PRODUCTION OF STEVIOL GLYCOSIDES THROUGH WHOLE-CELL BIOTRANSFORMATION

Abstract

In several aspects and embodiments, the invention provides microbial cells and methods for producing advanced glycosylation products from lower glycosylated intermediates. The microbial cell expresses one or more UDP-dependent glycosyltransferase enzymes in the cytoplasm for the glycosylation of the intermediates. When the microbial strain is incubated with a plant extract or a portion thereof comprising the intermediates, these glycosylated intermediates are available for further glycosylation by the cell, and the advanced glycosylation products can be recovered from the medium and / or the microbial cells.

Description

PRODUCTION OF STEVIOL GLYCOSIDES THROUGH WHOLE-CELL BIOTRANSFORMATION BACKGROUND Small-molecule glycosyltransferases are encoded by a large multigene family in the plant kingdom. These enzymes transfer sugars from nucleotide sugars to a wide range of secondary metabolites, thereby altering the physical and chemical properties of the receiving molecule. For example, steviol glycosides are a class of compounds found in the leaves of Stevia rebaudiana Bertoni, a perennial shrub in the Asteraceae (Compositae) family native to certain regions of South America. They are structurally characterized by a single base, steviol, which is distinguished by the presence of carbohydrate residues at the C13 and C19 positions. They accumulate in stevia leaves, comprising approximately 10% to 20% of the total dry weight. On a dry weight basis, the four main glycosides found in Stevia leaves typically include stevioside (9.1%), rebaudioside A (3.8%), rebaudioside C (0.6-1.0%) and dulcoside A (0.3%). Other steviol glycosides are present in small or trace amounts, including rebaudioside B, D, E, F, G, H, I, J, K, L, M and O, dulcoside B, steviolbioside and rubusoside. The minor glycosylation product rebaudioside M (RebM) is estimated to be about 200–350 times more potent than sucrose and is described as having a clean, sweet taste with a slightly bitter or licorice aftertaste. Prakash I. et al., Development of Next Generation Stevia Sweetener: Rebaudioside M, Foods 3(1), 162–175 (2014). While RebM is of great interest to the global food industry, its low prevalence in stevia extract necessitates innovative synthesis processes. There remains a need for economical methods to produce high-value glycosides that are minor products of natural plant extracts. SUMMARY OF THE INVENTION In various aspects and embodiments, the invention provides microbial cells and methods for producing advanced glycosylation products from lower glycosylated intermediates. The microbial cell expresses one or more UDP-dependent glycosyltransferase enzymes for the glycosylation of the intermediates. When the microbial strain is incubated with a plant extract or a fraction thereof comprising the glycoside intermediates, these intermediates are available for further glycosylation by the microbial cells. In various embodiments, the advanced glycosylation products are recovered from the medium and / or microbial cells. Glycoside intermediates can be any glycosylated secondary metabolite, such as those found naturally in plant extracts, including glycosylated terpenoids, flavonoids, cannabinoids, polyketides, stilbenoids, and polyphenols, among others. In some embodiments, the glycosylated intermediate is a glycosylated terpenoid, such as steviol glycoside or mogroside, and can be used as a sweetener. In some embodiments, the biosynthesis of the product RCLZbn / Lznza / YiAi involves at least two glycosylation reactions of the glycoside intermediate in the microbial cell. In some formulations, the glycoside intermediates are derived from stevia leaf extract. For example, RebM and other advanced glycation end products (AGEs) can be biosynthesized from steviol glycoside intermediates such as stevioside, steviolbioside, rebaudioside A (RebA), and rebaudioside C (RebC), among others. The microbial cell expresses one or more UDP-dependent glycosyltransferase enzymes in the cytoplasm for the glycosylation of lower-value intermediates. When the microbial strain is incubated with a stevia leaf extract or a fraction thereof comprising the steviol glycoside intermediates, these intermediates become available to the cells for further glycosylation, and these products can be recovered from the medium and / or microbial cells.Therefore, this process uses advanced intermediates in stevia leaf extract, namely steviol glycosides with one to five glycosylations (as well as, in some formulations, the aglycone core, steviol). These advanced intermediates from stevia leaf extract are readily available from existing industrial extraction of steviol glycosides. In several embodiments, the microbial cell expresses at least one, at least two, at least three, or at least four UGT enzymes that glycosylate a glycoside intermediate, and these may be selected from those listed in Table 1 and those provided herein as SEQ ID NO: 1-45, as well as their derivatives. In several embodiments, the UGT enzymes are independently selected from 1-2' glycosylating UGT enzymes, 1-3' glycosylating UGT enzymes, and O-glycosylating UGT enzymes. In several embodiments, these enzymes are expressed intracellularly, as they do not contain secretion or membrane translocation signals. In some modalities, particularly with regard to the glycosylation of steviol glycoside intermediates in stevia leaf extract, the microbial cell expresses four UGT enzymes, such as a UGT 13-0 glycosylating enzyme, a UGT 19-0 glycosylating enzyme, a UGT 1-2' glycosylating enzyme, and a UGT 1-3' glycosylating enzyme. In several forms, the microbial cell expresses a 1-3' glycosylating UGT enzyme. For example, the 1-3' glycosylating UGT enzyme can be selected from SrUGT76G1, MbUGT1-3 and their derivatives (e.g., UGT76G1_L200A or MbUGT1-3_1, MbUGT1-3_1.5 or MbUGT1-3_2). In these and other modalities, the microbial cell expresses a 1-2' glycosylating UGT enzyme. For example, the 1-2' glycosylating UGT enzyme can be selected from SrUGT91D2, SrUGT91D1, SrUGT91D2e, OsUGT1-2, MbUGT1,2, MbUGTI,2.2, and derivatives thereof. In these or other modalities, the microbial cell expresses a C13 UGT O-glycosylated enzyme. For example, the C13 UGT O-glycosylated enzyme can be selected from SrUGT85C2 and its derivatives (e.g., MbUGTC13). In these or other modalities, the microbial cell expresses a C19 O-glycosylated UGT enzyme. For example, the C19 O-glycosylated enzyme can be selected from SrUGT74G1, MbUGTc19 and derivatives thereof (e.g., MbUGTc19-2). RpLZbn / Lzzza / YiAi Whole-cell conversion requires intracellularly expressed enzymes to act on an externally fed substrate (e.g., glycosylated intermediates) and the cell to regenerate the UDP-glucose cofactor. This contrasts with processes that rely on enzymes purified or secreted outside the cell, which require an exogenous supply of UDP-glucose, a UDP-glucose precursor, a UDP-glucose regeneration mechanism, or a UDP-glucose regeneration enzyme system. In embodiments of the present invention, catalysis (glycosylation) is carried out in live microbial cells. Recycling of the UDP-glucose cofactor occurs using native cellular metabolism without requiring additional externally supplied enzymes or substrate feed. According to this disclosure, genetic modifications of the microbial cell allow glycoside intermediates to be available for further glycosylation by intracellularly expressed enzymes. In some embodiments, advanced glycosylated products can be recovered from the medium, as opposed to being extracted from lysed cells. In some embodiments, the microbial cell has one or more genetic modifications that increase the availability of UDP-glucose. In some embodiments, without intending to be limited to theory, these modifications may also stress the cell for glucose availability, leading to increased expression of endogenous transporters to import glycoside intermediates into the cell.In some modalities, without intending to limit ourselves to theory, cells are made permeable through genetic modification or components of the media, allowing the passive diffusion of products and substrates. In several modes, the microbial cell overexpresses one or more endogenous transporters (e.g., compared to a parental microbial strain), or in certain modes, it is modified to express a recombinant and / or genetically modified transport protein. In some modes, the microbial cell expresses one or more additional copies of an endogenous transport protein or a derivative thereof. For example, the expression or activity of transport proteins can be modified to increase the transport into the cell of steviol glycoside substrates (e.g., one or more steviosides, steviolbiosides, RebA, and / or RebC), while exporting products such as RebM and / or RebD, as well as other advanced glycation end products. Example transport proteins that can be overexpressed or manipulated to alter activity or substrate specificity in the microbial cell include E.coli acrAD, xylE, ascF, bglF, chbA, ptsG / crr, wzxE, rfbX, as well as orthologs or derivatives thereof. Other transport proteins include those selected from bacterial or endogenous transport proteins that transport the desired glycoside intermediate. For example, the transporter may be from the host species or another bacterial or yeast species, and may be engineered to adjust its affinity for particular glycoside intermediates or products. In several modalities, the method results in at least 40% conversion, or at least 50% conversion, or at least 75% conversion of the glycoside intermediates into RCLZbn / Lznza / YiAi the desired product (e.g., conversion of stevioside, steviolbioside, and RebA to RebD, RebM, and / or other highly glycosylated rebaudiosides). In the production of RebM, the product profile may strongly favor RebM over RebD. The method can be carried out by batch fermentation, fed-batch fermentation, continuous fermentation, or semi-continuous fermentation. For example, in some embodiments, the method is carried out by batch fermentation with incubation times of less than approximately 72 hours, or in some embodiments, less than approximately 48 hours or less than approximately 24 hours. In some aspects, the invention provides methods for preparing a product comprising an advanced steviol glycoside, such as RebM. The method comprises incorporating the target steviol glycoside into a product, such as a food, beverage, oral care product, sweetener, flavoring agent, or other product. In some aspects, the invention provides methods for preparing a sweetening composition comprising a plurality of high-intensity sweeteners, such as two or more of a steviol glycoside, a mogroside, sucralose, aspartame, neotame, Advantame, acesulfame potassium, saccharin, cyclamate, neohesperidin dihydrochalcone, gnetifolin E, and / or piceatannol 4'-O-pD-glucopyranoside. Other aspects and features of the invention will become evident from the detailed description below. DESCRIPTION OF THE FIGURES FIGURE 1. Structure of RebM. The steviol scaffold (a diterpenoid) is shown in an inset. RebM contains six glycosylations: (1) a C13 O-glycosylation, (2) a C13 1-2' glycosylation, (3) a C13 1-3' glycosylation, (4) a C19 O-glycosylation, (5) a C19 1-2' glycosylation, and (6) a C19 1-3' glycosylation. FIGURE 2. Glycosylation products produced by the action of UGT enzymes on steviol and steviol glycoside intermediates. FIGURE 3. Glycosylated products produced by the action of UGT enzymes on rhamnose-containing steviol glycoside intermediates. FIGURE 4. Conversion of steviolbioside to RebM. For five major compounds in the RebM pathway, compound concentrations are shown relative to the total concentration of these compounds in a control strain lacking the UGT enzymes. Strains were fed 0.2 mM steviolbioside and incubated with substrate for 48 hours before extraction. Strains 3 and 4 show no conversion of steviolbioside, while strain 5 shows substantial conversion to RebD and RebM. FIGURE 5. Conversion of stevioside and RebA to RebM. The bioconversion of two commercially available intermediates found in the leaf extract, stevioside and RebA, to RebM is shown using strains 1 to 5. The bioconversion of the steviol core is also shown. Values ​​are expressed as a percentage of the total steviol glycoside composition. The conversion of steviol to RebM by all strains (RCLZbn / Lznza / YiAi) shows that all strains are capable of forming RebM. As was the case with steviolbioside, both stevioside and RebA are converted to RebM, but only in strain 5. For the conversion of stevioside and RebA, the RebD:RebM ratio strongly favors RebM (1:20). FIGURE 6. Bioconversion of steviol glycosides in 96-well plates. For five major compounds in the RebM pathway, compound concentrations are shown with or without expressed UGT enzymes. Strains were fed 0.5 g / L of stevioside / RebA leaf extract and incubated with substrate for 48 hours before sampling. FIGURE 7. Bioconversion of a stevioside / RebA leaf extract at bioreactor scale. For five major compounds in the RebM pathway, compound concentrations in the medium were sampled at various time points during fermentation. The strains were fed 2 g / L of the stevioside / RebA leaf extract and incubated with substrate for a total of 30 hours. FIGURE 8. Genetic modifications to increase the flux of native E. coli to UDP-glucose. Metabolites: G6P, glucose-6-phosphate; G1P, glucose-1-phosphate; 6PGL, 6-phosphoglucono-1,5-lactone; 6PG, 6-phosphogluconate; R5P, ribose-5-phosphate; PRPP, 5-phosphoribosyl-1-pyrophosphate; UMP, uridine monophosphate; UDP, uridine diphosphate; UTP, uridine triphosphate; UDP-Glu, UDP-glucose; UDP-Gal, UDP-galactose; UDP-GIcNAc, UDP-A / β-acetylglucosamine; CTP, cytidine triphosphate; dUDP, deoxyuridine diphosphate; RebA, rebaudioside A; RebM, rebaudioside M. Enzymes: 1. glk, glucokinase; 2. pgi, glucose-6-phosphate isomerase; 3. zwf, glucose-6-phosphate-1-dehydrogenase; 4. pg!, 6-phosphogluconolactonase; 5. edd, phosphogluconate dehydratase; 6. gnd, 6-phosphogluconate dehydrogenase; 7. rpiA, ribose-5-phosphate isomerase or rpe, ribose-5-phosphate 3-epimerase; 8. prs, ribose-phosphate diphosphokinase; 9. pyrEF, orotate phosphoribosyltransferase and orotidine-5'-phosphate decarboxylase; 10. pyrK, uridylate kinase; 11.ndk, UDP kinase; 12. murG, N-acetylglucosaminyl transferase; 13. nrdABEF, ribonucleoside diphosphate reductase; 14. pyrG, CTP synthase; 15. pgm, phosphoglucomutase; 16. galU, UDP-glucose pyrophosphorylase; 17. galETKM, UDP-glucose-4-epimerase / galactose-1-phosphate uridyltransferase / galactokinase / galactose-1-epimerase; 18. ushA, 5'-nucleotidase / UDP-sugar hydrolase; 19. UGT, UDP-glycosyltransferase. The scheme illustrates the sequential genomic modifications leading from strain 1 to strain 5. Strain 1 is a base strain of E. coli. Strain 2 has a deletion of 17 and strain 3 has an additional deletion of 18. Strain 4 was constructed from strain 3 by removing 2. Strain 5 was constructed from strain 4 by overexpressing 15 and 16. DETAILED DESCRIPTION OF THE INVENTION In various aspects and embodiments, the invention provides microbial cells and methods for producing advanced glycosylation products from lower glycosylated intermediates. In various embodiments, the microbial cell expresses one or more UDP-dependent glycosyltransferase enzymes (e.g., intracellularly) for the glycosylation of the intermediates. In various embodiments, RCLZbn / Lznza / YiAi The microbial cell has one or more genetic modifications that increase the availability of UDP-glucose. When the microbial strain is incubated with the glycoside intermediates (e.g., from a plant extract or fraction thereof), these glycoside intermediates become available to the cell for further glycosylation by intracellularly expressed glycosyltransferase enzymes. In some embodiments, the advanced glycosylation products can be recovered from the medium, or in some embodiments, they are recovered from both the medium and the microbial cells.Glycoside intermediates can be any glycosylated secondary metabolite, such as those found naturally in plant extracts, including glycosylated terpenoids, glycosylated flavonoids, glycosylated cannabinoids, glycosylated polyketides, glycosylated stilbenoids, and glycosylated polyphenols, among others. In some embodiments, the glycoside intermediate is a glycosylated terpenoid, such as mogroside or steviol glycoside. In some embodiments, the glycoside intermediate has one, two, three, four, or