Glycosyl transferase for catalytic production of rebaudioside M and recombinant gene of glycosyl transferase

Abstract

The invention discloses glycosyl transferase for catalytic production of rebaudioside M and a recombinant gene of the glycosyl transferase. Compared with wild type glycosyl transferase, the glycosyl transferase variant disclosed by the invention has the advantages that the thermal stability and the enzyme activity are obviously improved. In a high-temperature environment of 70 DEG C, the wild type glycosyl transferase is completely inactivated, and the enzyme activity of the glycosyl transferase variant disclosed by the invention is more than 50U / mg. Meanwhile, in a high-temperature environment of 70 DEG C, the solubility of a reaction substrate rebaudioside D is improved, the thermal motion of enzyme molecules and substrate molecules is intensified, and the collision frequency and energy between the enzyme molecules and the substrate molecules are increased, so that the catalytic reaction speed can be further increased. Therefore, the glycosyl transferase variant disclosed by the invention is suitable for being used as a biocatalyst and is used for large-scale production of a sweetening agent rebaudioside M with a better taste.

Description

Technical Field

[0001] The present invention relates to the field of bioengineering, and more particularly to a glycosyltransferase for catalyzing the production of rebaudioside M and a recombinant gene thereof. Background Art

[0002] Stevioside, also known as steviol glycoside, boasts high sweetness (300-450 times that of sucrose) and low caloric value (1 / 300 of sucrose). It is a natural sucrose substitute and is known as the "world's third most potent sugar source." Stevioside is extracted from the stevia leaf, and manufacturers primarily extract stevioside (STV) and rebaudioside A (Reb A). However, STV and Reb A have a distinct bitterness and licorice aftertaste, which significantly limits the use of steviol glycosides as sweeteners in food.

[0003] Research has found that rebaudioside M (Reb M) retains the advantages of stevia's high sweetness and low caloric value while being virtually free of bitterness and licorice aftertaste, making it a high-quality sugar substitute. However, large-scale production of Reb M faces significant bottlenecks: Reb M accounts for less than 1% of stevia leaves, and the cost of producing Reb M through plant extraction is high. Furthermore, the catalytic activity of the glycosyltransferase required to catalyze the conversion of Reb D to Reb M is currently low, resulting in low production efficiency. This results in high costs for Reb M production through microbial fermentation or enzymatic methods. Therefore, improving the activity of glycosyltransferases is a pressing technical challenge. Summary of the Invention

[0004] In view of the shortcomings of the prior art, one of the objectives of the present invention is to provide a glycosyltransferase variant with an optimal reaction temperature of about 70°C and an enzyme activity of more than 40 U / mg.

[0005] To achieve the above objectives, the present invention provides the following technical solution: a thermostable glycosyltransferase, whose wild-type sequence is shown in SEQ ID NO. 1. This wild-type glycosyltransferase sequence is derived from the protein sequence library GI: 1777435608. Sequence design of the wild-type glycosyltransferase was performed using the Lésign platform, ultimately yielding a computationally optimized enzyme variant.

[0006] A glycosyltransferase for catalyzing the production of rebaudioside M. The glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO: 1 and is mutated with the following mutation set: K55N+N70T+S181P+S275R+K441E+T460K+S193R+M146R+E3V.

[0007] Another object of the present invention is to provide another glycosyltransferase variant with an optimal reaction temperature of about 70 °C and an enzyme activity of more than 40 U / mg.

[0008] To achieve the above object, the present invention provides the following technical solution: A glycosyltransferase for catalytic production of rebaudioside M, the glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO:1 and is mutated with the following mutation set:

[0009] K55N+N70T+S181P+S275R+K441E+T460K+M146R+S275G+D327E+K224E.

[0010] Another object of the present invention is to provide another glycosyltransferase variant with an optimal reaction temperature of about 70 °C and an enzyme activity of more than 40 U / mg.

