Coding gene of fucosyltransferase and application thereof

Abstract

The invention discloses a coding gene of fucosyltransferase and application of the coding gene. A nucleotide sequence of the fucosyltransferase coding gene comprises (1) a nucleotide sequence of SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16 or SEQ ID NO: 18; and (2) nucleotide sequences and the like for coding the amino acid sequences of SEQ ID NO: 1, SEQ ID NO: 3, SEQ ID NO: 5, SEQ ID NO: 7, SEQ ID NO: 9, SEQ ID NO: 11, SEQ ID NO: 13, SEQ ID NO: 15 or SEQ ID NO: 17. According to the invention, codon optimization is carried out on the coding gene of the fucosyltransferase, so that the fucosyltransferase with heterologous expression can be obtained, the fucosyltransferase effectively catalyzes the reaction, and the yield of the 3-fucosyllactose is increased.

Description

Technical Field

[0001] The present invention belongs to the field of biotechnology and relates to a coding gene of fucosyltransferase and its application. Background Art

[0002] Human milk oligosaccharides (HMOs) are the third largest solid component in human milk (with a content of 5 - 15 g / L), second only to lactose and lipids. In recent years, HMOs, with 3-fucosyllactose (3-FL) as one of the typical representatives, have received extensive attention. It has various physiological functions for infants, including preventing pathogens from adhering to epithelial cells, reducing infant diarrhea, and promoting the development of intestinal flora, etc. Compared with sialylated and neutral HMOs, fucosylated HMOs account for the highest proportion in total HMOs. Among them, 2'-fucosyllactose (2'-FL) accounts for about 31%, while 3-FL accounts for about 5%. As a form of fucosylated HMOs, 3-FL is composed of β-lactose and L-fucose, where L-fucose is linked to the glucose unit in lactose through an α-1,3 bond. Although the proportion of 3-FL in HMOs is relatively small, as a prebiotic, it still plays an important role in preventing infant infectious diseases and inhibiting the growth of harmful microorganisms.

[0003] The synthesis methods of 3-fucosyllactose mainly include chemical synthesis, enzymatic synthesis, and biosynthesis. Chemical synthesis methods usually face some problems due to their complex synthesis steps and harsh reaction conditions. In enzymatic synthesis, the synthesis of 3-FL usually requires guanosine 5'-diphosphate (GDP)-L-fucose as the donor substrate, β-lactose as the acceptor substrate, and is catalyzed by α-1,3-fucosyltransferase (α-1,3-FucT). However, the cost of GDP-L-fucose is higher than that of 3-FL itself, and the benefit of this method is lower than the cost. In recent years, the research on producing various human milk oligosaccharides using metabolic engineering technology has gradually emerged. The most successful example is the commercial production of 2'-fucosyllactose by engineered Escherichia coli. However, Escherichia coli cannot directly synthesize 3-FL because it lacks endogenous α-1,3-fucosyltransferase for 3-FL synthesis in the cell. By analyzing the metabolic network, Escherichia coli can synthesize the precursor substance GDP-L-fucose of 3-FL from mannose-6-phosphate through a series of reactions catalyzed by phosphomannomutase, mannose-1-phosphate guanylyltransferase, GDP-D-mannose-4,6-dehydratase, and GDP-L-fucose synthase. This discovery provides a potential possibility for producing 3-FL using metabolic engineering technology.