five glycosyl groups, including glycosyl groups selected from glycosyl, galactosyl, mannosyl, xylosyl, and rhamnosyl groups, among others. In several embodiments, the glycosylated product has at least four glycosyl groups, or at least five glycosyl groups, or at least six glycosyl groups. In other embodiments, the product has seven, eight, nine, or more glycosyl groups.In some forms, the biosynthesis of the product involves at least two glycosylations of the intermediate by the microbial cell. In some formulations, the glycoside intermediates are derived from stevia leaf extract. For example, steviol glycosides with five, six, or more glycosylations (such as RebD or RebM) can be biosynthesized from steviol glycoside intermediates such as stevioside, steviolbioside, rebaudioside A, dulcoside A, dulcoside B, rebaudioside C, and rebaudioside F, among others. The microbial cell expresses one or more UDP-dependent glycosyltransferase enzymes for the glycosylation of these lower-value precursors. When the microbial strain is incubated with a stevia leaf extract or a fraction thereof comprising steviol glycoside intermediates, these intermediates are available for further glycosylation to RebD or RebM or another advanced glycosylation product (e.g., Rebl), which can be recovered from the medium and / or microbial cells. In several forms, UDP-dependent glycosyltransferase enzymes are expressed intracellularly, meaning they lack membrane translocation or secretion domains or peptides. Therefore, UGT enzyme expression occurs in the cytoplasm, and these enzymes are not targeted out of the cell via a secretion or transport signal. In several forms, UGT enzymes also lack membrane anchoring domains. That is, in several forms, UGT enzymes do not possess a transmembrane domain. In some formulations, the process uses advanced intermediates in stevia leaf extract, namely steviol (the aglycone intermediate) and steviol glycosides with one to five glycosylations, which are available to the microbial cell for further glycosylation. These advanced intermediates from stevia leaf extract are readily available from the RpLZbn / Lznza / YiAi existing industrial extraction of steviol glycosides. As shown in Table 2, the leaf extract may contain mainly the intermediates of the stevioside and rebaudioside A (RebA) pathways. In several embodiments, the stevia leaf extract is an extract of steviol glycosides. In some embodiments, the extract comprises one or more of stevioside, steviolbioside, and rebaudioside A as prominent components. In some embodiments, the extract comprises one or more of dulcoside A, dulcoside B, RebC, and / or RebF as prominent components. A prominent component generally constitutes at least about 10% of the steviol glycosides in the extract or fraction thereof, but in some embodiments, it may constitute at least about 20%, or at least about 25%, or at least about 30% of the steviol glycosides in the extract or fraction thereof. RebM is illustrated in FIGURE 1. RebM contains six glycosylations of a steviol core: (1) a C13 O-glycosylation, (2) a C13 1-2' glycosylation, (3) a C13 1-3' glycosylation, (4) an O19 O-glycosylation, (5) an O19 1-2' glycosylation, and (6) a C19 1-3' glycosylation. Although several glycosylation products are possible (FIGURE 2), RebM can be synthesized from steviol by the action of four UGT enzymes, and each of the UGT glycosylation enzymes is capable of acting on several substrates. For example, both UGT91D2 and OsUGT1-2 are 1-2' glycosylating enzymes that can produce steviolbioside from steviolmonoside (by action at O13), as well as RebD from RebA (by action at O19). Furthermore, UGT76G1 is a 1-3' glycosylating enzyme that can produce RebA from stevioside (by action at O13), as well as RebM from RebD (by action at O19). UGT enzymes for the glycosylation of steviol and steviol glycosides (including for the biosynthesis of RebM) are described in US 2017 / 0332673, which is incorporated herein by reference in its entirety. Example UGT enzymes are listed in Table 1 below (the referenced patent applications are incorporated herein by reference in their entirety): Table 1: Example of UGT enzymes Glycosylation Type Enzyme Gene ID Protein ID Description C13 SrUGT85C2 AY345978.1 AAR06916.1 MbUGTC13 US 2017 / 0332673 C19 SrUGT74G1 AY345982.1 AAR06920.1 MbUGTc19 - - US 2017 / 0332673 MbUGTc19-2 US 2017 / 0332673 1-2' SrUGT91D1 AY345980.1 AAR06918.1 SrUGT91D2 ACE87855.1 ACE87855.1 SrUGT91D2e - - US 2011 / 038967 OsUGTI-2 NM_001057542.1 NP_001051007.2 WO 2013 / 022989 1-3' SrUGT76G1 FB917645.1 CAX02464.1 MbUGT1-3 US 2017 / 0332673 76G1_L200A US 2017 / 0332673 MbUGT1-3_1 US 62 / 866.148 MbUGT1-3_1.5 This disclosure MbUGT1-3_2 US 62 / 866.148 The amino acid sequences for example UGT enzymes are provided in this disclosure as SEQ IDs 1-17. SEQ ID 1 is Stevia rebaudiana UGT85C2. SEQ ID 13 is SrUGT85C2 with a P215T substitution and an Ala insertion at the 2aposition to increase stability. SEQ ID 2 is Stevia rebaudiana UGT74G1 (with an Ala insertion at the 2aposition). SEQ IDs 8 and 12 are circular permutants based on SrUGT74G1 (MbUGTC19 and MbUGTC19-2). SEQ ID 3 is Stevia rebaudiana UGT76G1. Circular permutants with 1-3' glycosylating activity are described as SEQ ID NO: 10 (MbUGT1-3) and SEQ ID NO: 15, 16, and 17 (MbUGT1-3_1, MbUGT1-3_1.5, and MbUGT1-3_2, respectively). UGT76G1 with an L200A substitution and Ala at position 2 is described as SEQ ID NO: 14. Stevia rebaudiana UGT91D1, UGT91D2, and UGT91D2e are described as SEQ ID NO: 4, 5, and 6. Oryza sativa UGT1-2 is described as SEQ ID NO: 7. MbUGT1,2 and MbUGT1,2.2 are circular permutant enzymes with 1-2' glycosylating activity (SEQ ID NO: 9 and 11). Additional UGT enzymes are provided, such as SEQ ID NO: 18 to 46, from the species Siraitia grosvenor (monk fruit), Momordica charantia (bitter melon), Cucumis sativa (cucumber), Cucurbita maxima (pumpkin), Cucurbita moschata (squash), Prunus persica (peach), Theobroma cacao (cacao), Corchorus capsularis (white jute), Ziziphus jujube (red date), Vitis vinifera (grapevine), Juglans regia (pecan), Hevea brasiliensis (rubber tree), Manihot esculenta (cassava), Cephalotus follicularis (pitcher plant), and Coffea arabica (coffee). UGT enzymes can be selected and, optionally, engineered based on the desired product and available intermediate. In several forms, the microbial cell expresses at least one, at least two, at least three, or at least four UGT enzymes. In some forms, the UGT enzymes glycosylate a steviol glycoside substrate, including those listed in Table 2. In some forms, the microbial cell expresses four UGT enzymes that glycosylate a steviol glycoside intermediate, such as a UGT 13-0 glycosylating enzyme, a UGT 19-0 glycosylating enzyme, a UGT 1-2' glycosylating enzyme, and a UGT 1-3' glycosylating enzyme. The action of these general classes of UGT enzymes on glycosylated intermediates of the stevia leaf is illustrated in Figures 2 and 3. In several forms, the microbial cell expresses a 1-3' glycosylating UGT enzyme. For example, the 1-3' glycosylating UGT enzyme can be selected from SrUGT76G1, MbUGT1-3_1, MbUGT1-3_1.5 and MbUGT1-3_2, and derivatives thereof. In these and other modalities, the microbial cell expresses a 1-2' glycosylating UGT enzyme. For example, the 1-2' glycosylating UGT enzyme can be selected from SrUGT91D2, SrUGT91D1, SrUGT91D2e, OsUGT1-2, MbUGTI,2, MbUGTI,2.2, and derivatives thereof. In these or other modalities, the microbial cell expresses a C13 UGT O-glycosylated enzyme. For example, the C13 UGT O-glycosylated enzyme can be selected from SrUGT85C2 and its derivatives (e.g., MbUGTC13). In these or other modalities, the microbial cell expresses a C19 O-glycosylated UGT enzyme. For example, the C19 O-glycosylated enzyme can be selected from SrUGT74G1, MbUGTc19 and derivatives thereof (e.g., MbUGTc19-2). In these and other forms, the microbial cell expresses a 1-3' UGT glycosylating enzyme and a 1-2' UGT glycosylating enzyme. In some forms, the microbial cell expresses a 1-3' UGT glycosylating enzyme, a 1-2' UGT glycosylating enzyme, and a UGT O-glycosylating enzyme. In some forms, the microbial cell expresses a 1-3' UGT glycosylating enzyme, a 1-2' UGT glycosylating enzyme, a C19 UGT O-glycosylating enzyme, and a C19 UGT O-glycosylating enzyme. In some embodiments, the microbial cell expresses a 13' glycosylating UGT enzyme, a 1-2' glycosylating UGT enzyme, a 019 O-glycosylating UGT enzyme, and a 013 O-glycosylating UGT enzyme. In some embodiments, the microbial cell expresses SrUGT85C2 or a derivative thereof (e.g., MbUGTC13), MbUGTI,2.2 or a derivative thereof, SrUGT74G1 or a derivative thereof (e.g., MbUGTc19 or MbUGTc19-2), and SrUGT76G1 or MbUGT1-3_1 or a derivative thereof (e.g., 76G1_L200A, MbUGT1-3_1.5, or MbUGT1-3_2).Beyond theory, genetically modified UGT enzymes can provide a greater carbon flux to RebM (as well as higher glycosylation products), particularly due to substrate-binding pockets that are better able to accommodate larger substrates without a substantial loss of activity at lower glycosylated intermediates. In these configurations, UGT enzymes (such as 1-3' and 1-2' glycosylating UGT enzymes) can exhibit increased activity rates (e.g., substrate binding and turnover rates) with more highly glycosylated steviol substrates such as RebA or RebD. UGT enzyme derivatives generally comprise an amino acid sequence that has at least approximately 50% identity, at least approximately 60% identity, at least approximately 70% identity, at least approximately 80% identity, at least approximately 85% identity, or at least approximately 90% identity, or at least approximately 95% identity, or at least 96%, 97%, 98%, or 99% identity with one or more of the SEQ IDs listed 1 to 46. In some embodiments, the derivative has 1 to 20, 1 to 10, or 1 to 5 amino acid modifications (independently selected from amino acid substitutions, insertions, and deletions) with respect to one of the SEQ IDs listed 1 to 46. In some embodiments, for example, with respect to the production of RebM and other glycosides of RCLZfrOI ZOZ-1 / YILI advanced steviol (e.g., having at least 5 glycosyl groups) from steviol glycoside intermediates, the derivatives of these UGT enzymes comprise an amino acid sequence having at least about 50% identity, at least about 60% identity, at least about 70% identity, at least about 80% identity, at least about 85% identity, or at least about 90% identity, or at least about 95% identity, or at least 96%, 97%, 98% or 99% identity with one or more of the SEQ ID NO: 1 to 17. In some embodiments, the derivative has 1 to 20 or 1 to 10, or 1 to 5 amino acid modifications (selected independently of substitutions, insertions and amino acid deletions), with respect to one of the SEQ ID NO: 1 to 17. In some forms, the microbial cell expresses a UGT 1-3' enzyme comprising an amino acid sequence that is at least about 75% identical to the amino acid sequence of SEQ ID NO:15, SEQ ID NO:16 or SEQ ID NO:17. In some embodiments, UGT 1-3' comprises an amino acid sequence that is at least approximately 80% identical to SEQ ID NO: 15, 16, or 17. In some embodiments, the amino acid sequence of the UGT 1-3' enzyme is at least approximately 85% identical to SEQ ID NO: 15, 16, or 17, or at least approximately 90% identical to SEQ ID NO: 15, 16, or 17, or at least approximately 95% identical to SEQ ID NO: 15, 16, or 17, or at least approximately 98% identical to SEQ ID NO: 15, 16, or 17. In some embodiments, the amino acid sequence of the UGT 1-3' enzyme comprises the amino acid of SEQ ID NO: 15, 16, or 17. For example, the amino acid sequence may have 1 to 20 amino acid modifications selected independently of substitutions, deletions, and insertions, with respect to the amino acid sequence of SEQ ID NO: 15, 16, or 17. In some modalities, the amino acid sequence has 1 to 10 amino acid modifications (e.g., 1 to 5) selected independently of substitutions, deletions, and insertions, with respect to the amino acid sequence of SEQ ID NO: 15, 16, or 17. The amino acid modifications in the amino acid sequence of SEQ ID NO: 15, 16, or 17 can be guided by available enzyme structures and the construction of homology models. Example structures are described in, for example, Li, et al., Crystal Structure of Medicao truncatula UGT85H2-insihts into the Structural Basis of a Multifunctional (so) Flavonoid Glvcosvltransferase, J. of Mol. Blol. 370.5 (2007): 951-963.Publicly available crystal structures (e.g., PDB entry: 2PQ6) can be used to report amino acid modifications. For example, one or more amino acid modifications can be made at or near the active site to enhance substrate binding and / or to improve the reaction geometries of these substrates with catalytic side chains. In some forms, the UGT 1-3' enzyme comprises an amino acid substitution at positions corresponding to positions 29, 200, 357, and 414 of SEQ ID NO:3 (Stevia rebaudiana UGT76G1). The substitutions at these positions, which are included in the RCLZbn / Lznza / YiAi enzymes from SEQ ID NO: 15 and 17 (positions 183, 354, 54, and 111, respectively, in SEQ ID NO: 15) can provide dramatic improvements in activity. In some modalities, the identity of the amino acids at the positions corresponding to positions 183, 354, 54, and 111 of SEQ ID NO: 15 allows for further modifications at other positions.For example, in some embodiments, the UGT 1-3' enzyme comprises an amino acid sequence that is at least about 60% identical to the amino acid sequence of SEQ ID NO: 15 or 17, wherein the UGT enzyme comprises: a glycine (G) or threonine (T) at the position corresponding to position 54 of SEQ ID NO: 15; a leucine (L) or isoleucine (I) at the position corresponding to position 111 of SEQ ID NO: 15; a methionine (M) or leucine (L) at the position corresponding to position 183 of SEQ ID NO: 15; and an alanine (A) or glycine (G) or serine (S) in the position corresponding to position 354 of SEQ ID NO: 15. In some embodiments, the UGT 1-3' enzyme comprises a methionine (M) in the position corresponding to position 183 of SEQ ID NO: 15. In some embodiments, the UGT 1-3' enzyme comprises a glycine (G) in the position corresponding to position 54 of SEQ ID NO: 15.In some embodiments, the UGT 1-3' enzyme comprises a leucine (L) at the position corresponding to position 111 of SEQ ID NO: 15. In some embodiments, the UGT 1-3' has two or three of a methionine (M) at the position corresponding to position 183 of SEQ ID NO: 15, a glycine (G) at the position corresponding to position 54 of SEQ ID NO: 15, and a leucine (L) at the position corresponding to position 111 of SEQ ID NO: 15. These modifications may provide substantial improvements to the enzyme activity. In some embodiments, the UGT 1-3' enzyme comprises an insertion of 5 to approximately 15 amino acids, such as 6 to 12 amino acids, or approximately 6 or approximately 11 amino acids, after the position corresponding to position 155 of SEQ ID NO:15. In some embodiments, the insertion is a flexible, hydrophilic sequence consisting predominantly of glycine and serine residues. In some embodiments, the sequence is GSGGSG (SEQ ID NO:47) or GSGGSGGSG (SEQ ID NO:48). In several configurations, the UGT 1-3' enzyme shows improved conversion of stevioside to Reb and improved conversion of RebD to RebM, compared to UGT76G1L200A (SEQ ID NO: 14). This improved conversion is shown in a bioconversion assay where stevioside or RebD substrate is fed to microbial cells expressing the UGT 1-3' enzyme. The improved conversion can be demonstrated in reactions with cells containing recombinantly expressed UGT 1-3', or in vitro reactions with purified or partially purified UGT 1-3'. Changes in the amino acid sequence of an enzyme may alter its activity or have no measurable effect. Silent changes with no measurable effect are often conservative substitutions and small insertions or deletions on solvent-exposed surfaces located far from active sites and substrate-binding sites. In contrast, enzyme activity is more likely to be affected by non-conservative substitutions, large insertions, or RpLZbn / Lznza / YiAi deletions and changes within active sites, substrate-binding sites, and hidden positions important for protein conformation or folding. Changes that alter enzyme activity can increase or decrease the reaction rate or increase or decrease the affinity or specificity for a particular substrate. For example, changes that increase the size of a substrate-binding site can allow an enzyme to act on larger substrates, and changes