[0011] To achieve the above object, the present invention provides the following technical solution: A glycosyltransferase for catalytic production of rebaudioside M, the glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO:1 and is mutated with the following mutation set:

[0012] K55N+N70T+S181P+S275R+K441E+T460K+K224E+I94F+R414H+A112S.

[0013] Another object of the present invention is to provide another glycosyltransferase variant with an optimal reaction temperature of about 70 °C and an enzyme activity of more than 40 U / mg.

[0014] To achieve the above object, the present invention provides the following technical solution: A glycosyltransferase for catalytic production of rebaudioside M, the glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO:1 and is mutated with the following mutation set:

[0015] K55N+N70T+S181P+S275R+K441E+T460K+I94F+R90L+S242I+H156Y+A180V+R175H.

[0016] Another object of the present invention is to provide another glycosyltransferase variant with an optimal reaction temperature of about 70 °C and an enzyme activity of more than 40 U / mg.

[0017] To achieve the above object, the present invention provides the following technical solution: A glycosyltransferase for catalytic production of rebaudioside M, the glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO:1 and is mutated with the following mutation set:

[0018] K55N+N70T+S181P+S275R+K441E+T460K+E3V+R414H+K224E+A112S+R90L+M146R+R298Q+A180V。

[0019] The sixth object of the present invention is to provide another glycosyltransferase variant with an optimal reaction temperature of about 70 °C and an enzyme activity of more than 40 U / mg.

[0020] To achieve the above object, the present invention provides the following technical solution: A glycosyltransferase for catalyzing the production of rebaudioside M, characterized in that the glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent, and is mutated with the following mutation set:

[0021] K55N+N70T+S181P+S275R+K441E+T460K+S193R+D100A+R298Q+K224E+T460R+R141P+R414H。

[0022] The seventh object of the present invention is to provide another glycosyltransferase variant with an optimal reaction temperature of about 70 °C and an enzyme activity of more than 40 U / mg.

[0023] To achieve the above object, the present invention provides the following technical solution: A glycosyltransferase for catalyzing the production of rebaudioside M, characterized in that the glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent, and is mutated with the following mutation set:

[0024] K55N+N70T+S181P+S275R+K441E+T460K+D100A+R414H+S275G+K224E+R405P+T264S+I94F+R90L。

[0025] The eighth object of the present invention is to provide another glycosyltransferase variant with an optimal reaction temperature of about 70 °C and an enzyme activity of more than 40 U / mg.

[0026] To achieve the above object, the present invention provides the following technical solution: A glycosyltransferase for catalyzing the production of rebaudioside M, the glycosyltransferase is based on the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent, and is mutated with the following mutation set:

[0027] K55N+N70T+S181P+S275R+K441E+T460K+R414H+T460R+R90L+A112S+N50K+S193R+E3V+H156Y+T264S+R405P+D327E+S275G。

[0028] A ninth object of the present invention is to provide a genetic material capable of expressing the above-mentioned glycosyltransferase variant.

[0029] To achieve the above object, the present invention provides the following technical solution: a recombinant genetic material of a glycosyltransferase, a DNA or RNA capable of expressing the above-mentioned glycosyltransferase.

[0030] A tenth object of the present invention is to provide a recombinant strain capable of expressing the above-mentioned glycosyltransferase variant.

[0031] To achieve the above object, the present invention provides the following technical solution: a recombinant strain of a glycosyltransferase, comprising a recombinant genetic material capable of expressing the above-mentioned glycosyltransferase.

[0032] Compared with the prior art, the advantages of the present invention are as follows: the substrate Reb D and the product Reb M catalyzed by this enzyme have poor solubility at low temperatures, and increasing the temperature can greatly promote dissolution, which is beneficial to the progress of the reaction.