[0004] α-1,3-fucosyltransferase (α-1,3-FucT) can transfer the fucosyl group in GDP-L-fucose to β-lactose, and then produce 3-FL. Dumon et al. evaluated α-1,3-FucT from Helicobacter pylori - FutA (GenBank: AAD07447.1), and found that when the gene encoding FutA was overexpressed in Escherichia coli, 0.5 g / L of 3-FL could be produced. To improve the soluble expression of α-1,3-FucT, the researchers systematically truncated and extended the C-terminus of three α-1,3-FucTs from Helicobacter pylori. The results showed that when the membrane-anchoring region of the transferase was removed and a longer repeating unit was retained at the C-terminus, the protein expression level could be significantly increased, and the yield of 3-FL could be increased by 10 to 20 times. In addition, the quadruple mutant of Helicobacter pylori FutA (S46F / A128N / H129E / Y132I) was also applied to metabolically engineered Escherichia coli to produce 3-FL, and the yield reached 4.6 g / L. FutM2 from Bacteroides gallinaceum (NCBI No.: WP_204430034.1), FucTa from Helicobacter pylori (UniProt ID: Q9L8S4), azoT from Azospirillum brasilense (NCBI No.: WP_040134538.1), Fut3Bc from Neobacillus cucumis (NCBI No.: WP_101650151.1), Amuc0760co from Akkermansia muciniphila (UniProt ID: B2UQ61) and other α-1,3-FucTs have also been successfully used in the production of 3-FL. However, there are still some deficiencies in the production of 3-FL: one is that the yield of 3-FL needs to be urgently improved, and the other is that there is still a lack of efficient α-1,3-FucT.

[0005] Therefore, there is an urgent need to provide a novel α-1,3-FucT to achieve the efficient production of 3-FL and effectively increase the yield of 3-FL. Summary of the Invention

[0006] In view of the deficiencies of the prior art and actual needs, the present invention provides a coding gene for fucosyltransferase and its application. By optimizing the codons of the coding gene for fucosyltransferase, the present invention can obtain a heterologously expressed fucosyltransferase, and the fucosyltransferase effectively catalyzes the reaction, increasing the yields of 3-fucosyllactose and 2'-fucosyllactose.

[0007] To achieve the object of this invention, the following technical solutions are adopted in the present invention:

[0008] In the first aspect, the present invention provides a coding gene for fucosyltransferase. The nucleic acid sequence of the coding gene for fucosyltransferase includes any one or a combination of at least two of the following sequences:

[0009] (1) Any one or a combination of at least two of the sequences shown in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16 or SEQ ID NO:18;

[0010] (2) A nucleic acid sequence encoding the amino acid sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 or SEQ ID NO:17;

[0011] (3) A nucleic acid sequence obtained by substituting, deleting or adding one or more nucleotides to the sequence shown in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16 or SEQ ID NO:18 and encoding a fucosyltransferase with activity;

[0012] (4) A nucleic acid sequence having at least 80% sequence homology with the nucleic acid sequence described in (1), (2) or (3) and having the same or similar function.

[0013] By optimizing the codons of the coding gene for fucosyltransferase, the present invention can obtain a heterologously expressed fucosyltransferase, and the fucosyltransferase effectively catalyzes the reaction, increasing the yield of 3-fucosyllactose.

[0014] Second aspect, the present invention provides a fucosyltransferase encoded by the fucosyltransferase encoding gene described in the first aspect, and the amino acid sequence of the fucosyltransferase includes any one or a combination of at least two of the following sequences:

[0015] (1) Any one or a combination of at least two of the sequences shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 or SEQ ID NO:17;

[0016] (2) An amino acid sequence with fucosyltransferase activity formed by substituting, deleting or adding one or more than two amino acids to the amino acid sequences shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 or SEQ ID NO:17;

[0017] (3) An amino acid sequence having at least 80% sequence homology with the amino acid sequence described in (1) or (2) and having the same or similar functions.

[0018] Third aspect, the present invention provides a genetically engineered strain that simultaneously expresses fucosyltransferase and lactose permease, and the genetically engineered strain contains the fucosyltransferase encoding gene described in the first aspect.

[0019] Preferably, the genetically engineered strain includes any one of Corynebacterium glutamicum, Escherichia coli, Bacillus subtilis or Saccharomyces cerevisiae.

[0020] Fourth aspect, the present invention provides a preparation method of the genetically engineered strain described in the third aspect, and the preparation method includes: transducing a recombinant vector containing the fucosyltransferase encoding gene described in the first aspect into a Corynebacterium glutamicum strain to obtain the genetically engineered strain.