that place a catalytic amino acid side chain closer to a target site on a substrate can increase the enzyme rate. Knowledge of an enzyme's three-dimensional structure and the location of relevant active sites, substrate-binding sites, and other interaction sites can facilitate the rational design of derivatives and provide a mechanistic view of the phenotype of specific changes. Plant UGTs share a highly conserved secondary and tertiary structure, while exhibiting relatively low amino acid sequence identity. (Osmani et al., Substrate specificity of plant UDP-dependent β-cosyltransferases predicted from crystal structures and homology modeling, Phytochemistry 70 (2009) 325-347). The sugar-acceptor and sugar-donating substrates of UGTs are housed in a cleft formed between the N- and C-terminus domains.Several regions of the primary sequence contribute to the formation of the substrate-binding pocket, including structurally conserved domains, as well as loop regions that differ both in their amino acid sequence and sequence length. The construction of UGT derivatives can be guided by homology models compared to known structures. For example, based on crystal structure analysis of SrUGT76G1_L200A and amino acid sequence alignment of SrUGT76G1 to MbUGT3-1_1, the steviol core of stevioside is predicted to be close (within 4 Λ) to the following residues of MbUGT3-1_1: I244, L280, W351, A354, I357, M362, and T438. Furthermore, the C19 1-2 glycosylation is predicted to be close (within 4 Λ) to T438. The steviol core of RebD is predicted to be close (within 4 Å) to the following hydrophobic side chains of MbUGT31_1: L239, M242, I244, L280, I353, A354, and I357. The 1-2' C13 glycosylation is predicted to be close (within 4 Å) to the following hydrogen-bonding side chains of MbUGT3-1_1: S301 and D77.The positioning and amino acid content of the V341-Q352 and K355-A367 helices of MbUGT3-1_1 may be important for catalysis, since the mutation corresponding to L200A is in a loop between these helices. The L76 and / or D77 positions of MbUGT3-1_1 may interact with the C13 glycosylation of stevioside. The amino acid sequence of one or more UGT enzymes may optionally include an inserted or substituted alanine at position 2 to decrease turnover in the cell. In several embodiments, one or more UGT enzymes comprise an inserted or substituted alanine amino acid residue at position 2 to provide additional stability in vivo. The identity of amino acid sequences, that is, the percentage of sequence identity, can be determined by sequence alignments. Such alignments are RCLZbn / Lznza / YiAi can be analyzed using several known algorithms, such as the one described by Karlin and Altschul (Karlin & Altschul (1993) Proc. Nati. Acad. Sci. USA 90: 5873-5877), with hmmalign (HMMER package), or with the CLUSTAL algorithm (Thompson, JD, Higgins, DG & Gibson, TJ (1994) Nucleic Acids Res. 22, 4673-80). The degree of sequence identity (sequence match) can be calculated using, for example, BLAST, BLAT, or BlastZ (or BlastX). A similar algorithm is incorporated into the BLASTN and BLASTP programs of Altschul et al. (1990) J. Mol. Biol. 215: 403-410. BLAST protein alignments can be performed using the BLASTP program, score = 50, word length = 3. To obtain alignments with gaps for comparative purposes, Gapped BLAST is used as described in Altschul et al. (1997) Nucleic Acids Res. 25: 3389-3402. When using the BLAST and Gapped BLAST programs, the default parameters of the respective programs are used. UGT enzymes can be integrated into the microbial cell chromosome or, alternatively, expressed extrachromosomally. For example, UGT enzymes can be expressed from a bacterial artificial chromosome (BAC) or a yeast artificial chromosome (YAC). The expression of UGT enzymes can be tuned for optimal activity using, for example, gene modules (e.g., operons) or independent expression of UGT enzymes. For instance, the expression of genes or operons can be regulated by selecting promoters, such as inducible or constitutive promoters, with varying strengths (e.g., strong, intermediate, or weak). Examples of promoters with different strengths include Trc, T5, and T7. Additionally, the expression of genes or operons can be regulated by manipulating the number of copies of the gene or operon in the cell. In some configurations, the cell expresses only one copy of each UGT enzyme. In others, the expression of genes or operons can be regulated by manipulating the order of genes within a module, where genes transcribed first are generally expressed at a higher level.In some modalities, the expression of genes or operons is regulated through the integration of one or more genes or operons into the chromosome. Optimization of UGT expression can also be achieved by selecting appropriate promoters and ribosome binding sites. In some approaches, this may involve selecting high-copy-count plasmids, or single-copy-count, low-copy-count, or medium-copy-count plasmids. The transcription termination stage can also be targeted for gene expression regulation through the insertion or removal of structures such as stem-loops. Whole-cell conversion requires that the substrate (e.g., glycoside intermediates) be available to the cell for glycosylation by expressed enzymes (e.g., intracellularly expressed enzymes), and preferably the product can be extracted from the extracellular environment. Whole-cell systems have advantages because the cell provides regeneration of the UDP-glucose cofactor. This contrasts with processes that use enzymes from cell lysis or secretion outside the cell, which require an exogenous supply of UDP-glucose. RCLZbn / Lznza / YiAi is a UDP-glucose precursor, a UDP-glucose regeneration mechanism, or a UDP-glucose regeneration enzyme system. In embodiments of the present invention, catalysis (glycosylation) is carried out using live microbial cells, and the recycling of the UDP-glucose cofactor takes place using cellular metabolism without requiring enzyme feeding or substrate feeding for UDP-glucose regeneration. US patent 2017 / 0332673 describes E. coli strains that overexpress MEP pathway enzymes, along with a downstream steviol biosynthesis pathway, and UGT enzymes to drive RebM production from glucose. However, these strains do not biocatalyze steviol glycoside intermediates fed to RebM, which may be due, in part, to the host cell's inability to access the steviol glycoside substrate. According to this disclosure, genetic modifications of the microbial cell allow the glycosylated intermediates to be available for further glycosylation in a whole-cell system. Furthermore, the product can be recovered from extracellular media, facilitating subsequent purification and processing. In some cases, the microbial cell has one or more genetic modifications that increase the availability of UDP-glucose. In some cases, and not just theoretically, these modifications can also stress the cell for glucose availability, leading to increased expression of endogenous transporters to import steviol glycosides into the cell. Wild-type UDP-glucose levels in exponentially growing E. coli are around 2.5 mM (Bennett BD, Kimball EH, Gao M, Osterhout R, Van dien SJ, Rabinowitz JD. Absolute metabolite concentrations and implied enzyme active site occupancy in Escherichia coli. Nat Chem Biol. 2009;5(8):593-9). In some modalities, the genetic modifications of the host cell are designed to increase UDP-glucose, for example, to at least 5 mM, or at least 10 mM, in exponentially growing cells (e.g., those that do not have recombinant expression of UGT enzymes).In some forms, the microbial cell has a deletion, inactivation, or reduced activity or expression of a gene encoding an enzyme that consumes UDP-glucose. For example, the microbial cell may have a deletion, inactivation, or reduced activity of ushA (UDP-sugar hydrolase) and / or one or more of galE, gaIT, galK, and galM (which are responsible for the biosynthesis of UDP-galactose from UDP-glucose), or their ortholog in the microbial species. In some forms, the galETKM genes are inactivated, deleted, or substantially reduced in expression. Alternatively or additionally, the microbial cell has a deletion, inactivation, or reduced activity or expression of E. coli otsA (trehalose-6-phosphate synthase), or an ortholog of this in the microbial species. Alternatively or additionally, the microbial cell has a deletion, inactivation, or reduced activity or expression of E. coli ugd (UDP-glucose 6-dehydrogenase), or an ortholog of this in the microbial species.Reducing or eliminating the activity of otsA and ugd can eliminate or reduce the sinks of UDP-glucose to trehalose or UDP-glucuronidate, respectively. RCLZbn / Lzzza / YiAi Other UDP-glucose sinks that can be reduced or eliminated include eliminating or reducing the activity or expression of genes responsible for lipid glycosylation and LPS biosynthesis, and genes responsible for glycosylating undecaprenyl diphosphate (UPP). Genes involved in lipid glycosylation or LPS biosynthesis include E. coli waaG (lipopolysaccharide glucosyltransferase 1), E. coli waaO (UDP-D-glucose:(glucosyl)LPS α-1,3-glucosyltransferase), and E. coli waaJ (UDP-glucose:(glucosyl)LPS α-1,2-glucosyltransferase). The genes responsible for undecaprenyl-diphosphate glycosylation (UPP) include E. coli yfdG (putative bactoprenol-bound glucose translocase), E. coli yfdH (bactoprenol glucosyltransferase), E. coli yfdl (serotype-specific glucosyltransferase), and E. coli wcaJ (undecaprenylphosphate glucose phosphotransferase).Deletion, inactivation, or reduction of the activity or expression of one or more of these gene products (or corresponding orthologs in the microbial cell) can increase the availability of UDP-glucose. In these and other modalities, the microbial cell has a deletion, inactivation, or reduced activity or expression of a gene that encodes an enzyme that consumes a UDP-glucose precursor. For example, in some modalities, the microbial cell has a deletion, inactivation, or reduced activity or expression of pgi (glucose-6-phosphate isomerase), or its ortholog in the microbial species of the host cell. In these or other modalities, the cell exhibits overexpression or increased activity of one or more genes encoding an enzyme involved in the conversion of glucose-6-phosphate to UDP-glucose. For example, pgm (phosphoglucomutase) and / or galU (UTP-glucose-1-phosphate uridyltransferase) (or their orthologs or derivatives) may be overexpressed or modified to increase enzyme productivity. Alternatively or additionally, E. coli ycjU (β-phosphoglucomutase), which converts glucose-6-phosphate to glucose-1-phosphate, and Bifidobacterium bifidum ugpA, which converts glucose-1-phosphate to UDP, or an ortholog or derivative of these enzymes, may be overexpressed or modified to increase enzyme productivity. Alternatively or additionally, the microbial cell has one or more genetic modifications that increase the flux to the pentose phosphate pathway (PPP), such as an overexpression or increased activity of E. coli zwf (or a genetically modified homolog or derivative thereof), which is an NADP+-dependent glucose-6-phosphate dehydrogenase. Alternatively or additionally, the microbial cell has one or more genetic modifications that increase glucose transport. Such modifications include increased expression or activity of E. coli galP (galactose: H+ symporter) and E. coli glk (glucokinase), or alternatively Zymomonas mobilis glf and E. coli glk, or genetically modified homologs, orthologs, or derivatives of these genes. Alternatively or additionally, the microbial cell has one or more genetic modifications that increase UTP production and recycling. Such modifications include increased expression or activity of E. coli pyrH (UMP kinase), E. coli cmk (cytidylate kinase), E. coli adk (adenylate kinase), or E. coli ndk (nucleoside diphosphate kinase), or their homologs, orthologs, or derivatives. RCLZbn / Lznza / YiAi genetically modified versions of these enzymes. Alternatively or additionally, the microbial cell has one or more genetic modifications that increase UDP production. Such modifications include the overexpression or increased activity of one or more of E. coli upp (uracil phosphoribosyltransferase), E. coli dctA (C4 dicarboxylate / orotate: H+ symporter), E. coli pyrE (orotate phosphoribosyltransferase), and E. coli pyrF (orotidine-5'-phosphate decarboxylase), including homologs, orthologs, or genetically modified derivatives of these. For example, in some forms, the microbial cell overexpresses or has increased activity of upp, pyrH, and cmk, or their homolog or genetically modified derivative. Alternatively, the microbial cell overexpresses or has increased activity of dctA, pyr, pyrH, and cmk, or their homolog or genetically modified derivative. Alternatively or additionally, the microbial cell may have one or more genetic modifications that eliminate or reduce the regulation of glucose uptake. For example, the microbial cell may have a deletion, inactivation, or reduced expression of sgrS, which is a small regulatory RNA in E. coli. Alternatively or additionally, the microbial cell may have one or more genetic modifications that reduce glucose-1-phosphate dephosphorylation. Example modifications include the deletion, inactivation, or reduced expression or activity of one or more of E. coli agp (glucose-1-phosphatase), E. coli yihX (αD-glucose-1-phosphate phosphatase), E. coli ybiV (sugar phosphatase), E. coli yidA (sugar phosphatase), E. coli yigL (phosphosugar phosphatase), and E. coli phoA (alkaline phosphatase), or an ortholog of these in the microbial cell. Alternatively or additionally, the microbial cell may have one or more genetic modifications that reduce the conversion of glucose-1-phosphate to TDP-glucose. Example modifications include the deletion, inactivation, or reduced expression or activity of one or more of E. coli rffH (dTDP-glucose pyrophosphorylase) and E. coli rfbA (dTDP-glucose pyrophosphorylase), or an ortholog of these in the microbial cell. Alternatively or additionally, the microbial cell may have one or more genetic modifications that reduce the conversion of glucose-1-phosphate to ADP-glucose. Example modifications include deletion, inactivation, or reduced expression or activity of E. coli gigC (glucose-1-phosphate adenylyltransferase), or an ortholog of this enzyme in the microbial cell. In some modalities, the microbial cell is a bacterial cell comprising the genetic modifications: ushA and galETKM are deleted, inactivated, or their expression is reduced; pgi is deleted, inactivated, or its expression is reduced; and pgm and galU are overexpressed or complemented. In some approaches, endogenous genes are edited to inactivate or reduce enzyme activity by changing the amino acid sequence of the encoded protein or to reduce expression by editing expression control sequences. Editing can modify endogenous promoters, ribosomal binding sequences, or other expression control sequences, and / or in some approaches, modify trans-acting and / or cis-acting factors in regulation. RCLZbn / Lznza / YiAi gene editing. Genome editing can take place using CRISPR / Cas genome editing techniques or similar techniques employing zinc finger nucleases and TALEN. In some modalities, endogenous genes are replaced by homologous recombination. The microbial cell in several embodiments does not express a recombinant biosynthesis pathway for the production of precursors (e.g., comprising one or more plant enzymes). For example, microbial cells producing RebM (and other advanced glycation products) according to the embodiments of the invention do not express a steviol biosynthesis pathway, such as copayl synthase, kaurene synthase, kaurene oxidase, and / or kaurenoic acid hydroxylase, so that the production of RebM and other advanced glycation products depends on feeding steviol glycoside intermediates to the cell. In various embodiments, the microbial cell is a bacterium selected from Escherichia spp., Bacillus spp., Rhodobacter spp., Zymomonas spp. The Pseudomonas spp. In some embodiments, the bacterial species is selected from Escherichia coli, Bacillus subtilis, Rhodobacter capsulatus, Rhodobacter sphaeroides, Zymomonas mobilis or Pseudomonas putida. In some embodiments, the bacterial cell is an E. coli cell. In other modes, the cell is a fungal cell, such as a yeast cell, including Saccharomyces spp., Schizosaccharomyces spp., Pichia spp., Phaffia spp., Kluyveromyces spp., Candida spp., Talaromyces spp., Brettanomyces spp., Pachysolen spp., Debaryomyces spp., Yarrowia spp., and industrial polyploid yeast strains. In one mode, the yeast may be a species of Saccharomyces, Pichia, or Yarrowia, including Saccharomyces cerevisiae, Pichia pastoris, and Yarrowia hypolytica. In some modes, the yeast cell expresses one or more bacterial transporters, or derivatives thereof, that import glycoside intermediates into the cell for further glycosylation. In several modes, the microbial cell overexpresses one or more endogenous transporters (e.g., compared to a parental microbial strain), or in certain modes, it is modified to express a recombinant and / or genetically modified transport protein. In some modes, the microbial cell expresses one or more additional copies of an endogenous transport protein or a derivative thereof. For example, the expression or activity of transport proteins can be modified to increase the transport into the cell of steviol glycoside intermediates (e.g., one or more of stevioside, steviolbioside, and RebA, among others), while exporting products such as RebM and / or RebD and / or other advanced glycosylated steviol glycosides. Example transport proteins that can be overexpressed or manipulated to alter substrate activity or specificity in the microbial cell include E. coli acrAD, xylE, ascF, bglF, chbA, ptsG / crr, wzxE, rfbX, as well as orthologs or derivatives of these. Derivatives and orthologs of these proteins generally comprise amino acid sequences that share at least approximately 30%, 40%, or 50% identity. RCLZbn / Lznza / YiAi at least around 60% identity, at least around 70% identity, at least around 80% identity, at least around 85% identity, or at least around 90% identity, or at least around 95% identity, or at least 96%, 97%, 98% or 99% identity with the E. coli transport protein. acrAD is a multidrug efflux pump of E. coli described in Aires JR and Nikaido H., Aminolipids are captured from both periplasm and cytoplasm by the AcrD multidrug efflux transporter of Escherichia coli, J Bacteriol. 