[0033] Compared with the wild-type glycosyltransferase, the thermal stability and enzyme activity of the glycosyltransferase variant of the present invention have been significantly improved. In a high-temperature environment of 70 °C, the wild-type glycosyltransferase is completely inactivated, and the enzyme activity of the glycosyltransferase variant of the present invention is above 50 U / mg. At the same time, in a high-temperature environment of 70 °C, the solubility of the reaction substrate rebaudioside D increases, and the thermal motion of the enzyme molecules and substrate molecules intensifies, increasing the collision frequency and energy between them. Therefore, the catalytic reaction rate can be further increased. Therefore, the glycosyltransferase variant of the present invention is suitable as a biocatalyst for the large-scale production of rebaudioside M with a better taste. Detailed implementation manners

[0034] The term "recombinant gene" refers to a DNA or RNA capable of expressing the glycosyltransferase of the present invention. Generally, the recombinant gene is initially synthesized in vitro by the solid-phase phosphoramidite triester method or the TdT biosynthetic method or other suitable techniques known in the art. After having the template sequence, it can be amplified by PCR or other suitable techniques known in the art. After having the recombinant strain, it can be further amplified on a large scale by culturing the strain. In certain embodiments, the recombinant gene may also include residual sequences of restriction enzyme sites, other accessory elements, such as control elements (such as promoters, etc.), labeling substances (such as fluorescent labels, etc.), and other sequences that do not affect the expression of the target gene.

[0035] The term "cloning scar" refers to a promoter sequence where protein expression depends on the initiation of transcription of the starting messenger ribonucleic acid (mRNA), followed by a ribosome-binding site (RBS) that attracts the translation machinery, followed by a signal peptide sequence that aids in transporting the protein to the periplasm. The mature protein is usually cloned after the signal peptide and is cleaved from the signal peptide by signal peptidase as it passes through the membrane. However, when cloning constructs after the signal peptide, restriction endonucleases usually require specific sequences to cut DNA, leaving a cloning scar after the signal peptide sequence.

[0036] The term "signal peptide" refers to a short peptide (usually 16 - 30 amino acids long) present at the N-terminus of most newly synthesized proteins that are destined to enter the secretory pathway. It can also be referred to as a signal sequence, targeting signal, localization signal, localization sequence, transit peptide, leader sequence, or leader peptide. The signal peptide is usually cleaved from the protein by signal peptidase.

[0037] Whether it is a cloning scar, a signal peptide, or other elements in the recombinant gene, they do not affect the realization of the glycosyltransferase function. Therefore, if the amino acid sequence of the finally obtained protein differs only by the amino acid sequence corresponding to the above DNA sequence from the amino acid sequence disclosed in the present invention, it still falls within the protection scope of the present invention.

[0038] The term "signal peptide cleavage site" refers to a dipeptide between which signal peptidase cleaves the signal peptide from the mature protein. In most (but not all) cases, the dipeptide is Ala - Ala. The signal peptide cleavage site can be calculated using algorithms such as SignalP 4.1, which is available online at http:

[0039] / / www.cbs.dtu.dk / services / SignalP / (Center for Biological Sequence Analysis, Technical University of Denmark).

[0040] The term "promoter" refers to a DNA region that initiates the transcription (writing to mRNA) of a specific gene. The promoter is usually located near the transcription start site of the gene, on the same strand and upstream of the DNA (pointing to the 5' region of the sense strand). The promoter can be inducible, meaning that the expression of the gene operably linked to the promoter can be turned on by the presence of an inducer substance. Alternatively, the promoter can be constitutive, i.e., it is not regulated by any inducer substance.

[0041] The abbreviation "RBS" refers to the ribosome-binding site, or the binding site of the ribosome. This is a sequence of nucleotides upstream of the start codon of the mRNA transcript that is responsible for recruiting ribosomes during the initiation of protein translation.

[0042] The term "expression" refers to the process by which DNA is transcribed into messenger RNA (mRNA) and then translated into protein. To achieve the smooth expression and screening of glycosyltransferase, the above-mentioned signal peptide, promoter, and RBS may be introduced into the recombinant gene. Therefore, some corresponding peptide segments may remain on the expressed glycosyltransferase protein. These peptide segments do not affect the function of glycosyltransferase. Therefore, even if the above-mentioned peptide segments are present in excess in the product, as long as the amino acid sequence of the main part is the same as the sequence of the present invention, the product is still an infringing product.