[0021] Fifth aspect, the present invention provides the application of the fucosyltransferase encoding gene described in the first aspect in the preparation of fucosyllactose.

[0022] Preferably, the fucosyllactose includes 3-fucosyllactose and / or 2'-fucosyllactose.

[0023] Sixth aspect, the present invention provides a method for preparing fucosyllactose, and the method includes: overexpressing the fucosyltransferase encoding gene described in the first aspect to obtain the fucosyllactose.

[0024] Preferably, the method for overexpressing the fucosyltransferase-encoding gene includes introducing the fucosyltransferase-encoding gene described in the first aspect into a free expression vector of a host cell or integrating the fucosyltransferase-encoding gene described in the first aspect into the chromosome of the host cell.

[0025] For Corynebacterium glutamicum, free vectors that can be used include but are not limited to pBL1, pEKEx1, pEKEx2, pXMJ19, pJC1, pHM1519, pVWEx1, pZ8-1, pECTAC-K99, pECTAC-XK99E, pECTAC-XC99E, pECTAC-XT99A, pNG2, pAPE12 plasmid vectors or plasmid vectors based on them. For integration into the chromosome of the host cell, it can be integrating the fucosyltransferase-encoding gene into the chromosome using a suicide plasmid vector based on pK18mobsacB or based on it, or integrating the fucosyltransferase-encoding gene into the chromosome using the CRISPR / Cas9 system.

[0026] The polypeptide sequence (SEQ ID NO:1) of α-1,3-fucosyltransferase derived from Candidatus Akkermansia timonensis:

[0027] MKTLKISFLQSTPDFGKESFRRLLQDRYHVEENDSDFDYLVATPWFYVNREAFYDFLERAPGHVTIMYGCHEAIAPDFMLFDYYIGVDAIPYYDRTVKLPSLRPHLQEVHGAKEGLNAENLLAAKTGFCNFIYANRKSHPNRDAMFHKLSGYKFVNSLGSHLNNTPGDGRRAEDWYASSIEMKKPYKFSIAFENAWYPGYTSEKIVTSMLAGTIPIYWGNPDIGKEFNTLSFINCHEFATLDDAVAYVKKVDENDDLWCEIMSRPWKTPEQETLFLEETARETEKLYRIFDRPPEEAHRKGDGTWISYYQRFLKRGHKGRLAWHSLKSRRRK.

[0028] The gene sequence (SEQ ID NO:2) of α-1,3-fucosyltransferase derived from Candidatus Akkermansia timonensis:

[0029] atgaagaccctgaagatttcatttttgcagtccactccggattttggtaaggagtcctttcgccgcctgctgcaggatcgctatcatgtggaggagaacgatagcgatttcgattacttggttgcgaccccgtggttctatgtcaaccgagaagccttttatgacttcctcgaacgcgctccaggccacgtgaccatcatgtatggctgccacgaggcgattgcgccggatttcatgctgttcgattattacatcggcgttgatgcaatcccgtattacgatcgaacggtgaagcttccaagccttcgccctcacctgcaggaggttcacggcgctaaagagggtctgaatgcagaaaacctccttgccgcgaagacaggcttttgcaacttcatttacgctaaccgcaagtctcaccccaaccgcgatgcgatgttccacaagctgtccggctacaagttcgttaactctctgggctcccatcttaacaatacccctggtgacggccgtcgcgcggaagattggtacgcaagctctatcgagatgaaaaagccgtataaattctctattgcatttgagaacgcgtggtatccaggctacacctccgagaaaatcgtcacatccatgcttgcaggtaccatcccgatttattggggaaaccccgacatcggcaaggaatttaacacgctttcctttatcaactgccacgaattcgcgaccctggacgacgccgttgcttacgttaaaaaagttgacgaaaatgatgacttgtggtgcgaaatcatgtctcgaccatggaaaaccccagagcaagagaccctcttcctggaagaaaccgcacgcgagacagagaaactgtaccgcatcttcgaccgtccccctgaagaggctcaccgaaagggtgatggcacctggatctcctactaccagcgattcctgaaacgaggtcacaagggtcgcttggcttggcattccctgaaatcccgccgccgtaagtaa。