2005; 187(6):1923-9. In addition, a homologous structure is described in Yu, EW, et al., Structural basis of multiple drug-binding capacity of the AcrB multidrug efflux pump, Science 2003; 300(5621):976-80. xylE is a xylose transporter from E. coli with homology to glucose transporters. xylE is described in Sumiya M, et al., Molecular genetics of a receptor protein for D-xylose, encoded by the gene xvlF, in Escherichia coli, Recept Channels 1995;3(2):117-28, with structure described by Sun L, et al., Crystal structure of a bacterial homologue of glucose transporters GLUT-1-4, Nature 2012;490(7420):361-6. ascF is described by Hall BG and Xu L, Nucleotide sequence, function, activation, and evolution of the cryptic ase operon of Escherichia coli K12, Mol. Biol. Evo!. 1992;9(4):688-706. The ase operon is considered to play a role in cellobiose metabolism in E. coli. bglF is described by Schnetz K, et al., Identification of catalytic residues in the beta-glucoside permease of Escherichia coli by site-specific mutagenesis and demonstration of interdomain cross-reactivity between the beta-glucoside and glucose systems, J. Biol. Chem. 1990;265(23):1346-471. A homologous structure is described in Herzberg O., An atomic model for protein-protein phosphoryl group transfer, J. Biol. Chem. 1992;276(34):24819-23. bglF catalyzes the transport and phosphorylation of beta-glucosides. chba is described in Keyhani NO, et al., The transport / phosphorvlation of N,N'diacetylchitobiose in Escheríchia coli. Characterization of phosphor-IIB(Chb') and of a potential transition state analogue in the phosphotransfer reaction between the proteins IIA(Chb) and IIBÍChb). J. Biol. Chem. 2000; 275 (42): 33102-9; with structure described in Tang C, et al., Solution structure of enzyme llAíChitobiose) from the N.N'-diacetvIchitobiose branch of the Escheríchia coli phosohotransferase svstem. J Biol Chem 2005;280(12):11770-80. The phosphoenolpyruvate-dependent sugar phosphotransferase (sugar PTS) system is an important carbohydrate active transport system that catalyzes the phosphorylation of incoming sugar substrates concomitantly with their translocation across the cell membrane. ptsG codifica la permeasa específica de glucosa del sistema de transporte de fosfotransferasa (PTS) y se describe en Meins M, et ál., Glucose permease of Escheríchia coli. Purification of the IIGIc subunit and functional characterization of its oligomeric forms. J. Biol. Chem. 1988;263(26):12986-93. Se describe una estructura en Cai M, et ál., Solution structure of the phosphorvl transfer complex between the signal-transducing protein IIAGlucose and the cytoplasmic RCLZbn / Lznza / YiAi domain of the qlucose transporter IICBGlucose of the Escheríchia coli qlucose phosphotransferase svstem. J. Biol. Chem. 2003;278(27):25191-206. wzxE y su papel en el transporte molecular es descrito por Rick PD, et áL, Evidence that the wzxE gene of Escheríchia coli K-12 encodes a protein involved in the transbilaver movement of a trisaccharide-lipid intermedíate in the assembly of enterobacterial common antiqen. J. Biol. Chem. 2003;278(19) :16534-42. rfbx es un transportador de lipopolisacáridos descrito en Hong Y, et ál., Proqress in our understanding of wzx flippase for translocation of bacterial membrane lipid-linked oliqosaccharide. J. Bacteríol. 2018;200(1). Other transport proteins include those selected from bacterial or endogenous transport proteins that transport the desired glycoside intermediate into the cell and / or transport the desired product out of the cell. For example, the transporter may be from the host species or another bacterial or yeast species, and it may be engineered to fine-tune its affinity for particular glycoside intermediates or products. For example, the host cell may overexpress a transporter that is at least 30%, 40%, or 50% identical to an E. coli transporter.coli seleccionado de ampG, araE, araJ, bcr, cynX, emrA, emrB, emrD, emrE, emrK, emrY, entS, exuT, fsr, fucP, galP, garP, glpT, gudP, gudT, hcaT, hsrA, kgtP, lacY, IgoT, IpIT, IptA, IptB, IptC, IptD, IptE, IptF, IptG, mdfA, mdtD, mdtG, mdtH, mdtM, ​​mdtL, mhpT, msbA, nanT, narK, narU, nepl, nimT, nupG, proP, setA, setB, setC, shiA, tfaP, tolC, tsgA, uhpT, xapB, xylE, yaaU, yajR, ybjJ, ycaD, ydeA, ydeF, ydfJ, ydhC, ydhP, ydjE, ydjK, ydiM, ydiN, yebQ, ydcO, yegT, yfaV, yfcJ, ygaY, ygcE, ygcS, yhhS, yhjE, yhjX, yidT, yihN, yjhB, yynfM. Algunas modalidades, the transportador is the menos rededor of 60%, the menus of rededor of 70%, the menus of rededor of 80%, the menus of rededor of 90%, or the menos of rededor of 95% idéntico of the transportador of E. coli. In some variants, the microbial cell expresses a transport protein that is at least 50% identical to a transporter from a eukaryotic cell, such as a yeast, fungus, or plant cell. In some variants, the transport protein is an ABC family transporter, which is optionally a PDR (pleiotropic drug resistance) subclass transporter, an MDR (multidrug resistance) transporter, an MFS (major facilitator superfamily) transporter, or a SWEET family transporter (also known as the PQloop, Saliva, or MtN3 family). In other embodiments, the transport protein is from a family selected from: AAAP, SulP, LCT, APC, MOP, ZIP, MPT, VIC, CPA2, ThrE, OPT, Trk, BASS, DMT, MC, AEC, Amt, Nramp, TRP-CC, ACR3, NCS1, PÍT, ArsAB, IISP, GUP, MIT, Ctr, and CDF. In some forms, the transporter is an ABC family transport protein (also known as an ATP-binding cassette transporter), which typically includes multiple subunits, one or two of which are transmembrane proteins and one or two of which are membrane-associated ATPases. The ATPase subunits utilize the energy from the binding of ATP. RCLZbn / Lznza / YiAi hydrolyzes adenosine triphosphate (ATP) to activate the translocation of various substrates across membranes, either for substrate uptake or export. The ABC family transporter can be of any subclass, including, but not limited to: ABCA, ABCB, ABCC, ABCD, ABCE, ABCF, and ABCG. In some cases, the transport protein is a Major Facilitator Superfamily (MFS) transport protein, which are single-polypeptide secondary carriers capable of transporting small solutes in response to chemiosmotic ion gradients. Compounds transported by MFS transport proteins can include simple sugars, oligosaccharides, inositols, drugs, amino acids, nucleosides, organophosphate esters, Krebs cycle metabolites, and a wide variety of organic and inorganic anions and cations. Examples of MFS transport proteins include XylE (from E. coli), QacA (from S. aureus), Bmr (from B. subtilis), UhpT (from E. coli), LacY (from E. coli), FucP (from E. coli), and ExtU (from E. coli). In some forms, the transporter belongs to the SWEET family of transport proteins (also known as the PQ-loop, Saliva, or MtN3 family), which is a family of sugar transporters and a member of the TOG superfamily. Eukaryotic SWEET family members have seven transmembrane segments (TMS) in a 3+1+3 repeating arrangement. Examples of SWEET transport proteins include SWEET1, SWEET2, SWEET9, SWEET12, SWEET13, and SWEET14. In some forms, the transport protein is at least 50% identical to a transport protein from S. cerevisiae. In some forms, the transporter is at least 60%, 70%, 80%, 90%, or 95% identical to the S. cerevisiae transporter. Examples of S. cerevisiae transport proteins include AC1, ADP1, ANT1, AQR1, AQY3, ARN1, ARN2, ARR3, ATG22, ATP4, ATP7, ATP19, ATR1, ATX2, AUS1, AVT3, AVT5, AVT6, AVT7, AZR1, CAF16, CCH1, COT1, CRC1, CTR3, DAL4, DNF1, DNF2, DTR1, DUR3, ECM3, ECM27, ENB1, ERS1, FEN2, FLR1, FSF1, FUR4, GAP1, GET3, GEX2, and GGC1. GUP1, HOL1, HCT10, HXT3, HXT5, HXT8, HXT9, HXT11, HXT15, KHA1, ITR1, LEU5, LYP1, MCH1, MCH5, MDL2, MME1, MNR2, MPH2, MPH3, MRS2, MRS3, MTM1, MUP3, NFT1, OAC1, ODC2, OPT1, ORT1, PCA1, PDR1, PDR3, PDR5, PDR8, PDR10, PDR11, PDR12, PDR15, PDR18, PDRI, PDRI 1, PET8, PHO89, PIC2, ​​PMA2, ​​PMC1, PMR1, PRM10, PUT4, QDR1, QDR2, QDR3, RCH1,SAL1, SAM3, SBH2, SEO1, SGE1, SIT1, SLY41, SMF1, SNF3, SNQ2, SPF1, SRP101, SSU1, STE6, STL1, SUL1, TAT2, THI7, THI73, TIM8, TIM13, TOK1, TOM7, TOM70, TPN1, TPO1, TPO2, TPO3, TPO4, TRK2, UGA4, VBA3, VBA5, VCX1, VMA1, VMA3, VMA4, VMA6, VMR1, VPS73, YEA6, YHK8, YIA6, YMC1, YMD8, YOR1, YPK9, YVC1, ZRT1; YBR241C, YBR287W, YDR061W, YDR338C, YFR045W, YGL114W, YGR125W, YIL166C, YKL050C, YMR253C, YMR279C, YNL095C, YOL075C, YPR003C, y YPR011C., RCLZbn / Lznza / YiAi In some modalities, the S. cerevisiae transport protein is selected from one or more of ADP1, AQR1, ARN1, ARN2, ATR1, AUS1, AZR1, DAL4, DTR1, ENB1, FLR1, GEX2, HOL1, HXT3, HXT8, HXT11, NFT1, PDR1, PDR3, PDR5, PDR8, PDR10, PDR11, PDR12, PDR15, PDR18, QDR1, QDR2, QDR3, SEO1, SGE1, SIT1, SNQ2, SSU1, STE6, THI7, THI73, TIM8, TPN1, TPO1, TPO2, TPO3, TPO4, YHK8, YMD8, YOR1, and YVC1. In some modalities, the S. cerevisiae transport protein is selected from one or more of FLR1, PDR1, PDR3, PDR5, PDR10, PDR15, SNQ2, TPO1, and YOR1. In some forms, the carrier is at least 50%, at least 60%, at least 70%, at least 80%, or at least 90 or 95% identical to XP_013706116.1 (from Brassica napus), NP_001288941.1 (from Brassica rapa), NEC1 (from Petunia hybrida) and SWEET13 (from Triticum urartu). In several embodiments with respect to the biosynthesis of RebM, the method results in at least 40%, 50%, or 75% conversion of stevioside, steviolbioside, and RebA to RebM. In some embodiments, the ratio of RebM to RebD is at least 2:1, 4:1, 6:1, 8:1, 9:1, 10:1, 15:1, or 20:1. The method can be carried out by batch fermentation, fed-batch fermentation, continuous fermentation, or semi-continuous fermentation. For example, in some embodiments, the method is carried out by batch fermentation or fed-batch fermentation with incubation times of less than approximately 72 hours, or in some embodiments, less than approximately 48 hours or less than approximately 24 hours. Although native UGT enzymes are generally plant enzymes (which often have optimum temperatures in the range of 20-24 °C) or derived from plant enzymes, the present disclosure in some modalities allows the production of the glycosylated product with high yield in microbial cells (e.g., bacterial cells such as E. coti), with enzyme productivity at temperatures of around 24 °C or higher, such as from around 24 °C to around 37 °C, or from around 27 °C to around 37 °C, or from around 30 °C to around 37 °C. In some formulations, the growth or production phase media may contain one or more detergents in sufficient quantity to enhance cell permeability without significantly impacting growth or viability. Examples of detergents include Tween 20, Triton X-100, and SDS, among others. In some variations, the process is scalable for large-scale production. For example, in some variations, the crop size is at least around 100 L, at least around 200 L, at least around 500 L, at least around 1,000 L, or at least around 10,000 L. In several modalities, the methods also include recovering the glycosylated product from the cell culture or from the cells used. In some modalities, the culture produces at least RCLZfrO / Ί ΖΟΖ -1 / ΥΙΛΙ around 100 mg / L, or at least around 200 mg / L, or at least around 500 mg / L, or at least around 1 g / L, or at least around 2 g / L, or at least around 5 g / L, or at least around 10 g / L, or at least around 20 g / L, or at least around 30 g / L, or at least around 40 g / L, or at least around 50 g / L of the glycosylated product, which in some forms is extracted from the culture medium. In some embodiments, glycosylated products (e.g., RebM) are purified from components of the medium. Therefore, in some embodiments, the methods involve separating the growth medium from the host cells and isolating the desired glycosylation products (e.g., RebM) from the growth medium. In some embodiments, a product such as RebM is further extracted from the cellular material. In some respects, the invention provides methods for preparing a product comprising a glycosylated product, such as RebM. The method comprises incorporating the target steviol glycoside (produced according to this description) into a product, such as a food, beverage, oral care product, sweetener, flavoring agent, or other product. The purified steviol glycosides, prepared according to the present invention, can be used in a variety of products, including, but not limited to, foods, beverages, texturizers (e.g., starches, fibers, gums, fats and fat mimetics, and emulsifiers), pharmaceutical compositions, tobacco products, nutraceutical compositions, oral hygiene compositions, and cosmetic compositions.Non-exhaustive examples of flavors for which RebM can be used in combination include lime, lemon, orange, fruit, banana, grape, pear, pineapple, mango, bitter almond, cola, cinnamon, sugar, cotton candy, and vanilla flavors. Non-exhaustive examples of other food ingredients include flavors, acidulants and amino acids, coloring agents, bulking agents, modified starches, gums, texturizers, preservatives, antioxidants, emulsifiers, stabilizers, thickeners, and gelling agents. In some aspects, the invention provides methods for preparing a sweetener product comprising a plurality of high-intensity sweeteners, such plurality including two or more of a steviol glucoside, a mogroside, sucralose, aspartame, neotame, Advantame, acesulfame potassium, saccharin, cyclamate, neohesperidin dihydrochalcone, gnetifolin E, and / or piceatannol 4'-O-β-O-glucopyranoside. The method may further comprise incorporating the sweetener product into a food, beverage, oral care product, sweetener, flavoring agent, or other product, including those described above. Target steviol glycosides, such as RebM, and the sweetener compositions comprising them, can be used in combination with various physiologically active substances or functional ingredients. Functional ingredients are generally classified into categories such as carotenoids, dietary fiber, fatty acids, saponins, antioxidants, nutraceuticals, flavonoids, isothiocyanates, phenols, plant sterols and stanols (phytosterols and phytostenols); polyols; prebiotics, probiotics; phytoestrogens; soy protein; sulfides / thiols; amino acids; and proteins. RCLZbn / Lznza / YiAi vitamins and minerals. Functional ingredients can also be classified according to their health benefits, such as cardiovascular, cholesterol-lowering, and anti-inflammatory. Furthermore, the target steviol glycosides, such as RebM, and the sweetening compositions obtained according to this invention can be applied as high-intensity