[0043] The term "expression vector" has the ability to incorporate and express a heterologous polynucleotide fragment in a host cell. Many prokaryotic and eukaryotic expression vectors are commercially available. Selecting a suitable expression vector is within the knowledge of a person skilled in the art.

[0044] The term "chassis cell" refers to a suitable host vector for expressing the DNA of the present invention. The host can include any organism capable of containing and expressing the nucleic acids or genes disclosed herein, but is not limited thereto. The chassis cell can be a prokaryote or a eukaryote, unicellular or multicellular, including mammalian cells, plant cells, fungi, etc. According to the prior art, a person skilled in the art can achieve the heterologous expression of the recombinant DNA of the present invention in different disclosed chassis cells by adjusting parameters through a limited number of experiments. The chassis cell can be selected from at least one of Escherichia coli, Pichia pastoris, Saccharomyces cerevisiae, Hansenula anomala, Candida, Rhodotorula, Bacillus, Escherichia, Salmonella, Clostridium, Streptomyces, Staphylococcus, Neisseria, Shigella. The present invention only lists the types of chassis cells and does not constitute a limitation on the types of chassis cells. The chassis cell is preferably Escherichia coli, and suitable Escherichia coli strains (including many other strains) include BL21(DE3), C600, DH5αF′, 1113101, JM83, JM101, JM103, JM105, JM107, JM109, JM110, MC1061, MC4100, MM294, NM522, NM554, TGI, χ1776, XL1-Blue, and Y1089 + and so on. All of the above Escherichia coli strains are commercially available strains.

[0045] The term "identity" means that the residues in two sequences are the same when the alignment is maximally corresponding, as measured using sequence comparison or analysis algorithms such as those described herein. For example, if the corresponding segments of two sequences have the same residues at 5 out of 10 positions when correctly aligned, the two sequences are said to have 50% identity. Most bioinformatics programs report the percentage identity of the aligned sequence regions, which are usually not the entire molecule. If the alignment is long enough and contains enough identical residues, an expectation value can be calculated, which indicates that the level of identity in the alignment is unlikely to occur randomly.

[0046] The present invention will be further described in detail below in conjunction with embodiments.

[0047] Example 1

[0048] Proteins are the material basis of life and an important component of human cells and tissues. All important components in the human body require the participation of proteins, which play a very important role in the life activities of cells and organisms. It can be said that without proteins, there is no life. There are very many types of proteins in the human body, and their functions are also different. Some form human tissues, some can provide energy, some can participate in material metabolism and transport, some promote growth and development, and regulate immune functions. Different proteins undertake different responsibilities and functions, and their functions are determined by the structure of the proteins. And the 3D structure of proteins is determined by the amino acid sequence of proteins. Therefore, the design of proteins depends on the correspondence between structure and sequence. To design proteins with specific functions, sequences that conform to the functional structure need to be designed. Understanding and designing proteins is of great significance for promoting innovation and progress in biology and medicine.

[0049] Designing protein sequences for a specific function is a very difficult task. It is difficult to predict what structure and function the designed sequences will finally present. Moreover, the sample space of protein sequences with a fixed length is also very large. To complete the above work, Liwen developed a protein design platform based on deep learning algorithms - Lésign. This platform realizes functions such as protein structure prediction, sequence design, and result evaluation. Each functional module collaborates through interfaces to form a computational pipeline integrating prediction, design, and evaluation.

[0050] Using the Lésign platform, sequence design was carried out on wild-type glycosyltransferase (the amino acid sequence is shown in SEQ ID NO.1), and finally the optimal enzyme variant at the computational level was obtained.

