further facilitate delivery of the drug compound to the desired location in the digestive tract.

[0087] Pharmaceutical compositions for oral use can also be presented as hard gelatine capsules wherein the active ingredient is mixed with an inert solid diluent, for example, calcium carbonate, calcium phosphate or kaolin, or as soft gelatine capsules wherein the active ingredient is mixed with water or an oil medium such as peanut oil, liquid paraffin or olive oil.

[0088] Pharmaceutical compositions formulated as aqueous suspensions contain the active compound(s) in admixture with one or more suitable excipients, for example, with suspending agents, such as sodium carboxymethylcellulose, methyl cellulose, hydropropylmethylcellulose, sodium alginate, polyvinylpyrrolidone, hydroxypropyl^-cyclodextrin, gum tragacanth and gum acacia; dispersing or wetting agents such as a naturally-occurring phosphatide, for example, lecithin, or condensation products of an alkylene oxide with fatty acids, for example, polyoxyethyene stearate, or condensation products of ethylene oxide with long chain aliphatic alcohols, for example, hepta-decaethyleneoxycetanol, or condensation products of ethylene oxide with partial esters derived from fatty acids and a hexitol for example, polyoxyethylene sorbitol monooleate, or condensation products of ethylene oxide with partial esters derived from fatty acids and hexitol anhydrides, for example, polyethylene sorbitan monooleate. The aqueous suspensions may also contain one or more preservatives, for example ethyl, or n- propyl p-hydroxy-benzoate, one or more colouring agents, one or more flavouring agents or one or more sweetening agents, such as sucrose, stevia, or saccharin.

[0089] Pharmaceutical compositions can be formulated as oily suspensions by suspending the active compound(s) in a vegetable oil, for example, arachis oil, olive oil, sesame oil or coconut oil, or in a mineral oil such as liquid paraffin. The oily suspensions may contain a thickening agent, for example, beeswax, hard paraffin or cetyl alcohol. Sweetening agents such as those set forth above, and/or flavouring agents may be added to provide palatable oral preparations. These compositions can be preserved by the addition of an anti-oxidant such as ascorbic acid.

[0090] The pharmaceutical compositions can be formulated as a dispersible powder or granules, which can subsequently be used to prepare an aqueous suspension by the addition of water. Such dispersible powders or granules provide the active ingredient in admixture with one or more dispersing or wetting agents, suspending agents and/or preservatives. Suitable dispersing or wetting agents and suspending agents are exemplified by those already mentioned above. Additional excipients, for example, sweetening, flavouring and colouring agents, can also be included in these compositions.

[0091] Pharmaceutical compositions of the invention can also be formulated as oil-in-water emulsions. The oil phase can be a vegetable oil, for example, olive oil or arachis oil, or a mineral oil, for example, liquid paraffin, or it may be a mixture of these oils. Suitable emulsifying agents for inclusion in these compositions include naturally-occurring gums, for example, gum acacia or gum tragacanth; naturally-occurring phosphatides, for example, soy bean, lecithin; or esters or partial esters derived from fatty acids and hexitol, anhydrides, for example, sorbitan monoleate, and condensation products of the said partial esters with ethylene oxide, for example, polyoxyethylene sorbitan monoleate. The emulsions can also optionally contain sweetening and flavouring agents.

[0092] Pharmaceutical compositions can be formulated as a syrup or elixir by combining the active ingredient(s) with one or more sweetening agents, for example glycerol, propylene glycol, sorbitol or sucrose. Such formulations can also optionally contain one or more demulcents, preservatives, flavouring agents and/or colouring agents.

If desired, other active ingredients may be included in the compositions. In one embodiment, the glycoside prodrugs may be combined with other ingredients or substances that have glycosidase activity, or that may in other ways alter drug metabolism and pharmacokinetic profile of various compounds in vivo, including ones in purified form as well as such compounds found within food, beverages, and other products. In one embodiment, the cannabinoid glycoside prodrug is administered in combination with, or formulated together with, substances that have direct glycosidase activity, or that contribute to modifications to the gut microflora that will alter the glycosidase activity in one or more regions of the intestines. Examples of such compositions include, but are not limited to, yogurt, prebiotics, probiotics, or fecal transplants.

[0093] In a further preferred embodiment, the pharmaceutical compositions are formulated for parenteral administration. The term "parenteral" as used herein includes subcutaneous injections, intravenous, intramuscular, intrathecal, intrasternal injection or infusion techniques.

[0094] Parenteral pharmaceutical compositions can be formulated as a sterile injectable aqueous or oleaginous suspension according to methods known in the art and using one or more suitable dispersing or wetting agents and/or suspending agents, such as those mentioned above. The sterile injectable preparation can be a sterile injectable solution or suspension in a non-toxic parentally acceptable diluent or solvent, for example, as a solution in 1 ,3-butanediol. Acceptable vehicles and solvents that can be employed include, but are not limited to, water, Ringer's solution, lactated Ringer's solution and isotonic sodium chloride solution. Other examples include, sterile, fixed oils, which are conventionally employed as a solvent or suspending medium, and a variety of bland fixed oils including, for example, synthetic mono- or diglycerides. Fatty acids such as oleic acid can also be used in the preparation of injectables.

[0095] Due to the highly lipophilic nature of cannabinoids, these molecules are typically poorly absorbed through membranes such as the skin of mammals, including humans, and the success of transdermally administering therapeutically effective quantities of cannabinoid to a subject in need thereof within a reasonable time frame and over a suitable surface area has been substantially limited. It is therefore proposed that the cannabinoid glycoside prodrugs of the present invention, through conjugation of the hydrophobic cannabinoid aglycone to the hydrophilic glycosidic moieties, provide a molecule having an amphiphilic character favourable for passive diffusion which should be more readily absorbed through the skin.

[0096] Accordingly, in one embodiment, the pharmaceutical compositions are formulated for topical administration. Such topical formulations may be presented as, for example, aerosol sprays, powders, sticks, granules, creams, liquid creams, pastes, gels, lotions, ointments, on sponges or cotton applicators, or as a solution or a suspension in an aqueous liquid, a nonaqueous liquid, an oil-in-water emulsion, or a water-in-oil liquid emulsion. [0097] Topical pharmaceutical compositions can be formulated with thickening (gelling) agents. The thickening agent used herein may include anionic polymers such as polyacrylic acid (CARBOPOL® by Noveon, Inc., Cleveland, Ohio), carboxypolymethylene, carboxymethylcellulose and the like, including derivatives of Carbopol® polymers, such as Carbopol® Ultrez 10, Carbopol® 940, Carbopol® 941 , Carbopol® 954, Carbopol® 980, Carbopol® 981 , Carbopol® ETD 2001 , Carbopol® EZ-2 and Carbopol® EZ-3, and other polymers such as Pemulen® polymeric emulsifiers, and Noveon® polycarbophils. Thickening agents or gelling agents are present in an amount sufficient to provide the desired rheological properties of the composition.

[0098] Topical pharmaceutical compositions can be formulated with a penetration enhancer. Non-limiting examples of penetration enhancing agents include C8-C22 fatty acids such as isostearic acid, octanoic acid, and oleic acid; C8-C22 fatty alcohols such as oleyl alcohol and lauryl alcohol; lower alkyl esters of C8-C22 fatty acids such as ethyl oleate, isopropyl myristate, butyl stearate, and methyl laurate; di(lower)alkyl esters of C6-C22 diacids such as diisopropyl adipate; monoglycerides of C8-C22 fatty acids such as glyceryl monolaurate; tetrahydrofurfuryl alcohol polyethylene glycol ether; polyethylene glycol, propylene glycol; 2-(2- ethoxyethoxyl)ethanol; diethylene glycol monomethyl ether; alkylaryl ethers of polyethylene oxide; polyethylene oxide monomethyl ethers; polyethylene oxide dimethyl ethers; dimethyl sulfoxide; glycerol; ethyl acetate; acetoacetic ester; N-alkylpyrrolidone; and terpenes.

[0099] The topical pharmaceutical compositions can further comprise wetting agents (surfactants), lubricants, emollients, antimicrobial preservatives, and emulsifying agents as are known in the art of pharmaceutical formations.

[00100] Transdermal delivery of the cannabinoid glycoside prodrug can be further facilitated through the use of a microneedle array drug delivery system.

[00101 ] Other pharmaceutical compositions and methods of preparing pharmaceutical compositions are known in the art and are described, for example, in "Remington: The Science and Practice of Pharmacy" (formerly "Remingtons Pharmaceutical Sciences"); Gennaro, A., Lippincott, Williams & Wilkins, Philadelphia, PA (2000).

[00102] The pharmaceutical compositions of the present invention described above include one or more cannabinoid glycoside prodrugs of the invention in an amount effective to achieve the intended purpose. Thus the term "therapeutically effective dose" refers to the amount of the cannabinoid glycoside prodrug that improves the status of the subject to be treated, for example, by ameliorating the symptoms of the disease or disorder to be treated, preventing the disease or disorder, or altering the pathology of the disease. Determination of a therapeutically effective dose of a compound is well within the capability of those skilled in the art. In one embodiment, cannabinoid glycosides can be combined to enable simultaneous delivery of multiple cannabinoids in a site-specific manner, including THC and CBD, whose effects in some ways may be synergistic (Russo 2006). Accordingly, in one embodiment, the pharmaceutical composition comprises one or more CBD-glycosides and one or more THC-glycosides formulated together in a single dosage form.

[00103] The exact dosage to be administered to a subject can be determined by the practitioner, in light of factors related to the subject requiring treatment. Dosage and administration are adjusted to provide desired levels of the cannabinoid glycoside prodrug and/or the cannabinoid drug compound obtained upon hydrolysis of the prodrug. Factors which may be taken into account when determining an appropriate dosage include the severity of the disease state, general health of the subject, age, weight, and gender of the subject, diet, time and frequency of administration, drug combination(s), reaction sensitivities, and tolerance/response to therapy. Dosing regimens can be designed by the practitioner depending on the above factors as well as factors such as the half-life and clearance rate of the particular formulation.

[00104] In accordance with the present invention, there is provided a method of producing a cannabinoid glycoside, comprising incubating a cannabinoid aglycone with one or more sugar donors in the presence of one or more glycosyltransferases.

[00105] In one embodiment, the one or more glycosyltransferases is a UGT76G1 or UGT76G1 -like glucosyltransferase. In one embodiment, the one or more glycosyltransferases comprise a UGT76G1 or UGT76G1 -like glucosyltransferase and a Os03g0702000 or Os03g0702000-like glucosyltransferase.

[00106] In one embodiment, the one or more sugar donors are selected from the group consisting of UDP-glucose, UDP-glucuronic acid, UDP-mannose, UDP-fructose, UDP-xylose, UDP-rhamnose, UDP-fluoro-deoxyglucose, and combinations thereof. In a preferred embodiment, the sugar donor is UDP-glucose. [00107] In accordance with the present invention, the cannabinoid aglycone is a cannabinoid, an endocannabinoid, or a vanilloid. In a preferred embodiment, the cannabinoid glycoside prodrug produced by the methods of the present invention is a compound of the Formula (I).

[00108] In one embodiment, the method of producing a cannabinoid glycoside comprises incubating a cannabinoid aglycone with UDP-glucose, in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase under conditions that allow for glycosylation.

[00109] In one embodiment, the method of producing a cannabinoid glycoside comprises incubating a cannabinoid aglycone with one or more sugar donors in the presence of a first glycosyltransferase and a second glycosyltransferase under conditions which allow for glycosylation. In one embodiment, sugar donor is UDP-glucose, the first glycosyltransferase is a UGT76G1 or UGT76G1 -like glucosyltransferase, and the second glycosyltransferase is a Os03g0702000 or Os03g0702000-like glucosyltransferase.

[00110] In one embodiment, the method of producing a cannabinoid glycoside comprises incubating a cannabinoid aglycone with UDP-glucose in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and Os03g0702000 or Os03g0702000-like glucosyltransferase under conditions which allow for glycosylation.

[00111 ] In one embodiment, the method of producing a cannabinoid glycoside comprises incubating a cannabinoid aglycone with maltodextrin, in the presence of a cyclodextrin glucanotransferase under conditions that allow for glycosylation.

[00112] In one embodiment, the method of producing a cannabinoid glycoside comprises incubating a cannabinoid aglycone with UDP-glucose and maltodextrin in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and cyclodextrin glucanotransferase under conditions which allow for glycosylation.

[00113] In a preferred embodiment, the glycosyltransferase employed in the methods of producing the cannabinoid glycoside is UGT76G1 or UGT76G1 -like glucosyltransferase. In one embodiment, the UGT76G1 or UGT76G1 -like glucosyltransferase comprises the sequence as set forth in SEQ ID NO:1 , 3, 5 or 7. [00114] In one embodiment, the glycosyltransferase employed in the methods of producing the cannabinoid glycoside is Os03g0702000 or Os03g0702000-like glucosyltransferase. In one embodiment, the Os03g0702000 or Os03g0702000-like glucosyltransferase comprises the sequence as set forth in SEQ ID NO:9.

[00115] In one embodiment, the method of producing the cannabinoid glycoside further comprises incubating with sucrose synthase. In one embodiment, the sucrose synthase comprises the sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25.

[00116] In one embodiment, the method for the production of a cannabinoid glycoside prodrug comprises expressing one or more of the glycosyltransf erases in a cell or plant which produces the cannabinoid aglycone and isolating the cannabinoid glycoside prodrug.

[00117] Glycosylation of cannabinoids improves solubility in aqueous solutions, as demonstrated by accelerated elution from an C18 analytical HPLC column, indicating that the new cannabinoid-glycosides require far less organic solvent for elution from the hydrophobic chromatography column. This improved solubility was further demonstrated by testing the aqueous solubility of purified solid cannabosides, where solutions were successfully prepared up to 500mg/ml (50% mass/volume) with a mixture of higher glycoside forms of cannabosides. Given the markedly improved solubility and novel secondary and tertiary glycosylation on cannabinoids, glycosylated cannabinoids can act as efficient prodrugs for selective delivery of cannabinoids to desired tissues where the glucose molecules can be hydrolyzed to release the aglycone cannabinoids. Additionally the glycosylations promote stability of CBD and CBDV by protecting them from oxidation and ring-closure of the C6'-hydroxyl group, which prevents degradation into A9-THC or A9-THCV, respectively, and subsequently into cannabinol (CBN) or cannabinavarin (CBNV), respectively

[00118] Increasing the diversity and complexity of sugar attachments to cannabinoids, and administration of a mixture of glycosides will provide altered prodrug delivery kinetics, thus providing an extended release formulation of the drug. The primary detoxification mechanism for cannabinoids in humans is CYP450 mediated hydroxylation of the C7 methyl group of CBD and CBDV, or the C1 1 methyl group of THC and CBN, glycosylation of the acceptor hydroxyl groups of the cannabinoid resorcinol ring may afford protection from C7/C1 1 hydroxylation and subsequent elimination from the body due to steric hindrance preventing the cannabinoid- glycoside from binding in the CYP450 active site. In fact, the hydroxyl groups of CBD are thought to facilitate the binding to the detoxification cytochrome P450 CYP3A4 in the epithelium of the small intestine (Yamaori 201 1 ). Reduced degradation or metabolism in the stomach and small intestine due to these effects could also lead to higher total bioavailability of any glycosylated product upon oral delivery.

[00119] In some cases, removal of the sugar from glycosides in the body may be required in order for the compounds to exert their primary biological activity. Therefore, glycoside prodrugs may enable stable drug formulations that are resistant to abuse, due to the potential for their primary biological effects to only occur after oral ingestion. As most abuse-deterrent compounds are simply mixing or formulation based deterrents, they can still be compromised by simple physical and chemical methods. As one example, the beta-glycosides described herein will only release the aglycone upon the action of beta-glycosidase enzymes. Beta- glycosidases are known to be secreted by microbes that occupy the large intestines of mammals, therefore upon oral ingestion the glycoside prodrugs will remain glycosylated until they reach the large intestine. A similar approach may be used for abuse-resistant, abuse- deterrent, and site-specific delivery of other compounds through glycosylation. It has been found that the UGT76G1 enzyme (SEQ ID N0.1 ) from Stevia rebaudiana transfers a glucose molecule from the sugar donor UDP-glucose (UDPG) to the hydroxyl groups of CBD to create novel CBD-O-glycosides (Table 1 , Figures 2 & 4). The UDPG is inverted by UGT76G1 to produce β-D-glucose residues covalently linked through the to the hydroxyl acceptor sites on CBD. To improve the catalytic efficiency UGT76G1 open reading frame (ORF) codon optimization was performed (SEQ ID NOs. 4 and 6) for expression in Pichia pastoris. Similar to its activity towards steviol glycosides, UGT76G1 is highly productive and has an equilibrium constant (Keq) for CBD of -24. Through experimentation and analysis it was determined that UGT76G1 has the unique ability to apply multiple glucose moieties to the CBD molecule. Upon prolonged incubation of CBD with UGT76G1 and UDPG, HPLC analysis of the reaction mixture yielded 8 glycoside product mobility groups, suggesting that UGT76G1 is able to glycosylate both the C2' and C6' hydroxyl groups on CBD, as well as glycosylating the primary glucose residues with a secondary and tertiary glucose moieties. The secondary and tertiary glycosylations by UGT76G1 occurs at the C3 hydroxyl group of the recipient sugar (3→1 connectivity), as would be suggested by its activity in Stevia, creating 0-(3-1 )-glycosides, and the subsequent products. The CBD-glycoside product mobility groups also suggest that CBD can dock in the UGT76G1 active site both forwards and backwards creating a cis-like- conformation for the glycosylations relative to the cannabinoid backbone (mechanism depicted in Figure 3), or possibly the rotational freedom about the bond at C1 ' (C6 described by Mazur 2009) allows the hydroxyl group to rotate after glycosylation, placing the other hydroxyl group adjacent to the UDPG in the active site and creating a trans-like-conformation for the glycosylations on the cannabinoid backbone (mechanism depicted in Figure 4). Potential CBD molecular docking in the active site of UGT76G1 is depicted in Figure 6 where CBD is superpositioned over the bi-functional substrate for UGT76G1 , Rebaudioside E (RebE) (Figure 6).

[00120] As CBD was successfully glycosylated by UGT76G1 , CBDV was incubated with UGT76G1 and UDPG to test for glycosylation activity. CBDV depletion was observed upon HPLC analysis, in addition to the appearance of four additional product peak mobility groups, which were dependent on addition of both UGT76G1 and UDPG. The four new products formed displayed the same absorbance characteristics as CDBV and were determined to be the primary glycosides CBDV-2'-0-glucopyranosides, CBDV-6'-0-glucopyranosides, and the secondary glycosides CBDV-2'-0-(3-1 )-diglucopyranoside, and CBDV-6'-0-(3-1 )- diglucopyranoside (compounds VB202, VB206, VB204 and VB208, respectively, Table 2). With additional reaction time it was determined that higher order glycoside products were also formed. CBDV-glycoside production was similar to CBD-glycosides from UGT76G1 (Table 2), and proceeded to completion with a Keq -24. Given the number of CBDV-glycoside products, UGT76G1 transfers multiple glucose molecules onto CBDV on both C2' and C6' hydroxyl groups, as well as onto the primary and secondary glycosylations.

[00121 ] When the cannabinoid A9-THC was incubated with UGT76G1 and UDPG, HPLC analysis of the reaction mixture showed three main product peak mobility groups. The three products were identified as A9-THC-1 -0-glucopyranoside, A9-THC-1 -0-(3-1 )- diglucopyranoside, and A9-THC-1 -0-(3-1 ,3-1 )-triglucopyranoside (formal pyran numbering, Table 3, Figure 7). Given that the rigid structure of A9-THC does not have the same rotational freedom as CBD around the C1 ' resorcinol ring attachment, the cannabinoid backbone is recognized in the active site of UGT76G1 with the A9-THC C1 hydroxyl group situated towards the UDPG sugar donor (pyran numbering, Figure 1 B).

[00122] As UGT76G1 demonstrated glycosylation activity for all other phytocannabinoids analyzed, it was also tested for glycosylation activity against cannabinol (CBN). Effective glycosylation of CBN by UGT76G1 was observed, in a similar pattern to A9-THC, as both share a single hydroxyl recipient group at the C1 position of the resorcinol ring. The activity seen with UGT76G1 is consistent with a broad recognition of cannabinoids by the enzyme active site. [00123] Alternative cannabinoid substrates may be inserted into this UGT76G1 glycosylation reaction infrastructure to generate novel cannabinoid-glycosides, given they possess hydroxyl groups in similar positions on the cannabinoid backbone. Ideal candidates are cannabigerol (CBG), cannabichromene (CBC), cannabidiol hydroxyquinone (CBDHQ), HU-331 , other isomers of A9-THC such as Δδ-THC, etc., and synthetic analogues of A9-THC such as HU- 210.

[00124] Similar to the secondary 3→1 glycosylation activity of UGT76G1 , it was determined that following a primary glycosylation by UGT76G1 , the UGT enzyme Os03g0702000 (SEQ ID NO.9) from Oryza sativa is also capable of transferring an additional glucose moiety from UDP- glucose onto the C2-hydroxyl of the primary sugar (Tables 1 - 1 1 , Figures 7- 9 & 12 - 14). This glycosylation activity is consistent with the activity of UGT Os03g0702000 towards steviol glycosides in establishing C2-hydroxyl secondary glycosylations (2→1 connectivity) on existing primary glucose residues. This secondary glycosylation was observed with CBDV (Table 2, Figure3), and THC (Table 2, Figure 7), generating novel CBDV and A9THC-1 -0-(2-1 )- diglucopyranoside species, respectively. Consistent with broad substrate recognition and reactivity, this activity of Os03g0702000 was further demonstrated for the remainder of the substrates identified in Figure 1 .

[00125] In addition to the UDPG-dependent glucosyltransferase activity, cyclodextrin- glucanotransferase (CGTase, Toruzyme 3.0L, trademark of Novozymes Inc.) is capable of transferring a short a-(1 -4)-maltodextrin chain onto the hydroxyl groups of cannabinoids. The CGTase is also capable of glycosylating primary and secondary glycosylations established by UGT76G1 and Os03g0702000, resulting in carbohydrate attachments that start with β-D- glucose molecules, but terminating in a-D-glucose molecules termed β-primed-a-glucosyl (Tables 1 -1 1 ). a-glycosylation by cyclodextrin glucanotransferase mediated maltodextrin transfer can occur on any of the hydroxyl groups of the primary or secondary sugars covalently linked to the cannabinoid. One skilled in the art will appreciate that this makes possible any number of conformations of a-glycosyl chains linked to the glycosides listed in Tables 1 -1 1 .

[00126] Alternative enzymes with homology to UGT76G1 and Os03g0702000 may be used to produce the same glycosylation of cannabinoids. Suitable enzymes for establishing the primary glycosylation similar to UGT76G1 are additional members of the UGT76 clade such as UGT76G2 or UGT76H1 . BLAST results with the UGT76G1 protein sequence yield a maximum homology of 49% identity, as much as 66% positives (similar identity). Ideal candidates may have low overall peptide identity or similarity, but will likely have conserved amino acids at the opening adjacent to the UDPG catalytic site. This sequence is exemplified by a leucine at position 379, and a broader peptide sequence of SDFGLDQ (AA's 375 to 381 of UGT76G1 ). Suitable enzymes for producing the secondary glycosylation of Os03g0702000 are members of the UGT91 clade, including UGT91 D1 and UGT91 D2.

[00127] The glycosylation reactions performed herein included UDP-glucose as the nucleotide sugar donor, however there is some cross-reactivity amongst UGTs that allows for use of alternative nucleotide sugars such as UDP-glucuronic acid, etc. Glucuronic acid is the predominant nucleotide sugar utilized by phase-ll detoxification UGTs in the liver, and cannabinoid-glucuronides are a common detoxification product. Additional nucleotide sugars which could be used to donate carbohydrate moieties to create novel glycosides with similar properties include UDP-glucuronic acid, UDP-mannose, UDP-fructose, UDP-xylose, UDP- rhamnose, UDP-fluorodeoxyglucose, etc. In addition, nucleotide sugars can also be used in combination to create glycosides that contain multiple types of residues on the same aglycone backbone. Alternative strategies to further improve the solubility and delivery of cannabinoids and other compounds described herein include their glycosylation and then functionalizing the sugar moieties with additional ligands or modifications. Examples of this include sulfation, myristoylation, phosphorylation, acetylation, etc.