[0030] Polypeptide sequence (SEQ ID NO: 3) of an α-1,3-fucosyltransferase from Akkermansia sp.:

[0031] MKTLKISFLQSTPDFGREGIYQLLKNRYRVVEDDSDFDYLIATPWFYVNREAFYDFLERAPGHITVMYGCHEAVAPDFMLFDYYIGLDTVPGSDRTVKLPFLRHHLQEVHGGKEGLDVRALLASKTGFCNFIYANRKSHPNRDAIFHKLSAFRFVNSLGPHLNNTPGDGHRSEDWYASSIRMKKPYKFSIAFENAWYPGYTSEKIVTSMLAGTIPIYWGNPDIGREFNSAAFINCHDFPTLDDAAAYVKKVDEDDGLWCEIMSRPWKTPEQEALFLEETERETAKLYRIFDQSPEEARRKGDGTWISYYQRFLKRGHRLRLAWRRLKNRLSR。

[0032] Gene sequence (SEQ ID NO: 4) of an α-1,3-fucosyltransferase from Akkermansia sp.:

[0033] atgaaaaccctcaagatctcgttcctgcagtccactccagatttcggtcgtgagggcatctaccaactgttgaagaatcgttaccgagtcgttgaagatgatagcgacttcgattacctcattgcaaccccctggttctatgtgaatcgcgaagccttctacgacttcctggaacgcgcaccaggccatatcaccgtcatgtacggttgccacgaggcggtagcaccagatttcatgctcttcgattactatattggtctggatacggttcccggttccgatcgcaccgtgaagctgcccttcctgcgtcatcacttgcaggaagtgcacggaggaaaggagggactggacgtccgcgcgttgctcgcgtcaaagaccggtttttgcaacttcatctatgcgaatcgtaagtctcatcccaaccgagacgcgatcttccataaactgtccgcgttccgattcgtcaactcgctcggtccgcacttgaacaataccccgggtgacggacaccgatcggaagattggtacgcatccagcattcgcatgaagaaaccgtataagttctccatcgcctttgaaaacgcgtggtatccgggttataccagcgagaagatcgtcacctctatgctggcgggaacaattccaatctactggggcaacccagatatcggccgcgagtttaactccgcggccttcattaattgtcacgactttcccaccctggacgatgccgcagcttacgtgaagaaggtggacgaagacgatggcctgtggtgtgagattatgtctcgtccatggaagacgcccgagcaggaagccctcttcctcgaggagaccgagcgcgaaaccgccaaactctaccgcatttttgaccagtcgccggaagaggcacgccgcaagggtgatggtacctggatttcatactaccaacgtttccttaaacgcggccaccgactccgacttgcttggcgccgacttaaaaatcgattgtcacgctaa。

[0034] Polypeptide sequence (SEQ ID NO: 5) of an α-1,3-fucosyltransferase derived from Akkermansia sp.:

[0035] MKTLKISFLQSTPDFGREGIHQLLKDRYHVVEDDSDFDYLVATPWFYVNREAFYDFLERAPGHITIMYGCHEAIAPDFMLFDYYIGLDTVPGSDRTVKLPFLRHHLQEVHGTKEGLDVRALLASKTAFCNFIYSNRKSHPNRDAIFHKLSACQFVNSLGPHLNNTPGDGHQEEDWYTSSIRMKKPYKFSIAFENAWYPGYTSEKIVTSMLAGTIPIYWGNPDIGLEFNSGAFINCHDFPTLDDAVAYVKKVDEDDDLWCEIMSRPWKTPEQEARFLEETARETAKLYRIFDQSPEEARRKGDGTWISYYQRFLKRGHRLRLAWRRLKNRLRRQ。