[0051] 1. Preparation of glycosyltransferase

[0052] 1.1 Construction of recombinant cells:

[0053] The nucleotide sequences of the target genes were all synthesized by Beijing Tsingke Biotechnology Co., Ltd., and the nucleotide sequence of the glycosyltransferase was inserted into an expression vector. Specifically, it was inserted into the plasmid pET28a(+) to obtain the corresponding plasmid. Subsequently, the synthesized plasmid was transferred into the chassis cell (E. coli BL21(DE3)), thereby constructing an E. coli strain containing the plasmid. There are many other available plasmids and chassis cells in the prior art, and this example only provides a specific scheme.

[0054] 1.2 Expression and purification of glycosyltransferase:

[0055] The recombinant bacteria were inoculated into a shake flask at 1% and cultured overnight, and then inoculated into a fermenter at 1%. The fermenter contained liquid TB medium and was cultured until the OD 600 value was about 6.0, then IPTG was added to a concentration of 100 μM, and the temperature was reduced to 16 °C and cultured overnight. After overnight culture, the cells were collected by centrifugation at 4000g for 10 min and washed with 0.1 M PB buffer (pH 7.0). Next, the cells were lysed by sonication, and then centrifuged at 12000 rpm for 1 h. The supernatant was the crude enzyme solution. There are many other available methods for inducing the expression of recombinant bacteria in the prior art, and this example only provides a specific scheme.

[0056] 1.3 The crude enzyme solution was purified by nickel column. The crude enzyme solution was eluted under different elution conditions to obtain the target protein. According to the prior art, those skilled in the art can adjust the parameters through a limited number of experiments to achieve the purification of glycosyltransferase, and specific descriptions are not provided here. There are many other available methods for purifying glycosyltransferase in the prior art, and this example only provides a specific scheme.

[0057] The wild-type glycosyltransferase (ORM, SEQ ID NO: 1) and glycosyltransferase variants were heterologously expressed and purified according to the above preparation method of glycosyltransferase. The prepared glycosyltransferase and mutants were verified by gel electrophoresis (SDS-PAGE) to determine that the sizes of the glycosyltransferase and mutants were consistent with the expectations.

[0058] 2. Enzyme activity assay:

[0059] 3.1 Catalytic reaction system: The total volume of the reaction system for enzyme activity determination is 0.5 mL, which contains 10 mM of substrate RD, 0.002 mg / mL of glycosyltransferase or variant, 1 mM of uridine diphosphate glucose (UDP-G) or adenosine diphosphate glucose (ADP-G) or guanosine diphosphate glucose (GDP-G) or cytidine diphosphate glucose (CDP-G), 3 mM of magnesium chloride, and the volume is made up to 0.5 mL with glycine-sodium hydroxide buffer (0.1 M) at pH 9.0.

[0060] 3.2 Method for determining the optimum reaction temperature:

[0061] The reaction is carried out according to the catalytic reaction system in 3.1. The reaction temperatures of the wild-type glycosyltransferase are 25 °C, 30 °C, 35 °C, 40 °C, 45 °C, 50 °C, 55 °C, 60 °C respectively, and the reaction temperatures of the glycosyltransferase variant are 50 °C, 55 °C, 60 °C, 65 °C, 70 °C, 75 °C respectively. The reaction time is 1 h for all, and the reaction conditions are 1000 rpm / min.

[0062] After the reaction, the concentration of RM in the reaction solution is detected by HPLC: LC-2030C HT system (SHIMADZU, Japan), mobile phase ratio: 25% CH3CN for 0 - 3 min, 25% - 50% CH3CN for 3 - 8 min, 50% - 100% CH3CN for 8 - 12 min, 100% CH3CN for 12 - 15 min, 25% CH3CN (acetonitrile contains 0.1% HCOOH) for 15 - 20 min, combined with another mobile phase H2O (containing 0.1% HCOOH). The flow rate is 1 ml / min, the wavelength is 210 nm, and the chromatographic column is Shim-pack GIST C18 (250×4.6 mm, 5 μm), and the column temperature is 30 °C. The liquid phase detection peak diagram is obtained, and the concentration of RM is calculated according to the peak area.