[00128] The endocannabinoid system has recently been the subject of intense research efforts due to its demonstrated role in and impact on a broad range of clinical pathologies. As UGT76G1 has been determined to recognize a broad class of phytocannabinoids, it was hypothesized that the same enzyme would also recognize and glycosylate endocannabinoids, which are the endogenous signaling molecules recognized by the cannabinoid receptors in Humans. Upon testing a sample of four prototypic endocannabinoids including arachidonoylethanolamide (anandamide, AEA), 2-arachidonoylethanolamide (2-AG), 1 - arachidonoylethanolamide (1 -AG), and docosahexaenoyl ethanolamide (DHEA, synaptamide), it was found that UGT76G1 effectively glycosylated each endocannabinoid (Tables 5-8, Figures 9-12). Glycosylation of endocannabinoids enables the creation of endocannabinoid-glycosides and other fatty acid neurotransmitter-glycosides, representing a new method of targeted delivery of endocannabinoids.

[00129] As endocannabinoids such as AEA, 2-AG, 1 -AG, and synaptamide are glycosylated by UGT76G1 , it is hypothesized that similar endocannabinoids will also be suitable substrates for glycosylation by UGT76G1 . Other endocannabinoid candidates that are likely to be glycosylated by UGT76G1 include oleoyl ethanolamide (OEA), eicsapentaenoyl ethanolamide, prostaglandin ethanolamide, docosahexaenoyl ethanolamide, linolenoyl ethanolamide, 5(Z),8(Z),1 1 (Z)- eicosatrienoic acid ethanolamide (mead acid ethanolamide), heptadecanoul ethanolamide, stearoyl ethanolamide, docosaenoyl ethanolamide, nervonoyl ethanolamide, tricosanoyl ethanolamide, lignoceroyl ethanolamide, myristoyl ethanolamide, pentadecanoyl ethanolamide, palmitoleoyl ethanolamide, docosahexaenoic acid (DHA), and similar compounds. These glycolipids may have a wide range of commercial uses, ranging from pharmaceutical use as a novel endocannabinoid drug with improved solubility and pharmacokinetic properties, to use as an antibacterial agent, to use as a detergent similar to other glycolipids, etc.

[00130] It has been characterized that AEA and CBD are full agonists of the toll-like vanilloid receptor type 1 (TRPV1 ), which is the receptor for capsaicin. In addition, other cannabinoids and botanical extracts, including but not limited to CBD, CBN, cannabigerol (CBG), and various propyl homologues of CBD, THC, and CBG have been demonstrated to bind and have activity towards transient receptor potential channels (TRPs) (De Petrocellis 201 1 ). This includes stimulating and desensitizing TRPV1 , as well as TRPA1 , TRPV2, and also antagonism of TRPM8. Although stimulation of TRPV1 leads to vasodilation and inflammation, capsaicin and its analogues act to desensitize the receptors to stimulants, and provide potent antiinflammatory effects (Bisogno 2001 ). Analogous effects may occur with TRPA1 in addition to other TRPs. For CBD, this may occur at concentrations that are lower than what is required for binding of cannabinoid receptors, and at concentrations that are within the range of those typically attained in human clinical testing and use. In addition to acting as a direct agonist of the TRPV1 receptor, CBD has been shown to inhibit fatty acid amide hydroxylase (FAAH), the enzyme responsible for facilitating the metabolism of the endocannabinoid anandamide (Watanabe, 1998; DE e Petrocellis 2010). Given that these phytocannabinoids act as ligands of diverse TRPs, it was postulated that UGT76G1 would be capable of glycosylating many different ligands of the same TRPs, including TRPM8, TRPV2, TRPA1 , and TRPV1 . Capsaicin is capable of contorting into a CBD-like structure (Bisogno 2001 ), therefore it was postulated that capsaicin was likely to be a suitable substrate for glycosylation by UGT76G1 . To this end, it was shown that UGT76G1 is capable of glycosylating the vanilloid moiety of capsaicin in a structurally identical way to PaGT3 from Phytolacca americana (Noguchi 2009). As the glycosylated structure of capsaicin is the vanilloid head, it was further hypothesized that UGT76G1 would be capable of glycosylation of the minimal vanilloid, i.e., vanillin, as well as many analogues. Consistent with this hypothesis, through HPLC analysis it was determined that UGT76G1 created multiple glycoside products of vanillin (Figure 14, Table 10). Seeking to test the ability of UGT76G1 to glycosylate vanilloids more broadly, curcumin, the well characterized vanilloid found in turmeric spice, isolated from the ginger Curcuma longa was applied as a substrate in the glycosylation reaction. Consistent with the glycosylation of vanillin, UGT76G1 effectively glycosylated curcumin, creating multiple glycoside product peaks, suggesting a bifunctional recognition and glycosylation by UGT76G1 similar to that seen with CBD and steviol glycosides (Figures 15A & 15B, Table 1 1 ).

[00131 ] Cannabinoid glycosides may also have direct bioactive and therapeutic effects, beyond their utility a prodrug for their aglycone form. Quercetin is an antioxidant flavonoid that is ubiquitous in vegetables and often present both in its aglyone and glycosylated forms. It has been demonstrated through in vitro studies that quercetin glucuronides act as a bioactive agent as well as a precursor molecule to aglycone quercetin (Terao 201 1 ). In many cases, including with glycosides that exert antibacterial and antitumor effects, the glycosidic residues are crucial to activity (Kren & Rezanka 2008).

[00132] Glycosides have also been demonstrated to receive facilitated transport across the blood brain barrier (BBB) by the glucose transporter GLUT1 . A prime example is the glycoside of ibuprofen achieving a significant increase of ibuprofen aglycone concentration in the brain (Chen 2009). Similar to these glycosides, glycosides of cannabinoids and other compounds described herein may benefit from enhanced facilitated transport across the BBB or other barriers. Glucose transporters are a wide group of membrane proteins encoded by the human genome and that are found not only in the BBB but across many different cells and tissues, including brain, erythrocytes, fat, muscle, kidney, liver, intestine, and pancreas, so glycosylation will be tailored to provide site-specific delivery to any of these tissues. Accordingly, in one embodiment, there is provided a method for facilitating the transport of a cannabinoid drug across the blood brain barrier of a subject comprising administering to the subject a cannabinoid glycoside prodrug in accordance with the present invention.

[00133] Delivery of cannabinoids and cannabidiol to the brain may be especially useful because of oligodendrocyte protective (oligoprotective) and general neuroprotective effects. It has been demonstrated that cannabinoid signaling is involved with both oligodendrocyte differentiation (Gomez 2010) and that cannabinoids promote oligodendrocyte progenitor survival (Molina-Holgado 2002). Drug formulations that include cannabidiol as a major ingredient have been approved to treat muscle spasticity and pain from multiple sclerosis, a neurodegenerative disorder that causes loss of myelin and oligodendrocyte progenitor cells. The effects of cannabidiol have been demonstrated to mediate oligoprotective effects through attenuation of endoplasmic reticulum stress pathways (Mecha 2012). Cannabidiol has also been studied extensively for its antipsychotic effects, however the exact role in protection of oligodendroctyes and promotion of remyelination has not yet been described (Zuardi 2012). Despite the correlation between the clinical symptoms of psychosis with neuropathological analysis that indicates dysmyelination is involved, the role of dysmyelination as a driver or cause of schizophrenia and other psychoses remains controversial (Mighdoll 2015). Remyelination has also been described as potentially useful for treatment of Alzheimer's disease and other forms of dementia (Bartzokis 2004). Therefore, delivery of cannabinoids to the brain may be especially useful for its established neuroprotective and oligoprotective effects. Cannabinoid glycoside drug formulations co-administered in combination with other agents that influence other aspects of repair or regeneration, such as oligodendrocyte progenitor differentiation or remyelination, may also prove to be beneficial. This includes compounds such as anti-LINGO-1 monoclonal antibodies, guanabenz, sephinl , benzatropine, clemastine, polyunsaturated fatty acids, etc.

[00134] In the course of the present work, it was discovered that UGT76G1 , Os03g0702000 and cyclodextrin glucanotransferase (CGTase) were capable of primary, secondary and tertiary glycosylations of steviol glycosides and aglycone products of diverse chemical structure, including cannabinoids, endocannabinoids, vanillin, curcumin, and capsaicin.

[00135] In the screening and analysis methods described by Dewitte 2016, a 50mm HPLC separation column combined with a high solvent flow rate was used limiting the separation and overall detection of glycoside products. Thus, the interpretation of the glycosylation reaction products for many compounds is speculative, yet still reinforces the significance of the present finding that UGT76G1 has broad substrate specificity. Clearly, the work described herein demonstrate that UGT76G1 can glycosylate not only steviol glycosides, but other forms of glycosides, and novel aglycone compounds such as cannabidiol as well. Internal studies that used an improved separation methodology involving a 150mm length C18 column coupled with a low solvent flowrate also enabled the clear detection of secondary and tertiary glycosides. These compounds were unable to be detected by the methods described in Dewitte 2016, and provide additional verification of the ability of UGT76G1 to not only glycosylate compounds with diverse chemical structures, but also to perform multiple higher order glycosylations on glycosides of these same compounds.

[00136] The reactions described herein take place in vitro using recombinant enzymes and all necessary cofactors, and the expression of UGT76G1 enzyme within the cells of a Cannabis plant is possible for the in vivo biotransformation of cannabinoids prior to extraction of cannabinoids from plant tissue. As UGT76G1 is an enzyme from the plant Stevia rebaudiana, it will be compatible with expression in the genus Cannabis. The ideal strategy for expression of UGT76G1 within the Cannabis plant is to genetically engineer the UGT76G1 open reading frame under a promoter element that is specific for the same tissue that cannabinoids are produced in, namely the secretory trichomes of the plant. Suitable promoter elements include the promoter for the cytosolic 0-acetylserine(thiol)lyase (OASA1 ) enzyme from Arabidopsis thaliana (Gutierrez-Alcala 2005). Candidates for transformation with UGT76G1 include Cannabis sativa, Cannabis indica, and Cannabis ruderalis. A similar approach may be used with UGT76G1 and similar enzymes for in planta production of glycosylated secondary metabolites within many other different plant species, and may be especially useful when plant species already produce large quantities of the desired aglycone product or known enzyme substrate.

[00137] In the course of performing phytocannabinoid glycosylation reactions CBD and THC displayed noticeable antimicrobial activity, even preventing large-scale reaction mixtures from becoming contaminated after failure of the sterile filter apparatus. Prior pilot-scale glycosylation reaction utilizing steviol glycosides as substrates during enzymatic processing were quite susceptible to infection in the absence of strict sanitation techniques. CBD and THC pilot-scale reactions remained aseptic for over a week in the same reaction vessels with very limited ongoing maintenance or care. To this end, the use of the aglycone cannabinoids and their respective glycosides is proposed as efficient antimicrobial agents. Accordingly, in one embodiment, there is provided an antimicrobial agent comprising an effective amount of a cannabinoid glycoside prodrug in accordance with the present invention.

[00138] Similarly, upon the production of large quantities of cannabinoid-glycosides and formulation in aqueous solutions, it was observed that multiple cannabinoid-glycosides in water had foaming properties similar to detergents. This is consistent with other glycoside detergents like 8-octylglycoside, 8-octylthioglycoside, and similar, and establishes a potential use for cannabinoid-glycosides as a detergent. Accordingly, in one embodiment, there is provided a detersive agent comprising an effective amount of a cannabinoid glycoside prodrug in accordance with the present invention. Nucleic Acids

[00139] The present invention provides for nucleic acids comprising nucleotide sequences encoding a glycosyltransferase. The glycosyltransf erases of the present invention are capable of primary, secondary, tertiary glycosylations or a combination thereof. In certain embodiments, the glycosyltransf erases are capable of primary, secondary and tertiary glycosylations. In other embodiments, the glycosyltransf erases are capable of secondary and tertiary glycosylations. In certain embodiments, the nucleic acids encode a glucosyltransferase, including but not limited to a UDP-glucosyltransferase. The glucosyltransferases include but are not limited to a Stevia rebaudiana UDP-glucosyltransferase, such as UGT76G1 or UGT74G1 or an Oryza sativa glucosyltrasferase, such as Os03g0702000. In other embodiments, the invention provides for nucleic acids comprising nucleotide sequences encoding a cyclodextrin glucanotransferase. Also provided are nucleic acids comprising nucleotide sequences that encode a sucrose synthase.

[00140] Nucleic acids include, but are not limited to, genomic DNA, cDNA, RNA, fragments and modified versions, including but not limited to codon optimized versions thereof. For example, the nucleotide sequences may be codon optimized for expression in Pichia pastoris or E. coli. The nucleic acids may include the coding sequence of the glycosyltransferase or sucrose synthase, in isolation, in combination with additional coding sequences (e.g., including but not limited to a purification tag).

[00141 ] In certain embodiments, the nucleic acid comprises a sequence encoding UGT76G1 or UGT76G1 -like glucosyltransferase. UGT76G1 -like glucosyltransferase include for example, other members of the UGT76G1 clade such as UGT76G2 or UGT76H1 . In certain embodiments, the nucleic acid comprises a sequence encoding an UGT76G1 glucosyltransferase having the amino acid sequence as set forth in any one of SEQ ID NOs:1 , 3, 5 and 7 and listed below or fragments and variants thereof.

[00142] SEQ ID NO:1 (UGT76G1 (native protein sequence))

MENKTETTVRRRRRIILFPVPFQGHINPILQLANVLYSKGFSITIFHTNFNKPKTSNYPHFTFRFIL

DNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELLMLASEEDEEVSCLITDALWYFAQS

VADSLNLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVKDIKSAYSN

WQILKEILGKMIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLDHDRT

VFQWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSKQSFLWVVRPGFVKGSTWVEPLP

DGFLGERGRIVKWVPQQEVLAHGAIGAFWTHSGWNSTLESVCEGVPMIFSDFGLDQPLNARY

MSDVLKVGVYLENGWERGEIANAIRRVMVDEEGEYIRQNARVLKQKADVSLMKGGSSYESLE

SLVSYISSL [00143] SEQ ID NO:3 (UGT76G1 with a 6x Histidine tag at the N-terminus)

MHHHHHHGSGENKTETTVRRRRRIILFPVPFQGHINPILQLANVLYSKGFSITIFHTNFNKPKTS

NYPHFTFRFILDNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELLMLASEEDEEVSCLI

TDALWYFAQSVADSLNLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPML

KVKDIKSAYSNWQILKEILGKMIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTA

SSSSLLDHDRTVFQWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSKQSFLWVVRPGFV

KGSTWVEPLPDGFLGERGRIVKWVPQQEVLAHGAIGAFWTHSGWNSTLESVCEGVPMIFSDF

GLDQPLNARYMSDVLKVGVYLENGWERGEIANAIRRVMVDEEGEYIRQNARVLKQKADVSLM

KGGSSYESLESLVSYISSL

[00144] SEQ ID N0:5 (UGT76G1 with a 6x Histidine-Glutamine tag at the N-terminus)

MHQHQHQSGSMENKTETTVRRRRRIILFPVPFQGHINPILQLANVLYSKGFSITIFHTNFNKPKT

SNYPHFTFRFILDNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELLMLASEEDEEVSCL

ITDALWYFAQSVADSLNLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPML

KVKDIKSAYSNWQILKEILGKMIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTA

SSSSLLDHDRTVFQWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSKQSFLWVVRPGFV

KGSTWVEPLPDGFLGERGRIVKWVPQQEVLAHGAIGAFWTHSGWNSTLESVCEGVPMIFSDF

GLDQPLNARYMSDVLKVGVYLENGWERGEIANAIRRVMVDEEGEYIRQNARVLKQKADVSLM

KGGSSYESLESLVSYISSL

[00145] SEQ ID N0:7

MENKTETTVRRRRRIILFPVPFQGHINPILQLANVLYSKGFSITIFHTNFNKPKTSNYPHFTFRFIL

DNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELLMLASEEDEEVSCLITDALWYFAQS

VADSLNLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVKDIKSAYSN

WQILKEILGKMIKQTRASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLDHDRT

VFQWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSKQSFLWVVRPGFVKGSTWVEPLP

DGFLGERGRIVKWVPQQEVLAHGAIGAFWTHSGWNSTLESVCEGVPMIFSDFGLDQPLNARY

MSDVLKVGVYLENGWERGEIANAIRRVMVDEEGEYIRQNARVLKQKADVSLMKGGSSYESLE

SLVSYISSLGSHHHHHH

[00146] In certain embodiments, the nucleic acid comprises a sequence encoding UGT76G1 having the amino acid sequence as set forth in AAR06912.1 . In certain embodiments, the nucleic acid molecule comprises a sequence encoding UGT76G1 glucosyltransferase and comprising the nucleotide sequence as set forth in any one of SEQ ID NOs: 2, 4, 6 and 8 and listed below, or fragments and variants thereof. [00147] SEQ ID NO:2 (UGT76G1 native nucleic acid sequence)

ATGGAAAATAAAACGGAGACCACCGTTCGCCGGCGCCGGAGAATAATATTATTCCCGGTA

CCATTTCAAGGCCACATTAACCCAATTCTTCAGCTAGCCAATGTGTTGTACTCTAAAGGATT

CAGTATCACCATCTTTCACACCAACTTCAACAAACCCAAAACATCTAATTACCCTCACTTCA

CTTTCAGATTCATCCTCGACAACGACCCACAAGACGAACGCATTTCCAATCTACCGACTCA

TGGTCCGCTCGCTGGTATGCGGATTCCGATTATCAACGAACACGGAGCTGACGAATTACG

ACGCGAACTGGAACTGTTGATGTTAGCTTCTGAAGAAGATGAAGAGGTATCGTGTTTAATC

ACGGATGCTCTTTGGTACTTCGCGCAATCTGTTGCTGACAGTCTTAACCTCCGACGGCTTG

TTTTGATGACAAGCAGCTTGTTTAATTTTCATGCACATGTTTCACTTCCTCAGTTTGATGAG

CTTGGTTACCTCGATCCTGATGACAAAACCCGTTTGGAAGAACAAGCGAGTGGGTTTCCTA

TGCTAAAAGTGAAAGACATCAAGTCTGCGTATTCGAACTGGCAAATACTCAAAGAGATATT

AGGGAAGATGATAAAACAAACAAGAGCATCTTCAGGAGTCATCTGGAACTCATTTAAGGAA

CTCGAAGAGTCTGAGCTCGAAACTGTTATCCGTGAGATCCCGGCTCCAAGTTTCTTGATAC

CACTCCCCAAGCATTTGACAGCCTCTTCCAGCAGCTTACTAGACCACGATCGAACCGTTTT

TCAATGGTTAGACCAACAACCGCCAAGTTCGGTACTGTATGTTAGTTTTGGTAGTACTAGT

GAAGTGGATGAGAAAGATTTCTTGGAAATAGCTCGTGGGTTGGTTGATAGCAAGCAGTCG

TTTTTATGGGTGGTTCGACCTGGGTTTGTCAAGGGTTCGACGTGGGTCGAACCGTTGCCA

GATGGGTTCTTGGGTGAAAGAGGACGTATTGTGAAATGGGTTCCACAGCAAGAAGTGCTA

GCTCATGGAGCAATAGGCGCATTCTGGACTCATAGCGGATGGAACTCTACGTTGGAAAGC

GTTTGTGAAGGTGTTCCTATGATTTTCTCGGATTTTGGGCTCGATCAACCGTTGAATGCTA

GATACATGAGTGATGTTTTGAAGGTAGGGGTGTATTTGGAAAATGGGTGGGAAAGAGGAG

AGATAGCAAATGCAATAAGAAGAGTTATGGTGGATGAAGAAGGAGAATACATTAGACAGAA

TGCAAGAGTTTTGAAACAAAAGGCAGATGTTTCTTTGATGAAGGGTGGTTCGTCTTACGAA

TCATTAGAGTCTCTAGTTTCTTACATTTCATCGTTGTAA

[00148] SEQ ID N0:4 (Sequence encoding SEQ ID NO:3 codon optimized for expression in Pichia pastoris)

ATGCACCACCATCACCACCATGGTTCTGGTGAAAACAAAACTGAAACTACTGTTAGAAGAA

GAAGAAGAATCATTTTGTTTCCAGTACCATTTCAAGGCCATATCAATCCAATTCTTCAATTG

GCCAATGTTTTGTACTCCAAAGGATTCTCCATCACCATTTTTCACACCAATTTCAACAAACC

AAAGACTTCCAACTATCCTCACTTCACTTTCAGATTTATTTTGGATAATGATCCTCAAGATG

AAAGAATTTCCAATCTTCCGACTCATGGTCCTTTGGCTGGTATGAGAATTCCAATCATCAAT

GAACATGGTGCTGATGAATTAAGAAGAGAATTGGAACTTTTGATGTTGGCTTCTGAAGAAG

ATGAAGAAGTTTCATGTTTAATCACTGATGCTTTATGGTATTTTGCTCAATCTGTTGCTGAT

TCTTTGAATTTGCGACGGTTGGTTTTGATGACTTCTTCTTTGTTCAACTTTCATGCTCATGT

TTCTTTACCTCAGTTTGATGAACTTGGATATTTGGATCCAGATGACAAAACTAGATTGGAAG

AACAAGCTAGTGGGTTTCCTATGTTGAAAGTCAAAGATATCAAATCTGCTTACTCCAACTG

GCAAATTCTCAAAGAAATTTTGGGAAAAATGATCAAACAAACAAAAGCTTCTTCTGGAGTCA

TTTGGAACTCATTCAAAGAATTGGAAGAATCTGAATTGGAAACTGTTATTAGAGAAATTCCT

GCTCCAAGTTTTTTGATTCCTTTGCCAAAACATTTGACTGCTTCTTCTTCTTCTTTATTGGAT

CACGATAGAACTGTTTTTCAATGGTTAGATCAACAACCTCCATCTTCTGTTTTGTATGTTAG

TTTTGGATCTACTTCTGAAGTTGATGAAAAAGATTTTTTGGAAATTGCTAGAGGTTTGGTTG

ATTCCAAACAAAGTTTTTTATGGGTTGTTAGACCAGGATTTGTCAAAGGATCTACTTGGGTC

GAACCTTTGCCAGATGGATTTTTGGGAGAAAGAGGAAGAATTGTCAAATGGGTTCCACAG

CAAGAAGTTTTGGCTCATGGTGCTATTGGTGCTTTTTGGACTCATTCTGGATGGAACTCTA

CTTTGGAATCTGTTTGTGAAGGTGTTCCAATGATTTTTTCTGATTTTGGTTTGGATCAACCA

TTGAATGCTAGATACATGTCTGATGTTTTGAAAGTTGGTGTTTATTTGGAAAATGGGTGGG

AAAGAGGTGAAATTGCCAATGCTATTAGAAGAGTCATGGTTGATGAAGAAGGAGAATACAT TAGACAAAATGCTAGAGTTTTGAAACAAAAAGCTGATGTTTCTTTGATGAAGGGTGGATCTT CTTATGAATCTTTGGAATCTTTGGTTTCTTACATTTCTTCTCTTTAA

[00149] SEQ ID NO:6 (Sequence encoding SEQ ID NO:5 codon optimized for expression in Pichia pastoris)