sweeteners to produce calorie-free, reduced-calorie, or diabetic-friendly beverages and food products with improved flavor characteristics. They can also be used in beverages, foods, pharmaceuticals, and other products where sugar cannot be used. Moreover, RebM and the sweetening compositions can be used as sweeteners not only for beverages, foods, and other products intended for human consumption, but also in animal feed and animal fodder with enhanced characteristics. Examples of products in which target steviol glycosides and sweetening compositions may be used include, but are not limited to, alcoholic beverages such as vodka, wine, beer, liquor, and sake; natural juices; soft drinks; carbonated beverages; diet drinks; calorie-free beverages; reduced-calorie foods and beverages; yogurt drinks; instant juices; instant coffee; powdered instant beverages; canned goods; syrups; fermented soybean paste; soy sauce; vinegar; dressings; mayonnaise; ketchup; curry; soup; instant broth; powdered soy sauce; powdered vinegar; types of cookies; rice crackers; crackers; bread; chocolates; caramel; sweets; chewing gum; gelatin; pudding; preserved fruits and vegetables; crème fraîche; jam; flower paste; powdered milk; ice cream; sorbet; bottled fruits and vegetables; canned and boiled beans; meat and foods boiled in sugary sauce;agricultural plant-based food products; seafood; ham; sausage; fish ham; fish sausage; fish paste; fried fish products; dried seafood; frozen food products; preserved seaweed; preserved meat; tobacco; medicinal products; and many others. During the manufacture of products such as food, beverages, pharmaceuticals, cosmetics, tableware and chewing gum, conventional methods such as mixing, kneading, dissolving, pickling, permeation, percolation, spraying, atomizing, infusion and other methods may be used. The modalities of the invention are demonstrated in the following non-exhaustive examples. EXAMPLES Table 2 shows the steviol glycoside content of three batches of stevia leaf extract. Two intermediates, RebA and stevioside, on the pathway to RebM, are the two main glycosides in the batches. Table 2: Steviol glycoside composition of available stevia leaf extract RpLZbn / Lzzza / YiAi % Batch 1 Batch 2 Batch 3 Rebaudioside A 38.2 10.5 30.3 Stevioside 8.5 9.0 18.4 Rebaudioside C 12.9 4.2 16.6 Rebaudioside B 4.3 7.1 1.2 Rubusoside 5.0 2.2 2.0 Rebaudioside F 2.0 2.7 2.1 Steviolbioside 0.3 3.7 0.3 Rebaudioside D 0.2 2.1 0.9 Dulcoside A 0.9 0.4 0.5 RCLZbn / Lzzza / YiAi Figure 4 shows the bioconversion of glycosylated steviol glycoside intermediates. In the experiment, 0.2 mM steviolbioside was fed to genetically modified E. coli strains in 96-well plates. Product formation was examined after 48 hours. Whole-cell bioconversion is possible even from an early intermediate such as steviolbioside, but for native E. coli, the glycosylated substrate is not available to UGT enzymes (strain 5 shows substantial activity, while strains 3 and 4 show negligible activity). Details of the strains are described in Figure 8. The values ​​shown in Figure 4 are the compound concentration relative to the control (the concentration of each compound is divided by the total concentration of these compounds for the empty vector control). The modifications contained in strain 5 demonstrate the bioconversion of steviolbioside to RebM. Each of strains 1-5 contained a BAC (bacterial artificial chromosome), which is a single-copy plasmid, expressing four separate UGT enzymes: MbUGTC13, MbUGTI.2_2, MbUGTc19_2, and 76G1_L200A. The control contained the same BAC structure but without the UGT enzymes. Figure 8 shows genetic modifications to increase the flux of native E. coli to UDP-glucose, a critical substrate for UGTs in the RebM pathway. A higher-than-native flux to UDP-glucose may allow for greater RebM formation by increasing the amount of substrate available to UGTs. The modifications resulting in strain 5 also result in the cell's ability to convert fed steviol glycosides into RebM. For strains 2 and 3, the UDP-glucose-consuming enzymes (ushA, galETKM) were deleted. Strain 4 was constructed from strain 3 by deleting an enzyme (pgi) that consumes a UDP-glucose precursor, glucose-6-phosphate (G6P). Strain 5 was constructed from strain 4 by overexpressing two enzymes used to convert G6P to UDP-glucose (pgm, galU) via glucose-1-phosphate (G1P). Figure 5 shows the conversion of two commercially available intermediates found in the leaf extract, stevioside and RebA, to RebM. The bioconversion of the steviol core is also shown. Values ​​are expressed as a percentage of the total steviol glycoside composition. The conversion of steviol to RebM by all strains shows that all strains are capable of forming RebM. As was the case with steviolbioside, both stevioside and RebA are converted to RebM, but only in strain 5. It is likely that only strain 5 makes the steviol glycosides available to the UGT enzymes. For the conversion of stevioside and RebA, the RebD:RebM ratio strongly favors RebM (1:20). RebM was secreted into the media. Figure 6 shows the bioconversion of steviol glycosides in 96-well plates. For these studies, 76G1_L200A was replaced with MbUGT1-3_1.5. For five major compounds in the RebM pathway, compound concentrations are shown with and without expressed UGT enzymes. Strains were fed 0.5 g / L of stevioside / RebA leaf extract and incubated with substrate for 48 hours before sampling. As shown, the cells produce almost exclusively RebM, with only small amounts of Rebl and RebD. The strain was used for initial bioreactor experiments. Specifically, the RebM-producing strain was inoculated from a working cell bank into a 50 mL centrifuge tube containing 10 mL of LB (Luria-Bertani) broth with appropriate antibiotics. The first seed culture was incubated for 20 hours in a shaking incubator at 37 °C. After 20 hours, a second seed culture was initiated by inoculating 100 mL of fermentation medium into a 500 mL Erlenmeyer flask using the first seed. This second seed culture was incubated for 10 hours before being transferred to the bioreactors, each containing 170 mL of fermentation medium. Sampling was performed every 10 hours. Relevant metabolites in the medium were quantified using LC-MS / MS and YSI. Cell density was measured using a spectrophotometer at an absorbance of 600 nm. FIGURE 7 shows the bioconversion of a stevioside / RebA leaf extract at bioreactor scale.For five major compounds in the RebM pathway, compound concentrations in the medium were sampled at various time points during fermentation. The strains were fed 2 g / L of the stevioside / RebA leaf extract and incubated with substrate for a total of 30 hours. As shown in Figure 7, stevioside and RebA are lost over time, with a corresponding increase in highly glycosylated products such as RebM, Rebl, and RebD. The UGT enzymes that convert pathway intermediates such as steviolbioside, stevioside, and RebA into RebM are expressed in the cytoplasm of E. coli and therefore require that the intermediates be available to the UGT enzymes, probably through the action of a transporter or through increased membrane permeability. Steviol glycosides are large molecules that are unlikely to be absorbed by native strains of E. coli. This would explain the negligible conversion of steviolbioside to RebM by strain 1. Strains 1-4 can take up an earlier non-glycosylated intermediate (steviol) and convert it to RebM, suggesting that the reason for the negligible conversion is the lack of absorption of steviol glycosides into the cytoplasm, not the inactivity of UGT enzymes on pathway intermediates under these conditions. RpLZbn / Lzzza / YiAi RCLZbn / Lzzza / YiAi SEQUENCES >SrUGT85C2 [Stevia rebaudiana] (SEQ ID NO:1) MDAMATTEKKPHVIFIPFPAQSHIKAMLKLAQLLHHKGLQITFVNTDFIHNQFLESSGPHCLDGA PGFRFETIPDGVSHSPEASIPIRESLLRSIETNFLDRFIDLVTKLPDPPTCIISDGFLSVFTIDAAKKLGIPVM MYWTLAACGFMGFYHIHSLIEKGFAPLKDASYLTNGYLDTVIDWVPGMEGIRLKDFPLDWSTDLNDKVL MFTTEAPQRSHKVSHHIFHTFDELEPSIIKTLSLRYNHIYTIGPLQLLLDQIPEEKKQTGITSLHGYSLVKEE PECFQWLQSKEPNSWYVNFGSTTVMSLEDMTEFGWGLANSNHYFLWIIRSNLVIGENAVLPPELEEHI KKRGFIASWCSQEKVLKHPSVGGFLTHCGWGSTIESLSAGVPMICWPYSWDQLTNCRYICKEWEVGLE MGTKVKRDEVKRLVQELMGEGGHKMRNKAKDWKEKARIAIAPNGSSSLNIDKMVKEITVLARN >SrUGT74G1 [Stevia rebaudiana] (SEQ ID NO:2) MAEQQKIKKSPHVLLIPFPLQGHINPFIQFGKRLISKGVKTTLVTTIHTLNSTLNHSNTTTTSIEIQAI SDGCDEGGFMSAGESYLETFKQVGSKSLADLIKKLQSEGTTIDAIIYDSMTEWVLDVAIEFGIDGGSFFT QACWNSLYYHVHKGLISLPLGETVSVPGFPVLQRWETPLILQNHEQIQSPWSQMLFGQFANIDQARW VFTNSFYKLEEEVIEWTRKIWNLKVIGPTLPSMYLDKRLDDDKDNGFNLYKANHHECMNWLDDKPKESV VYVAFGSLVKHGPEQVEEITRALIDSDVNFLWVIKHKEEGKLPENLSEVIKTGKGLIVAWCKQLDVLAHE SVGCFVTHCGFNSTLEAISLGVPWAMPQFSDQTTNAKLLDEILGVGVRVKADENGIVRRGNLASCIKMI MEEERGVIIRKNAVKWKDLAKVAVHEGGSSDNDIVEFVSELIKA >SrUGT76G1 [Stevia rebaudiana] (SEQ ID NO:3) MENKTETTVRRRRRIILFPVPFQGHINPILQLANVLYSKGFSITIFHTNFNKPKTSNYPHFTFRFIL DNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELLMLASEEDEEVSCLITDALWYFAQSVADSL NLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVKDIKSAYSNWQILKEILGKM IKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLDHDRTVFQWLDQQPPSSVLY VSFGSTSEVDEKDFLEIARGLVDSKQSFLWWRPGFVKGSTVWEPLPDGFLGERGRIVKVWPQQEVLA HGAIGAFWTHSGWNSTLESVCEGVPMIFSDFGLDQPLNARYMSDVLKVGVYLENGWERGEIANAIRRV MVDEEGEYIRQNARVLKQKADVSLMKGGSSYESLESLVSYISSL >SrUGT91D1 [Stevia rebaudiana] (SEQ ID NO:4) MYNVTYHQNSKAMATSDSIVDDRKQLHVATFPWLAFGHILPFLQLSKLIAEKGHKVSFLSTTRNI QRLSSHISPLINWQLTLPRVQELPEDAEATTDVHPEDIQYLKKAVDGLQPEVTRFLEQHSPDWIIYDFTH YWLPSIAASLGISRAYFCVITPWTIAYLAPSSDAMINDSDGRTTVEDLTTPPKWFPFPTKVCWRKHDLAR MEPYEAPGISDGYRMGMVFKGSDCLLFKCYHEFGTQWLPLLETLHQVPWPVGLLPPEIPGDEKDETW VSIKKWLDGKQKGSWYVALGSEALVSQTEWELALGLELSGLPFVWAYRKPKGPAKSDSVELPDGFV ERTRDRGLVWTSWAPQLRILSHESVCGFLTHCGSGSIVEGLMFGHPLIMLPIFCDQPLNARLLEDKQVGI EIPRNEEDGCLTKESVARSLRSVWENEGEIYKANARALSKIYNDTKVEKEYVSQFVDYLEKNARAVAID HES >SrUGT91 D2 [Stevia rebaudíana] (SEQ ID NO:5) MATSDSIVDDRKQLHVATFPWLAFGHILPYLQLSKLIAEKGHKVSFLSTTRNIQRLSSHISPLINW QLTLPRVQELPEDAEATTDVHPEDIPYLKKASDGLQPEVTRFLEQHSPDWIIYDYTHYWLPSIAASLGISR AHFSVTTPWAIAYMGPSADAMINGSDGRTTVEDLTTPPKWFPFPTKVCWRKHDLARLVPYKAPGISDG YRMGLVLKGSDCLLSKCYHEFGTQWLPLLETLHQVPWPVGLLPPEVPGDEKDETWVSIKKWLDGKQK GSWYVALGSEVLVSQTEWELALGLELSGLPFVWAYRKPKGPAKSDSVELPDGFVERTRDRGLVWTS WAPQLRILSHESVCGFLTHCGSGSIVEGLMFGHPLIMLPIFGDQPLNARLLEDKQVGIEIPRNEEDGCLT KESVARSLRSVWEKEGEIYKANARELSKIYNDTKVEKEYVSQFVDYLEKNTRAVAIDHES >SrUGT91 D2e [Stevia rebaudíana] (SEQ ID NO:6) MATSDSIVDDRKQLHVATFPWLAFGHILPYLQLSKLIAEKGHKVSFLSTTRNIQRLSSHISPLINW QLTLPRVQELPEDAEATTDVHPEDIPYLKKASDGLQPEVTRFLEQHSPDWIIYDYTHYWLPSIAASLGISR AHFSVTTPWAIAYMGPSADAMINGSDGRTTVEDLTTPPKWFPFPTKVCWRKHDLARLVPYKAPGISDG YRMGLVLKGSDCLLSKCYHEFGTQWLPLLETLHQVPWPVGLLPPEIPGDEKDETWVSIKKWLDGKQK GSWYVALGSEVLVSQTEWELALGLELSGLPFVWAYRKPKGPAKSDSVELPDGFVERTRDRGLVWTS WAPQLRILSHESVCGFLTHCGSGSIVEGLMFGHPLIMLPIFGDQPLNARLLEDKQVGIEIPRNEEDGCLT KESVARSLRSVWEKEGEIYKANARELSKIYNDTKVEKEYVSQFVDYLEKNARAVAIDHES > OsUGT1-2 [Oryza sativa] (SEQ ID NO:7) MDSGYSSSYAAAAGMHWICPWLAFGHLLPCLDLAQRLASRGHRVSFVSTPRNISRLPPVRPA LAPLVAFVALPLPRVEGLPDGAESTNDVPHDRPDMVELHRRAFDGLAAPFSEFLGTACADWVIVDVFH HWAAAAALEHKVPCAMMLLGSAHMIASIADRRLERAETESPAAAGQGRPAAAPTFEVARMKLIRTKGS SGMSLAERFSLTLSRSSLWGRSCVEFEPETVPLLSTLRGKPITFLGLMPPLHEGRREDGEDATVRWLD AQPAKSWYVALGSEVPLGVEKVHELALGLELAGTRFLWALRKPTGVSDADLLPAGFEERTRGRGWA TRWVPQMSILAHAAVGAFLTHCGWNSTIEGLMFGHPLIMLPIFGDQGPNARLIEAKNAGLQVARNDGDG SFDREGVAAAIRAVAVEEESSKVFQAKAKKLQEIVADMACHERYIDGFIQQLRSYKD >MbUGTC19 [permutante circular] (SEQ ID NO:8) MAECMNWLDDKPKESWYVAFGSLVKHGPEQVEEITRALIDSDVNFLWVIKHKEEGKLPENLSE VIKTGKGLIVAWCKQLDVLAHESVGCFVTHCGFNSTLEAISLGVPWAMPQFSDQTTNAKLLDEILGVGV RVKADENGIVRRGNLASCIKMIMEEERGVIIRKNAVKWKDLAKVAVHEGGSSDNDIVEFVSELIKAGSGE QQKIKKSPHVLLIPFPLQGHINPFIQFGKRLISKGVKTTLVTTIHTLNSTLNHSNTTTTSIEIQAISDGCDEG GFMSAGESYLETFKQVGSKSLADLIKKLQSEGTTIDAIIYDSMTEWVLDVAIEFGIDGGSFFTQACWNSL YYHVHKGLISLPLGETVSVPGFPVLQRWETPLILQNHEQIQSPWSQMLFGQFANIDQARWVFTNSFYKL RCLZbn / Lznza / YiAi EEEVIEWTRKIWNLKVIGPTLPSMYLDKRLDDDKDNGFNLYKANHH >MbUGT1,2 [permutante circular] (SEQ ID NO:9) MAGSSGMSLAERFSLTLSRSSLWGRSCVEFEPETVPLLSTLRGKPITFLGLMPPLHEGRREDG EDATVRWLDAQPAKSWYVALGSEVPLGVEKVHELALGLELAGTRFLWALRKPTGVSDADLLPAGFEE RTRGRGWATRWVPQMSILAHAAVGAFLTHCGWNSTIEGLMFGHPLIMLPIFGDQGPNARLIEAKNAGL QVARNDGDGSFDREGVAAAIRAVAVEEESSKVFQAKAKKLQEIVADMACHERYIDGFIQQLRSYKDDS GYSSSYAAAAGMHWICPWLAFGHLLPCLDLAQRLASRGHRVSFVSTPRNISRLPPVRPALAPLVAFVA LPLPRVEGLPDGAESTNDVPHDRPDMVELHRRAFDGLAAPFSEFLGTACADWVIVDVFHHWAAAAALE H KVPCAM M LLGS AH ΜI AS IADRRLERAETESP AAAGQG RPAAAPTFEVARM KLIRT K >MbUGT1-3 [permutante circular] (SEQ ID NO: 10) MANWQILKEILGKMIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLDH DRTVFQWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSKQSFLWWRPGFVKGSTWVEPLPDGF LGERGRIVKWVPQQEVLAHGAIGAFWTHSGWNSTLESVCEGVPMIFSDFGLDQPLNARYMSDVLKVG VYLENGWERGEIANAIRRVMVDEEGEYIRQNARVLKQKADVSLMKGGSSYESLESLVSYISSLENKTET TVRRRRRIILFPVPFQGHINPILQLANVLYSKGFSITIFHTNFNKPKTSNYPHFTFRFILDNDPQDERISNLP THGPLAGMRIPIINEHGADELRRELELLMLASEEDEEVSCLITDALWYFAQSVADSLNLRRLVLMTSSLFN FHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVKDIKSAYS >MbUGT1,2.2 [permutante circular] (SEQ ID NO:11) MATKGSSGMSLAERFWLTLSRSSLWGRSCVEFEPETVPLLSTLRGKPITFLGLMPPLHEGRRE DGEDATVRWLDAQPAKSWYVALGSEVPLGVEKVHELALGLELAGTRFLWALRKPTGVSDADLLPAGF EERTRGRGWATRWVPQMSILAHAAVGAFLTHCGWNSTIEGLMFGHPLIMLPIFGDQGPNARLIEAKNA GLQVARNDGDGSFDREGVAAAIRAVAVEEESSKVFQAKAKKLQEIVADMACHERYIDGFIQQLRSYKDD SGYSSSYAAAAGMHWICPWLAFGHLLPCLDLAQRLASRGHRVSFVSTPRNISRLPPVRPALAPLVAFV ALPLPRVEGLPDGAESTNDVPHDRPDMVELHRRAFDGLAAPFSEFLGTACADWVIVDVFHHWAAAAAL EHKVPCAMMLLGSAEMIASIADERLEHAETESPAAAGQGRPAAAPTFEVARMKLIR >MbUGTC19-2 [permutante circular] (SEQ ID NO:12) MANHHECMNWLDDKPKESWYVAFGSLVKHGPEQVEEITRALIDSDVNFLWVIKHKEEGKLPE NLSEVIKTGKGLIVAWCKQLDVLAHESVGCFVTHCGFNSTLEAISLGVPWAMPQFSDQTTNAKLLDEIL GVGVRVKADENGIVRRGNLASCIKMIMEEERGVIIRKNAVKWKDLAKVAVHEGGSSDNDIVEFVSELIKA GSGEQQKIKKSPHVLLIPFPLQGHINPFIQFGKRLISKGVKTTLVTTIHTLNSTLNHSNTTTTSIEIQAISDG CDEGGFMSAGESYLETFKQVGSKSLADLIKKLQSEGTTIDAIIYDSMTEWVLDVAIEFGIDGGSFFTQAC WNSLYYHVHKGLISLPLGETVSVPGFPVLQRWETPLILQNHEQIQSPWSQMLFGQFANIDQARWVFTN SFYKLEEEVIEWTRKIWNLKVIGPTLPSMYLDKRLDDDKDNGFNLYKA RCLZbn / Lznza / YiAi >MbUGTC13 (Stevia rebaudiana UGT85C2, P215T) (SEQ ID NO: 13) MADAMATTEKKPHVIFIPFPAQSHIKAMLKLAQLLHHKGLQITFVNTDFIHNQFLESSGPHCLDG APGFRFETIPDGVSHSPEASIPIRESLLRSIETNFLDRFIDLVTKLPDPPTCIISDGFLSVFTIDAAKKLGIPV MMYWTLAACGFMGFYHIHSLIEKGFAPLKDASYLTNGYLDTVIDWVPGMEGIRLKDFPLDWSTDLNDKV LMFTTEATQRSHKVSHHIFHTFDELEPSIIKTLSLRYNHIYTIGPLQLLLDQIPEEKKQTGITSLHGYSLVKE EPECFQWLQSKEPNSWYVNFGSTTVMSLEDMTEFGWGLANSNHYFLWIIRSNLVIGENAVLPPELEEH IKKRGFIASWCSQEKVLKHPSVGGFLTHCGWGSTIESLSAGVPMICWPYSWDQLTNCRYICKEWEVGL EMGTKVKRDEVKRLVQELMGEGGHKMRNKAKDWKEKARIAIAPNGSSSLNIDKMVKEITVLARN >76G1_L200A [Stev / a rebaudiana, L200A] (SEQ ID NO:14) MAENKTETTVRRRRRIILFPVPFQGHINPILQLANVLYSKGFSITIFHTNFNKPKTSNYPHFTFRFIL DNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELLMLASEEDEEVSCLITDALWYFAQSVADSL NLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVKDIKSAYSNWQIAKEILGK MIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLDHDRTVFQWLDQQPPSSVL YVSFGSTSEVDEKDFLEIARGLVDSKQSFLWWRPGFVKGSTWVEPLPDGFLGERGRIVKWVPQQEVL AHGAIGAFWTHSGWNSTLESVCEGVPMIFSDFGLDQPLNARYMSDVLKVGVYLENGWERGEIANAIRR VMVDEEGEYIRQNARVLKQKADVSLMKGGSSYESLESLVSYISSL >MbUGT1-3_1 [permutante circular] (SEQ ID NO: 15) MAFLWWRPGFVKGSTVWEPLPDGFLGERGRIVKWVPQQEVLAHGAIGAFWTHGGWNSTLE SVCEGVPMIFSDFGLDQPLNARYMSDVLKVGVYLENGWERGEIANAIRRLMVDEEGEYIRQNARVLKQ KADVSLMKGGSSYESLESLVSYISSLGSGGSGGSGRRRRIILFPVPFQGHINPMLQLANVLYSKGFSITIF HTNFNKPKTSNYPHFTFRFILDNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELLMLASEEDEE VSCLITDALWYFAQSVADSLNLRRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLK VKDIKSAYSNWQIAKEILGKMIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLD HDRTVFQWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSQS >MbUGT1-3_1.5 [permutante circular] (SEQ ID NO:16) MAFLWWRPGFVKGSTWVEPLPDGFLGERGRIVKWVPQQEVLAHGAIGAFWTHGGWNSTLE SVCEGVPMIFSDFGLDQPLNARYMSDVLKVGVYLENGWERGEIANAIRRLMVDEEGEYIRQNARVLKQ KADVSLMKGGSSYESLESLVSYISSLGSGGSGGSGRRRRIILFPVPFQGHINPMLQLANVLYSKGFSITIF HTNFNKPKTSNYPHFTFRFILDNDPQTTHGPLAGMRIPIINEHGADELRRELELQMLASEEDEEVSCLITD ALWYFAQSVADSLNLPRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVKDIKSA YSNWQIAKEILGKMIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLDHDRTVF QWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSQS >MbUGT1-3_2 [permutante circular] RCl ZfrO I 70Ζ -1 / ΥΙΛΙ (SEQ ID NO: 17) MAFLWWRPGFVKGSTWVEPLPDGFLGERGRIVKWVPQQEVLAHGAIGAFWTHGGWNSTLE SVCEGVPMIFSDFGLDQPLNARYMSDVLKVGVYLENGWERGEIANAIRRLMVDEEGEYIRQNARVLKQ KADVSLMKGGSSYESLESLVSYISSLGSGGSGRRRRIILFPVPFQGHINPMLQLANVLYSKGFSITIFHTN FNKPKTSNYPHFTFRFILDNDPQDERISNLPTHGPLAGMRIPIINEHGADELRRELELQMLASEEDEEVS