[0036] Gene sequence (SEQ ID NO: 6) of an α-1,3-fucosyltransferase derived from Akkermansia sp.:

[0037]

[0038] Polypeptide sequence (SEQ ID NO: 7) of an α-1,3-fucosyltransferase from Akkermansia sp.:

[0039] MKTLKISFLQSTPDFGKEGIRHLLEDRYHIEESDSDFDYLIATPWFYVNREAFYDFLERSPGHLTIMYGCHEAIAPDFMLFDYYIGVDAIPASERTVKLPSLRHHLQEVHGAKKGLNAENLLAAKTGFCNFIYANRKSHPNRDAMFHKLSSYKFVNSLGSHLNNTPGDGLRTGDWYASSIRMKKPYKFSIAFENAWYPGYTSEKLVTSMLAGTIPIYWGNPDIGREFNSSSFINCHDFPTLDDAVAYVKKVDEDNGLWCEIMSRPWKTPEQEILFLEETARETEKLYRIFDQAPEEAHRKGDGTWISYYQRFLKRGHKGRLAWHSLKSRLHQ。

[0040] Gene sequence (SEQ ID NO: 8) of an α-1,3-fucosyltransferase from Akkermansia sp.:

[0041] atgaaaacgcttaagatctccttcctccagtctacaccagactttggcaaagaaggtatccgccacttgcttgaagatcgttaccacattgaagaatccgactcggactttgactaccttattgcaacaccctggttttatgtcaatcgcgaggctttctacgacttcctggaacgcagcccgggccacctgaccattatgtacggctgtcatgaggcaatcgcgcccgactttatgcttttcgactactacattggagtggacgcgattcccgcatcagaacgaacggtgaagctgccgtcgttgcgccaccatctccaagaggtccacggcgcaaagaagggtctcaacgctgaaaacctcttggctgctaaaaccggtttctgtaacttcatttacgccaaccgcaagtcccaccccaaccgtgacgccatgttccataaactctcgtcctataagttcgttaattcccttggctctcatctgaacaatacccccggagacggcctccgaaccggtgattggtacgcgtccagcattcgcatgaaaaaaccatataaattttcgatcgccttcgaaaatgcgtggtacccaggttatacttcagaaaagcttgtcacatctatgcttgcaggcactatcccaatttactggggcaaccctgatattggacgtgaatttaattcatcatcttttatcaactgccacgacttcccaacgctggatgatgcggtcgcctatgttaaaaaggtcgatgaggacaacggcctctggtgcgaaatcatgtcgcgtccgtggaagacccccgaacaggagattctgtttctcgaggaaaccgctcgcgagaccgaaaagctctaccgcattttcgatcaggctcctgaggaagcacaccgcaagggagacggtacctggatttcttactaccagcgcttcctcaaacgcggccataagggtcgcttggcttggcactctctcaagtcccgcctgcaccagtaa。

[0042] Polypeptide sequence of α-1,3-fucosyltransferase derived from Akkermansiaceae bacterium (SEQ ID NO:9):

[0043] MKTLKISFLQSTPDFGKEDFLRLLQDRYHVEENDSDFDYLVATPWFYINREAFYDFLERSPGHVTIMYGCHEAIAPDFMLFDYYIGVDAIPSSERTVKLPSLRHHLQEVHGAKEGLNAENLLAAKTGFCNFIYANRKSHPNRDAMFHKLSGYKFVNSLGSHLNNTPGDGHRAGDWYASSIQMKKPYKFSIAFENAWYPGYTSEKIVTSMLAGTIPIYWGNPDIGREFNTLSFINCHEFPTLDHAVAYVKKVDENDDLWCEIMSRPWKTPEQEALFLEETARETEKLYRIFDRLPEKAHRKGNGTWISYYQRFLKRGHKSRLAWHSLKSRRRK。

[0044] Gene sequence of α-1,3-fucosyltransferase derived from Akkermansiaceae bacterium (SEQ ID NO:10):