ATGCATCAACATCAACACCAATCTGGATCTATGGAGAACAAGACCGAGACTACAGTTAGAA

GAAGAAGAAGAATAATCCTGTTTCCAGTACCATTCCAAGGACACATCAACCCAATCTTGCA

GTTAGCAAATGTACTTTATTCTAAAGGCTTTAGTATTACGATTTTTCACACTAATTTTAATAA

GCCAAAAACATCCAATTACCCTCACTTCACATTCAGATTTATCTTGGATAACGATCCTCAAG

ATGAACGTATCTCCAACCTGCCAACACATGGACCATTGGCCGGTATGCGTATTCCTATAAT

CAACGAGCATGGTGCTGATGAGCTTAGACGTGAACTGGAACTGTTGATGCTGGCATCGGA

GGAAGATGAAGAGGTTAGTTGCTTGATAACGGATGCCCTCTGGTATTTCGCACAATCAGTC

GCTGACTCCTTGAACCTTAGGAGATTGGTATTGATGACTAGTTCGTTGTTCAACTTCCATG

CCCATGTTTCTTTGCCTCAATTTGATGAGCTGGGTTATTTGGATCCTGACGATAAGACTCG

TTTAGAAGAACAGGCGTCAGGCTTCCCCATGTTAAAGGTTAAAGATATTAAGTCCGCCTAT

TCTAACTGGCAAATTCTCAAAGAGATTCTAGGGAAAATGATTAAACAAACCAAGGCCTCTTC

AGGAGTAATCTGGAACAGTTTCAAAGAACTAGAAGAATCCGAGTTGGAAACTGTTATTCGT

GAAATCCCTGCTCCATCTTTCCTTATCCCATTACCAAAGCACCTCACTGCCTCCTCTAGTTC

TCTTCTGGACCATGATAGAACAGTCTTTCAGTGGCTCGATCAGCAACCTCCATCTTCTGTC

TTGTACGTTAGTTTTGGTTCCACCTCGGAAGTAGATGAAAAAGACTTTCTGGAAATTGCTC

GAGGACTAGTTGACTCCAAGCAATCCTTTCTGTGGGTTGTTAGACCTGGATTCGTAAAAGG

ATCCACCTGGGTAGAACCCCTCCCAGATGGATTTTTGGGCGAAAGGGGAAGAATTGTTAA

ATGGGTGCCTCAACAAGAAGTTTTAGCTCATGGGGCCATTGGAGCTTTTTGGACTCATAGT

GGATGGAATTCTACCTTAGAATCTGTTTGTGAAGGAGTTCCAATGATTTTTTCTGATTTTGG

ATTGGATCAGCCTCTTAATGCCAGATATATGTCCGATGTCCTCAAGGTCGGAGTGTACCTG

GAAAATGGTTGGGAGAGAGGTGAGATTGCAAATGCTATACGTAGAGTCATGGTTGATGAA

GAGGGCGAGTATATTAGACAAAACGCTAGAGTGCTAAAGCAGAAGGCCGATGTTTCCCTT

ATGAAGGGGGGAAGTTCATATGAGAGTTTGGAATCCCTAGTGTCCTACATTTCTTCGCTAT

AA

[00150] SEQ ID NO:8 (Sequence encoding SEQ ID NO:7 codon optimized for expression in Escherichia coli)

ATGGAAAATAAAACCGAAACCACCGTCCGTCGCCGTCGTCGTATCATTCTGTTCCCGGTCC

CGTTCCAAGGTCACATCAACCCGATTCTGCAGCTGGCCAACGTGCTGTATAGCAAAGGTTT

CTCTATCACCATCTTCCATACGAACTTCAACAAACCGAAAACCTCTAACTACCCGCACTTTA

CGTTCCGTTTTATTCTGGATAACGACCCGCAGGATGAACGCATCAGTAATCTGCCGACCCA

TGGTCCGCTGGCGGGTATGCGTATTCCGATTATCAACGAACACGGCGCAGATGAACTGCG

TCGCGAACTGGAACTGCTGATGCTGGCCTCTGAAGAAGATGAAGAAGTTAGTTGCCTGAT

CACCGACGCACTGTGGTATTTTGCCCAGAGTGTTGCAGATTCCCTGAACCTGCGTCGCCT

GGTCCTGATGACGAGCTCTCTGTTCAATTTTCATGCCCACGTTTCCCTGCCGCAGTTCGAT

GAACTGGGTTATCTGGACCCGGATGACAAAACCCGCCTGGAAGAACAAGCTTCAGGCTTT

CCGATGCTGAAAGTCAAAGATATTAAAAGTGCGTACTCCAACTGGCAGATTCTGAAAGAAA

TCCTGGGTAAAATGATCAAACAAACCCGTGCAAGTTCCGGCGTCATCTGGAATTCCTTCAA

AGAACTGGAAGAATCAGAACTGGAAACGGTGATTCGCGAAATCCCGGCTCCGTCTTTTCT

GATTCCGCTGCCGAAACATCTGACCGCGTCATCGAGCTCTCTGCTGGATCACGACCGTAC

GGTGTTTCAGTGGCTGGATCAGCAACCGCCGAGTTCCGTGCTGTACGTTAGCTTCGGTAG CACCTCTGAAGTGGATGAAAAAGACTTTCTGGAAATCGCTCGTGGCCTGGTTGATTCAAAA

CAATCGTTCCTGTGGGTGGTTCGCCCGGGTTTTGTGAAAGGCAGCACGTGGGTTGAACC

GCTGCCGGATGGCTTCCTGGGTGAACGTGGTCGCATTGTCAAATGGGTGCCGCAGCAAG

AAGTGCTGGCACATGGTGCTATCGGCGCGTTTTGGACCCACTCAGGTTGGAACTCGACGC

TGGAAAGCGTTTGTGAAGGTGTCCCGATGATTTTCTCGGATTTTGGCCTGGACCAGCCGC

TGAATGCACGTTATATGAGCGATGTTCTGAAAGTCGGTGTGTACCTGGAAAACGGTTGGG

AACGCGGCGAAATTGCGAATGCCATCCGTCGCGTTATGGTCGATGAAGAAGGCGAATATA

TCCGTCAGAATGCTCGCGTCCTGAAACAAAAAGCGGACGTTAGTCTGATGAAAGGCGGTT

CATCGTACGAATCCCTGGAATCACTGGTCTCCTACATTTCTTCTCTGGGCTCGCATCATCA

TCATCATCATTAA

[00151 ] In certain embodiments, the nucleic acid molecule encodes an UGT76G1 glucosyltransferase and comprises the nucleotide sequence as set forth in GenBank Accession number AY345974.1 or a variant or fragment thereof.

[00152] In certain embodiments, the nucleic acid comprises a sequence encoding UGT76G2 glucosyltransferase. In specific embodiments, the nucleic acid comprises a sequence encoding UGT76G2 glucosyltransferase having the amino acid sequence as set forth in SEQ ID NO:27 and listed below or variants and fragments thereof.

[00153] SEQ ID NO:27

MENKTETTVRRRRRIILFPVPVQGHINPILQLANVLYSKGFSITIFHTNFNKPKTSNYPHFTFRFIL

DNDPQDVRISNLPTHGPLTVMRILIINEHGADELQRELELLMLASEEDGEVSCLITDQIWYFTQS

VADSLNLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVKDIKCGFS

MWKQGKEIFENITKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLDHD

RTVFPWLDQQPSRSVLYVSFGSATEVDAKDFLEIARGLVDSKQSFLWVVRPGFVKGSTWVEP

LPDGFLGERGRIVKWVPQQEVLAHGAIGAFWTHSGWNSTLESVCEGVPMIFSAFAFDQPLNA

RYMSDVLKVGVYLENGWERGEIANAIRRVMVDEEGGYIRQNASVLKQKADVSLMKGGSSYES

LESLVAYISSL

[00154] In specific embodiments, the nucleic acid comprises a sequence encoding UGT76G2 glucosyltransferase and having the nucleic acid sequence as set forth in SEQ ID NO:28 and listed below or variants and fragments thereof.

[00155] SEQ ID NO:28

ATGGAAAATAAAACGGAGACCACCGTTCGCCGGCGCCGGAGAATAATATTATTCCCGGTA

CCAGTTCAAGGCCACATTAACCCAATTCTTCAGCTAGCCAATGTGTTGTACTCCAAAGGAT

TCAGTATCACCATCTTTCACACCAACTTCAACAAACCCAAAACATCTAATTACCCTCACTTC

ACTTTCAGATTCATCCTCGACAACGACCCACAAGACGTACGCATTTCCAATCTACCGACTC

ATGGTCCGCTCACTGTTATGCGGATTCTGATTATCAACGAACACGGAGCTGACGAATTACA

ACGCGAACTGGAACTGTTGATGTTAGCTTCTGAAGAAGATGGAGAGGTATCGTGTTTAATC

ACCGATCAGATTTGGTACTTCACGCAATCTGTTGCTGACAGTCTTAACCTCCGACGGCTTG

TTTTGATGACAAGCAGCTTGTTTAATTTTCATGCACATGTTTCACTTCCTCAGTTTGATGAG CTTGGTTACCTCGATCCTGATGACAAAACCCGTTTGGAAGAACAAGCGAGTGGGTTTCCTA

TGCTGAAAGTGAAAGATATCAAGTGTGGTTTTTCGATGTGGAAACAAGGCAAAGAGATATT

CGAGAACATTACGAAACAAACAAAAGCATCTTCAGGAGTCATCTGGAACTCATTTAAGGAA

CTCGAAGAGTCTGAGCTCGAAACTGTTATCCGTGAGATCCCGGCTCCAAGTTTCTTGATAC

CACTCCCCAAGCATTTGACAGCCTCTTCCAGCAGCTTACTAGACCACGATCGAACCGTTTT

TCCATGGTTAGACCAACAACCGTCACGTTCGGTACTGTATGTTAGTTTTGGTAGTGCTACT

GAAGTGGATGCGAAAGATTTCTTGGAAATAGCTCGTGGGTTGGTTGATAGCAAGCAGTCG

TTTTTATGGGTGGTTCGACCTGGTTTTGTCAAGGGTTCGACGTGGGTCGAACCGTTGCCA

GATGGGTTCTTGGGTGAAAGAGGACGTATTGTGAAATGGGTTCCGCAGCAAGAAGTGCTA

GCTCATGGAGCAATAGGCGCATTCTGGACTCATAGCGGATGGAACTCTACGTTGGAAAGC

GTTTGTGAAGGTGTTCCTATGATTTTCTCGGCTTTTGCGTTCGATCAACCGTTGAATGCTA

GATACATGAGTGATGTTTTGAAGGTAGGGGTGTATTTGGAAAATGGGTGGGAAAGAGGAG

AGATAGCAAATGCAATAAGAAGAGTTATGGTGGATGAAGAAGGAGGATACATTAGACAGAA

TGCAAGTGTTTTGAAACAAAAGGCAGATGTTTCTTTGATGAAGGGTGGTTCGTCTTACGAA

TCATTAGAGTCTCTAGTTGCTTACATTTCATCGTTGTAA

[00156] In certain embodiments, the nucleic acid comprises a sequence encoding UGT76H1 glucosyltransferase. In specific embodiments, the nucleic acid comprises a sequence encoding UGT76H1 glucosyltransferase having the amino acid sequence as set forth in SEQ ID NO:29 and listed below or variants and fragments thereof.

[00157] SEQ ID NO:29

MLQLATYLHSQGISITIAQYPNFNSPDSSNHPELTFLPLSSGNLSVADISGGFFKFIQTLNHNCKP

HFREYLVQNMSSDDKESIVIIRDNLMFFAGEIAGELGLPSIILRGSNAVMLTASDIIPQLHQEGRF

PPPDSLLQETIPELVPFRYKDLPFIGYPIHQTLEFSITMMTPKSPASAILINTLEFLEQSALTQIRDH

YKVPVFTIGPLHKIVTTRSTSILEEDTSCINWLDKQSPKSVVYVSLGSLAKLDEKVASEMACGLA

MSNHKFLWVVRPGMVHGFEWVEFLPDSLVGEMKARGLIVKWAPQTTVLAHNAVGGFWSHC

GWNSTIECLAEGVPMMCQPFFADQLLNARYVSDVWKTGFEIVIEKGEIACAIKRVLVDEEGEEM

RQRAMEIKEKVKIAINDGGSSYDSFKDLVAFISSL

[00158] In specific embodiments, the nucleic acid comprises a sequence encoding UGT76H1 glucosyltransferase and having the nucleic acid sequence as set forth in SEQ ID NO:30 and listed below or variants and fragments thereof.

ATGCTTCAGCTTGCAACTTACCTCCATTCTCAAGGGATTTCAATAACCATCGCTCAGTACCC

CAACTTCAACTCGCCGGATTCTTCCAACCATCCAGAACTAACCTTCCTCCCACTATCCTCC

GGCAACTTATCCGTCGCCGACATCTCCGGCGGCTTTTTCAAGTTCATCCAAACTCTTAACC

ATAACTGCAAACCCCATTTCCGGGAATACCTTGTTCAGAACATGAGTTCTGATGATAAGGA

ATCAATCGTTATCATCCGTGATAATCTCATGTTTTTCGCCGGAGAAATCGCCGGCGAGCTG

GGTCTGCCTTCGATCATTTTACGTGGCAGCAATGCTGTCATGTTGACTGCTAGCGACATCA

TCCCTCAACTTCATCAAGAAGGTCGTTTTCCGCCACCAGATTCTTTGTTGCAGGAAACAAT

TCCAGAACTGGTTCCATTCAGATACAAAGATCTACCATTTATTGGCTATCCAATACATCAAA

CCCTTGAATTTAGTATCACCATGATGACCCCCAAATCACCTGCTTCCGCCATTCTTATCAAC ACCCTCGAATTTCTTGAACAATCGGCATTAACCCAGATCCGTGATCATTACAAAGTTCCAGT

TTTTACAATCGGACCATTGCACAAAATAGTCACAACTCGTTCCACTAGCATTCTTGAAGAAG

ATACAAGTTGCATCAATTGGTTAGATAAACAATCACCCAAATCAGTGGTTTATGTGAGTTTA

GGAAGCTTAGCAAAGTTGGATGAAAAGGTTGCATCTGAAATGGCATGTGGTTTAGCCATG

AGTAACCATAAGTTCCTATGGGTGGTTCGACCCGGTATGGTTCATGGGTTTGAATGGGTC

GAGTTTTTGCCGGATAGTTTGGTGGGTGAAATGAAGGCTAGAGGTTTGATTGTGAAATGG

GCACCCCAGACGACGGTTTTGGCGCATAACGCGGTTGGTGGATTTTGGAGTCATTGCGGT

TGGAACTCGACCATAGAATGCTTAGCTGAAGGGGTCCCGATGATGTGTCAACCGTTTTTTG

CTGATCAGTTGTTGAATGCTAGGTATGTGAGTGATGTTTGGAAGACGGGTTTTGAGATTGT

TATCGAGAAAGGTGAGATTGCGTGCGCGATTAAACGAGTTTTGGTGGATGAAGAAGGCGA

AGAAATGAGGCAGAGAGCTATGGAGATTAAAGAAAAGGTTAAAATTGCAATCAACGATGGT

GGTTCTTCTTATGACTCGTTCAAGGACTTGGTGGCGTTTATTTCATCACTCTAA

[00159] In certain embodiments, the nucleic acid comprises a sequence encoding Oryza sativa Os03g0702000 or Os03g0702000-like glucosyltransferase. Os03g0702000-like glucosyltransferase include for example, other members of the UGT91 clade such as UGT91 D1 or UGT91 D2. In certain embodiments, the nucleic acid comprises a sequence encoding Os03g0702000 glucosyltransferase having the amino acid sequence as set forth in SEQ ID NO: 9 and listed below or a variant or fragment thereof.

[00160] SEQ ID NO:9

MHQHQHQSGSMDSGYSSSYAAAAGMHVVICPWLAFGHLLPCLDLAQRLASRGHRVSFVSTP

RNISRLPPVRPALAPLVAFVALPLPRVEGLPDGAESTNDVPHDRPDMVELHRRAFDGLAAPFSE

FLGTACADWVIVDVFHHWAAAAALEHKVPCAMMLLGSAHMIASIADRRLERAETESPAAAGQG

RPAAAPTFEVARMKLIRTKGSSGMSLAERFSLTLSRSSLVVGRSCVEFEPETVPLLSTLRGKPIT

FLGLMPPLHEGRREDGEDATVRWLDAQPAKSVVYVALGSEVPLGVEKVHELALGLELAGTRFL

WALRKPTGVSDADLLPAGFEERTRGRGVVATRWVPQMSILAHAAVGAFLTHCGWNSTIEGLM

FGHPLIMLPIFGDQGPNARLIEAKNAGLQVARNDGDGSFDREGVAAAIRAVAVEEESSKVFQAK

AKKLQEIVADMACHERYIDGFIQQLRSYKD

[00161 ] In certain embodiments, the nucleic acid molecule encodes Os03g0702000 glucosyltransferase and comprises a nucleotide sequence as set forth in SEQ ID NO: 10 and as detailed below or a variant or fragment thereof.

[00162] SEQ ID NO:10

ATGCATCAGCACCAACATCAGAGCGGTTCTATGGACTCCGGCTACTCCTCCTCCTACGCC

GCCGCCGCCGGGATGCACGTCGTGATCTGCCCGTGGCTCGCCTTCGGCCACCTGCTCCC

GTGCCTCGACCTCGCCCAGCGCCTCGCGTCGCGGGGCCACCGCGTGTCGTTCGTCTCCA

CGCCGCGGAACATATCCCGCCTCCCGCCGGTGCGCCCCGCGCTCGCGCCGCTCGTCGC

CTTCGTGGCGCTGCCGCTCCCGCGCGTCGAGGGGCTCCCCGACGGCGCCGAGTCCACC

AACGACGTCCCCCACGACAGGCCGGACATGGTCGAGCTCCACCGGAGGGCCTTCGACGG

GCTCGCCGCGCCCTTCTCGGAGTTCTTGGGCACCGCGTGCGCCGACTGGGTCATCGTCG

ACGTCTTCCACCACTGGGCCGCAGCCGCCGCTCTCGAGCACAAGGTGCCATGTGCAATG

ATGTTGTTGGGCTCTGCACATATGATCGCTTCCATAGCAGACAGACGGCTCGAGCGCGCG GAGACAGAGTCGCCTGCGGCTGCCGGGCAGGGACGCCCAGCGGCGGCGCCAACGTTCG

AGGTGGCGAGGATGAAGTTGATACGAACCAAAGGCTCATCGGGAATGTCCCTCGCCGAG

CGCTTCTCCTTGACGCTCTCGAGGAGCAGCCTCGTCGTCGGGCGGAGCTGCGTGGAGTT

CGAGCCGGAGACCGTCCCGCTCCTGTCGACGCTCCGCGGTAAGCCTATTACCTTCCTTGG

CCTTATGCCGCCGTTGCATGAAGGCCGCCGCGAGGACGGCGAGGATGCCACCGTCCGCT

GGCTCGACGCGCAGCCGGCCAAGTCCGTCGTGTACGTCGCGCTAGGCAGCGAGGTGCC

ACTGGGAGTGGAGAAGGTCCACGAGCTCGCGCTCGGGCTGGAGCTCGCCGGGACGCGC

TTCCTCTGGGCTCTTAGGAAGCCCACTGGCGTCTCCGACGCCGACCTCCTCCCCGCCGG

CTTCGAGGAGCGCACGCGCGGCCGCGGCGTCGTGGCGACGAGATGGGTTCCTCAGATG

AGCATACTGGCGCACGCCGCCGTGGGCGCGTTCCTGACCCACTGCGGCTGGAACTCGAC

CATCGAGGGGCTCATGTTCGGCCACCCGCTTATCATGCTGCCGATCTTCGGCGACCAGG

GACCGAACGCGCGGCTAATCGAGGCGAAGAACGCCGGATTGCAGGTGGCAAGAAACGAC

GGCGATGGATCGTTCGACCGAGAAGGCGTCGCGGCGGCGATTCGTGCAGTCGCGGTGG

AGGAAGAAAGCAGCAAAGTGTTTCAAGCCAAAGCCAAGAAGCTGCAGGAGATCGTCGCG

GACATGGCCTGCCATGAGAGGTACATCGACGGATTCATTCAGCAATTGAGATCTTACAAG

GATTGA

[00163] In certain embodiments, the nucleic acid molecule encodes Os03g0702000 glucosyltransferase and comprises the sequence as set forth in GenBank Accession number XM 015773655 or a variant or fragment thereof.

[00164] In certain embodiments, the nucleic acid comprises a sequence encoding UGT91 D1 glucosyltransferase. In certain embodiments, the nucleic acid comprises a sequence encoding UGT91 D1 glucosyltransferase having the amino acid sequence as set forth in SEQ ID NO:31 and listed below or a variant or fragment thereof.

[00165] SEQ ID NO:31

MYNVTYHQNSKAMATSDSIVDDRKQLHVATFPWLAFGHILPFLQLSKLIAEKGHKVSFLSTTRNI

QRLSSHISPLINVVQLTLPRVQELPEDAEATTDVHPEDIQYLKKAVDGLQPEVTRFLEQHSPDWI

IYDFTHYWLPSIAASLGISRAYFCVITPWTIAYLAPSSDAMINDSDGRTTVEDLTTPPKWFPFPTK

VCWRKHDLARMEPYEAPGISDGYRMGMVFKGSDCLLFKCYHEFGTQWLPLLETLHQVPVVPV

GLLPPEIPGDEKDETWVSIKKWLDGKQKGSVVYVALGSEALVSQTEVVELALGLELSGLPFVW

AYRKPKGPAKSDSVELPDGFVERTRDRGLVWTSWAPQLRILSHESVCGFLTHCGSGSIVEGL

MFGHPLIMLPIFCDQPLNARLLEDKQVGIEIPRNEEDGCLTKESVARSLRSVVVENEGEIYKANA

RALSKIYNDTKVEKEYVSQFVDYLEKNARAVAIDHES

[00166] In certain embodiments, the nucleic acid molecule encodes UGT91 D1 glucosyltransferase and comprises a nucleotide sequence as set forth in SEQ ID NO: 32 and as detailed below or a variant or fragment thereof.

[00167] SEQ ID NO:32 ATGTACAACGTTACTTATCATCAAAATTCAAAAGCAATGGCTACCAGTGACTCCATAGTTGA

CGACCGTAAGCAGCTTCATGTTGCGACGTTCCCATGGCTTGCTTTCGGTCACATCCTCCCT

TTCCTTCAGCTTTCGAAATTGATAGCTGAAAAGGGTCACAAAGTCTCGTTTCTTTCTACCAC

CAGAAACATTCAACGTCTCTCTTCTCATATCTCGCCACTCATAAATGTTGTTCAACTCACAC

TTCCACGTGTCCAAGAGCTGCCGGAGGATGCAGAGGCGACCACTGACGTCCACCCTGAA

GATATTCAATATCTCAAGAAGGCTGTTGATGGTCTTCAACCGGAGGTCACCCGGTTTCTAG

AACAACACTCTCCGGACTGGATTATTTATGATTTTACTCACTACTGGTTGCCATCCATCGCG

GCTAGCCTCGGTATCTCACGAGCCTACTTCTGCGTCATCACTCCATGGACCATTGCTTATT

TGGCACCCTCATCTGACGCCATGATAAATGATTCAGATGGTCGAACCACGGTTGAGGATC

TCACGACACCGCCCAAGTGGTTTCCCTTTCCGACCAAAGTATGCTGGCGGAAGCATGATC

TTGCCCGAATGGAGCCTTACGAAGCTCCGGGGATATCTGATGGATACCGTATGGGGATGG

TTTTTAAGGGATCTGATTGTTTGCTTTTCAAATGTTACCATGAGTTTGGAACTCAATGGCTA

CCTCTTTTGGAGACACTACACCAAGTACCGGTGGTTCCGGTGGGATTACTGCCGCCGGAA

ATACCCGGAGACGAGAAAGATGAAACATGGGTGTCAATCAAGAAATGGCTCGATGGTAAA

CAAAAAGGCAGTGTGGTGTACGTTGCATTAGGAAGCGAGGCTTTGGTGAGCCAAACCGAG

GTTGTTGAGTTAGCATTGGGTCTCGAGCTTTCTGGGTTGCCATTTGTTTGGGCTTATAGAA

AACCAAAAGGTCCCGCGAAGTCAGACTCGGTGGAGTTGCCAGACGGGTTCGTGGAACGA

ACTCGTGACCGTGGGTTGGTCTGGACGAGTTGGGCACCTCAGTTACGAATACTGAGCCAC

GAGTCAGTTTGTGGTTTCTTGACTCATTGTGGTTCTGGATCAATTGTGGAAGGGCTAATGT

TTGGTCACCCTCTAATCATGCTACCGATTTTTTGTGACCAACCTCTGAATGCTCGATTACTG

GAGGACAAACAGGTGGGAATCGAGATACCAAGAAATGAGGAAGATGGTTGCTTGACCAAG

GAGTCGGTTGCTAGATCACTGAGGTCCGTTGTTGTGGAAAACGAAGGGGAGATCTACAAG

GCGAACGCGAGGGCGCTGAGTAAAATCTATAACGACACTAAGGTGGAAAAAGAATATGTA

AGCCAATTCGTAGACTATTTGGAAAAGAATGCGCGTGCGGTTGCCATCGATCATGAGAGTT

AA

[00168] In certain embodiments, the nucleic acid comprises a sequence encoding UGT91 D2 glucosyltransferase. In certain embodiments, the nucleic acid comprises a sequence encoding UGT91 D2 glucosyltransferase having the amino acid sequence as set forth in SEQ ID NO: 33 and listed below or a variant or fragment thereof.