CLITDALWYFAQSVADSLNLPRLVLMTSSLFNFHAHVSLPQFDELGYLDPDDKTRLEEQASGFPMLKVK DIKSAYSNWQIAKEILGKMIKQTKASSGVIWNSFKELEESELETVIREIPAPSFLIPLPKHLTASSSSLLEHD RTVFQWLDQQPPSSVLYVSFGSTSEVDEKDFLEIARGLVDSQS >SgUGT720-269-1 (Siraitia grosvenorii) (SEQ ID NO: 18) MEDRNAMDMSRIKYRPQPLRPASMVQPRVLLFPFPALGHVKPFLSLAELLSDAGIDWFLSTEY NHRRISNTEALASRFPTLHFETIPDGLPPNESRALADGPLYFSMREGTKPRFRQLIQSLNDGRWPITCIIT DIMLSSPIEVAEEFGIPVIAFCPCSARYLSIHFFIPKLVEEGQIPYADDDPIGEIQGVPLFEGLLRRNHLPGS WSDKSADISFSHGLINQTLAAGRASALILNTFDELEAPFLTHLSSIFNKIYTIGPLHALSKSRLGDSSSSAS ALSGFWKEDRACMSWLDCQPPRSWFVSFGSTMKMKADELREFWYGLVSSGKPFLCVLRSDWSGG EAAELIEQMAEEEGAGGKLGMWEWAAQEKVLSHPAVGGFLTHCGWNSTVESIAAGVPMMCWPILGD QPSNATWIDRVWKIGVERNNREWDRLTVEKMVRALMEGQKRVEIQRSMEKLSKLANEKWRGINLHPT ISLKKDTPTTSEHPRHEFENMRGMNYEMLVGNAIKSPTLTKK >SgUGT94-289-3 (Siraitia grosvenorii) (SEQ ID NO: 19) MTIFFSVEILVLGIAEFAAIAMDAAQQGDTTTILMLPWLGYGHLSAFLELAKSLSRRNFHIYFCSTS VNLDAIKPKLPSSFSDSIQFVELHLPSSPEFPPHLHTTNGLPPTLMPALHQAFSMAAQHFESILQTLAPHL LIYDSLQPWAPRVASSLKIPAINFNTTGVFVISQGLHPIHYPHSKFPFSEFVLHNHWKAMYSTADGASTE RTRKRGEAFLYCLHASCSVILINSFRELEGKYMDYLSVLLNKKWPVGPLVYEPNQDGEDEGYSSIKNW LDKKEPSSTVFVSFGSEYFPSKEEMEEIAHGLEASEVNFIWWRFPQGDNTSGIEDALPKGFLERAGER GMWKGWAPQAKILKHWSTGGFVSHCGWNSVMESMMFGVPIIGVPMHVDQPFNAGLVEEAGVGVEA KRDPDGKIQRDEVAKLIKEWVEKTREDVRKKAREMSEILRSKGEEKFDEMVAEISLLLKI >SgUGT74-345-2 (Siraitia grosvenorii) (SEQ ID NO:20) MDETTVNGGRRASDWVFAFPRHGHMSPMLQFSKRLVSKGLRVTFLITTSATESLRLNLPPSSS LDLQVISDVPESNDIATLEGYLRSFKATVSKTLADFIDGIGNPPKFIVYDSVMPWVQEVARGRGLDAAPF FTQSSAVNHILNHVYGGSLSIPAPENTAVSLPSMPVLQAEDLPAFPDDPEWMNFMTSQFSNFQDAKWI FFNTFDQLECKKQSQWNWMADRWPIKTVGPTIPSAYLDDGRLEDDRAFGLNLLKPEDGKNTRQWQW LDSKDTASVLYISFGSLAILQEEQVKELAYFLKDTNLSFLWVLRDSELQKLPHNFVQETSHRGLWNWCS QLQVLSHRAVSCFVTHCGWNSTLEALSLGVPMVAIPQWVDQTTNAKFVADVWRVGVRVKKKDERIVTK EELEASIRQWQGEGRNEFKHNAIKWKKLAKEAVDEGGSSDKNIEEFVKTIA >SgUGT75-281-2 (Siraitia grosvenorii) RCLZbn / Lznza / YiAi (SEQ ID NO:21) MGDNGDGGEKKELKENVKKGKELGRQAIGEGYINPSLQLARRLISLGVNVTFATTVI_AGRRMK NKTHQTATTPGLSFATFSDGFDDETLKPNGDLTHYFSELRRCGSESLTHLITSAANEGRPITFVIYSLLLS WAADIASTYDIPSALFFAQPATVLALYFYYFHGYGDTICSKLQDPSSYIELPGLPLLTSQDMPSFFSPSGP HAFILPPMREQAEFLGRQSQPKVLVNTFDALEADALRAIDKLKMLAIGPLIPSALLGGNDSSDASFCGDL FQVSSEDYIEWLNSKPDSSWYISVGSICVLSDEQEDELVHALLNSGHTFLWVKRSKENNEGVKQETDE EKLKKLEEQGKMVSWCRQVEVLKHPALGCFLTHCGWNSTIESLVSGLPWAFPQQIDQATNAKLIEDV WKTGVRVKANTEGIVEREEIRRCLDLVMGSRDGQKEEIERNAKKWKELARQAIGEGGSSDSNLKTFLW EIDLEI >SgUGT720-269-4 (Siraitia grosvenoríi) (SEQ ID NO:22) MAEQAHDLLHVLLFPFPAEGHIKPFLCLAELLCNAGFHVTFLNTDYNHRRLHNLHLLAARFPSLH FESISDGLPPDQPRDILDPKFFISICQVTKPLFRELLLSYKRISSVQTGRPPITCVITDVIFRFPIDVAEELDIP VFSFCTFSARFMFLYFWIPKLIEDGQLPYPNGNINQKLYGVAPEAEGLLRCKDLPGHWAFADELKDDQL NFVDQTTASSRSSGLILNTFDDLEAPFLGRLSTIFKKIYAVGPIHSLLNSHHCGLWKEDHSCLAWLDSRA AKSWFVSFGSLVKITSRQLMEFWHGLLNSGKSFLFVLRSDWEGDDEKQWKEIYETKAEGKWLWG WAPQEKVLAHEAVGGFLTHSGWNSILESIAAGVPMISCPKIGDQSSNCTWISKVWKIGLEMEDRYDRVS VETMVRSIMEQEGEKMQKTIAELAKQAKYKVSKDGTSYQNLECLIQDIKKLNQIEGFINNPNFSDLLRV >SgUGT94-289-2 (Siraitia grosvenoríi) (SEQ ID NO:23) MDAQQGHTTTILMLPWVGYGHLLPFLELAKSLSRRKLFHIYFCSTSVSLDAIKPKLPPSISSDDSI QLVELRLPSSPELPPHLHTTNGLPSHLMPALHQAFVMAAQHFQVILQTLAPHLLIYDILQPWAPQVASSL NIPAINFSTTGASMLSRTLHPTHYPSSKFPISEFVLHNHWRAMYTTADGALTEEGHKIEETLANCLHTSC GWLVNSFRELETKYIDYLSVLLNKKWPVGPLVYEPNQEGEDEGYSSIKNWLDKKEPSSTVFVSFGTE YFPSKEEMEEIAYGLELSEVNFIVWLRFPQGDSTSTIEDALPKGFLERAGERAMWKGWAPQAKILKHW STGGLVSHCGWNSMMEGMMFGVPIIAVPMHLDQPFNAGLVEEAGVGVEAKRDSDGKIQREEVAKSIK EWIEKTREDVRKKAREMDTKHGPTYFSRSKVSSFGRLYKINRPTTLTVGRFWSKQIKMKRE >SgUGT94-289-1 (Siraitia grosvenoríi) (SEQ ID NO:24) MDAQRGHTTTILMFPWLGYGHLSAFLELAKSLSRRNFHIYFCSTSVNLDAIKPKLPSSSSSDSIQ LVELCLPSSPDQLPPHLHTTNALPPHLMPTLHQAFSMAAQHFAAILHTLAPHLLIYDSFQPWAPQLASSL NIPAINFNTTGASVLTRMLHATHYPSSKFPISEFVLHDYWKAMYSAAGGAVTKKDHKIGETLANCLHASC SVILINSFRELEEKYMDYLSVLLNKKWPVGPLVYEPNQDGEDEGYSSIKNWLDKKEPSSTVFVSFGSEY FPSKEEMEEIAHGLEASEVHFIWWRFPQGDNTSAIEDALPKGFLERVGERGMWKGWAPQAKILKHW STGGFVSHCGWNSVMESMMFGVPIIGVPMHLDQPFNAGLAEEAGVGVEAKRDPDGKIQRDEVAKLIKE VWEKTREDVRKKAREMSEILRSKGEEKMDEMVAAISLFLKI > RCLZbn / Lznza / YiAi charantia] (SEQ ID NO:25) MAQPQTQARVLVFPYPTVGHIKPFLSLAELLADGGLDWFLSTEYNHRRIPNLEALASRFPTLHF DTIPDGLPIDKPRVIIGGELYTSMRDGVKQRLRQVLQSYNDGSSPITCVICDVMLSGPIEAAEELGIPWTF CPYSARYLCAHFVMPKLIEEGQIPFTDGNLAGEIQGVPLFGGLLRRDHLPGFWFVKSLSDEVWSHAFLN QTLAVGRTSALIINTLDELEAPFLAHLSSTFDKIYPIGPLDALSKSRLGDSSSSSTVLTAFWKEDQACMSW LDSQPPKSVIFVSFGSTMRMTADKLVEFWHGLVNSGTRFLCVLRSDIVEGGGAADLIKQVGETGNGIW EWAAQEKVLAHRAVGGFLTHCGWNSTMESIAAGVPMMCWQIYGDQMINATWIGKVWKIGIERDDKWD RSTVEKMIKELMEGEKGAEIQRSMEKFSKLANDKWKGGTSFENLELIVEYLKKLKPSN >XP_022151546.1 Ácido 7-desoxiloganetico similar a la glucosiltransferasa [Momordica charantia] (SEQ ID NO:26) MAQPRVLLFPFPAMGHVKPFLSLAELLSDAGVEWFLSTEYNHRRIPDIGALAARFPTLHFETIP DGLPPDQPRVLADGHLYFSMLDGTKPRFRQLIQSLNGNPRPITCIINDVMLSSPIEVAEEFGIPVIAFCPC SARFLSVHFFMPNFIEEAQIPYTDENPMGKIEEATVFEGLLRRKDLPGLWCAKSSNISFSHRFINQTIAAG RASALILNTFDELESPFLNHLSSIFPKIYCIGPLNALSRSRLGKSSSSSSALAGFWKEDQAYMSWLESQP PRSVIFVSFGSTMKMEAWKLAEFWYGLVNSGSPFLFVFRPDCVINSGDAAEVMEGRGRGMWEWASQ EKVLAHPAVGGFLTHCGWNSTVESIVAGVPMMCCPIVADQLSNATWIHKVWKIGIEGDEKWDRSTVEM MIKELMESQKGTEIRTSIEMLSKLANEKWKGGTSLNNFELLVEDIKTLRRPYT >XP_022151514.1 Ácido 7-desoxiloganetico similar a la glucosiltransferasa [Momordica charantia] (SEQ ID NO:27) MEQSDSNSDDHQHHVLLFPFPAKGHIKPFLCLAQLLCGAGLQVTFLNTDHNHRRIDDRHRRLLA TQFPMLHFKSISDGLPPDHPRDLLDGKLIASMRRVTESLFRQLLLSYNGYGNGTNNVSNSGRRPPISCVI TDVIFSFPVEVAEELGIPVFSFATFSARFLFLYFWIPKLIQEGQLPFPDGKTNQELYGVPGAEGIIRCKDLP GSWSVEAVAKNDPMNFVKQTLASSRSSGLILNTFEDLEAPFVTHLSNTFDKIYTIGPIHSLLGTSHCGLW KEDYACLAWLDARPRKSWFVSFGSLVKTTSRELMELWHGLVSSGKSFLLVLRSDWEGEDEEQWKE ILESNGEGKWLWGWAPQEEVLAHEAIGGFLTHSGWNSTMESIAAGVPMVCWPKIGDQPSNCTWVSR VWKVGLEMEERYDRSTVARMARSMMEQEGKEMERRIAELAKRVKYRVGKDGESYRNLESLIRDIKITK SSN >XP_004147933.2 PREDECIDO: ácido 7-desoxiloganetico similar a la glucosiltransferasa [Cucumis sativus] (SEQ ID NO:28) MGLSPTDHVLLFPFPAKGHIKPFFCLAHLLCNAGLRVTFLSTEHHHQKLHNLTHLAAQIPSLHFQ SISDGLSLDHPRNLLDGQLFKSMPQVTKPLFRQLLLSYKDGTSPITCVITDLILRFPMDVAQELDIPVFCFS TFSARFLFLYFSIPKLLEDGQIPYPEGNSNQVLHGIPGAEGLLRCKDLPGYWSVEAVANYNPMNFVNQTI ATSKSHGLILNTFDELEVPFITNLSKIYKKVYTIGPIHSLLKKSVQTQYEFWKEDHSCLAWLDSQPPRSVM RCLZbn / Lznza / YiAi FVSFGSIVKLKSSQLKEFWNGLVDSGKAFLLVLRSDALVEETGEEDEKQKELVIKEIMETKEEGRWVIVN WAPQEKVLEHKAIGGFLTHSGWNSTLESVAVGVPMVSWPQIGDQPSNATWLSKVWKIGVEMEDSYDR STVESKVRSIMEHEDKKMENAIVELAKRVDDRVSKEGTSYQNLQRLIEDIEGFKLN >XP_022978164.1 Ácido 7-desoxiloganetico similar a la glucosiltransferasa [Cucúrbita maxíma] (SEQ ID NO:29) MELSHTHHVLLFPFPAKGHIKPFFSLAQLLCNAGLRVTFLNTDHHHRRIHDLNRLAAQLPTLHFD SVSDGLPPDEPRNVFDGKLYESIRQVTSSLFRELLVSYNNGTSSGRPPITCVITDVMFRFPIDIAEELGIP VFTFSTFSARFLFLIFWIPKLLEDGQLRYPEQELHGVPGAEGLIRWKDLPGFWSVEDVADWDPMNFVN QTLATSRSSGLILNTFDELEAPFLTSLSKIYKKIYSLGPINSLLKNFQSQPQYNLWKEDHSCMAWLDSQP RKSWFVSFGSWKLTSRQLMEFWNGLVNSGMPFLLVLRSDVIEAGEEWREIMERKAEGRWVIVSWA PQEEVLAHDAVGGFLTHSGWNSTLESLAAGVPMISWPQIGDQTSNSTWISKVWRIGLQLEDGFDSSTIE TMVRSIMDQTMEKTVAELAERAKNRASKNGTSYRNFQTLIQDITNIIETHI >XP_022950128.1 Ácido 7-desoxiloganetico similar a la glucosiltransferasa [Cucúrbita moschata] (SEQ ID NQ:30) MELSPTHHLLLFPFPAKGHIKPFFSLAQLLCNAGARVTFLNTDHHHRRIHDLDRLAAQLPTLHFD SVSDGLPPDESRNVFDGKLYESIRQVTSSLFRELLVSYNNGTSSGRPPITCVITDCMFRFPIDIAEELGIP VFTFSTFSARFLFLFFWIPKLLEDGQLRYPEQELHGVPGAEGLIRCKDLPGFLSDEDVAHWKPINFVNQI LATSRSSGLILNTFDELEAPFLTSLSKIYKKIYSLGPINSLLKNFQSQPQYNLWKEDHSCMAWLDSQPPK SWFVSFGSWKLTNRQLVEFWNGLVNSGKPFLLVLRSDVIEAGEEWRENMERKAEGRWMIVSWAPQ EEVLAHDAVGGFLTHSGWNSTLESLAAGVPMISWTQIGDQTSNSTWVSKVWRIGLQLEDGFDSFTIET MVRSVMDQTMEKTVAELAERAKNRASKNGTSYRNFQTLIQDITNIIETHI >XP_020422423.1 Ácido 7-desoxiloganetico glucosiltransferasa [Prunus pérsica] (SEQ ID NO:31) MAMKQPHVIIFPFPLQGHMKPLLCLAELLCHAGLHVTYVNTHHNHQRLANRQALSTHFPTLHFE SISDGLPEDDPRTLNSQLLIALKTSIRPHFRELLKTISLKAESNDTLVPPPSCIMTDGLVTFAFDVAEELGL PILSFNVPCPRYLWTCLCLPKLIENGQLPFQDDDMNVEITGVPGMEGLLHRQDLPGFCRVKQADHPSLQ FAINETQTLKRASALILDTVYELDAPCISHMALMFPKIYTLGPLHALLNSQIGDMSRGLASHGSLWKSDLN CMTWLDSQPSKSIIYVSFGTLVHLTRAQVIEFWYGLVNSGHPFLWVMRSDITSGDHQIPAELENGTKER GCIVDWVSQEEVLAHKSVGGFLTHSGWNSTLESIVAGLPMICWPKLGDHYIISSTVCRQWKIGLQLNEN CDRSNIESMVQTLMGSKREEIQSSMDAISKLSRDSVAEGGSSHNNLEQLIEYIRNLQHQN >EOY07351.1 UDP-glucosil transferasa 85A3, putativo [Theobroma cacao] (SEQ ID NO:32) MRQPHVLVLPFPAQGHIKPMLCLAELLCQAGLRVTFLNTHHSHRRLNNLQDLSTRFPTLHFESV SDGLPEDHPRNLVHFMHLVHSIKNVTKPLLRDLLTSLSLKTDIPPVSCIIADGILSFAIDVAEELQIKVIIFRTI SSCCLWSYLCVPKLIQQGELQFSDSDMGQKVSSVPEMKGSLRLHDRPYSFGLKQLEDPNFQFFVSET QAMTRASAVIFNTFDSLEAPVLSQMIPLLPKVYTIGPLHALRKARLGDLSQHSSFNGNLREADHNCITWL RCl ZfrO I 70Ζ -1 / ΥΙΛΙ DSQPLRSWYVSFGSHWLTSEELLEFWHGLVNSGKRFLWVLRPDIIAGEKDHNQIIAREPDLGTKEKGL LVDWAPQEEVLAHPSVGGFLTHCGWNSTLESMVAGVPMLCWPKLPDQLVNSSCVSEVWKIGLDLKDM CDRSTVEKMVRALMEDRREEVMRSVDGISKLARESVSHGGSSSSNLEMLIQELET >XP_022155979.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Momordica charantia] (SEQ ID NO:33) MDAHQQAEHTTTILMLPWVGYGHLTAYLELAKALSRRNFHIYYCSTPVNIESIKPKLTIPCSSIQF VELHLPSSDDLPPNLHTTNGLPSHLMPTLHQAFSAAAPLFEEILQTLCPHLLIYDSLQPWAPKIASSLKIPA LNFNTSGVSVIAQALHAIHHPDSKFPLSDFILHNYWKSTYTTADGGASEKTRRAREAFLYCLNSSGNAILI NTFRELEGEYIDYLSLLLNKKVIPIGPLVYEPNQDEDQDEEYRSIKNWLDKKEPCSTVFVSFGSEYFPSN EEMEEIAPGLEESGANFIWWRFPKLENRNGIIEEGLLERAGERGMVIKEWAPQARILRHGSIGGFVSHC GWNSVMESIICGVPVIGVPMRVDQPYNAGLVEEAGVGVEAKRDPDGKIQRHEVSKLIKQVWEKTRDD VRKKVAQMSEILRRKGDEKIDEMVALISLLPKG >XP_022986080.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Cucúrbita maxíma] (SEQ ID NO:34) MDAQKAVDTPPTTVLMLPWIGYGHLSAYLELAKALSRRNFHVYFCSTPVNLDSIKPNLIPPPSSI QFVDLHLPSSPELPPHLHTTNGLPSHLKPTLHQAFSAAAQHFEAILQTLSPHLLIYDSLQPWAPRIASSLN IPAINFNTTAVSIIAHALHSVHYPDSKFPFSDFVLHDYWKAKYTTADGATSEKIRRGAEAFLYCLNASCDV VLVNSFRELEGEYMDYLSVLLKKKWSVGPLVYEPSEGEEDEEYWRIKKWLDEKEALSTVLVSFGSEYF PSKEEMEEIAHGLEESEANFIWWRFPKGEESCRGIEEALPKGFVERAGERAMWKKWAPQGKILKHG SIGGFVSHCGWNSVLESIRFGVPVIGVPMHLDQPYNAGLLEEAGIGVEAKRDADGKIQRDQVASLIKRV WEKTREDIWKTVREMREVLRRRDDDMIDEMVAEISWLKI >XP_022156002.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Momordica charantia] (SEQ ID NO:35) MDARQQAEHTTTILMLPWVGYGHLSAYLELAKALSRRNFHIYYCSTPVNIESIKPKLTIPCSSIQF VELHLPFSDDLPPNLHTTNGLPSHLMPALHQAFSAAAPLFEAILQTLCPHLLIYDSLQPWAPQIASSLKIP ALNFNTTGVSVIARALHTIHHPDSKFPLSEIVLHNYWKATHATADGANPEKFRRDLEALLCCLHSSCNAIL INTFRELEGEYIDYLSLLLNKKVTPIGPLVYEPNQDEEQDEEYRSIKNWLDKKEPYSTIFVSFGSEYFPSN EEMEEIARGLEESGANFIWWRFHKLENGNGITEEGLLERAGERGMVIQGWAPQARILRHGSIGGFVSH CGWNSVMESIICGVPVIGVPMGLDQPYNAGLVEEAGVGVEAKRDPDGKIQRHEVSKLIKQVWEKTRD DVRKKVAQMSEILRRKGDEKIDEMVALISLLLKG >XP_022943327.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Cucúrbita moschatá] (SEQ ID NO:36) MDAQKAVDTPPTTVLMLPWIGYGHLSAYLELAKALSRRNFHVYFCSTPVNLDSIKPNLIPPPPSI RCLZbn / Lznza / YiAi QFVDLHLPSSPELPPHLHTTNGLPSHLKPTLHQAFSAAAQHFEAILQTLSPHLLIYDSLQPWAPRIASSLN IPAINFNTTAVSIIAHALHSVHYPDSKFPFSDFVLHDYWKAKYTTADGATSEKTRRGVEAFLYCLNASCDV VLVNSFRELEGEYMDYLSVLLKKKWSVGPLVYEPSEGEEDEEYWRIKKWLDEKEALSTVLVSFGSEYF PPKEEMEEIAHGLEESEANFIWWRFPKGEESSSRGIEEALPKGFVERAGERAMWKKWAPQGKILKHG SIGGFVSHCGWNSVLESIRFGVPVIGAPMHLDQPYNAGLLEEAGIGVEAKRDADGKIQRDQVASLIKQV WEKTREDIWKKVREMREVLRRRDDDDMMIDEMVAVISWLKI >XP_022996307.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Cucúrbita maxíma] (SEQ ID NO:37) MSSNLFLKISIPFGRLRDSALNCSVFHCKLHLAIAIAMDAQQAANKSPTATTIFMLPWAGYGHLS AYLELAKALSTRNFHIYFCSTPVSLASIKPRLIPSCSSIQFVELHLPSSDEFPPHLHTTNGLPSRLVPTFHQ AFSEAAQTFEAFLQTLRPHLLIYDSLQPWAPRIASSLNIPAINFFTAGAFAVSHVLRAFHYPDSQFPSSDF VLHSRWKIKNTTAESPTQAKLPKIGEAIGYCLNASRGVILTNSFRELEGKYIDYLSVILKKRVFPIGPLVYQ PNQDEEDEDYSRIKNWLDRKEASSTVLVSFGSEFFLSKEETEAIAHGLEQSEANFIWGIRFPKGAKKNAI EEALPEGFLERAGGRAMWEEWVPQGKILKHGSIGGFVSHCGWNSAMESIVCGVPIIGIPMQVDQPFN AGILEEAGVGVEAKRDSDGKIQRDEVAKLIKEWVERTREDIRNKLEKINEILRSRREEKLDELATEISLLS RN >XP_022957664.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Cucúrbita moschata] (SEQ ID NO:38) MDAQQAANKSPTASTIFMLPWVGYGHLSAYLELAKALSTRNFHVYFCSTPVSLASIKPRLIPSCS SIQFVELHLPSSDEFPPHLHTTNGLPAHLVPTIHQAFAAAAQTFEAFLQTLRPHLLIYDSLQPWAPRIASS LNIPAINFFTAGAFAVSHVLRAFHYPDSQFPSSDFVLHSRWKIKNTTAESPTQVKIPKIGEAIGYCLNASR GVILTNSFRELEGKYIDYLSVILKKRVLPIGPLVYQPNQDEEDEDYSRIKNWLDRKEASSTVLVSFGSEFF LSKEETEAIAHGLEQSEANFIWGIRFPKGAKKNAIEALPEGFLERVGGRAMWEEVWPQGKILKHGNIG GFVSHCGWNSAMESIMCGVPVIGIPMQVDQPFNAGILEEAGVGEAKRDSDGKIQRDEVAKLIKEWVE RTREDIRNKLEEINEILRTRREEKLDELATEISLLCKN >OMO57892.1 UDP-glucuronosyl / UDP-glucosyltransferase [Corchorus capsulaos] (SEQ ID NO:39) MDSKQKKMSVLMFPWLAYGHISPFLELAKKLSKRNFHTFFFSTPINLNSIKSKLSPKYAQSIKFV ELHLPSLPDLPPHYHTTNGLPPHLMNTLKKAFDMSSLQFSKILKTLNPDLLVYDFIQPWAPLLALSNKIPA VHFLCTSAAMSSFSVHAFKKPCEDFPFPNIYVHGNFMNAKFNNMENCSSDDSISDQDRVLQCFERSTKI ILVKTFEELEGKFMDYLSVLLNKKIVPTGPLTQDPNEDEGDDDERTKLLLEWLNKKSKSSTVFVSFGSEY FLSKEEREEIAYGLELSKVNFIWVIRFPLGENKTNLEEALPQGFLQRVSERGLWENWAPQAKILQHSSI GGFVSHCGWSSVMESLKFGVPIIAIPMHLDQPLNARLWDVGVGLEVIRNHGSLEREEIAKLIKEWLGN GNDGEIVRRKAREMSNHIKKKGEKDMDELVEELMLICKMKPNSCHLS >XP_015886141.1 PREDECIDO: beta-D-glucosil crocetina similar a la beta-1,6RCl ZfrO I 70Ζ -1 / ΥΙΛΙ glucosiltransferasa ¡soforma X1 [Ziziphusjujuba] (SEQ ID NQ:40) MMERQRSIKVLMFPWLAHGHISPFLELAKRLTDRNFQIYFCSTPVNLTSVKPKLSQKYSSSIKLV ELHLPSLPDLPPHYHTTNGLALNLIPTLKKAFDMSSSSFSTILSTIKPDLLIYDFLQPWAPQLASCMNIPAV NFLSAGASMVSFVLHSIKYNGDDHDDEFLTTELHLSDSMEAKFAEMTESSPDEHIDRAVTCLERSNSLILI KSFRELEGKYLDYLSLSFAKKWPIGPLVAQDTNPEDDSMDIINWLDKKEKSSTVFVSFGSEYYLTNEEM EEIAYGLELSKVNFIWWRFPLGQKMAVEEALPKGFLERVGEKGMWEDWAPQMKILGHSSIGGFVSHC GWSSLMESLKLGVPIIAMPMQLDQPINAKLVERSGVGLEVKRDKNGRIEREYLAKVIREIWEKARQDIEK KAREMSNIITEKGEEEIDNWEELAKLCGM >XP_002271587.3 PREDECIDO: beta-D-glucosil crocetina similar a la beta-1,6glucosiltransferasa [Vitis vinifera] (SEQ ID NO:41) MDARQSDGISVLMFPWLAHGHISPFLQLAKKLSKRNFSIYFCSTPVNLDPIKGKLSESYSLSIQLV KLHLPSLPELPPQYHTTNGLPPHLMPTLKMAFDMASPNFSNILKTLHPDLLIYDFLQPWAPAAASSLNIPA VQFLSTGATLQSFLAHRHRKPGIEFPFQEIHLPDYEIGRLNRFLEPSAGRISDRDRANQCLERSSRFSLIK TFREIEAKYLDYVSDLTKKKMVTVGPLLQDPEDEDEATDIVEWLNKKCEASAVFVSFGSEYFVSKEEME EIAHGLELSNVDFIWWRFPMGEKIRLEDALPPGFLHRLGDRGMWEGWAPQRKILGHSSIGGFVSHCG WSSVMEGMKFGVPIIAMPMHLDQPINAKLVEAVGVGREVKRDENRKLEREEIAKVIKEWGEKNGENVR RKARELSETLRKKGDEEIDWVEELKQLCSY >XP_018840205.1 PREDECIDO: beta-D-glucosil crocetina similar a la beta-1,6glucosiltransferasa [Juglans regía] (SEQ ID NO:42) MDTARKRIRWMLPWLAHGHISPFLELSKKLAKRNFHIYFCSTPVNLSSIKPKLSGKYSRSIQLVE LHLPSLPELPPQYHTTKGLPPHLNATLKRAFDMAGPHFSNILKTLSPDLLIYDFLQPWAPAIAASQNIPAIN FLSTGAAMTSFVLHAMKKPGDEFPFPEIHLDECMKTRFVDLPEDHSPSDDHNHISDKDRALKCFERSSG FVMMKTFEELEGKYINFLSHLMQKKIVPVGPLVQNPVRGDHEKAKTLEWLDKRKQSSAVFVSFGTEYFL SKEEMEEIAYGLELSNVNFIWWRFPEGEKVKLEEALPEGFLQRVGEKGMWEGWAPQAKILMHPSIGG FVSHCGWSSVMESIDFGVPIVAIPMQLDQPVNAKWEQAGVGVEVKRDRDGKLEREEVATVIREWMG NIGESVRKKEREMRDNIRKKGEEKMDGVAQELVQLYGNGIKNV >XP_021652171.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Hevea brasiliensis] (SEQ ID NO:43) METLQRRKISVLMFPWLAHGHLSPFLELSKKLNKRNFHVYFCSTPVNLDSIKPKLSAEYSFSIQL VELHLPSSPELPLHYHTTNGLPPHLMKNLKNAFDMASSSFFNILKTLKPDLLIYDFIQPWAPALASSLNIPA VNFLCTSMAMSCFGLHLNNQEAKFPFPGIYPRDYMRMKVFGALESSSNDIKDGERAGRCMDQSFHLIL AKTFRELEGKYIDYLSVKLMKKIVPVGPLVQDPIFEDDEKIMDHHQVIKWLEKKERLSTVFVSFGTEYFLS TEEMEEIAYGLELSKAHFIWWRFPTGEKINLEESLPKRYLERVQERGKIVEGWAPQQKILRHSSIGGFV RCLZbn / Lznza / YiAi SHCGWSSIMESMKFGVPIIAMPMNLDQPVNSRIVEDAGVGIEVRRNKSGELEREEIAKTIRKWVEKDGK NVSRKAREMSDTIRKKGEEEIDGWDELLQLCDVKTNYLQ >XP_021619073.1 beta-D-glucosil crocetina similar a la beta-1,6-glucosiltransferasa [Manihot esculenta] (SEQ ID NO:44) MATAQTRKISVLMFPWLAHGHLSPFLELSKKLANRNFHVYFCSTPVNLDSIKPKLSPEYHFSIQF VELHLPSSPELPSHYHTTNGLPPHLMKTLKKAFDMASSSFFNILKTLNPDLLIYDFLQPWAPALASSLNIP AVNFLCSSMAMSCFGLNLNKNKEIKFLFPEIYPRDYMEMKLFRVFESSSNQIKDGERAGRCIDQSFFML AKTFRELEGKYIDYVSVKCNKKIVPVGPLVEDTIHEDDEKTMDHHHHHHDEVIKWLEKKERSTTVFVSFG SEYFLSKEEMEEIAHGLELSKVNFIWWRFPKGEKINLEESLPEGYLERIQERGKIVEGWAPQRKILGHSS IGGFVSHCGWSSIMESMKLGVPIIAMPMNLDQPINSRIVEAAGVGIEVSRNQSGELEREEMAKTIRKWV EREGVYVRRKAREMSDVLRKKGEEEIDGWDELVQLCDMKTNYL >GAV83746.1 Proteína que contiene el dominio UDPGT [Cephalotus foílicularís] (SEQ ID NO:45) MDLKRRSIRVLMLPWLAHGHISPFLELAKKLTNRNFLIYFCSTPINLNSIKPKLSSKYSFSIQLVEL HLPSLPELPPHYHTTNGLPLHLMNTLKTAFDMASPSFLNILKTLKPDLLICDHLQPWAPSLASSLNIPAIIF PTNSAIMMAFSLHHAKNPGEEFPFPSININDDMVKSINFLHSASNGLTDMDRVLQCLERSSNTMLLKTFR QLEAKYVDYSSALLKKKIVLAGPLVQVPDNEDEKIEIIKWLDSRGQSSTVFVSFGSEYFLSKEEREDIAHG LELSKVNFIWWRFPVGEKVKLEEALPNGFAERIGERGLWEGWAPQAMILSHSSIGGFVSHCGWSSM MESMKFGVPIIAMPMHIDQPLNARLVEDVGVGLEIKRNKDGRFEREELARVIKEVLVYKNGDAVRSKAR EMSEHIKKNGDQEIDGVADALVKLCEMKTNSLNQD >Coffea Arábica UGT_1,6 (SEQ ID NO:46) MENHATFNVLMLPWLAHGHVSPYLELAKKLTARNFNVYLCSSPATLSSVRSKLTEKFSQSIHLV ELHLPKLPELPAEYHTTNGLPPHLMPTLKDAFDMAKPNFCNVLKSLKPDLLIYDLLQPWAPEAASAFNIP AWFISSSATMTSFGLHFFKNPGTKYPYGNAIFYRDYSVFVENLTRRDRDTYRVINCMERSSKIILIKGF NEIEGKYFDYFSCLTGKKWPVGPLVQDPVLDDEDCRIMQWLNKKEKGSTVFVSFGSEYFLSKKDMEEI AHGLEVSNVDFIWWRFPKGENMEETLPKGFFERVGERGLWNGWAPQAKILTHPNVGGFVSHCGW NSVMESMKFGLPIIAMPMHLDQPINARLIEEVGAGVEVLRDSKGKLHRERMAETINKVMKEASGESVRK KARELQEKLELKGDEEIDDWKELVQLCATKNKRNGLHYY > Circular permutant linker sequence (SEQ ID NO:47) GSGGSG > Circular permutant linker sequence (SEQ ID NO:48) GSGGSGGSG