[0063] Definition of enzyme activity: The amount of enzyme required to catalyze the formation of 1 μM Reb M per minute. The enzyme activities of the wild-type glycosyltransferase and the glycosyltransferase variant measured are shown in Tables 1 - 9:

[0064] Table 1 Enzyme activities of wild-type glycosyltransferase at different reaction temperatures

[0065]

[0066]

[0067] Table 2 Enzyme activities of glycosyltransferase variant ORM-2 at different reaction temperatures

[0068]

[0069] Table 3 Enzyme Activity of Glycosyltransferase Variant ORM-3 at Different Reaction Temperatures

[0070]

[0071] Table 4 Enzyme Activity of Glycosyltransferase Variant ORM-4 at Different Reaction Temperatures

[0072]

[0073] Table 5 Enzyme Activity of Glycosyltransferase Variant ORM-5 at Different Reaction Temperatures

[0074]

[0075] Table 6 Enzyme Activity of Glycosyltransferase Variant ORM-6 at Different Reaction Temperatures

[0076]

[0077] Table 7 Enzyme Activity of Glycosyltransferase Variant ORM-7 at Different Reaction Temperatures

[0078]

[0079] Table 8 Enzyme Activity of Glycosyltransferase Variant ORM-8 at Different Reaction Temperatures

[0080]

[0081]

[0082] Table 9 Enzyme Activity of Glycosyltransferase Variant ORM-9 at Different Reaction Temperatures

[0083]

[0084] As can be seen from Tables 1-9, the optimal reaction temperature of the wild-type glycosyltransferase is about 30 °C, and its thermal stability is poor. When the reaction temperature reaches 50 °C, the enzyme activity is only about 60% of the highest value, and when the environmental temperature reaches 60 °C, it is completely inactivated. Even at the optimal reaction temperature, the enzyme activity of the wild-type glycosyltransferase is low, only 0.24 U / mg, far from meeting the enzyme activity required for industrial production. The optimal reaction temperature of the glycosyltransferase variant is about 70 °C, and its thermal stability is very good. At a reaction temperature of about 70 °C, the enzyme activity of the glycosyltransferase variant of the present invention is above 50 U / mg, and the enzyme activity has been significantly improved, which is suitable for industrial production.

[0085] The above are only the preferred embodiments of the present invention, and the protection scope of the present invention is not limited to the above embodiments. All technical solutions falling within the concept of the present invention belong to the protection scope of the present invention. It should be noted that for those of ordinary skill in the art, several improvements and refinements made without departing from the principle of the present invention should also be regarded as within the protection scope of the present invention.

Claims

1. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+S193R+M146R+E3V.

2. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+M146R+S275G+D327E+K224E.

3. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+K224E+I94F+R414H+A112S.

4. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+I94F+R90L+S242I+H156Y+A180V+R175H.

5. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+E3V+R414H+K224E+A112S+R90L+M146R+R298Q+A180V.

6. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+S193R+D100A+R298Q+K224E+T460R+R141P+R414H.

7. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+D100A+R414H+S275G+K224E+R405P+T264S+I94F+R90L.

8. A glycosyltransferase for catalytic production of rebaudioside M, characterized in that The glycosyltransferase uses the wild-type glycosyltransferase described in SEQ ID NO:1 as the parent and is mutated with the following mutation sets: K55N+N70T+S181P+S275R+K441E+T460K+R414H+T460R+R90L+A112S+N50K+S193R+E3V+H156Y+T264S+R405P+D327E+S275G.

9. A recombinant genetic material of a glycosyltransferase, characterized in that DNA or RNA capable of expressing the glycosyltransferase according to any one of claims 1 to 8.

10. A recombinant strain of a glycosyltransferase, characterized in that Recombinant genetic material comprising the glycosyltransferase according to claim 9.

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