[00169] SEQ ID NO:33

MATSDSIVDDRKQLHVATFPWLAFGHILPYLQLSKLIAEKGHKVSFLSTTRNIQRLSSHISPLINV

VQLTLPRVQELPEDAEATTDVHPEDIPYLKKASDGLQPEVTRFLEQHSPDWIIYDYTHYWLPSIA

ASLGISRAHFSVTTPWAIAYMGPSADAMINGSDGRTTVEDLTTPPKWFPFPTKVCWRKHDLAR

LVPYKAPGISDGYRMGLVLKGSDCLLSKCYHEFGTQWLPLLETLHQVPVVPVGLLPPEIPGDEK

DETWVSIKKWLDGKQKGSVVYVALGSEVLVSQTEVVELALGLELSGLPFVWAYRKPKGPAKS

DSVELPDGFVERTRDRGLVWTSWAPQLRILSHESVCGFLTHCGSGSIVEGLMFGHPLIMLPIFG

DQPLNARLLEDKQVGIEIPRNEEDGCLTKESVARSLRSVVVEKEGEIYKANARELSKIYNDTKVE

KEYVSQFVDYLEKNARAVAIDHES

[00170] In certain embodiments, the nucleic acid molecule encodes UGT91 D2 glucosyltransferase and comprises a nucleotide sequence as set forth in SEQ ID NO: 34 and as detailed below or a variant or fragment thereof.

[00171 ] SEQ ID NO:34 ATGGCCACATCTGACTCTATCGTTGATGACAGAAAACAATTGCATGTTGCTACTTTCCCAT

GGTTGGCCTTTGGACACATTCTGCCCTACTTGCAATTGTCAAAGCTGATTGCAGAAAAAGG

TCATAAGGTGTCCTTTTTGTCTACCACAAGAAACATCCAGAGACTAAGTTCTCATATTTCTC

CATTGATTAATGTGGTTCAGTTGACCTTGCCTAGAGTCCAAGAACTTCCCGAAGACGCAGA

AGCTACTACTGATGTTCACCCTGAAGATATCCCATATCTAAAGAAGGCATCTGATGGACTT

CAACCAGAAGTAACCAGGTTTTTGGAGCAGCACAGTCCTGACTGGATTATCTATGATTATA

CTCATTACTGGCTTCCATCCATCGCAGCTAGTCTAGGCATTTCCAGAGCTCATTTCTCTGT

CACTACCCCATGGGCAATTGCATATATGGGTCCTTCTGCTGATGCAATGATCAACGGTTCT

GATGGTAGGACCACTGTTGAAGATTTAACTACACCTCCAAAGTGGTTCCCATTTCCTACTA

AAGTTTGTTGGCGAAAACACGATCTGGCACGTTTGGTCCCATATAAGGCTCCAGGTATCTC

CGATGGATATCGAATGGGTCTGGTGCTAAAGGGTTCTGATTGTCTGTTATCTAAGTGTTAC

CACGAATTTGGAACTCAATGGCTTCCTCTATTAGAGACTCTGCATCAAGTTCCAGTTGTTC

CTGTCGGTCTGCTACCACCTGAAATTCCCGGTGACGAAAAGGACGAAACTTGGGTTTCCA

TAAAAAAATGGCTGGATGGTAAGCAGAAGGGTAGTGTTGTATATGTCGCTTTAGGCTCCGA

GGTTTTGGTATCCCAGACTGAAGTTGTGGAACTTGCCTTAGGATTGGAGTTGTCCGGTTTG

CCATTCGTCTGGGCATATAGAAAGCCAAAGGGACCAGCTAAGTCAGACTCAGTTGAATTG

CCAGATGGTTTCGTAGAAAGGACAAGAGACAGAGGATTGGTTTGGACATCATGGGCCCCA

CAATTGAGAATTCTGAGTCATGAAAGTGTGTGTGGATTCTTGACTCACTGTGGCTCTGGCA

GTATTGTTGAAGGACTGATGTTTGGACACCCACTGATAATGTTGCCAATCTTCGGTGACCA

ACCTCTGAATGCAAGATTGCTGGAGGATAAACAAGTTGGTATCGAAATCCCAAGAAACGAG

GAAGACGGCTGCCTGACTAAGGAATCAGTTGCACGTAGTTTAAGATCTGTAGTTGTTGAAA

AAGAAGGTGAAATATATAAGGCTAACGCTAGAGAACTTTCAAAGATATACAATGATACCAA

GGTGGAGAAAGAATATGTTTCACAGTTTGTGGACTATTTGGAGAAAAACGCTAGAGCCGTT

GCTATCGATCACGAATCATAG

[00172] In certain embodiments, the nucleic acid comprises a sequence encoding Stevia rebaudiana UDP-glycosyltransferase 74G1 . In certain embodiments, the nucleic acid comprises a sequence encoding Stevia rebaudiana UDP-glycosyltransferase 74G1 which comprises the amino acid sequence as set forth in SEQ ID NO: 13 and as listed below or a variant or fragment thereof.

[00173] SEQ ID NO:13

MAEQQKIKKSPHVLLIPFPLQGHINPFIQFGKRLISKGVKTTLVTTIHTLNSTLNHSNTTTTSIEIQA

ISDGCDEGGFMSAGESYLETFKQVGSKSLADLIKKLQSEGTTIDAIIYDSMTEWVLDVAIEFGID

GGSFFTQACVVNSLYYHVHKGLISLPLGETVSVPGFPVLQRWETPLILQNHEQIQSPWSQMLF

GQFANIDQARWVFTNSFYKLEEEVIEWTRKIWNLKVIGPTLPSMYLDKRLDDDKDNGFNLYKA

NHHECMNWLDDKPKESVVYVAFGSLVKHGPEQVEEITRALIDSDVNFLWVIKHKEEGKLPENL

SEVIKTGKGLIVAWCKQLDVLAHESVGCFVTHCGFNSTLEAISLGVPVVAMPQFSDQTTNAKLL

DEILGVGVRVKADENGIVRRGNLASCIKMIMEEERGVIIRKNAVKWKDLAKVAVHEGGSSDNDI

VEFVSELIKA

[00174] In certain embodiments, the nucleic acid molecule encodes Stevia rebaudiana UDP-glycosyltransferase 74G1 and comprises a nucleotide sequence as set forth in SEQ ID NO: 14 and as listed below or a variant or fragment thereof. [00175] SEQ ID NO:14

ATGGCGGAACAACAAAAGATCAAGAAATCACCACACGTTCTACTCATCCCATTCCCTTTAC

AAGGCCATATAAACCCTTTCATCCAGTTTGGCAAACGATTAATCTCCAAAGGTGTCAAAACA

ACACTTGTTACCACCATCCACACCTTAAACTCAACCCTAAACCACAGTAACACCACCACCAC

CTCCATCGAAATCCAAGCAATTTCCGATGGTTGTGATGAAGGCGGTTTTATGAGTGCAGGA

GAATCATATTTGGAAACATTCAAACAAGTTGGGTCTAAATCACTAGCTGACTTAATCAAGAA

GCTTCAAAGTGAAGGAACCACAATTGATGCAATCATTTATGATTCTATGACTGAATGGGTTT

TAGATGTTGCAATTGAGTTTGGAATCGATGGTGGTTCGTTTTTCACTCAAGCTTGTGTTGTA

AACAGCTTATATTATCATGTTCATAAGGGTTTGATTTCTTTGCCATTGGGTGAAACTGTTTC

GGTTCCTGGATTTCCAGTGCTTCAACGGTGGGAGACACCGTTAATTTTGCAGAATCATGAG

CAAATACAGAGCCCTTGGTCTCAGATGTTGTTTGGTCAGTTTGCTAATATTGATCAAGCAC

GTTGGGTCTTCACAAATAGTTTTTACAAGCTCGAGGAAGAGGTAATAGAGTGGACGAGAAA

GATATGGAACTTGAAGGTAATCGGGCCAACACTTCCATCCATGTACCTTGACAAACGACTT

GATGATGATAAAGATAACGGATTTAATCTCTACAAAGCAAACCATCATGAGTGCATGAACT

GGTTAGACGATAAGCCAAAGGAATCAGTTGTTTACGTAGCATTTGGTAGCCTGGTGAAACA

TGGACCCGAACAAGTGGAAGAAATCACACGGGCTTTAATAGATAGTGATGTCAACTTCTTG

TGGGTTATCAAACATAAAGAAGAGGGAAAGCTCCCAGAAAATCTTTCGGAAGTAATAAAAA

CCGGAAAGGGTTTGATTGTAGCATGGTGCAAACAATTGGATGTGTTAGCACACGAATCAGT

AGGATGCTTTGTTACACATTGTGGGTTCAACTCAACTCTTGAAGCAATAAGTCTTGGAGTC

CCCGTTGTTGCAATGCCTCAATTTTCGGATCAAACTACAAATGCCAAGCTTCTAGATGAAAT

TTTGGGTGTTGGAGTTAGAGTTAAGGCTGATGAGAATGGGATAGTGAGAAGAGGAAATCT

TGCGTCATGTATTAAGATGATTATGGAGGAGGAAAGAGGAGTAATAATCCGAAAGAATGCG

GTAAAATGGAAGGATTTGGCTAAAGTAGCCGTTCATGAAGGTGGTAGCTCAGACAATGATA

TTGTCGAATTTGTAAGTGAGCTAATTAAGGCTTAA

[00176] In certain embodiments, the nucleic acid molecule encodes Stevia rebaudiana UDP-glycosyltransferase 74G1 and comprises the sequence as set forth in GenBank Accession number AY345982 or a variant or fragment thereof.

[00177] In other embodiments, the invention provides for nucleic acids comprising nucleotide sequences encoding a cyclodextrin glucanotransferase (W01996033267; US6271010).

[00178] Also provided are nucleic acids comprising nucleotide sequences that encode a sucrose synthase. Accordingly, in certain embodiments, the nucleic acid comprises a sequence encoding sucrose synthase which comprises the amino acid sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25 and listed below or a variant or fragment thereof.

[00179] SEQ ID NO:15 {Stevia rebaudiana SUS1 isoform) MAERVLTRVHSLRERLDSTLATHRNEILLFLSRIESHGKGILKPHQVMTEFEAICKEDQSKLSDG

AFYEVLKCTQEAIVQPPWVALAIRLRPGVWEYVRVNVNVLVVEELSVPEYLHFKEELVNGTSN

GNFVLELDFEPFTASFPRPTLTKSIGNGVEFLNRHLSAKMFHDKDSMHPLLDFLRTHHYKGKTM

MLNDRIQNLNALQSVLRKASEYLSTLDAATPYSEFEHKFQEIGLERGWGDKAEVVMEMIHMLL

DLLEAPDACTLEKFLGRIPMVFNVVILSPHGYFAQENVLGYPDTGGQVVYILDQVPALEREMLK

RIKEQGLDIIPRILIVTRLLPDAVGTTCGQRLEKVFGAEHSHILRVPFRTEKGILRKWISRFEVWP

YIETFTEDVAKEVTAELQAKPDLIIGNYSEGNLVASLLAHKLGVTQCTIAHALEKTKYPDSDIYWK

NFEEKYHFSSQFTADLIAMNHTDFIITSTFQEIAGSKDTVGQYESHTAFTMPGLYRVVHGIDVFD

PKFNIVSPGADMGIYYSYTEKEKRLTALHPEIDELLFSSVENEEHLCVLKDKSKPILFTMARLDNV

KNLTGLVEWYAKNDRLRELVNLVVVGGDRRKESKDLEEQAQMQKMHELIETYKLNGQFRWIS

SQMNRVRNGELYRVIADTRGAFIQPAFYEAFGLTVVEAMTCGLPTFATLHGGPAEIIVHGKSGF

HIDPYHGDQVTELLVNFFEKTKQDPGHWEAISKGGLQRIQEKYTWQIYSDRLLTLAGVYGFWK

HVSKLDRLEIRRYLEMFYALKYRKLAESVPLAVDE

[00180] SEQ ID N0:17 {Stevia rebaudiana SUS2 isoform)

MATSKLSRTHSMRERVEETLSAHRNEIVSLLSRYVAQGKAILQPHQILHELENIIGDVTSRQKLT

DGPFGDALKTAQECIVLPPFVALAVRPRPGVWEYVRVDAYQLSVEQLTVSEYLTFKEELVGES

NSSLMLELDFEPFNASFPRPTRSSSIGNGVQFLNRHLSSSMFRSKDCLEPLLDFLRTHRHNGH

VMMLNDRITSMTRLQSSLVKAEEYLSKLPSDTDYSEFQYELQGMGFERGWGNNAERIIEMMHL

LSDILQAPDPSILESFLARIPMVFNVVILSIHGYFGQANVLGLPDTGGQIVYILDQVRALENEMLLK

LKHQGLDIKPRILIVTRLIPDAKGTSCNQRLERVSGTEHTHILRVPFRTEKGILRKWISRFDVWPF

LEKFTQDAASEISAELHGTPDLIIGNYSDGNLVASLLSYKMGVTQCNIAHALEKTKYPDSDLYWK

KFDEKYHFSCQFTADLLAMNNADFIITSTYQEIAGTKNTVGQYESHSSFTLPGLYRVVHGIDVFD

PKFNIVSPGADMSIYFSYTEKEKRLTSLHTTIEKLLFDPTQTEDYIGNLSDKSKPIIFSMARLDHVK

NITGLVEWYAKNEKLRGLANLVVVAGYNNVKRSSDREEIAEIEKMHQLIKKYKLDGQMRWISAQ

TNRAQNGELYRYIADGRGIFVQPAIYEAFGLTVVEAMTCGLPTFATCHGGPGEIIENGVSGFHID

PYHPDTASATMADFFQKCKEDPSYWFKISEAGLKRIYERYTWKIYSERLMTLAGVYSFWKYVS

KLERRETRRYLEMFYILKFRDLVKSVPVATDDEA

[00181 ] SEQ ID NO:19 {Stevia rebaudiana SUS3 isoform)

MATPKLTRTPSMRERLEETLSAHRNDIVSLLSRYVDQGKAILQPHHLLDEIDNFIGDQNCRQKLA

DSLFGEILKSAQEGIILPPYVTLAVRPRPGVWDFLRVNVDELSVEQLTVSEYLSFKEELVDGQSR

NPFVLELDLEPFNATFPRMSRSSSIGNGVQFLNRHLSSIMFRNKDCMDPFLDFLRAHKHKGYA

MMLNDRIQTMSRLESSLAKAEDHLSKLPPETPYSEFEYVLQGMGFERGWGDNCERVLGMMHL

LSDILQAPDPSILEKFLGKMPMIFNVVVLSIHGYFGQANVLGLPDTGGQVVYILDQVRSLENEML

LKLRHQGLDIKPKILIVTRLIPNAKGTSCNQRLEKVSGTEYTYILRVPFRTEKGILGKWLSRFDIW

PYLEAFTTDAASEIAAELHGVPDLLIGNYSDGNLVASLLSNKLGVTQCNIAHALEKTKYPDSDLY

WKKFEDKYHFSCQFTADLLAMNNADFIITSTYQEIAGTKNTVGQYENHSSFTLPGLYRVVHGID

VFDPKFNIVSPGADMAIYFSYADKERRLTSLHPTIEKLLFDTEQNDVHIGNINDPSKPMIFTMARL

DHVKNITGFVECYAKNNKLREHANLVVIAGYNDAKKSSDREEIAEIEKMHNLIKQYKLDGQMRW

ISAQTNRARNGEFYRYIADGRGVFVQPAFYEAFGLTVVEAMTCGLPTFATCHGGPAEIIEDGVS

GFHIDPYHPDKMSTTLADFFQKCKEEPSYWGKISDGGLKRISERYTWKIYSERLMTLAGVYSF

WKYVSKLERRETRRYLEMFYILKFRQLVKSVPLAVDEEP

[00182] SEQ ID NO:21 {Stevia rebaudiana SUS4 isoform) MASASSSIMKRSESIVDTMPEALKQSRYHMKKCFLKYVEKGIRMMKRHHLIQEMETAIEDKDEK

AQLLDGLLGYILCTTQEAAVVPPCVAFAIRPNPGFWEFVKVNSNDLSVDGITATDYLKFKEMIVD

ETWAKDENALEIDFGSMDFNLPNMSLSCSIGNGVNFTSKFITCKLYAQSSCQQLLVDYLLSLNH

QGENLMINDALNSVSKLRAALIVAHASLSSLPNDTPYQSFELRFKEWGFEKGWGDNAERARET

IRFLLEVLQAPDPINLEALFSRIPNIFNVVLFSIHGYFGQSNVLGLPDTGGQVVYVLDQVVAMEEE

LLMRIKQQGLNFKPQILVVTRLLPDAKGTKCNQVLEPVLNTKHSHILRVPFRTDKGVLRKWVSR

FDIYPYLENFTQDASAKIIEMMEGKPDLIIGNYTDGNLVASLMANKLGTTLGTIAHALEKTKYEDS

DMNWKQFDPKYHFSCQFTADMIAMNSADFIITSTFQEIAGSKDRPGQYESHEAFTLPGLYRVV

SGINVFDPKFNIASPGADQTVYFPYTETKKRFTAFQPAIEELLFSKVENEEHIGYLEDKTKPIIFS

MARLDTVKNITGLTEWFGENKRLRSLVNLVIVAGFFDPSKSKDREEMAEIKKMHLLIEKYQLKG

QIRWIAAQTDKNRNSELYRFIADSKGAFVQPALYEAFGLTVIEAMNCGLPTFATNQGGPAEIIVD

GVSGFQIDPNFGDQSSNKIADFFQKCKEDPGYWNNISEGGLKRIYECYTWKIYANKVLNMGNI

YSFWKRLNKEQKEAKQRYIELFYNLHYKNLVRTVPIASDEAQPAPVSRAKLATQPTRRTQSRL

QRLFGA

[00183] SEQ ID NO:23 {Stevia rebaudiana SUS5 isoform)

MAASSSPIMKRSESVLDTMPEALRQSRYHMKKCFLKYVGKGKRMVKLHHLMQEMETVIEDKD

EKAQLLEGLLGYILCTTQEAAVVPPYVAFAIRPNPGFWEFVKVNSNDLSVKGITSTDYLKFKEMI

VDETWANDENALEIDFGAMDFNLPTMSLSSSIGNGVNFTSKFIISKLYAHSGSQLQSLVDYLLSL

NHQGEKLMINDKLNTVSKLQAALIVAHSFLSSLPNDTPYQSFELRFKEWGFEKGWGDYAERVQ

ETIRFLLEVLQAPDPVNLEAFFSRVPNIFNIVLFSIHGYFGQSNVLGLPDTGGQVVYVLDQVVAM

EEELLLRIKQQGLSFKPHILVVTRLLPDAKGTECSQVLEPVLNTKHSHILRVPFRTEKGVLRKWV

SRFDIYPYLEKFTQDASAKITEMMEGKPDLIIGNYTDGNLVASLMANKLGSTLGTIAHALEKTKYE

DSDMKWKHLDTKYHFSCQFTADMIAMNSADFIITSTFQEIAGSKDRPGQYESHEAFTLPGLYRV

VSGINVFDPKFNIASPGADQTVYFPYTETPKRFTTFQPAIQELLFSKVENDEHIGYLEDKNKPIIF

SMARLDMVKNITGLTEWFGENKRLRSLVNLVIVAGFFDPSKSKDREEMEEIKKMHLLIEKYELK

GQIRWIVAQTDKNRNSELYRCIADSKGAFVQPALYEAFGLTVIEAMNCGLPTFATNQGGPAEIIV

DGVSGFQIDPNYGDESSNKIADFFQKCKQDPGYWNRISDGGLMRIYECYTWKIYANKVLNMG

NIYTFWKQLNKEQKDAKQRYIELFYNQHYKNLVRTVPIVSDEDDQVTRAKPATQPSTRRTQSA

LQRLLGA

[00184] SEQ ID NO:25 {Stevia rebaudiana SUS6 isoform)

MDFGIAETLAEALKQNRYHARRCFERFTSRGKRMVKPQELLHMIEKTIDDKLERTKVLEGSMG

QILSSTQEAIVIPPYVILGLRANPGQWAYVKINADDVTVESLTPSQYLKFKESIYDQEWAKDENA

LELDFGAFDFDTPRLILPSSIGNGLGYISKFMTSRIGGDLENAKPLLDHLLALKYHGEKLMINETID

TVSKLQKALIVADVYLSAHPKDEQYQTLEPKLKEWGFEKGWGDTAERVRETMKMLSEILQAPD

PINMQSFFSRLPVVFNIVIFSIHGYFGQSDVLGLPDTGGQVVYILDQVKALEEEILLRIKMQGLNA

KPRILVVSRLIPDAQGTKCNEEMEPILNTMHSHILRVPFRTSKGVVPQWVSRFDIYPYLERFSQD

AASKILEVMECKPDLILGNYTDGNIVASLIAKKFGVTQGTIAHALEKTKYEDSDVNWKNFEKKYH

FSCQFTADLISMNAADFIITSTYQEIVGSKQRPGQYETHGAFSMPGLCRVVSGINVFDPKFNIAS

PGAEQSVYFPYTEKEKRLTDFHPAIKELLFNEQDNDEHMGYLADVTKPIIFSMARLDTVKNITGL

TEWFGKNKRLRSLVNLVVVAGFFDPSKSKDREEMEEIKKMHELIEKYKLKGQMRWIAAQNDRT

RNGELYRCISDTKGAFVQPALYEAFGLTVIEAMNCGLPTFATNQGGPAEIIVDGVSGFHIDPVN

GDESSNKIADFFTKCKVDGEYWDRVSQAGLQRIYECYTWKMYANKALNMGSMYGFWRQLNK

ETKQAKQRYIDILYNLQFKNLAKTIEIPDFVTPKLQEPVKTEPTKPLQEARPREPVQKLVPEETRL

PKLELTKLGQPNLMSNARKPLIVLVSVLIVAYASKNLYRRYFK [00185] In certain embodiments, the nucleic acid molecule encodes sucrose synthase and comprises a nucleotide sequence as set forth in SEQ ID NO: 16,18, 20, 22, 24 or 26 and listed below or a fragment or variant thereof.