Claims

1. A method for producing a glycosylated product from glycoside intermediates, comprising: providing a microbial cell that intracellularly expresses one or more UDP-dependent glycosyltransferase enzymes, incubating the microbial strain with the glycoside intermediates, and recovering the glycosylated product.

2. The method of claim 1, wherein the glycoside intermediates are provided as a plant extract or a fraction thereof.

3. The method of claim 1, wherein the glycoside intermediate is a glycosylated secondary metabolite, optionally selected from a glycosylated terpenoid, a glycosylated flavonoid, a glycosylated cannabinoid, a glycosylated polyketide, a glycosylated stilbenoid, and a glycosylated polyphenol.

4. The method of claim 3, wherein the glycoside intermediate is a glycosylated terpenoid.

5. The method of claim 4, wherein the glycoside intermediate is mogroside or steviol glycoside.

6. The method of any of claim 1, wherein the glucoside intermediate has one, two, three, four or five glucosyl groups.

7. The method of claim 6, wherein the glycosyl groups are independently selected from glucosyl, galactosyl, mannosyl, xylosyl, and rhamnosyl groups.

8. The method of claim 6, wherein the glycosylated product has at least five glycosyl groups, or at least six glycosyl groups.

9. The method of claim 8, wherein the glycosylated product has seven, eight, or nine glycosyl groups.

10. The method of claim 8, wherein the biosynthesis of the product involves at least two glycosylation reactions of the intermediate in the microbial cell.

11. The method of any of claim 1, wherein the glycoside intermediates are steviol glycosides from stevia leaf extract.

12. The method of claim 11, wherein the steviol glycoside intermediates comprise one or more of stevioside, steviolbioside, rebaudioside A, dulcoside A, dulcoside B, rebaudioside C, and rebaudioside F.

13. The method of claim 12, wherein the extract comprises stevioside, steviolbioside, and rebaudioside A as prominent components. RCLZbn / Lznza / YiAi 14. The method of claim 13, wherein the glycosylated product comprises RebM.

15. The method of claim 12, wherein the glycosylated product comprises RebK, RebC+1 and / or RebC+2.

16. The method of any of claims 1 to 15, wherein the microbial cell is a bacterial cell.

17. The method of claim 16, wherein the bacterial cell is Escherichia spp., Bacillus spp., Rhodobacter spp., Zymomonas spp., or Pseudomonas spp.

18. The method of claim 17, wherein the bacterial cell is Escherichiacoli, Bacillus subtilis, Rhodobacter capsulatus, Rhodobacter sphaeroides, Zymomonas mobiliso Pseudomonas putida.

19. The method of claim 18, wherein the bacterial cell is E. coli.

20. The method of claim 19, wherein the bacterial cell comprises the genetic modifications: ushA and galETKM are deleted, inactivated or reduced in expression; pgi is deleted, inactivated or reduced in expression; and pgm and galU are overexpressed.