[00186] SEQ ID NO:16 (encodes SUS1 isoform)

ATGGCGGAACGTGTACTCACTCGTGTTCACAGTCTTCGTGAGCGTCTCGATTCAACTCTCG

CAACTCATCGTAATGAAATCCTCTTGTTTCTTTCAAGGATTGAAAGCCATGGAAAAGGAATA

TTGAAGCCTCATCAAGTTATGACTGAATTTGAAGCTATCTGCAAAGAAGATCAGAGCAAAC

TCTCTGATGGTGCTTTTTATGAAGTTCTTAAATGCACACAGGAAGCAATAGTGCAACCTCC

ATGGGTTGCACTCGCGATCCGTCTTCGACCCGGTGTTTGGGAATATGTTAGAGTCAATGTT

AATGTTTTGGTGGTTGAAGAATTAAGTGTTCCTGAATATCTTCACTTCAAAGAAGAATTGGT

TAATGGAACATCGAATGGCAACTTCGTGTTGGAACTGGATTTTGAACCTTTTACCGCATCG

TTTCCTCGACCAACTTTAACCAAGTCTATTGGTAATGGTGTTGAGTTTCTAAACAGACATTT

ATCTGCTAAAATGTTTCATGATAAGGATAGCATGCACCCTCTTCTTGATTTCCTACGGACTC

ACCACTATAAGGGAAAGACAATGATGTTGAATGATAGAATCCAAAACCTCAATGCTCTACAA

TCGGTGTTGCGAAAGGCGTCAGAGTACTTATCAACACTCGACGCAGCAACACCGTACTCT

GAGTTTGAACATAAGTTTCAAGAAATCGGGTTGGAGAGAGGTTGGGGTGATAAAGCGGAG

GTCGTAATGGAGATGATCCACATGCTTCTAGACCTTCTAGAAGCACCCGACGCATGCACA

CTCGAGAAGTTTCTCGGAAGAATCCCAATGGTTTTCAATGTTGTCATTCTTTCGCCTCACG

GCTACTTCGCCCAAGAAAATGTGTTGGGATATCCCGACACTGGCGGTCAGGTTGTTTACAT

CTTGGATCAAGTTCCCGCTCTGGAACGCGAGATGCTCAAAAGGATTAAGGAGCAAGGACT

CGATATCATTCCTCGTATATTGATTGTTACGAGGCTTCTTCCCGACGCGGTTGGGACCACA

TGCGGGCAACGTTTAGAGAAAGTGTTTGGAGCCGAACACTCGCATATTCTTCGGGTCCCG

TTTAGAACCGAAAAGGGTATTCTTCGTAAATGGATCTCTCGTTTTGAGGTGTGGCCTTACA

TCGAGACTTTCACCGAGGATGTTGCTAAAGAAGTTACAGCAGAGTTGCAAGCAAAACCAGA

TTTGATCATTGGAAACTATAGTGAAGGAAATTTGGTTGCATCTTTGCTAGCTCACAAGTTGG

GTGTCACTCAGTGTACCATTGCTCATGCTTTGGAGAAAACTAAATACCCGGATTCTGATAT

CTACTGGAAGAACTTTGAGGAGAAATATCATTTCTCTTCGCAGTTTACCGCTGATCTTATCG

CTATGAACCATACCGACTTCATCATCACCAGTACTTTCCAAGAAATTGCTGGAAGTAAGGA

CACGGTTGGACAGTACGAGAGTCATACCGCGTTCACAATGCCGGGATTGTATCGGGTGGT

TCACGGGATCGATGTTTTTGACCCCAAATTCAATATTGTTTCACCCGGGGCCGATATGGGA

ATTTACTACTCGTATACCGAGAAAGAAAAGAGGCTCACTGCGCTTCACCCTGAAATCGATG

AACTTCTCTTTAGTTCCGTCGAAAACGAAGAACACTTATGTGTGTTGAAGGATAAGAGTAAA

CCAATCTTGTTCACAATGGCGCGATTGGATAATGTGAAGAATTTAACCGGACTGGTTGAAT

GGTACGCTAAAAACGACCGCCTTCGTGAGCTCGTGAACCTCGTGGTCGTCGGTGGTGAC

CGAAGGAAAGAGTCGAAAGATCTTGAAGAACAAGCTCAGATGCAGAAGATGCATGAACTT

ATCGAAACCTACAAACTCAACGGTCAGTTCAGGTGGATATCCTCACAAATGAACCGCGTGA

GGAACGGTGAGTTGTATCGCGTTATTGCTGACACACGAGGTGCGTTTATCCAGCCTGCGT

TTTACGAGGCGTTTGGGTTGACGGTTGTGGAGGCCATGACTTGTGGCCTGCCGACATTCG

CGACACTTCATGGTGGGCCCGCTGAGATTATTGTTCACGGGAAATCCGGGTTCCATATTG

ACCCGTATCACGGTGACCAGGTCACCGAGTTGCTGGTCAATTTCTTTGAGAAAACTAAACA

AGACCCGGGTCATTGGGAGGCCATTTCCAAGGGTGGTCTGCAACGTATTCAGGAGAAATA

CACGTGGCAGATTTATTCAGATAGGTTGTTGACGCTTGCCGGAGTTTATGGATTCTGGAAG

CATGTGTCGAAGCTTGACAGGCTCGAGATCCGTCGTTATCTTGAAATGTTTTACGCGCTCA

AGTATCGCAAACTGGCTGAATCTGTTCCATTGGCTGTTGATGAGTGA

[00187] SEQ ID NO:18 (encodes SUS2 isoform) ATGGCGACAAGTAAGTTGAGCAGAACGCATAGTATGCGTGAGCGTGTTGAAGAAACTCTT

TCCGCTCATCGCAACGAAATCGTTTCTCTTCTTTCTAGGTATGTGGCTCAGGGGAAGGCGA

TATTGCAGCCGCATCAGATACTCCATGAACTTGAGAATATCATCGGTGATGTTACTTCGCG

CCAAAAGCTTACAGATGGTCCGTTTGGAGATGCGTTGAAGACAGCACAGGAATGTATAGTT

CTACCTCCATTTGTAGCTTTAGCAGTTCGTCCAAGACCTGGTGTTTGGGAATACGTGCGCG

TGGATGCATATCAACTAAGTGTGGAACAACTAACTGTTTCAGAGTATCTTACCTTCAAAGAA

GAACTTGTTGGAGAGTCTAATAGTTCTTTAATGCTCGAGTTGGATTTTGAGCCATTTAATGC

TTCGTTTCCTAGACCAACCCGTTCTTCATCCATTGGCAATGGAGTTCAGTTCCTGAATCGC

CACCTGTCGTCAAGCATGTTTCGCAGCAAAGATTGTTTAGAACCGCTTCTGGATTTCCTAC

GCACACACAGACATAATGGACATGTAATGATGTTAAATGACCGCATAACAAGCATGACTAG

ACTTCAATCTTCTTTGGTCAAAGCAGAGGAATATCTTTCTAAACTACCATCTGATACAGACT

ACTCTGAGTTTCAATATGAATTGCAAGGAATGGGTTTTGAAAGAGGATGGGGAAACAATGC

TGAAAGAATCATTGAGATGATGCATCTTCTCTCAGACATTCTACAAGCTCCAGATCCTTCCA

TTTTGGAATCTTTTCTTGCTAGAATACCTATGGTGTTTAATGTTGTTATATTATCAATACATG

GCTACTTTGGGCAAGCAAATGTTTTGGGTTTGCCAGATACTGGTGGCCAGATTGTATATAT

ATTGGATCAAGTCCGTGCATTGGAAAATGAGATGCTTCTTAAATTAAAGCACCAAGGACTG

GATATCAAACCTAGGATTCTGATTGTGACTCGGTTAATACCTGATGCAAAAGGTACTTCAT

GTAACCAACGACTGGAAAGAGTCAGTGGAACTGAACACACACATATACTTCGTGTTCCTTT

TAGAACCGAGAAAGGAATTCTTCGTAAATGGATCTCAAGGTTTGATGTATGGCCTTTTTTG

GAGAAATTTACACAGGATGCAGCAAGTGAAATTTCTGCTGAGTTGCATGGTACTCCAGATC

TTATAATTGGAAATTATAGTGATGGCAATCTTGTTGCCTCTTTATTATCTTACAAAATGGGA

GTAACCCAGTGTAACATTGCTCATGCTTTAGAGAAAACAAAGTATCCAGATTCTGATTTATA

TTGGAAGAAATTTGATGAGAAATATCACTTTTCTTGTCAATTTACTGCTGATCTTTTAGCCAT

GAACAATGCAGATTTTATCATCACCAGCACATACCAAGAAATCGCGGGAACGAAAAATACT

GTCGGACAATACGAGAGTCATTCGTCTTTCACTCTCCCGGGGCTCTACAGGGTTGTTCAT

GGTATTGACGTTTTTGACCCTAAGTTCAACATTGTGTCTCCAGGGGCAGATATGTCTATAT

ACTTCTCATACACCGAGAAGGAAAAAAGACTTACATCTCTTCATACTACAATTGAGAAGTTA

TTGTTTGACCCTACACAAACTGAAGATTACATTGGAAATCTGAGTGATAAATCAAAACCGAT

AATTTTTTCAATGGCAAGACTTGATCATGTGAAGAACATTACGGGTCTGGTTGAGTGGTAC

GCTAAGAATGAGAAGCTTAGAGGACTAGCAAACCTTGTTGTGGTTGCTGGTTATAATAATG

TGAAGAGGTCTAGTGACAGAGAAGAAATTGCAGAAATTGAAAAAATGCATCAACTTATTAA

GAAATACAAATTAGATGGTCAGATGAGATGGATTTCAGCACAAACAAACCGCGCACAAAAT

GGTGAACTTTATCGCTATATTGCTGATGGAAGGGGAATCTTTGTACAGCCCGCTATTTATG

AAGCTTTTGGGCTGACAGTGGTGGAGGCCATGACTTGTGGGCTTCCAACATTTGCAACTT

GCCATGGTGGGCCAGGAGAGATAATTGAAAATGGTGTTTCGGGCTTCCATATCGACCCGT

ATCATCCGGATACTGCATCAGCCACAATGGCTGATTTTTTTCAGAAATGCAAGGAGGACCC

GAGTTATTGGTTCAAGATATCTGAAGCAGGGCTTAAAAGAATATATGAAAGGTACACATGG

AAAATTTACTCTGAACGGTTGATGACATTAGCTGGAGTTTATAGCTTCTGGAAGTATGTCTC

GAAACTTGAGAGACGTGAAACAAGACGATATCTTGAGATGTTTTATATTCTTAAGTTCCGTG

ATCTGGTAAAATCTGTTCCAGTGGCTACTGATGATGAGGCTTAG

[00188] SEQ ID NO:20 (encodes SUS3 isoform)

ATGGCGACACCTAAGCTTACGCGAACACCAAGCATGCGAGAGCGTCTTGAAGAAACTTTA

TCAGCTCATCGCAACGATATCGTCTCTCTTCTTTCCAGGTATGTAGATCAAGGTAAGGCCA

TATTGCAGCCCCACCACCTACTTGACGAAATCGATAACTTCATCGGAGATCAAAATTGCCG

CCAAAAGCTTGCTGATAGTCTATTCGGTGAAATCCTCAAGTCCGCACAGGAAGGTATAATT

CTTCCTCCATATGTAACGCTTGCTGTTCGTCCAAGACCTGGTGTTTGGGACTTTTTGCGTG

TGAATGTCGATGAATTGAGTGTCGAGCAACTTACTGTTTCTGAGTATTTAAGCTTCAAGGA

GGAGCTTGTAGATGGCCAGAGTAGGAACCCGTTTGTGTTGGAACTGGATCTGGAACCGTT

TAATGCAACATTTCCCCGGATGTCACGATCTTCATCCATCGGCAATGGAGTTCAGTTTCTC AACCGTCATCTCTCGTCAATTATGTTTCGCAACAAAGATTGTATGGATCCGTTTCTTGATTT

CCTTCGTGCTCATAAACATAAAGGATACGCGATGATGTTGAATGATCGGATACAAACAATG

TCTAGACTTGAATCTTCTTTAGCAAAAGCGGAGGATCATCTCTCTAAACTACCACCCGAAA

CACCGTACTCCGAATTCGAATACGTATTGCAAGGAATGGGGTTTGAAAGAGGTTGGGGGG

ATAATTGTGAAAGAGTTCTTGGTATGATGCATCTTCTTTCTGACATTCTTCAAGCTCCAGAT

CCTTCGATTCTTGAAAAGTTTCTTGGAAAGATGCCGATGATCTTCAATGTTGTTGTGTTATC

GATTCATGGTTACTTTGGTCAGGCTAATGTTTTGGGTTTGCCGGATACCGGTGGTCAGGTT

GTATATATATTGGATCAAGTACGTTCTTTGGAGAATGAAATGTTACTTAAATTAAGGCATCA

AGGACTTGATATCAAACCCAAGATTCTAATTGTAACTCGATTGATACCAAATGCCAAAGGTA

CTTCATGCAACCAACGATTGGAGAAAGTAAGTGGAACCGAATACACGTATATATTACGTGT

CCCTTTTAGGACAGAGAAAGGGATTCTTGGTAAATGGTTATCAAGGTTTGATATATGGCCT

TATTTGGAGGCGTTTACAACGGATGCAGCAAGTGAAATTGCTGCTGAGTTACACGGTGTTC

CGGATCTTTTAATAGGAAACTACAGTGATGGGAATCTCGTTGCCTCCTTGCTATCTAACAA

ATTGGGCGTAACCCAGTGCAACATTGCACACGCGTTAGAGAAAACAAAGTATCCAGATTCC

GACTTATATTGGAAGAAATTTGAGGACAAATATCACTTTTCATGTCAATTTACCGCCGACCT

TCTAGCAATGAACAATGCAGATTTTATCATCACTAGCACATACCAAGAGATTGCAGGAACG

AAAAACACCGTTGGACAATACGAGAATCATTCATCGTTCACTCTTCCGGGTCTATACAGGG

TTGTTCACGGTATCGATGTCTTTGACCCGAAGTTCAACATCGTGTCACCAGGGGCAGATAT

GGCAATTTACTTCTCATATGCCGATAAAGAGAGACGACTTACATCTCTACATCCCACAATTG

AGAAGCTATTGTTCGACACTGAGCAGAACGATGTACACATTGGAAATATAAATGACCCGTC

TAAACCCATGATTTTCACAATGGCGAGGCTTGATCATGTGAAGAATATAACTGGATTCGTC

GAGTGTTATGCTAAAAATAATAAGTTGAGGGAACACGCAAATCTTGTGGTTATTGCTGGTT

ATAATGACGCGAAGAAATCAAGTGATCGAGAAGAAATTGCGGAAATTGAAAAGATGCATAA

TCTTATCAAGCAATACAAACTTGATGGTCAGATGAGATGGATATCAGCCCAAACAAACCGG

GCCCGAAATGGGGAATTTTATCGGTATATCGCTGATGGTAGGGGCGTTTTCGTCCAGCCC

GCTTTCTATGAAGCATTTGGGCTTACGGTTGTGGAGGCGATGACATGTGGGCTCCCAACA

TTTGCCACGTGTCATGGTGGGCCTGCTGAGATCATTGAGGATGGTGTGTCGGGGTTCCAT

ATTGATCCATATCATCCTGATAAGATGTCGACTACGTTAGCTGATTTTTTTCAAAAGTGCAA

AGAGGAACCTAGTTACTGGGGTAAAATATCCGATGGCGGGCTGAAAAGAATAAGTGAAAG

GTACACATGGAAGATATATTCGGAACGGTTGATGACGTTGGCGGGCGTATATAGCTTTTG

GAAATATGTGTCAAAACTCGAGAGGCGTGAAACCCGTCGATACCTTGAGATGTTCTACATT

TTAAAGTTTCGTCAACTGGTGAAGTCGGTTCCGCTAGCTGTTGATGAGGAGCCGTAA

[00189] SEQ ID NO:22 (encodes SUS4 isoform)

ATGGCATCTGCTTCAAGTTCTATCATGAAACGGTCTGAATCAATAGTTGACACCATGCCAG

AAGCCTTAAAGCAGAGCCGCTATCATATGAAAAAATGTTTTCTAAAATATGTAGAAAAAGGA

ATTCGCATGATGAAAAGACATCATTTGATACAAGAAATGGAGACCGCAATTGAAGACAAGG

ATGAAAAGGCTCAGCTTCTAGATGGCTTACTTGGCTACATCTTGTGCACAACTCAGGAAGC

AGCCGTTGTTCCTCCTTGTGTTGCATTTGCTATAAGACCGAATCCTGGATTCTGGGAGTTT

GTTAAAGTCAACTCTAATGATCTATCGGTTGATGGGATAACTGCCACAGATTACTTGAAGTT

CAAGGAAATGATCGTAGATGAGACATGGGCTAAAGATGAAAATGCATTGGAGATTGACTTT

GGATCGATGGACTTTAACCTACCAAACATGAGTTTATCTTGTTCGATTGGAAATGGTGTTAA

CTTCACATCAAAATTCATTACTTGTAAACTTTACGCACAATCTAGTTGCCAACAACTGCTTG

TTGATTACTTGCTCTCATTGAATCATCAAGGAGAAAATCTTATGATCAATGATGCATTAAAC

TCAGTCTCAAAACTTCGAGCGGCTTTAATTGTAGCTCATGCGTCGCTATCTTCGTTGCCCA

ACGATACTCCATATCAAAGCTTCGAGCTTAGATTCAAAGAATGGGGATTTGAGAAGGGATG

GGGAGATAACGCGGAACGCGCGAGGGAAACAATTCGGTTTCTTTTGGAGGTTCTTCAAGC

ACCCGATCCGATAAACCTCGAGGCTTTATTCAGCAGGATTCCAAACATATTCAACGTTGTTT

TATTCTCGATTCATGGGTATTTTGGTCAATCCAATGTTCTTGGATTGCCCGATACTGGTGG

CCAAGTGGTTTATGTTTTGGATCAAGTGGTAGCTATGGAAGAAGAACTACTCATGAGGATC AAACAACAAGGACTCAACTTCAAGCCTCAAATTCTTGTGGTGACCCGACTTCTTCCTGATG

CTAAAGGGACCAAGTGTAATCAGGTGTTGGAACCAGTTCTGAACACGAAACATTCGCATAT

TCTTAGGGTTCCATTCAGGACTGATAAAGGTGTTCTTCGTAAATGGGTATCTCGATTTGATA

TCTATCCATATCTCGAAAACTTCACTCAGGATGCAAGTGCGAAAATCATTGAAATGATGGAA

GGGAAACCGGATCTTATCATCGGAAACTATACCGATGGAAACCTTGTTGCATCACTCATGG

CTAACAAACTCGGAACGACATTGGGAACAATTGCACATGCTTTGGAGAAAACCAAATACGA

AGATTCAGACATGAATTGGAAGCAATTCGACCCAAAATATCACTTCTCCTGCCAATTTACAG

CCGATATGATTGCAATGAACTCAGCTGATTTCATCATCACAAGTACTTTCCAAGAAATCGCT

GGAAGTAAAGATAGACCCGGACAATATGAAAGCCATGAAGCATTTACACTTCCAGGATTAT

ACAGAGTTGTTTCAGGCATCAACGTGTTCGATCCCAAATTCAATATCGCGTCTCCAGGAGC

CGATCAAACCGTTTATTTCCCGTACACCGAAACAAAGAAACGATTCACTGCATTTCAACCC

GCCATAGAGGAATTACTCTTCAGTAAAGTTGAAAACGAAGAACACATTGGATACTTAGAAG

ACAAAACCAAACCGATCATATTCTCAATGGCGCGTCTCGACACAGTTAAGAACATAACAGG

ACTAACCGAATGGTTTGGAGAGAACAAACGGCTCCGAAGCTTGGTTAATCTTGTAATCGTG

GCGGGTTTCTTTGACCCGTCAAAGTCAAAAGACAGAGAAGAAATGGCGGAAATAAAGAAA

ATGCATTTATTGATTGAAAAATATCAGCTTAAAGGTCAAATAAGATGGATTGCTGCACAAAC

TGATAAGAACCGAAACAGTGAGCTTTACCGGTTTATTGCTGACTCAAAAGGCGCGTTTGTG

CAGCCCGCTTTGTATGAGGCGTTTGGGCTCACGGTTATTGAGGCGATGAACTGTGGTTTA

CCGACTTTTGCAACTAATCAAGGTGGTCCAGCTGAGATTATCGTTGATGGTGTTTCTGGGT

TCCAGATTGATCCTAATTTTGGTGATCAGTCTAGTAATAAGATTGCTGATTTCTTCCAGAAG

TGTAAGGAAGATCCTGGTTATTGGAATAATATTTCAGAAGGCGGTTTGAAGCGTATATACG

AATGTTATACTTGGAAGATTTATGCGAATAAAGTGTTGAATATGGGGAACATATACTCGTTT

TGGAAGCGGTTAAACAAGGAACAAAAAGAAGCAAAACAAAGATACATTGAACTATTCTACA

ATCTACACTACAAGAACTTGGTTAGGACTGTACCAATTGCTAGTGATGAAGCTCAACCTGC

ACCAGTGTCAAGGGCAAAACTTGCAACACAACCCACAAGACGTACGCAATCCAGGTTGCA

AAGGCTGTTTGGAGCTTAA

[00190] SEQ ID NO:24 (encodes SUS5 isoform)

ATGGCAGCTTCTTCAAGTCCCATTATGAAACGGTCTGAGTCAGTACTCGACACCATGCCAG

AAGCTTTGAGGCAAAGTCGGTATCATATGAAAAAATGCTTTCTAAAATATGTAGGGAAAGG

AAAGCGGATGGTGAAACTCCACCATTTGATGCAAGAAATGGAGACCGTCATTGAGGACAA

GGACGAAAAGGCTCAGCTCTTGGAAGGCTTACTTGGTTACATCTTGTGCACCACTCAGGA

AGCAGCAGTTGTTCCTCCTTATGTCGCCTTTGCAATAAGGCCAAACCCTGGATTTTGGGAG

TTTGTTAAAGTCAACTCTAATGATCTCTCGGTTAAAGGGATCACTTCCACCGATTACTTGAA

GTTCAAGGAAATGATCGTTGACGAAACATGGGCTAATGATGAAAATGCATTGGAGATCGAC

TTTGGAGCAATGGACTTTAACTTGCCAACAATGAGCTTATCTTCTTCAATTGGAAATGGAGT

TAACTTCACATCAAAGTTTATTATTTCTAAACTTTATGCTCATTCTGGCAGCCAATTACAATC

TCTAGTTGATTACTTACTTTCATTAAATCATCAAGGAGAAAAACTTATGATAAATGACAAACT

AAACACAGTTTCAAAACTTCAAGCCGCTCTAATAGTAGCTCATTCTTTCCTTTCTTCATTGC

CCAACGACACACCGTATCAAAGCTTTGAACTTAGATTTAAAGAGTGGGGTTTTGAAAAAGG

ATGGGGAGATTATGCAGAAAGGGTGCAAGAAACAATTCGGTTTTTGTTGGAGGTTCTTCAA

GCACCCGACCCCGTAAACCTAGAGGCCTTTTTTAGCAGGGTTCCAAACATATTCAATATTG

TTTTATTCTCGATTCATGGGTATTTTGGTCAATCCAATGTTCTTGGCTTGCCCGATACCGGA

GGTCAGGTAGTTTATGTTTTGGATCAAGTTGTGGCAATGGAAGAAGAATTGCTACTTAGGA

TTAAGCAACAAGGACTCAGCTTCAAGCCTCATATTCTTGTGGTGACTCGACTTCTTCCCGA

TGCCAAAGGGACCGAGTGTAGCCAAGTTTTGGAACCAGTTCTCAACACGAAACACTCACA

CATTCTTAGAGTCCCATTTAGGACAGAAAAAGGTGTTCTTCGTAAATGGGTGTCTCGATTT

GATATCTATCCATACCTCGAAAAGTTTACTCAGGATGCAAGTGCAAAAATAACTGAAATGAT

GGAAGGAAAACCTGATCTTATCATTGGAAACTACACTGACGGAAACTTGGTTGCATCTCTC

ATGGCTAACAAACTCGGAAGCACATTGGGAACGATTGCACACGCGTTAGAGAAGACTAAA TACGAAGATTCAGACATGAAATGGAAACATTTGGACACAAAATATCACTTTTCTTGTCAATT

TACAGCTGATATGATAGCAATGAATTCAGCAGATTTCATCATCACTAGTACTTTCCAAGAAA

TTGCTGGAAGTAAAGATAGACCCGGTCAGTATGAAAGCCATGAAGCATTTACACTCCCGG

GTTTATATAGAGTTGTTTCGGGCATCAACGTGTTTGATCCCAAATTCAACATTGCATCTCCG

GGAGCTGATCAAACCGTTTATTTCCCTTACACGGAAACACCAAAACGATTCACTACTTTTCA

ACCCGCTATACAAGAATTACTCTTTAGTAAAGTTGAAAACGACGAACACATTGGATATTTAG

AAGATAAGAATAAACCAATCATCTTCTCAATGGCAAGACTCGACATGGTTAAGAACATAACG

GGGCTAACCGAATGGTTTGGGGAAAACAAGCGGTTAAGAAGTTTGGTTAATCTTGTAATTG

TGGCGGGGTTTTTTGATCCGTCAAAATCAAAAGATAGAGAAGAAATGGAAGAAATAAAGAA

AATGCATTTGTTGATTGAGAAATATGAACTTAAAGGTCAAATAAGATGGATAGTAGCACAAA

CTGATAAAAACAGAAATAGTGAACTTTATCGTTGTATCGCTGACTCAAAGGGGGCGTTTGT

GCAACCGGCTTTATATGAAGCGTTTGGGTTAACCGTTATTGAGGCTATGAATTGTGGGTTA

CCAACTTTTGCAACTAACCAAGGTGGTCCGGCTGAGATTATTGTTGATGGTGTTTCTGGGT

TCCAAATCGATCCTAATTATGGCGACGAGTCTAGCAACAAGATCGCTGATTTTTTTCAAAAA

TGCAAACAGGATCCAGGATACTGGAATAGGATTTCAGACGGTGGTTTGATGCGTATATACG

AATGCTACACATGGAAGATTTATGCAAATAAAGTGTTGAATATGGGGAACATTTACACATTT

TGGAAGCAGTTAAACAAGGAACAGAAAGATGCGAAACAAAGATACATTGAGCTATTCTACA

ATCAACATTACAAGAATTTGGTTAGGACTGTGCCGATTGTAAGTGATGAAGATGACCAAGT

TACAAGGGCAAAACCGGCAACACAACCTTCAACAAGGCGCACACAATCTGCCTTGCAAAG

GCTGCTTGGAGCTTAA

[00191 ] SEQ ID NO:26 (encodes SUS6 isoform)

ATGGATTTCGGTATAGCAGAGACTTTGGCCGAGGCATTGAAGCAAAACCGGTACCATGCA

AGGAGATGCTTTGAGCGTTTTACATCACGTGGAAAAAGGATGGTGAAGCCTCAAGAGTTAT

TACACATGATTGAAAAAACCATTGACGACAAGCTTGAAAGAACGAAGGTCTTGGAGGGCTC

AATGGGACAAATCTTGAGTTCCACACAGGAGGCAATCGTTATTCCACCATATGTTATTTTAG

GATTGAGAGCGAATCCAGGACAATGGGCATACGTTAAGATCAATGCTGATGACGTCACTG

TTGAGTCACTCACACCTTCACAATATCTAAAGTTCAAAGAATCCATCTACGATCAAGAATGG

GCAAAGGACGAAAATGCCCTTGAACTAGATTTCGGAGCGTTCGACTTTGATACGCCTCGAT

TAATCCTCCCGTCATCTATCGGCAACGGACTCGGTTACATTTCAAAGTTCATGACTTCAAG

AATTGGTGGTGATCTAGAAAACGCGAAGCCGTTGCTTGACCACTTGCTTGCTCTAAAATAT

CATGGAGAGAAGCTTATGATCAATGAGACAATAGATACAGTTTCAAAGCTCCAGAAAGCAT

TAATTGTTGCTGATGTCTACTTATCTGCACACCCGAAAGACGAACAATATCAAACCTTAGAG

CCCAAGCTTAAAGAATGGGGATTTGAGAAAGGATGGGGAGATACTGCTGAAAGAGTTAGA

GAGACAATGAAAATGCTTTCGGAGATTCTTCAAGCACCCGACCCGATTAACATGCAATCGT

TCTTTAGCAGGCTTCCGGTGGTCTTCAATATTGTCATATTTTCTATTCATGGGTATTTTGGT

CAATCAGATGTTCTTGGATTACCTGATACCGGAGGGCAGGTTGTTTACATTCTTGATCAAG

TTAAAGCATTAGAGGAAGAGATATTGCTAAGAATAAAAATGCAAGGATTGAATGCAAAGCC

TCGGATTCTTGTGGTGAGTCGACTCATTCCCGACGCACAAGGAACAAAGTGTAACGAGGA

AATGGAACCGATCTTGAACACAATGCATTCACACATCCTTCGGGTTCCTTTCAGAACCTCA

AAAGGCGTTGTTCCTCAATGGGTATCGCGGTTTGACATCTACCCGTATCTTGAAAGATTCT

CACAGGACGCTGCCTCTAAAATACTTGAAGTAATGGAATGTAAACCAGATCTCATACTTGG

AAACTACACAGATGGAAACATTGTTGCATCACTTATAGCCAAAAAGTTTGGAGTAACACAG

GGGACGATTGCACACGCGTTAGAGAAGACAAAGTACGAAGATTCGGATGTTAACTGGAAA

AACTTTGAAAAAAAGTATCATTTCTCATGTCAATTTACCGCGGATTTGATCTCAATGAACGC

TGCAGATTTCATAATCACAAGCACTTATCAAGAAATTGTGGGAAGCAAACAAAGACCCGGA

CAGTATGAGACCCACGGGGCGTTTAGTATGCCCGGACTTTGTAGAGTCGTGTCGGGCATC

AACGTGTTTGATCCTAAGTTCAACATTGCTTCACCCGGTGCGGAACAATCGGTTTATTTTC

CGTACACCGAGAAGGAGAAACGGTTAACGGATTTTCATCCCGCAATTAAAGAACTACTTTT

CAACGAACAAGACAATGACGAGCATATGGGATACCTCGCGGATGTAACCAAACCGATAATA TTCTCAATGGCGAGGCTCGATACGGTGAAGAACATAACAGGGTTAACCGAGTGGTTCGGT

AAGAACAAACGACTTAGAAGTCTTGTAAACTTGGTTGTTGTCGCGGGGTTCTTCGATCCAT

CAAAATCTAAAGACCGTGAAGAGATGGAGGAAATCAAGAAAATGCATGAACTAATAGAGAA

ATACAAACTCAAGGGTCAGATGAGATGGATCGCGGCTCAAAACGATAGGACCCGCAATGG

TGAATTGTATCGGTGTATTTCCGATACGAAGGGAGCGTTTGTGCAGCCCGCGTTGTATGA

GGCTTTTGGGCTCACGGTTATCGAGGCAATGAACTGCGGTCTCCCGACTTTTGCAACCAA

TCAAGGCGGGCCCGCGGAGATCATAGTTGACGGAGTTTCGGGATTTCATATTGATCCCGT

TAACGGAGATGAATCAAGCAACAAGATTGCTGATTTCTTCACGAAATGCAAAGTCGATGGC

GAGTATTGGGACCGCGTGTCGCAAGCGGGACTTCAACGTATTTACGAGTGCTACACATGG

AAGATGTATGCTAACAAAGCATTGAACATGGGTTCGATGTATGGTTTTTGGAGGCAATTAA

ACAAAGAAACTAAGCAAGCGAAGCAACGATACATCGATATCTTGTATAACTTACAATTCAAG

AATTTGGCAAAAACCATTGAAATCCCTGATTTTGTGACTCCTAAACTTCAAGAACCGGTCAA

AACCGAACCAACAAAACCATTACAAGAAGCAAGACCTCGAGAACCGGTGCAAAAACTGGTA

CCGGAAGAAACCCGACTGCCAAAACTAGAGTTGACCAAGCTTGGTCAACCGAATTTGATG

AGCAATGCAAGAAAACCATTGATTGTTCTTGTTTCTGTGTTGATAGTTGCATATGCATCCAA

GAACTTGTATAGGAGGTATTTCAAATAG

[00192] In other embodiments, there is provided a nucleic acid comprising a sequence having at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% identity to any one of the sequences set forth in SEQ ID NOs: 2, 4, 6, 8, 10, 14, 16,18, 20, 22, 24, 26, 28, 30, 32 and 34 and fragments thereof or the complement thereof.

[00193] In other embodiments, there is provided a nucleic acid encoding a polypeptide comprising a sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% percent identity to any one of the sequences set forth in SEQ ID NOs: 1 , 3, 5, 7, 9, 13, 15, 17, 19, 21 , 23, 25, 27, 29, 30, 31 and 33 and fragments thereof. A worker skilled in the art would readily appreciate that overall sequence identity or similarity may be less than 50% but regions of the enzyme (such as the catalytic site or areas adjacent to the catalytic site) may have conserved amino acids. For example, there are conserved amino acids at the opening adjacent to the UDPG catalytic site. In particular, a leucine at position 379 of UGT76G1 is conserved. In certain embodiments, the nucleic acid encodes an UDP- glucosyltransferase having the sequence SDFGLDQ at a position corresponding to amino acid residues 375 to 381 of the UGT76G1 set forth in SEQ ID NO:1 .

[00194] In certain embodiments, fragments are at least 10, at least 20, at least 50 nucleotides in length. The fragments may be used, for example, as primers or probes.

[00195] Also provided are nucleic acids that hybridize to the nucleic acids of the present invention or the complement thereof. In certain embodiments, there is provided a nucleic acid that hybridizes to any one of the sequences set forth in SEQ ID NOs: 2, 4, 6, 8, 10, 14, 16, 18, 20, 22, 24,26, 28, 30, 32 and 34 or the complement thereof under conditions of low, moderate or high stringency. A worker skilled in the art readily appreciates that hybridization and the strength of hybridization (i.e., the strength of the association between the nucleic acids) is impacted by such factors as the degree of complementary between the nucleic acids, stringency of the conditions involved, the Tm of the formed hybrid, and the G:C ratio within the nucleic acids. Such a worker could readily determine appropriate stringent (see, for example, Sambrook, et al., Molecular Cloning: A Laboratory Manual, 2nd ed., Cold Spring Harbor Laboratory Press, New York (1989) pp. 9.50-51 , 1 1 .48-49 and 1 1 .2-1 1 .3).

[00196] Typically under high stringency conditions only highly similar sequences will hybridize under these conditions (typically >95% identity). With moderate stringency conditions typically those sequence having greater than 80% identity will hybridize and with low stringency conditions those sequences having greater than 50% identity will hybridize.

[00197] A non-limiting example of "high stringency conditions" when used in reference to nucleic acid hybridization comprise conditions equivalent to binding or hybridization at 42 °C in a solution consisting of 5XSSPE (43.8 g/l NaCI, 6.9 g/l NaH2P04H20 and 1 .85 g/l EDTA, pH adjusted to 7.4 with NaOH), 0.5% SDS, 5X Denhardt's reagent and 100 μg/ml denatured salmon sperm DNA followed by washing in a solution comprising 0.1 XSSPE, 1 .0% SDS at 42°C when a probe of about 500 nucleotides in length is employed. A non-limiting example of "medium stringency conditions" when used in reference to nucleic acid hybridization comprise conditions equivalent to binding or hybridization at 42 °C in a solution consisting of 5XSSPE (43.8 g/l NaCI, 6.9 g/l NaH2P04H20 and 1 .85 g/l EDTA, pH adjusted to 7.4 with NaOH), 0.5% SDS, 5X Denhardt's reagent and 100 μg/ml denatured salmon sperm DNA followed by washing in a solution comprising 1 .0XSSPE, 1 .0% SDS at 42 °C when a probe of about 500 nucleotides in length is employed. A non-limiting example "Low stringency conditions" when used in reference to nucleic acid hybridization comprise conditions equivalent to binding or hybridization at 42.degree. C. in a solution consisting of 5XSSPE (43.8 g/l NaCI, 6.9 g/l NaH2P04H20 and 1 .85 g/l EDTA, pH adjusted to 7.4 with NaOH), 0.5% SDS, 5X Denhardt's reagent and 100 μg/ml denatured salmon sperm DNA followed by washing in a solution comprising 5XSSPE, 0.1 % SDS at 42 °C when a probe of about 500 nucleotides in length is employed.

[00198] The polynucleotides include the coding sequence polypeptide, in isolation, in combination with additional coding sequences (e.g., a purification tag, a localization signal, as a fusion-protein, as a pre-protein, or the like), in combination with non-coding sequences (e.g., introns or inteins, regulatory elements such as promoters (including inducible promoters, tissue- specific promoters (such as root-specific or leaf specific promoters), enhancers, terminators, and the like), and/or in a vector or host environment in which the polynucleotide encoding a transcription factor or transcription factor homologue polypeptide is an endogenous or exogenous gene.

[00199] Appropriate additional coding sequences (e.g., a purification tag, a localization signal, as a fusion-protein, as a pre-protein, or the like), non-coding sequences (e.g. regulatory elements such as promoters (including inducible promoters, tissue-specific promoters (such as root-specific or leaf specific promoters), enhancers, terminators, and the like), and vectors for use in prokaryotic such as E. coli and eukaryotic cells, including but not limited to yeast and plant cells are known in the art.

Polypeptides

[00200] The present invention provides for glycosyltransferases. The glycosyltransferases of the present invention are capable of primary, secondary and/or tertiary glycosylations. In certain embodiments, the glycosyltransferases are capable of primary, secondary and tertiary glycosylations. In other embodiments, the glycosyltransferases are capable of secondary and/or tertiary glycosylations. In certain embodiments, the glycosyltransferases is a glucosyltransferase, including but not limited to a UDP- glycotransferase. The glucosyltransferases include but are not limited to a Stevia rebaudiana UDP-glucosyltransferase, such as UGT76G1 or UGT74G1 or an Oryza sativa glucosyltrasferase, such as Os03g0702000. In other embodiments, the invention provides for a cyclodextrin glucanotransferase. Also provided are sucrose synthases.

[00201 ] In certain embodiments, there is provided an UGT76G1 or UGT76G1 -like glucosyltransferase. UGT76G1 -like glucosyltransferase include for example, other members of the UGT76G1 clade such as UGT76G2 or UGT76H1 . Accordingly, in certain embodiments, there is provided an UGT76G1 comprising the amino acid sequence as set forth in any one of SEQ ID NOs: 1 , 3, 5 and 7 or fragments and variants thereof. In certain embodiments, there is provided an UGT76G1 encoded by the nucleic acid molecule comprising the sequence as set forth in any one of SEQ ID NOs: 2, 4, 6 and 8.

[00202] In certain embodiments, there is provided an UGT76G2 comprising the amino acid sequence as set forth in SEQ ID NO: 27 or fragments and variants thereof. In certain embodiments, there is provided an UGT76G1 encoded by the nucleic acid molecule comprising the sequence as set forth in SEQ ID NO: 28. [00203] In certain embodiments, there is provided an UGT76H1 comprising the amino acid sequence as set forth in SEQ ID NO: 29 or fragments and variants thereof. In certain embodiments, there is provided an UGT76G1 encoded by the nucleic acid molecule comprising the sequence as set forth in SEQ ID NO: 30.

[00204] In certain embodiments, there is provided an Os03g0702000 or Os03g0702000- like glucosyltransferase. Os03g0702000-like glucosyltransferase include for example, other members of the UGT91 clade such as UGT91 D1 or UGT91 D2. Accordingly, in certain embodiments, there is provided an Os03g0702000 comprising an amino acid sequence as set forth in SEQ ID NO: 9 or fragments and variants thereof. In certain embodiments, there is provided an Os03g0702000 encoded by the nucleic acid molecule comprising the sequence as set forth in SEQ ID NO: 10.

[00205] In certain embodiments, there is provided an UGT91 D1 comprising the amino acid sequence as set forth in SEQ ID NO: 31 or fragments and variants thereof. In certain embodiments, there is provided an UGT91 D1 encoded by the nucleic acid molecule comprising the sequence as set forth in SEQ ID NO: 32.

[00206] In certain embodiments, there is provided an UGT91 D2 comprising the amino acid sequence as set forth in SEQ ID NO: 33 or fragments and variants thereof. In certain embodiments, there is provided an UGT76G1 encoded by the nucleic acid molecule comprising the sequence as set forth in SEQ ID NO: 34.

[00207] In certain embodiments, there is provided a Stevia rebaudiana UGT74G1 . Accordingly, in certain embodiments, the UGT74G1 comprises the amino acid sequence as set forth in SEQ ID NO: 13 or fragments and variants thereof. In certain embodiments, the UGT74G1 is encoded by the nucleic acid molecule comprising the sequence as set forth in SEQ ID NO: 14.

[00208] In other embodiments, the invention provides for a cyclodextrin glucanotransferase. Cyclodextrin-glucanotransferase is commercially available (CGTase, Toruzyme 3.0L, trademark of Novozymes Inc.).

[00209] In certain embodiments, there is provided sucrose synthase. Accordingly, in certain embodiments, the sucrose synthase comprises the amino acid sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25 or fragments and variants thereof. In certain embodiments, the polypeptide comprises an amino acid sequence encoded by the nucleic acid molecule comprises comprising the sequence as set forth in SEQ ID NO: 16,18, 20, 22, 24 or 26.

[00210] In other embodiments, there is provided a polypeptide comprising a sequence at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 96%, 97%, 98%, 99% or 100% percent identity to any one of the sequences set forth in SEQ ID NOs: 1 , 3, 5, 7, 9, 13, 15, 17, 19, 21 , 23, 25, 27, 29, 31 and 33 and fragments thereof. A worker skilled in the art would readily appreciate that overall sequence identity or similarity of related enzymes may be less than 50% but regions of the enzyme (such as the catalytic site or areas adjacent to the catalytic site) may have conserved amino acids and therefore the related enzymes have similar activity. For example, there are conserved amino acids at the opening adjacent to the UDPG catalytic site. In particular, a leucine at position 379 of UGT76G1 is conserved. In certain embodiments, the nucleic acid encodes an UDP-glucosyltransferase having the sequence SDFGLDQ at a position In certain embodiments, fragments are at least 10, at least 20, at least 50 amino acids in length. In certain embodiments, the polypeptide sequences contain heterologous sequences including but not limited to purification tags such as a HIS tag. In a certain embodiments, there is provided a polypeptide comprising a 6X HIS tag at the N-terminus. In other embodiments, there is provided a polypeptide comprising a 6X HIS tag at the C-terminus.

[00211 ] Methods for screening the activity of glycosyltransf erases including glucosyltransferases and cyclodextrin glucanotransferases are known in the art. As such, a worker skilled in the art could readily determine if the glycosyltransf erases are capable of primary, secondary and/or tertiary glycosylations (see, for example Dewitte et al., J Biotechnol. 2016 Sep 10;233:49-55. doi: 10.1016/j.jbiotec.2016.06.034; Grubb et al., Plant J. (2014) 79, 92- 105; Richman et al., Plant J. (2005) 41 , 56-67; Tanaka et al., Plant Cell Rep. (1996) 15, 819- 823; Tanaka et al., J. Nat. Prod (1993) 56(12), 2068-2072.. In addition, methods for screening the activity of sucrose synthase are also known in the art.(Baroja-Fernandez et al., PNAS. (2012) 109(1 ), 321 -326. doi: 10.1073/pnas.1 1 17099109; Barratt et al., Plant Physiol. (2001 ) 127, 655-664; Huber and Akazawa, Plant Physiol. (1986) 81 , 1008-1013.

Cells and Plants

[00212] The present invention further provides cells and plants which express one or more of the polypeptides of the present invention. The cells and plants may naturally express one or more of the polypeptides of the present invention or have been modified to express one or more the polypeptides of the present invention. The cells may be prokaryotic or eukaryotic cells and include but are not limited to, E. coli, yeast such as Pichia pastoris, Stevia rebaudiana, Phytolacca Americana, Cannabis including but not limited to Cannabis sativa, Cannabis indica and Cannabis ruderalis.

[00213] In certain embodiments, there is provided a cell which expresses an UGT76G1 or UGT76G1 -like glucosyltransferase (such as UGT76G2 and UGT76H1 ). Accordingly, in certain embodiments, there is provided a cell which expresses an UGT76G1 glucosyltransferase comprising a sequence encoding the amino acid sequence as set forth in any one of SEQ ID NOs: 1 , 3, 5 and 7. In certain embodiments, there is provided a cell which expresses an UGT76G1 -like glucosyltransferase comprising a sequence encoding the amino acid sequence as set forth in SEQ ID NO: 27 or 29. The cell may further express further glucosyltransferases, such as Os03g0702000 or Os03g0702000-like glucosyltransferase (such as UGT91 D1 and UGT91 D2) and/or a sucrose synthase, such as the sucrose synthase comprising the sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25.

[00214] Accordingly, in certain embodiments, there is provided a cell which expresses UGT76G1 glucosyltransferase comprising a sequence encoding the amino acid sequence as set forth in any one of SEQ ID NOs: 1 , 3, 5 and 7 and Os03g0702000 glucosyltransferase comprising the sequence as set forth in SEQ ID NO:10. The cell may further express a sucrose synthase comprising the sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25.

[00215] In certain embodiments, there is provided a cell which expresses an Os03g0702000 or Os03g0702000-like glucosyltransferase. Accordingly, in certain embodiments, there is provided a cell which expresses Os03g0702000 glucosyltransferase comprising a sequence encoding the amino acid sequence as set forth in SEQ ID NO: 10. The cell may further express a sucrose synthase, such as the sucrose synthase comprising the sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25.

[00216] Transgenic cells and plants (including plant cells, or plant explants, or plant tissues) can be produced by a variety of well established techniques. Following construction of a vector, most typically an expression cassette, including a polynucleotide of the invention, standard techniques can be used to introduce the polynucleotide into cell or a plant. Optionally, the plant cell, explant or tissue can be regenerated to produce a transgenic plant.

[00217] In a certain embodiments, there is provided Cannabis plants genetically engineered to express one or more of the proteins of the invention. A worker skilled in the art would readily appreciate appropriate vectors and promoters for genetically engineering Cannabis plats. For example, a tissue specific promoter, such as a secretory trichomes specific promoter may be used such that the proteins of the invention are expressed in the same tissue that cannabinoids are produced in, namely the secretory trichomes of the plant. Suitable promoter elements include the promoter for the cytosolic 0-acetylserine(thiol)lyase (OASA1 ) enzyme from Arabidopsis thaliana (Gutierrez-Alcala 2005).

[00218] Transformation and regeneration of plant cells is now routine, and the selection of the most appropriate transformation technique will be determined by the practitioner. Suitable methods can include, but are not limited to: electroporation of plant protoplasts; liposome- mediated transformation; polyethylene glycol (PEG) mediated transformation; transformation using viruses; micro-injection of plant cells; micro-projectile bombardment of plant cells; vacuum infiltration; and Agrobacterium tumeficiens mediated transformation. Transformation means introducing a nucleotide sequence into a plant in a manner to cause stable or transient expression of the sequence.

[00219] Successful examples of the modification of plant characteristics by transformation with cloned sequences which serve to illustrate the current knowledge in this field of technology, and which are herein incorporated by reference, include: U.S. Pat. Nos. 5,571 ,706; 5,677,175; 5,510,471 ; 5,750,386; 5,597,945; 5,589,615; 5,750,871 ; 5,268,526; 5,780,708; 5,538,880; 5,773,269; 5,736,369 and 5,610,042.

[00220] Following transformation, plants may be selected using a dominant selectable marker incorporated into the transformation vector. Typically, such a marker will confer antibiotic or herbicide resistance on the transformed plants, and selection of transformants can be accomplished by exposing the plants to appropriate concentrations of the antibiotic or herbicide.

Methods

[00221 ] The present invention further provides methods for the production of cannabinoid glycoside prodrugs and the cannabinoid glycosides prodrugs produced by the methods. The methods may be in vitro or in vivo (in a cell system or in planta). In certain embodiments, there is provided a method of producing cannabinoid glycoside prodrugs, said method comprising incubating a cannabinoid aglycone with one or more sugar donors in the presence of one or more glycosyltransferases. [00222] The aglycones include but are not limited to: cannabinoids, including but not limited to cannabidiol, cannabidivarin, cannabigerol, tetrahydrocannabinol, cannabinol and cannabidiolic acid, endocannabinoids including but not limited to arachidonoylethanolamide (anandamide, AEA), 2-arachidonoylethanolamide (2-AG), 1 -arachidonoylethanolamide (1 -AG), and docosahexaenoyl ethanolamide (DHEA, synaptamide); and vanilloids including but not limited to vanillin, curcumin, and capsaicin.