21. The method of any of claims 1 to 15, wherein the microbial cell is a yeast cell.

22. The method of claim 21, wherein the yeast is Saccharomyces spp., Schizosaccharomyces spp., Pichia spp., Phaffia spp., Kluyveromyces spp., Candida spp., Talaromyces spp., Brettanomyces spp., Pachysolen spp., Debaryomyces spp., or Yarrowia spp.

23. The method of claim 22, wherein the yeast is Saccharomyces cerevisiae, Pichia pastoris or Yarrowia lipolytica.

24. The method of any of claims 1 to 23, wherein the microbial cell expresses one or more UGT enzymes comprising the amino acid sequence of any of SEQ ID NO: 1 to 45, or a derivative thereof.

25. The method of claim 24, wherein the microbial cell expresses a 1-3' glycosylating UGT enzyme.

26. The method of claim 25, wherein the 1-3' glycosylation UGT enzyme is selected from SrUGT76G1, MbUGT1-3, MbUGT1-3_1, MbUGT1-3_1.5, MbUGT1-3_2, and derivatives thereof.

27. The method of claim 26, wherein the 1-3' glycosylation UGT enzyme is SrUGT76G1 comprising an L200A substitution.

28. The method of claim 26, wherein the 1-3' glycosylation UGT enzyme comprises an amino acid sequence that is at least 70% identical to the amino acid sequence of SEQ ID NO:15, SEQ ID NO:16 or SEQ ID NO:

17.

29. The method of any of claims 1 to 28, wherein the microbial cell expresses a 1-2' glycosylating UGT enzyme.

30. The method of claim 29, wherein the 1-2' glycosylating UGT enzyme RpLZbn / Lznza / YiAi 40 is selected from SrUGT91D2, SrUGT91D1, SrUGT91D2e, OsUGT1-2, MbUGT1,2, MbUGT1,2.2, and derivatives thereof.

31. The method of any of claims 1 to 30, wherein the microbial cell expresses a C13 O-glycosylating UGT enzyme.

32. The method of claim 31, wherein the C13 O-glycosylating UGT enzyme is SrUGT85C2 or a derivative thereof, optionally comprising a P215T substitution.

33. The method of any of claims 1 to 32, wherein the microbial cell expresses a C19 O-glycosylating UGT enzyme.

34. The method of claim 33, wherein the UGT O-glycosylating enzyme C19 is SrUGT74G1, MbUGTc19 or a derivative thereof.

35. The method of claim 24, wherein the microbial cell expresses a 1-3' glycosylating UGT enzyme and a 1-2' glycosylating UGT enzyme and, optionally, a C13 O-glycosylating UGT enzyme and a C19 O-glycosylating enzyme.

36. The method of claim 35, wherein the microbial cell expresses a 1-3' glycosylating UGT enzyme, a 1-2' glycosylating UGT enzyme, a C19 O-glycosylating UGT enzyme, and a C13 O-glycosylating UGT enzyme.

37. The method of claim 36, wherein the microbial cell expresses: an SrUGT85C2 or a derivative thereof; MbUGT1,2 or a derivative thereof; SrUGT74G1 or a derivative thereof; and an enzyme selected from SrUGT76G1 or a derivative thereof, MbUGT1-3_1 or a derivative thereof, MbUGT1-3_1.5 or a derivative thereof, and MbUGT 1-3_2 or a derivative thereof.

38. The method of any of claims 1 to 37, wherein the UGT enzymes are integrated into the chromosome of the microbial cell.

39. The method of any of claims 1 to 37, wherein the UGT enzymes are expressed extrachromosomally.

40. The method of any of claims 1 to 39, wherein the microbial cell has one or more genetic modifications that increase the availability of UDP-glucose.

41. The method of claim 40, wherein the microbial cell has a deletion, inactivation or reduced activity or expression of a gene encoding an enzyme that consumes UDP-glucose.

42. The method of claim 41, wherein the microbial cell has a deletion, inactivation or reduced activity of ushA or ortholog thereof and / or one or more of galE, gaIT, galK and galM or ortholog.

43. The method of claim 42, wherein the galETKM genes are inactivated, eliminated, or reduced in expression.

44. The method of claim 40, wherein the microbial cell has a deletion, inactivation, or reduced activity or expression of otsA (trehalose-6-phosphate synthase) or an ortholog thereof. RpLZbn / Lznza / YiAi 45. The method of claim 40, wherein the microbial cell has a deletion, inactivation or reduced activity or expression of ugd (UDP-glucose 6-dehydrogenase) or ortholog thereof.

46. ​​The method of claim 40, wherein the microbial cell has a deletion, inactivation or reduced expression of waaG, waaO, waaJ, yfdG, yfdl and / or wcaJ, or orthologs thereof.

47. The method of claim 40, wherein the microbial cell has a deletion, inactivation or reduced activity or expression of pgi (glucose-6 phosphate isomerase) or an ortholog thereof.

48. The method of claim 40, wherein the microbial cell has an overexpression or increased activity of one or more genes encoding an enzyme involved in the conversion of glucose-6-phosphate to UDP-glucose, which is optionally pgm (phosphoglucomutase) and / or galU (UTPglucose-1-phosphate uridyltransferase), or an ortholog or derivative thereof.

49. The method of claim 40, wherein the microbial cell has an overexpression or increased activity or expression of ycjU (β-phosphoglucomutase) and Bifidobacterium bifidum ugpA, or an ortholog or derivative thereof.

50. The method of claim 40, wherein the microbial cell has one or more genetic modifications that increase flux to the pentose phosphate pathway (PPP), and is optionally an overexpression or activity of E. coli zwf or a genetically modified homolog or derivative thereof.

51. The method of claim 40, wherein the microbial cell has one or more genetic modifications that increase glucose transport and optionally includes increased expression or activity of E. coli galP and E. coli glk (glucokinase), or alternatively Zymomonas mobills glf. and E. coli glk, or genetically modified homologs, orthologs, or derivatives thereof.

52. The method of claim 40, wherein the microbial cell has one or more genetic modifications that increase the production and recycling of UTP, and which optionally include increased expression or activity of E. coli pyrH (UMP kinase), E. coli cmk (cytidylate kinase), E. coli adk (adenylate kinase) or E. coli ndk (nucleoside diphosphate kinase), or genetically modified homologs, orthologs or derivatives of these enzymes.

53. The method of claim 40, wherein the microbial cell has one or more genetic modifications that increase UDP production, and which optionally include: overexpression or increased activity of upp (uracilphosphoribosyltransferase), pyrH (UMP kinase) and cmk (cytidylate kinase), or genetically modified homologs or derivatives thereof; or overexpression or increased activity of dctA (C4 dicarboxylate / orotate: H+ symporter), pyrE (orotate phosphoribosyltransferase), pyrH (UMP kinase) and cmk (cytidylate kinase), or genetically modified homologs or derivatives thereof.

54. The method of claim 40, wherein the microbial cell has one or more genetic modifications to eliminate or reduce the regulation of glucose uptake, which APIZfrO / Ί 70Z -1 / YILI 42 optionally include deletion, inactivation or reduced expression of sgrS small regulatory RNA.

55. The method of claim 40, wherein the microbial cell has one or more genetic modifications that reduce glucose-1-phosphate dephosphorylation, optionally including deletion, inactivation, or reduced expression or activity of one or more of E. coli agp (glucose-1 phosphatase), E. coli yihX (aD-glucose-1-phosphate phosphatase), E. coli ybiV (sugar phosphatase), E. coli yidA (sugar phosphatase), E. coli yigL (phosphosugar phosphatase), and E. coli phoA (alkaline phosphatase), or an ortholog thereof.

56. The method of claim 40, wherein the microbial cell has one or more genetic modifications that reduce the conversion of glucose-1-phosphate to TDP-glucose, optionally including a deletion, inactivation, or reduced expression or activity of one or more of E. coli rffH (dTDP-glucose pyrophosphorylase) and E. coli rfbA (dTDP-glucose pyrophosphorylase), or an ortholog thereof.

57. The method of claim 40, wherein the microbial cell has one or more genetic modifications that reduce the conversion of glucose-1-phosphate to ADP-glucose, and which optionally include deletion, inactivation, or reduced expression or activity of E. coli gigC (glucose-1-phosphate adenylyltransferase), or an ortholog thereof.

58. The method of claim 40, wherein the microbial cell is a bacterial cell comprising the genetic modifications: ushA and galETKM are deleted, inactivated or reduced in expression; pgi is deleted, inactivated or reduced in expression; and pgm and galU are overexpressed.

59. The method of any of claims 1 to 58, wherein the microbial cell expresses an endogenous transport protein that transports the glycoside intermediate(s) into the cell.

60. The method of any of claims 1 to 58, wherein the microbial cell overexpresses a recombinant transport protein that transports the glycoside intermediate(s) into the cell.

61. The method of any of claims 59 or 60, wherein the microbial strain expresses a transport protein selected from E. coli acrAD, xylE, ascF, bglF, chbA, ptsG / crr, wzxE, rfbX, or an ortholog or derivative thereof.

62. The method of any of claims 1 to 61, wherein the microbial cell expresses an endogenous or recombinant transporter that transports the glycosylated product out of the cell.

63. The method of any of claims 1 to 62, wherein the method results in at least a 40% conversion of the glucoside intermediate into glycosylated product by weight.

64. The method of claim 63, wherein the method results in at least a 50% conversion of the glucoside intermediate into a glycosylated product by weight.

65. The method of claim 64, wherein the method results in at least a 75 RCLZbn / Lznza / YiAi 43% conversion of the glucoside intermediate to glycosylated product by weight.

66. The method of any of claims 1 to 65, wherein a RebM to RebD ratio of at least 5:1 is recovered as a glycosylated product.

67. The method of any of claims 1 to 66, wherein the method is carried out by batch fermentation, continuous fermentation or semi-continuous fermentation.

68. The method of claim 67, wherein the method is carried out by fed-batch fermentation.

69. The method of claim 68, wherein the glycoside intermediates are incubated with the cell for about 72 hours or less.

70. A microbial cell comprising: one or more recombinant genes encoding one or more UDP-dependent glycosyltransferase (UGT) enzymes for intracellular expression; and one or more genetic modifications increasing the availability of UDP-glucose.

71. The microbial cell of claim 70, wherein the microbial cell converts the fed glycoside intermediates into a glycosylated product.

72. The microbial cell of claim 71, wherein the microbial cell is a bacterial cell.

73. The microbial cell of claim 72, wherein the bacterial cell is Escherichia spp., Bacillus spp., Rhodobacter spp., Zymomonas spp. or Pseudomonas spp.

74. The microbial cell of claim 73, wherein the bacterial cell is selected from Escheríchia cotí, Bacillus subtilis, Rhodobacter capsulatus, Rhodobacter sphaeroídes, Zymomonas mobílis or Pseudomonas putida.

75. The microbial cell of claim 74, wherein the bacterial cell is E. coli.

76. The microbial cell of claim 75, wherein the bacterial cell comprises the genetic modifications: ushA and galETKM are deleted, inactivated or reduced in expression; pgi is deleted, inactivated or reduced in expression; and pgm and galU are overexpressed.

77. The microbial cell of claim 70, wherein the microbial cell is a yeast cell.

78. The microbial cell of claim 77, wherein the yeast is Saccharomyces spp., Schizosaccharomyces spp., Pichia spp., Phaffia spp., Kluyveromyces spp., Candida spp., Talaromyces spp., Brettanomyces spp., Pachysolen spp., Debaryomyces spp., or Yarrowia spp.

79. The microbial cell of claim 78, wherein the yeast cell is selected from Saccharomyces cerevisiae, Pichia pastoris, and Yarrowia lipolytica.

80. The microbial cell of any of claims 70 to 79, wherein the microbial cell does not express a plant biosynthetic pathway that produces a glycoside intermediate.

81. The microbial cell of any of claims 70 to 80, wherein the microbial cell expresses an endogenous or recombinant transport protein that transports the RpLZbn / Lznza / YiAi 44 glucoside intermediate into the cell.

82. The microbial cell of any of claims 70 to 81, wherein the microbial cell expresses a 1-3' glycosylating UGT enzyme, which is optionally selected from SrUGT76G1, MbUGT1-3, MbUGT1-3_1, MbUGT1-3_1.5, MbUGT1-3_2, or a derivative thereof.

83. The microbial cell of claim 82, wherein the microbial cell expresses a 1-2' glycosylating UGT enzyme, which is optionally selected from SrUGT91D2, SrUGT91D1, SrUGT91D2e, OsUGT1-2, MbUGTI,2, MbUGTI,2.2 and derivatives thereof.

84. The microbial cell of any of claim 83, wherein the microbial cell expresses a C13 O-glycosylated UGT enzyme, which is optionally selected from SrUGT85C2 and its derivatives.

85. The microbial cell of claim 84, wherein the microbial cell expresses a UGT O-glycosylating C19 enzyme, which is optionally selected from SrUGT74G1, MbUGTc19 and derivatives thereof.

86. The microbial cell of claim 82, wherein the microbial cell expresses a 1-3' glycosylating UGT enzyme, a 1-2' glycosylating UGT enzyme, a C19 O-glycosylating UGT enzyme, and a C13 O-glycosylating UGT enzyme.

87. The microbial cell of claim 86, wherein the microbial cell expresses SrUGT85C2 or a derivative thereof, MbUGTI,2 or a derivative thereof, SrUGT74G1 or a derivative thereof, and SrUGT76G1 or a derivative thereof.

88. The microbial cell of claim 86 or 87, wherein the UGT enzymes are integrated into the chromosome of the microbial cell.

89. The microbial cell of claim 86 or 87, wherein the UGT enzymes are expressed extrachromosomally.

90. The microbial cell of claim 70, wherein the microbial cell has a deletion, inactivation or reduced activity or expression of a gene encoding an enzyme that consumes UDP-glucose.

91. The method of claim 90, wherein the microbial cell has a deletion, inactivation or reduced activity of ushA or ortholog thereof and / or one or more of galE, gaIT, galK and galM or ortholog.

92. The method of claim 91, wherein the galETKM genes are inactivated, eliminated, or reduced in expression.

93. The method of claim 70, wherein the microbial cell has a deletion, inactivation or reduced activity or expression of otsA (trehalose-6-phosphate synthase) or an ortholog thereof.

94. The method of claim 70, wherein the microbial cell has a deletion, inactivation, or reduced activity or expression of ugd (UDP-glucose 6-dehydrogenase) or an ortholog thereof. RCLZbn / Lznza / YiAi 95. The method of claim 70, wherein the microbial cell has a deletion, inactivation or reduced expression of waaG, waaO, waaJ, yfdG, yfdl and / or wcaJ, or orthologs thereof.

96. The method of claim 70, wherein the microbial cell has a deletion, inactivation or reduced activity or expression of pgi (glucose-6 phosphate isomerase) or an ortholog thereof.

97. The method of claim 70, wherein the microbial cell has an overexpression or increased activity of one or more genes encoding an enzyme involved in the conversion of glucose-6-phosphate to UDP-glucose, which is optionally pgm (phosphoglucomutase) and / or galU (UTPglucose-1-phosphate uridyltransferase), or an ortholog or derivative thereof.

98. The method of claim 70, wherein the microbial cell has an overexpression or increased activity or expression of ycjU (β-phosphoglucomutase) and Bifidobacterium bifidum ugpA, or an ortholog or derivative thereof.

99. The method of claim 70, wherein the microbial cell has one or more genetic modifications that increase flux to the pentose phosphate pathway (PPP), and is optionally an overexpression or increased activity of E. coli zwf or a genetically modified homolog or derivative thereof.

100. The method of claim 70, wherein the microbial cell has one or more genetic modifications that increase glucose transport and optionally includes increased expression or activity of E. coli galP and E. coli glk (glucokinase), or alternatively Zymomonas mobilis glf. and E. coli glk, or genetically modified homologs, orthologs, or derivatives of these genes.

101. The method of claim 70, wherein the microbial cell has one or more genetic modifications that increase the production and recycling of UTP, and which optionally include increased expression or activity of E. coli pyrH (UMP kinase), E. coli cmk (cytidylate kinase), E. coli adk (adenylate kinase) or E. coli ndk (nucleoside diphosphate kinase), or genetically modified homologs, orthologs or derivatives of these enzymes.

102. The method of claim 70, wherein the microbial cell has one or more genetic modifications that increase UDP production, and which optionally include: overexpression or increased activity of upp (uracilphosphoribosyltransferase), pyrH (UMP kinase) and cmk (cytidylate kinase), or genetically modified homologs or derivatives thereof; or overexpression or increased activity of dctA (C4 dicarboxylate / orotate: H+ symporter), pyrE (orotate phosphoribosyltransferase), pyrH (UMP kinase) and cmk (cytidylate kinase), or genetically modified homologs or derivatives thereof.

103. The method of claim 70, wherein the microbial cell has one or more genetic modifications to eliminate or reduce the regulation of glucose uptake, optionally including deletion, inactivation, or reduced expression of sgrS small regulatory RNA.

104. The method of claim 70, wherein the microbial cell has one or more genetic modifications that reduce glucose-1-phosphate dephosphorylation, optionally including deletion, inactivation, or reduced expression or activity of one or more of E. coli agp (glucose-1 phosphatase), E. coli yihX (aD-glucose-1-phosphate phosphatase), E. coli ybiV (sugar phosphatase), E. coli yidA (sugar phosphatase), E. coli yigL (phosphosugar phosphatase), and E. coli phoA (alkaline phosphatase), or an ortholog thereof in the microbial cell.

105. The method of claim 70, wherein the microbial cell has one or more genetic modifications that reduce the conversion of glucose-1-phosphate to TDP-glucose, optionally including a deletion, inactivation, or reduced expression or activity of one or more of E. coli rffH (dTDP-glucose pyrophosphorylase) and E. coli rfbA (dTDP-glucose pyrophosphorylase), or an ortholog thereof.

106. The method of claim 70, wherein the microbial cell has one or more genetic modifications that reduce the conversion of glucose-1-phosphate to ADP-glucose, and which optionally include deletion, inactivation, or reduced expression or activity of E. coli gigC (glucose-1-phosphate adenylyltransferase), or an ortholog thereof.