[00223] A worker skilled in the art would readily appreciate that the one or more sugar donors will be dependent on the one or more glycosyltransf erases used in the method and/or the desired end products. For example, for UDP-glucosyltransferases, the sugar donors include but are not limited to UDP-glucose, UDP-glucuronic acid, UDP-mannose, UDP-fructose, UDP-xylose, UDP-fluorodeoxyglucose, and UDP-rhamnose. For cyclodextrin glucanotransferase, the sugar donor includes maltodextrin.

[00224] In certain embodiments, there is provided a method of producing a cannabinoid glycoside, said method comprising incubating an aglycone with a sugar donor in the presence of a glycosyltransferase. Also provided are the cannabinoid glycosides produced by the above method. In specific embodiments, there is provided a method of producing a cannabinoid glycoside, said method comprising incubating an aglycone with UDP-glucose, in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase under conditions that allow for glycosylation. In other specific embodiments, there is provided a method of producing a glycoside prodrug, said method comprising incubating an aglycone with maltodextrin, in the presence of a cyclodextrin glucanotransferase under conditions that allow for glycosylation.

[00225] An exemplary method for producing cannabinoid-glycosides comprises incubating a cannabinoid, with UDP-glucose in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase under conditions which allow for glycosylation. Also provided are cannabinoid-glycosides produced by the above method.

[00226] A further exemplary method for producing cannabinoid-glycosides comprises incubating a cannabinoid with maltodextrin in the presence of a cyclodextrin glucanotransferase under conditions which allow for glycosylation. Also provided are cannabinoid-glycosides produced by the above method.

[00227] In certain embodiments, there is provided a method of producing a cannabinoid glycoside, said method comprising incubating an aglycone with one or more sugar donors in the presence of a first glycosyltransf erase and a second glycosyltransferase under conditions which allow for glycosylation. Also provided are cannabinoid glycosides produced by the above method.

[00228] A worker skilled in the art would readily appreciate that the first glycosyltransferase and a second glycosyltransferase may be provided concurrently or added sequentially. In addition, if more than one sugar donor is used, the sugar donors may be provided concurrently or added sequentially. Such a worker would further appreciate that the structure of the resulting cannabinoid glycoside may be dependent on the order the glycosyltransferases are provided. In addition, the ratio of first to second glycosyltransferase may impact the resulting products. A worker skilled in the art would further appreciate that the activity levels of the glycosyltransferases may dictate the ratios and the ratios could be readily determined by a worker skilled in the art. For example, the ratios first to second glycosyltransferase include but are not limited to 1 :1 , 1 :2, 1 :10, 1 :50 and vice versa.

[00229] In specific embodiments, there is provided a method of producing a cannabinoid glycoside, said method comprising incubating an aglycone with UDP-glucose in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and Os03g0702000 or Os03g0702000-like glucosyltransferase under conditions which allow for glycosylation. In alternative specific embodiments, there is provided a method of producing a cannabinoid glycoside, said method comprising incubating an aglycone with UDP-glucose and maltodextrin in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and cyclodextrin glucanotransferase under conditions which allow for glycosylation. Also provided are cannabinoid glycosides produced by the above methods.

[00230] An exemplary method for producing cannabinoid-glycosides comprises incubating cannabinoid, including but not limited to cannabidiol, cannabidivarin, canabigerol, tetrahydrocannabinol, cannabinol and cannabidiolic acid, with UDP-glucose in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and Os03g0702000 or Os03g0702000-like glucosyltransferase under conditions which allow for glycosylation. Also provided are cannabinoid-glycosides produced by the above method.

[00231 ] A further exemplary method for producing cannabinoid-glycosides comprises incubating cannabinoids with UDP-glucose and maltodextrin in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and and cyclodextrin glucanotransferase under conditions which allow for glycosylation. Also provided are cannabinoid-glycosides produced by the above method.

[00232] It is within the scope of the present invention that each of the above described glycosylation methods may be applied to a lower order cannabinoid glycoside to form a higher order cannabinoid glycoside. For example, a cannabinoid monoglycoside may be glycosylated using any of the glycosylation methods of the present invention to form a diglycoside, or a cannabinoid diglycoside may be glycosylated to form a triglycoside, etc.

[00233] Methods of purifying the cannabinoid glycosides are known in the art and include for example solid phase extraction, such as column purification.

[00234] The invention also provides cell culture and in planta methods for the production of cannabinoid glycosides. The methods comprise expressing one or more of the glycosyltransferases in a cell or plant which produces the aglycone and isolating the cannabinoid glycosides. In certain embodiments, one or more sucrose synthases are also expressed. Appropriate vectors and genetic engineering methods are known in the art.

[00235] The invention also provides methods for the conversion of UDP to UDPG utilizing the sucrose synthases of the present invention. Accordingly, in certain embodiments of the methods of producing cannabinoid glycosides which utilize UDP-glucose as a sugar donor, the methods further comprise the use of sucrose synthase to recycle UDP. In certain embodiments, there is provided a method of producing a cannabinoid glycoside, said method comprising incubating aglycone with UDP-glucose, in the presence of a UGT76G1 glucosyltransferase and a sucrose synthase under conditions that allow for glycosylation.

[00236] The invention will now be described with reference to specific examples. It will be understood that the following examples are intended to describe embodiments of the invention and are not intended to limit the invention in any way.

EXAMPLES

Example 1 : Conversion of cannabinoids to cannabinoid glycoside prodrugs

[00237] Glycosylation reactions consisted of 50mM KP04 pH 7.2, 3mM MgCI2, 0.005% CBD, 2.5% UGT76G1 purified enzyme preparation, and 2.5mM UDP-glucose. Buffers were degassed and tubes were purged with nitrogen, reactions were protected from light and incubated at 28°C with 180rpm agitation for 18 hours. Reactions were then extracted 3x with an equal volume of ethyl acetate, evaporated to dryness, and dissolved in a half volume of HPLC grade methanol. 50 microliters was injected on a reverse phase C18 column and eluted with a gradient of acetonitrile starting at 10% and increasing to 99%. UGT76G1 was produced through expression in Pichia pastoris and purified through standard molecular biology techniques. The UGT76G1 enzyme was found to glycosylate CBD in a UDP-glucose dependent manner. This activity was also proportional to the amount of UDP-glucose present. Incubation temperature was 28 °C, and an acceptable range would be 20 °C to 30 °C as high temperatures can cause significant degradation of CBD. Reactions were carried out in the dark to prevent photo- degradation of the substrates. Gentle agitation from 120 to 200rpm were used to mix the reactions in an inert atmosphere.

[00238] Substrate CBD in the reactions was replaced with A9THC and CBDV and performed in an identical fashion with similar results. Enzyme combinations needed to create various products are listed in Table 4 for CBD-glycosides, Table 5 for CBDV-glycosides, and Table 6 for A9THC-glycosides.

[00239] Other enzymes screened for activity towards CBD were the Stevia rebaudiana UGT74G1 , UGT85C2, UGPase, E.coli Maltodextrin phosphotransferase (MalP), and O.sativa Os03g0702000 (SEQ ID NO. 9). No primary glycosylation activity was seen with any other tested enzyme other than UGT76G1 .

Example 2: 2-0 glycosylation of of CBD-monoglycoside

[00240] Enzymatic reactions are performed as described in Example 1 but with the inclusion of recombinant Os03g0702000 enzyme at a 1 :2 ratio relative to UGT76G1 . Samples were extracted and analyzed as in Example 1 . Recombinant Os03g0702000 enzyme was codon optimized and expressed in E. coli BL21 -DE3 cells and purified by immobilized metal ion chromatography.

Example 3: Conversion of CBD to alpha-glycoside linked CBD compounds.

[00241 ] Recombinant cyclodextrin glucanotransferase (CGTase, Toruzyme 3.0L trade name, Novozymes Inc.) was added to reactions as indicated in Example 1 but without UDPG or UGT76G1 . Maltodextrin was used at 0.05% final concentration, and Toruzyme 3.0L was used at 0.1 %. Samples were extracted and analyzed as in example 1 . Additionally, reactions from Example 1 were carried out to convert cannabinoids to cannabinoid-glycosides, and then CGTase and maltodextrin were added and given adequate time to incubate with the cannabinoid-glycosides. The resulting products contain a β-glycosylation on the cannabinoid backbone, and a-glycosylations emanating from the primary sugar. This additional treatment created a new category of compounds termed β-primed, a-glycosylated cannabinoids.

Example 4: Purification of cannabinoid glycosides

[00242] Glycoside products were generated through the aforementioned biocatalytic reactions and purified to homogeneity by C18 solid phase extraction. 100mg Hypersep C18 columns (Thermo) were hydrated in methanol, rinsed with 50% methanol in water, rinsed with water, glycosylation reaction passed through the column, washed with water, washed with 10%, 20%, and 30% methanol, and the glycoside products were eluted with 45 and 60% methanol in water. Eluates were dried and extracted with ethyl acetate, and dried to completion to yield >95% pure cannabinoid -glycosides for further analysis and testing.

Example 5: HPLC analysis of cannabinoid glycoside prodrugs

The HPLC linetraces of the reaction products of glycosylation reactions of the cannabinoid aglycones CBD, CBDV, A9-THC, CBN, 1 -AG and 2-AG, DHEA, AEA, capsaicin, and vanillin, are provided in Figures 16 to 24, respectively. Enzymatic reactions were performed as described in Example 1 . The solid lines indicate the elution profile of the starting aglycone and the dashed lines indicate the elution profile of the glycosylation reaction product mixture.

[00243] In Figure 16, the CBD aglycone retention time is 13.65 minutes, and product peaks are observed at 8.87, 9.02, 9.97, 10.33, and 10.37 min.

[00244] In Figure 17, the CBDV aglycone retention time is 12.75 minutes, and product peaks are observed at 8.53, 9.70, and 10.01 min.

[00245] In Figure 18, the THC aglycone retention time is 14.45 minutes, and product peaks are observed at 9.46, 10.67, 10.97, 1 1 .28, 1 1 .67, and 12.49 min.

[00246] In Figure 19, the CBN aglycone retention time is 14.32 minutes, and product peaks are observed at 10.87, 1 1 .50, and 12.25 min. [00247] In Figure 20, the 1 -AG aglycone retention time is 14.18 minutes and the 2-AG aglycone retention time is 14.32 minutes, and product peaks are observed at 1 1 .40, 1 1 .78, 1 1 .83, 1 1 .97, 12.53, 12.92, 13.07, and 13.35 min.

[00248] In Figure 21 , the DHEA aglycone retention time is 13.78 minutes, and product peaks are observed at 10.09 and 12.43 min.

[00249] In Figure 22, the AEA aglycone retention time is 13.87 minutes, and product peaks are observed at 12.47 min.

[00250] In Figure 23, the vanillin aglycone retention time is 1 .95 minutes and product peaks are observed from 1 .25 to 1 .35 min.

[00251 ] In Figure 24, the capsaicin aglycone retention time is 1 1 .73 minutes, and product peaks are observed at 10.23 min.

Example 6A: LCMS analysis of CBD glycosides

As shown in the HPLC linetrace of Figure 16, input CBD aglycone (VB101 , 13.65') has been depleted to 5% of original quantity after +65 hours of incubation time. The CBD-glycosides elute off the HPLC column at 8.87, 9.02, 9.97, 10.33, and 10.37 min. The glycosylated products were identified by LCMS analysis. The glycosylated product "g1 " is a monoglycoside, "g2" is a diglycoside, "g3" is a triglycoside, and "g4" is a tetraglycoside. LC-LRMS was performed on a Shimadzu LC-MS 2010 EV instrument. The LC column used was a Silia Chrom XDB C18 5um, 150A, 4.6X50 mm. The method was 12 min 5 to 95 H20:ACN gradient . For LRMS electrospray ionization (ESI) was performed in positive mode.

[00252] VB101 (CBD aglycone) MS data: LC/ESI-LRMS. [M + H]+ (C21 H31 O2) Calcd: m/z = 315. Found: m/z = 315.

[00253] (CBDgl) MS data: LC/ESI-LRMS. [M + H]+ (C27H41 O7) Calcd: m/z = All. Found: m/z = All.

[00254] VB104 (CBDg2) MS data: LC/ESI-LRMS. [M + H]+ (C33H51O12) Calcd: m/z = 639. Found: m/z = 639.

[00255] VB110 (CBDg2) MS data: LC/ESI-LRMS. [M + H]+ (C33H51O12) Calcd: m/z = 639. Found: m/z = 639. [00256] (CBDg3) MS data: LC/ESI-LRMS. [M + H]+ (C39H61O17) Calcd: m/z = 801 . Found: m/z = 801 . [M + K + H]+ (C39H61O17K) Calcd: m/z = 420. Found: m/z = 420. [M + ACN + H20 + H]+ (C4iH63NOi7) Calcd: m/z = 860. Found: m/z = 860.

[00257] (CBDg4) MS data: LC/ESI-LRMS. [M + H]+ (C45H71 O22) Calcd: m/z = 964. Found: m/z = 964. [M + H20 + H]+ (C45H73O18) Calcd: m/z = 983. Found: m/z = 983.

[00258] (CBDg3) MS data: LC/ESI-LRMS. [M + H]+ (C39H61O17) Calcd: m/z = 801 . Found: m/z = 801 . [M + Na]+ (C39H6oOi7Na) Calcd: m/z = 823. Found: m/z = 823. [M + K + H]2+(C39H6iOi7K) Calcd: m/z = 420. Found: m/z = 420.

Example 6B: LCMS analysis of ΔΘ-THC glycosides

[00259] In a manner similar to that carried out in Example 6A, the products of the glycosylation reaction of A9-THC (shown in the HPLC linetrace of Figure 18) were identified by LCMS analysis.

[00260] VB301 (THC aglycone) MS data: LC/ESI-LRMS. [M + H]+ (C21 H31 O2) Calcd: m/z = 315. Found: m/z = 315. [M + 3ACN + 2H]2+ (C27H41 N3O2) Calcd: m/z = 314. Found: m/z = 314.

[00261 ] VB304 (THCg2) MS data: LC/ESI-LRMS. [M + H]+ (C33H51 O12) Calcd: m/z = 639. Found: m/z = 639.

[00262] VB308 (THCg3) MS data: LC/ESI-LRMS. [M + H]+ (C39H61O17) Calcd: m/z = 801 . Found: m/z = 801 . [M + Na]+ (C39H6oOi7Na) Calcd: m/z = 823. Found: m/z = 823. [M + K + H]+ (C39H6iOi7K) Calcd: m/z = 420. Found: m/z = 420.

Example 7: NMR analysis of cannabinoid glycosides

[00263] Figure 27 depicts the 1NMR spectra of isolated VB104 and Figure 28 depicts the 1 H MR spectra of isolated VB1 10. Each of these products was isolated from the reaction mixture produced by the glycosylation reaction of CBD. The 1 H NMR spectra of 10 mg/ml solutions of each compound prepared in CD3OD were obtained on a Bruker Avance II 400 MHz instrument using TopSpin acquisition and processing software.

Example 8: Solubility Analysis [00264] C18 retention times were empirically determined on a linear ramp of increasing acetonitrile on a Phenomenex Kinetex 2.6u 100A C18 column, on a Dionex HPLC equipped with Diode Array Detector. CLogP values in Table A were predicted by ChemDraw (CambridgeSoft). Reference cannabinoids were analyzed by HPLC and established logP values (http://pubchem.ncbi.nlm.nih.gov/) and used to create a calibration line as depicted in Figure 29. The predicted cLogP values correlated with the reference calibration line. C18 reverse phase HPLC retention times were plotted against the cLogP values presented in Table A, as depicted in Figure 29. Data point numbering correlates with table numbering. Open diamonds indicate novel cannabinoid glycosides, filled diamonds indicate reference cannabinoids and derivatives. ClogP values were predicted by ChemDraw (CambridgeSoft). Linear regression was performed on all data points (R2 = 0.9455).

Table A: CLogP values for select cannabinoid glycosides and reference cannabinoids

Figure imgf000082_0001

Example 9: Bioavailability Assay

[00265] In order to investigate the effectiveness of glycosylation to effect site-specific drug delivery, VB1 10 was administered to three mice by oral gavage and the animals sacrificed at 30, 60, and 90 minutes. Eight week old male Swiss mice were fasted for 12 hours prior to administration of 120mg/kg VB1 10 in 10% Ethanol USP, 10% Propylene Glycol USP, 0.05% Sodium Deoxycholate USP, 79.95% Saline USP. Following termination and tissue harvest, the intestinal contents were then extracted and analyzed by C18 reverse phase HPLC. As shown in Figure 30A, the small intestinal contents showed intact VB1 10, but no decoupled CBD. As shown in Figure 30B, the large intestinal contents contained both VB1 10 and CBD in the 60 and 90 minute time points. This decoupling of VB1 10 is consistent with the large intestinal decoupling seen for sennoside beta-glycosides, and is the result of secreted beta-glycosidases from the large intestinal microflora.

Example 10: Analysis of Large Intestine Contents Upon Administration of CBD and CBD Glycosides

[00266] In order to investigate the metabolism and decoupling of CBD-glycosides in the large intestine, an aqueous solution of a mixture of CBD-glycosides was administered to a mouse by oral gavage. As a control, a solution of CBD in cremophor, ethanol, and saline was administered to a second mouse. The animals were each sacrificed at 2 hours. Following termination and tissue harvest, the intestinal contents were then extracted and analyzed by C18 reverse phase HPLC. The mice employed in this example were eight week old male Swiss mice fasted for 12 hours prior to administration of the solutions.

[00267] The resulting extracts were analyzed by LCMS performed using a Shimadzu LC- MS 2010 EV. LC separation was carried out using a Silia Chrom XDB C18 5um, 150A, 4.6X50 mm. The method was 12 min, 5 to 95 H20:ACN gradient elution. Low resolution MS was performed in negative mode via electrospray ionization (ESI). Acetic acid and formic acid were used as sample additives during analysis, and the injection volume was 20 μΙ.

[00268] Analysis of the large intestinal contents of animals administered a mixture of oral CBD-glycosides indicated that both aglycone and glycosides were present, along with hydroxy metabolites of each:

[CBD - H], [2CBD - H] and [CBD*20H + Formic acid - H] MS data: LC/ESI-LRMS. [M - H] (C21 H29O2) Calcd: m/z = 313. Found: m/z = 313. [2M - H]- (C42H59O4) Calcd: m/z = 627. Found: m/z = 627. [IVhoH + Formic acid - H]- (C22H31O6 ) Calcd: m/z = 391 . Found: m/z = 391 .

[CBDgl - H], [CBDgl + CI] and [2CBDg1 - H] MS data: LC/ESI-LRMS. [Mgi - H] (C27H39O7) Calcd: m/z = 475. Found: m/z = 475. [Mgi + CI]" (C27H40O7CI) Calcd: m/z = 51 1 . Found: m/z = 51 1 . [2Mgi - H]- (C54H790i4) Calcd: m/z = 951 . Found: m/z = 951 .

[CBDg2 - H] and [CBDg2 + Acetic acid - H] MS data: LC/ESI-LRMS. [Mg2 - H] (C33H49O12) Calcd: m/z = 637. Found: m/z = 637. [Mg2 + Acetic acid - H]- (C35H53O14 ) Calcd: m/z = 697. Found: m/z = 697. [CBDg3 - H], [CBDg3*OH - H] and [CBDg3*OH - 2H] MS data: LC/ESI-LRMS. [Mg3 - H] (C39H59O17) Calcd: m/z = 799. Found: m/z = 799. [Μ93ΌΗ - H]- (C39H59O18) Calcd: m/z = 815. Found: m/z = 815. [MG3OH - 2H] 2 (CsgHssOis) Calcd: m/z = 407. Found: m/z = 407.

[00269] Analysis of the large intestinal contents of animals administered oral CBD indicated that hydroxy metabolites of CBD were present:

[CBD*20H + Formic acid - H] and [2CBD*30H + Acetic acid - H] MS data: LC/ESI-LRMS. [M *2OH + Formic acid— H]~ (C22H31O6") Calcd: m/z = 391 . Found: m/z = 391 . [2M*3OH + Acetic acid - H]- (C44H630i2 ) Calcd: m/z = 783.9. Found: m/z = 784.

[00270] The plasma and brains from the same animals were also extracted and analyzed by HPLC for the presence of CBD-glycosides and CBD. CBD was only present in the control animal that received CBD aglycone (data not shown). The contents of the small intestines from the same animals were also extracted and analyzed by HPLC for the presence of CBD- glycosides and CBD, but no CBD aglycone was present in the small intestines (data not shown, consistent with THC decoupling data shown in example 1 1 ). The presence of the CBD aglycone in the large intestinal contents indicates the successful delivery of CBD-glycosides, and the subsequent hydrolysis of the glycosides by beta-glycosidase enzymes only present in the large intestine. The presence of decoupled CBD in the large intestine, but not in the small intestine, indicates that glycoside decoupling only occurs upon transit to the large intestine. The presence of CBD detoxification metabolite CBD-20H is also consistent with delivery of CBD and absorption into the intestinal epithelium where CBD begins to be metabolized. This example illustrates the potential to administer CBD-glycosides, safely transit the CBD-glycosides through the small intestine without absorption, transit to the large intestine where the sugars can be decoupled to release CBD locally, avoiding systemic absorption and delivery of the CBD to other tissues where it can have unwanted effects.

Example 11 : Analysis of Large Intestine Contents Upon Administration of THC- Glycosides

[00271 ] In order to investigate the metabolism and decoupling of THC-glycosides in the large intestine, an aqueous solution of a mixture of THC-glycosides was administered to two mice by oral gavage. The first animal was sacrificed at 2 hours and the second animal was sacrificed at 4 hours. Following termination and tissue harvest, the intestinal contents were then extracted and analyzed by C18 reverse phase HPLC. The mice employed in this example were eight week old male Swiss mice fasted for 12 hours prior to administration of the solutions.

[00272] The resulting extracts were analyzed by LCMS under the same conditions employed in Example 10.

[00273] Analysis of the large intestinal contents from mice administered THC glycosides after 2 hours indicated that both THC aglycone and THC glycosides were present, along with hydroxy metabolites of each:

[THC - H], [THC*OH - H], [2THC*30H + Acetic acid - H] and [THC*20H + Formic acid - H] MS data: LC/ESI-LRMS. [M - H]- (C21 H29O2) Calcd: m/z = 313. Found: m/z = 313. [ΜΌΗ - H]- (C21 H29O3) Calcd: m/z = 329. Found: m/z = 329. [2M*3OH + Acetic acid - H]- (C44H630i2 ) Calcd: m/z = 783.9. Found: m/z = 783. [IVhoH + Formic acid - H]~ (C22H31 Cv) Calcd: m/z = 391 . Found: m/z = 391 .

[THCgl + CI], [THCgl + Acetic acid - H], [2THCg1 - H], and [2THCg1 + Acetic acid - H] MS data: LC/ESI-LRMS. [Mgi + CI]" (C27H40O7CI ) Calcd: m/z = 51 1 . Found: m/z = 51 1 . [Mgi + Acetic acid - H]- (C29H43O9 ) Calcd: m/z = 535. Found: m/z = 535. [2Mgi - H]- (C54H79O14) Calcd: m/z = 951 . Found: m/z = 951 . [2Mgi + Acetic acid - H]- (CseHssOie ) Calcd: m/z = 101 1 . Found: m/z = 101 1 .

[THCg2 - H], [THCg2 + Acetic acid - H] and [THCg2*OH + Formic acid - H] MS data:

LC/ESI-LRMS. [Mg2 - H]" (C33H49O12) Calcd: m/z = 637. Found: m/z = 637. [Mg2 + Acetic acid - H]- (C35H53O14 ) Calcd: m/z = 697. Found: m/z = 697. [MG2*0H + Acetic acid - H]- (C34H51O15 ) Calcd: m/z = 699. Found: m/z = 699.

[THCg3 - H], [THCg3 + Acetic acid - H], [CBDg3*OH - H] and [CBDg3*OH - 2H] MS data:

LC/ESI-LRMS. [Mg3 - HKC39H59O17) Calcd: m/z = 799. Found: m/z = 799. [Mg3 + Acetic acid - H]- (C4i H630i9-) Calcd: m/z = 859. Found: m/z = 859. [MG3OH - H]- (C39H59O18 ) Calcd: m/z = 815. Found: m/z = 815. [MG3OH - 2H] 2 (C39H58O182 ) Calcd: m/z = 407. Found: m/z = 407.

[00274] Analysis of the THC glycosides mixture extract after 4 hours indicated that both THC aglycone and THC glycosides were confirmed, along with hydroxy metabolites of each:

[THC - H], [THC*OH + Acetic acid - H], [2THC*30H + Acetic acid - H] and [THC*20H + Formic acid - H] MS data: LC/ESI-LRMS. [M - H]- (C21 H29O2) Calcd: m/z = 313. Found: m/z = 313. [ΜΌΗ + Acetic acid - H]- (C23H33O5 ) Calcd: m/z = 389. Found: m/z = 389. [2M*3OH + Acetic acid - H]- (C44H63O12 ) Calcd: m/z = 783.9. Found: m/z = 784. [IVhoH + Formic acid - H]- (C22H31 O6 ) Calcd: m/z = 391 . Found: m/z = 391 .

[THCgl + CI], [THCgl + Acetic acid - H], [2THCg1 - H], and [2THCg1 + Acetic acid - H] MS data: LC/ESI-LRMS. [Mgi + CI]- (C27H40O7CI ) Calcd: m/z = 51 1 . Found: m/z = 51 1 . [Mgi + Acetic acid - H]- (C29H43O9 ) Calcd: m/z = 535. Found: m/z = 535. [2Mgi - H]- (C54H79O14) Calcd: m/z = 951 . Found: m/z = 951 . [2Mgi + Acetic acid - H]- (CseHssOie ) Calcd: m/z = 101 1 . Found: m/z = 101 1 .

[THCg2 - H] and [THCg2 + Acetic acid - H] MS data: LC/ESI-LRMS. [Mg2 - H] (C33H49O12) Calcd: m/z = 637. Found: m/z = 637. [Mg2 + Acetic acid - H]- (C35H53O14 ) Calcd: m/z = 697. Found: m/z = 697.

[THCg3 - H], [THCg3 + Acetic acid - H], [CBDg3*OH - H], [CBDg3*OH - 2H] and [CBDg3*OH + Acetic acid - 2H] MS data: LC/ESI-LRMS. [Mg3 - H]-(C39H590i7) Calcd: m/z = 799. Found: m/z = 799. [Mg3 + Acetic acid - H]- (C41 H63O19 ) Calcd: m/z = 859. Found: m/z = 859. [MG3OH - H]- (C39H59O18-) Calcd: m/z = 815. Found: m/z = 815. [MG3OH - 2H] 2 (C39H58O182 ) Calcd: m/z = 407. Found: m/z = 407. [MG3OH + Acetic acid - 2H] 2 (C4i H6202o2 ) Calcd: m/z = 467. Found: m/z = 467.

[00275] The plasma and brains from the same animals were also extracted and analyzed by HPLC for the presence of THC-glycosides and THC, but neither compound was seen in these tissues (data not shown). The contents of the small intestines from the same animals were also extracted and analyzed by HPLC for the presence of THC-glycosides and THC, but no THC aglycone was observed (data not shown, consistent with CBD decoupling data shown in Example 10). The presence of the THC aglycone in the large intestinal contents at 2 and 4 hours indicates the successful delivery of THC-glycosides, and their subsequent hydrolysis of the glycosides by beta-glycosidases in the large intestine. The presence of decoupled THC in the large intestine, but not in the small intestine, indicates that glycoside decoupling only occurs upon transit to the large intestine. The presence of THC detoxification metabolites in the large intestine is further proof that the THC aglycone is present and being absorbed by the intestinal epithelium where it begins to be metabolized. This example illustrates the potential to administer THC-glycosides orally, transit the THC-glycosides through the small intestine without absorption, transit to the large intestine where the sugars can be decoupled to release THC locally, avoiding systemic absorption and delivery of the THC to the central nervous system where it can have unwanted psychoactivity.

Example 12: Discovery of novel sucrose synthase isoforms from Stevia rebaudiana

[00276] A number of research groups have utilized simple UDP to UDPG recycling systems to decrease the amount of UDPG needed for product formation (Hardin 2004, Bungarang 2013). These studies have characterized the primary sucrose synthase isoforms found in leaf tissue, which presumably carry out the synthesis of sucrose by reacting fructose with UDPG, producing sucrose and spent UDP.

[00277] As plants are known to contain numerous isoforms of the sucrose synthase enzyme, identification of alternative SUS enzymes from the Stevia rebaudiana plant with enhanced activity for the back reaction of UDP + sucrose UDPG + fructose was carried out. As steviol glycosides occur at a high level in Stevia leaves, it was postulated that a sucrose synthase from the leaves of Stevia would have improved ability to catalyze the back reaction that recycles UDP to UDPG. Six sucrose synthase isoforms were identified within the stevia transcriptome, all having similar homology to the 6 isoforms found in Arabidopsis thaliana and named in conjunction with their homologues. These transcripts were cloned as described in materials and methods with the corresponding sequence ID information listed herein.

[00278] Enzymatic activities were tested and assayed for their ability to enhance UGT reactions with decreased UDPG input. The best isoform, SrSUS4, was capable of recycling UDP to UDPG with sucrose, in concert with the steviol 19-O-glucosyltransferase SrUGT74G1 mediated glycosylation of steviol bioside to stevioside.

[00279] Targeted mutagenesis was performed to mutate a serine residue at the N-terminus that is commonly phosphorylated in planta to prevent dimerization (Hardin 2004). SrSUSI - S13D mutants were created by mutating serine at position 13 to an aspartic acid residue (S13D), thus forming a phospho-mimetic protein. Additionally, the creation of SrSusl - S13R,L14I was created to replace the serine with an arginine, a large charged residue, also to prevent dimerization and inactivation of the enzyme. Sucrose synthase mutants showed improved UDPG production activity compared to their native counterparts. SrSUS5 (SEQ ID NOs. 19 and 20) was identified in the Stevia transcriptome and primers designed (SEQ ID NOs. 67 and 68), but was not able to be amplified from cDNA. SrSus4 showed an impressive UDPG recycling activity with a 20% improvement over the activity seen in SrSusl . It is proposed that SrSus4 is the ideal isoform for carrying out the back reaction of converting of UDP to UDPG in the presence of sucrose. For midi-scale purification of cannabinoid glycosides the use of C18 flash chromatography columns were employed. Biotage flash C18 columns with 33g of resin were washed, loaded, washed, and eluted using peristaltic pumps to achieve the similar separation and purification as the gravity fed Hypersep columns listed previously.

[00280] Relative activity for UDPG production with SUS isoforms is as follows:

SrSus4 > SrSusl -Untagged > SrSus6 > SrSus2 > SrSusl > 6xHis-SrSus1 > SrSus3

Example 13: Improved in vitro catalysis of cannabinoid-glycosides

[00281 ] As the formation of cannabinoid glycosides via UGT enzyme requires the nucleotide sugar donor UDPG in stoichiometric amounts, it is advantageous to recycle or recapture the spent UDP following a glycosylation reaction. Utilizing the SUS4 isoform from Stevia rebaudiana, cannabinoid glycosides were successfully produced using only UMP as the input nucleotide.

[00282] A two step reaction took place, first to produce UDP from UMP, and second to produce UDPG from the UDP in tandem with the UGT reaction. First, a 5L reaction containing 50mM KP04 pH7.2, 200mM UMP disodium salt, 200mM ATP disodium salt, 1 M MgCI2, 10% UMPK recombinant enzyme in 50% glycerol was prepared. The reaction was incubated at 28C with stirring for > 24hours. The 5L reaction 1 was filtered at 0.45microns to remove precipitate then applied to a 50L reaction containing 50mM KP04 pH7.2, 50mM MgCI2, 300mM Sucrose, 200mg of CBD in 200ml DMSO, 5L UGT76G1 in 50% glycerol, 2.5L SrSUS4 in 50% glycerol. The main 50L reaction was then mixed and allowed to react. An additional 200mg of CBD in 200ml DMSO was added after the reaction went to completion, and allowed to continue incubating at the same conditions. After the remaining CBD was consumed by the reaction, the mixture was filtered by tangential flow filtration with a ultrafiltration membrane at 5kDa to remove enzymes and particulate, and then concentrated using nanofiltration membrane at 500Da. The nanofiltration retentate containing the cannabosides was then applied to hydrated C18 flash columns, washed with 10-30% methanol, and eluted with 40-65% methanol. The eluate was then concentrated by rotary evaporation to remove all solvent, shell-frozen in a vacuum beaker and lyophilized to dryness. The powdered cannabosides produced were then collected and stored at -20C in sealed vials. Sucrose should be sterile filtered to avoid carmelization or sugar breakdown, as autoclaving sucrose stock solutions greatly decreases reaction activity.

Figure imgf000090_0002

Figure imgf000090_0001
Figure imgf000091_0002

Figure imgf000091_0001
Table 3: AS-Tetrahydrocannabtnoi-giycoside compositions by R-group

R-group iocafjon is as depicted in Figure 18

Figure imgf000092_0001

4: -Cannabinof-glycoside compositions by R-group

-group iocaiioo is as depicted in Figure 18

Figure imgf000092_0002

Table 5: Anan amide (AEA) glycoside Compositions by -group

R-group location is as depicted in Figure 1 S

Figure imgf000093_0001

Table 6: 2-Arachidonoylg!ycero! (2-AG)-giycoside compositions by R-group

R-group location is as in Figure 18

Figure imgf000093_0002

Tabie 7: 1 -Arachidonoyiglycero! (1-AGJ-giycoside compositions by R-group

R-group location is as depicted in Figure 1B

Figure imgf000094_0001

Table 8: Docosahexaenoyi et anoioamtde (DHEA) glycoside compositions by R-group

R-group location is as depicted in Figure 18

Figure imgf000095_0001

Table 9: Capsiacin glycoside compositions by R-group

R-group location is as depicted in Figure 18 ^

Figure imgf000095_0002

TabielO: Vanillin glycoside compositions by R-group

R-grou location is as depicted in Figure 1B

Figure imgf000096_0001

Figure imgf000097_0001

[00283] It is obvious that the foregoing embodiments of the invention are examples and can be varied in many ways. Such present or future variations are not to be regarded as a departure from the spirit and scope of the invention, and all such modifications as would be obvious to one skilled in the art are intended to be included within the scope of the following claims.

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Claims

A cannabinoid glycoside prodrug compound having formula (I)
Figure imgf000102_0001
wherein
R is H, β-D-glucopyranosyl, or 3-0^-D-glucopyranosyl^-D-glucopyranosyl;
R' is H or β-D-glucopyranosyl, or 3-0^-D-glucopyranosyl^-D-glucopyranosyl; and
A is an aglycone moiety formed through reaction of a hydroxyl group on a cannabinoid compound, an endocannabinoid compound, or a vanilloid compound,
or a pharmaceutically compatible salt thereof.
2. A compound according to claim 1 , wherein A is A', A" or A'";
wherein ' is:
Figure imgf000102_0002
wherein A" is:
Figure imgf000103_0001
Figure imgf000104_0001
wherein G is H, β-D-glucopyranosyl, 3-0^-D-glucopyranosyl^-D-glucopyranosyl, glucopyranosyl-(1→3)^-D-glucopyranosyl-(1→3)-D-glucopyranosyl.
3. A compound according to claim 2, wherein A is A'.
4. A compound according to claim 3, wherein A' is:
Figure imgf000104_0002
wherein G is as defined above. 5. A compound according to claim 4, selected from:
Figure imgf000105_0001
104
Figure imgf000106_0001
Figure imgf000106_0002
wherein G is as defined above.
7. A compound according to claim 6, selected from:
Figure imgf000107_0001
106
Figure imgf000108_0001
Figure imgf000108_0002
Figure imgf000108_0003
107
Figure imgf000109_0001
Figure imgf000109_0002
compound according to claim 10, selected from:
Figure imgf000109_0003
Figure imgf000110_0001
VB405 , and
12. A compound according to claim 2, wherein A is A".
13. A compound according to claim 12, wherein A" is:
Figure imgf000110_0002
A compound according to claim 13, selected from
Figure imgf000110_0003
VB502 VB503
Figure imgf000111_0001
VB506
A compound according to claim 12, wherein A" is:
Figure imgf000111_0002
wherein G is as defined above.
A compound according to claim 15, selected from:
Figure imgf000111_0003
Figure imgf000112_0001
Figure imgf000113_0001
A compound according to claim 12, wherein A" is: r
Figure imgf000113_0002
wherein G is as defined above.
A compound according to claim 17, selected from:
Figure imgf000113_0003
Figure imgf000114_0001
Figure imgf000114_0002
Figure imgf000115_0001
A compound according to claim 12, wherein A" is:
H
Figure imgf000115_0002
A compound according to claim 19, selected from:
Figure imgf000115_0003
Figure imgf000116_0001
Figure imgf000116_0002
Figure imgf000116_0003
, and
Figure imgf000117_0001
VB806
. A compound according to claim 2, wherein A is A'".
Figure imgf000117_0002
A compound according to claim 22, selected from
Figure imgf000117_0003
Figure imgf000117_0004
Figure imgf000118_0001
24. A compound according to claim 21 , wherein A'" is:
Figure imgf000118_0002
A compound according to claim 24, selected from
Figure imgf000118_0003
Figure imgf000119_0001
Figure imgf000119_0002
Figure imgf000119_0003
Figure imgf000120_0001
Figure imgf000120_0002
Figure imgf000121_0001
Figure imgf000122_0001
Figure imgf000123_0001
Figure imgf000124_0001
28. A compound selected from: VB102, VB103, VB104, VB105, VB106, VB107, VB108, VB109, VB110, VB111, VB112, VB113, VB114, VB115, VB116, VB117, VB118, VB119, VB120, VB121, VB122, VB123, VB124, VB125, VB126, VB127, VB128, VB129, VB130, VB131, VB132, VB133, VB134, VB202, VB203, VB204, VB205, VB206, VB207, VB208, VB209, VB210, VB211, VB212, VB213, VB214, VB215, VB216, VB217, VB218, VB219, VB220, VB221, VB222, VB223, VB224, VB225, VB226, VB227, VB228, VB229, VB230, VB231, VB232, VB233, VB234, VB301, VB302, VB303, VB304, VB305, VB306, VB307, VB308, VB402, VB403, VB404, VB405, VB406, VB407, VB408, VB502, VB503, VB504, VB505, VB506, VB507, VB508, VB602, VB603, VB604, VB605, VB606, VB607, VB608, VB609, VB610, VB611, VB612, VB613, VB614, VB615, VB616, VB617, VB618, VB619, VB620, VB621, VB622, VB623, VB702, VB703, VB704, VB705, VB706, VB707, VB708, VB709, VB710, VB711, VB712, VB713, VB714, VB715, VB716, VB717, VB718, VB719, VB720, VB721, VB722, VB723, VB802, VB803, VB804, VB805, VB806, VB807, VB808, VB902, VB903, VB904, VB905, VB906, VB907, VB908, VB1002, VB1003, VB1004, VB1005, VB1006, VB1007, VB1008, VB1102, VB1103, VB1104, VB1105, VB1106, VB1107, VB1108, VB1109, VB1110, VB1111, VB1112, VB1113, VB1114, VB1115, VB1116, VB1117, VB1118, VB1119, VB1120, VB1121, VB1122, VB1123, VB1124, VB1125, VB1126, VB1127, VB1128, VB1129, VB1130, VB1131 , VB1132, VB1133, VB1134, VB1135, and VB1136.
29. A pharmaceutical composition comprising a compound as defined in any one of claims 1 to 28 and a pharmaceutically acceptable carrier, diluent, excipient, or adjuvant.
30. A method for the site-specific delivery of a cannabinoid drug to a subject, comprising the step of administering a cannabinoid glycoside prodrug as defined in any one of claims 1 to 28 to a subject in need thereof.
31 . The method of claim 30, wherein the cannabinoid glycoside prodrug is formulated for oral administration.
32. The method of claim 30, wherein the cannabinoid glycoside prodrug is formulated for parenteral administration.
33. The method of claim 30, wherein the cannabinoid glycoside is formulated for transdermal administration.
34. A method for the site-specific delivery of a cannabinoid drug to a subject, comprising the step of administering a pharmaceutical composition as defined in claim 29 to a subject in need thereof.
35. The method of claim 34, wherein the pharmaceutical composition is formulated for oral administration.
36. The method of claim 34, wherein the pharmaceutical composition is formulated for parenteral administration.
37. The method of claim 34, wherein the pharmaceutical composition is formulated for transdermal administration.
38. A method for facilitating the transport of a cannabinoid drug across the blood brain barrier of a subject comprising administering a cannabinoid glycoside prodrug as defined in any one of claims 1 to 28 to a subject in need thereof.
39. An antimicrobial agent comprising an effective amount of a cannabinoid glycoside prodrug as defined in any one of claims 1 to 28.
40. Use of an effective amount of a cannabinoid glycoside prodrug as defined in any one of claims 1 to 28 as an antimicrobial agent.
41 . A detersive agent comprising an effective amount of a cannabinoid glycoside prodrug as defined in any one of claims 1 to 28.
42. Use of an effective amount of a cannabinoid glycoside prodrug as defined in any one of claims 1 to 28 as a detersive agent.
43. A method of producing a cannabinoid glycoside, comprising incubating a cannabinoid aglycone with one or more sugar donors in the presence of one or more glycosyltransferases.
44. The method of claim 43, wherein the one or more glycosyltransferases is a UGT76G1 or UGT76G1 -like glucosyltransferase.
45. The method of claim 44, wherein the one or more glycosyltransferases further comprise a Os03g0702000 or Os03g0702000-like glucosyltransferase.
46. The method of any one of claims 43 to 45, wherein the one or more sugar donors are selected from the group consisting of UDP-glucose, UDP-glucuronic acid, UDP-mannose, UDP- fructose, UDP-xylose, UDP-rhamnose, UDP-fluoro-deoxyglucose and combinations thereof.
47. The method of claim 46, wherein the sugar donor is UDP-glucose.
48. A method of producing a cannabinoid glycoside comprising incubating a cannabinoid aglycone with UDP-glucose, in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase under conditions that allow for glycosylation.
49. A method of producing a cannabinoid glycoside comprising incubating a cannabinoid aglycone with one or more sugar donors in the presence of a first glycosyltransferase and a second glycosyltransferase under conditions which allow for glycosylation.
50. The method of claim 49, wherein the sugar donor is UDP-glucose, the first glycosyltransferase is a UGT76G1 or UGT76G1 -like glucosyltransferase, and the second glycosyltransferase is a Os03g0702000 or Os03g0702000-like glucosyltransferase.
51 . A method of producing a cannabinoid glycoside comprising incubating a cannabinoid aglycone with UDP-glucose in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and Os03g0702000 or Os03g0702000-like glucosyltransferase under conditions which allow for glycosylation.
52. The method of any one of claims 43 to 51 , wherein the cannabinoid aglycone is a cannabinoid, an endocannabinoid, or a vanilloid.
53. The method of any one of claims 43 to 51 , wherein the cannabinoid glycoside produced by the method is a compound of the Formula (I) as defined in any one of claims 1 to 28.
54. A method of producing a cannabinoid glycoside comprising incubating a cannabinoid aglycone with maltodextrin, in the presence of a cyclodextrin glucanotransferase under conditions that allow for glycosylation.
55. A method of producing a cannabinoid glycoside comprising incubating a cannabinoid aglycone with UDP-glucose and maltodextrin in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and cyclodextrin glucanotransferase under conditions which allow for glycosylation.
56. The method of any one of claims 44, 48, 51 , 52 and 55, wherein the UGT76G1 or UGT76G1 -like glucosyltransferase comprises the sequence as set forth in SEQ ID NO: 1 , 3, 5 or 7.
57. The method of any one of claims 45, 50 and 51 , wherein the Os03g0702000 or Os03g0702000-like glucosyltransferase comprises the sequence as set forth in SEQ ID NO: 9.
58. The method of any one of claims 43 to 57, further comprising incubating with sucrose synthase.
59. The method of claim 58, wherein the sucrose synthase comprises the sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25.
60. A method for the production of a cannabinoid glycoside comprising expressing one or more of the glycosyltransferases in a cell or plant which produces a cannabinoid aglycone and isolating the cannabinoid glycoside.
61 . A method of producing a higher order cannabinoid glycoside, comprising incubating a lower order cannabinoid glycoside with one or more sugar donors in the presence of one or more glycosyltransferases.
62. The method of claim 61 , wherein the one or more glycosyltransferases is a UGT76G1 or UGT76G1 -like glucosyltransferase.
63. The method of claim 62, wherein the one or more glycosyltransferases further comprise a Os03g0702000 or Os03g0702000-like glucosyltransferase.
64. The method of any one of claims 61 to 63, wherein the one or more sugar donors are selected from the group consisting of UDP-glucose, UDP-glucuronic acid, UDP-mannose, UDP- fructose, UDP-xylose, UDP-rhamnose, UDP-fluoro-deoxyglucose and combinations thereof.
65. The method of claim 64, wherein the sugar donor is UDP-glucose.
66. A method of producing a higher order cannabinoid glycoside comprising incubating a lower order cannabinoid glycoside with UDP-glucose, in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase under conditions that allow for glycosylation.
67. A method of producing a higher order cannabinoid glycoside comprising incubating a lower order cannabinoid glycoside with one or more sugar donors in the presence of a first glycosyltransferase and a second glycosyltransferase under conditions which allow for glycosylation.
68. The method of claim 67, wherein the sugar donor is UDP-glucose, the first glycosyltransferase is a UGT76G1 or UGT76G1 -like glucosyltransferase, and the second glycosyltransferase is a Os03g0702000 or Os03g0702000-like glucosyltransferase.
69. A method of producing a higher order cannabinoid glycoside comprising incubating a lower order cannabinoid glycoside with UDP-glucose in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and Os03g0702000 or Os03g0702000-like glucosyltransferase under conditions which allow for glycosylation.
70. The method of any one of claims 61 to 69, wherein the lower order cannabinoid glycoside is a lower order cannabinoid glycoside, a lower order endocannabinoid glycoside, or a lower order vanilloid glycoside.
71 . The method of any one of claims 61 to 70, wherein the higher order cannabinoid glycoside produced by the method is a compound of the Formula (I) as defined in any one of claims 1 to 28.
72. A method of producing a higher order cannabinoid glycoside comprising incubating a lower order cannabinoid glycoside with maltodextrin, in the presence of a cyclodextrin glucanotransferase under conditions that allow for glycosylation.
73. A method of producing a higher order cannabinoid glycoside comprising incubating a lower order cannabinoid glycoside with UDP-glucose and maltodextrin in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase and cyclodextrin glucanotransferase under conditions which allow for glycosylation.
74. The method of any one of claims 62, 66, 69, 70 and 73, wherein the UGT76G1 or UGT76G1 -like glucosyltransferase comprises the sequence as set forth in SEQ ID NO:1 , 3, 5 or 7.
75. The method of any one of claims 63, 68 and 69, wherein the Os03g0702000 or Os03g0702000-like glucosyltransferase comprises the sequence as set forth in SEQ ID NO:9.
76. The method of any one of claims 61 to 75, further comprising incubating with sucrose synthase.
77. The method of claim 76, wherein the sucrose synthase comprises the sequence as set forth in SEQ ID NO: 15, 17, 19, 21 , 23 or 25.
78. A method for the production of a higher order glycoside comprising incubating a lower order glycoside with UDP-glucose, in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase under conditions that allow for glycosylation, wherein the lower order glycoside is other than a steviol glycoside.
79. A method for the production of a glycoside comprising incubating an aglycone with UDP- glucose, in the presence of a UGT76G1 or UGT76G1 -like glucosyltransferase under conditions that allow for glycosylation.
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