[0045] atgaagactttgaaaatttctttccttcaatccacgcccgatttcggcaaggaagatttcttgcgtctgctccaagatcgttaccacgtggaggagaatgattcggatttcgattaccttgtggcgacgccctggttctacattaatcgtgaggctttctatgatttcctcgaacgctcacccggtcacgtgacaattatgtatggttgccacgaggccatcgcccctgactttatgctgttcgattactacattggcgtggatgcgatcccgtcctccgaacgaacggttaagttgccgtccctgcgtcaccaccttcaagaggtccatggagctaaggagggtctgaatgctgaaaacctgctggcagccaagaccggcttctgtaactttatctacgcgaatcgaaagtcccacccgaatcgcgatgcaatgtttcacaagctctcaggatataaattcgtaaattctctcggtagccacttgaacaacacccccggtgatggtcaccgagcaggcgattggtacgcatcgtcgatccaaatgaagaagccatataagttctcaattgcgttcgagaacgcgtggtacccaggttacacctcggaaaaaatcgtgactagcatgttggcaggtaccattcccatttactggggaaacccagacattggccgcgagtttaacaccctgtccttcattaactgccatgaattcccaacccttgatcacgcagttgcttacgttaagaaagtggatgagaacgatgatctctggtgcgagatcatgtcgcgaccgtggaagactcctgaacaggaagccctgttcctggaagaaacggcccgcgagacggagaaactgtatcgaatctttgaccgcttgcctgagaaagcccatcgaaagggcaacggcacctggatctcctattaccagcgtttcttgaagcgtggtcacaagtcacgcctcgcttggcacagcctcaaatcacgccgccgcaaataa。

[0046] Polypeptide sequence (SEQ ID NO: 11) of an α-1,3-fucosyltransferase derived from Akkermansia sp.:

[0047] MKTLKISFLQSTPDFGREGILQLLKDRYHVVEDNTDFDYLIATPWFYVNREAFYDFLERAPGHITVMYACHEAIAPDFMLFDYYIGLDTVPGSGRTVKLPFLRHHLQEVHGSKEGLDARALLASKTGFCNFIYSNRKSHSNRDAIFHKLSAFRFVNSLGPHLNNTPGDGHQSEDWYSSSIRMKKPYKFSIAFENAWYPGYTSEKIVTSMLAGTIPIYWGNPDIGREFNSAAFINCHDFPTLDDAVAYVKKVDEDDGLWCEIMSRPWKTPEQEALFLEETERETAKLYRIFDQSPEEARRKGDGTWISYYQRFLKRGHRLRLAWRRLKKRLRK。

[0048] Gene sequence (SEQ ID NO: 12) of an α-1,3-fucosyltransferase derived from Akkermansia sp.:

[0049] atgaaaacgcttaaaatctccttccttcagagcacgccagatttcggacgagaaggtatccttcagcttttgaaagatcgctaccatgtcgttgaagacaacaccgattttgattacttgatcgccactccttggttttacgtgaaccgagaagcgttctacgatttccttgagcgcgccccaggccacattaccgtcatgtacgcgtgccatgaagccatcgctcctgatttcatgctgttcgattactatattggactcgacactgttcccggttccggtcgaaccgttaaactcccgttcttgcgtcatcacttgcaggaggtccatggctccaaagaaggattggacgcacgagcacttcttgcgtctaagaccggcttctgcaacttcatttactctaaccgaaagtcccattcaaaccgcgatgcaatctttcacaagttgtcggccttccgctttgtaaacagcctcggccctcaccttaacaacaccccaggcgatggacaccaaagcgaagactggtacagctcatccatccgtatgaaaaaaccatacaagttctccatcgcctttgaaaacgcttggtatcctggatatacctccgagaagattgtcacctccatgctggccggcactatcccaatctactggggcaacccggatatcggccgtgaatttaattcagcggctttcattaactgtcatgacttcccaactttggacgacgcagtggcatacgtgaaaaaagttgatgaagatgatggcttgtggtgcgaaattatgtctcgtccatggaagacaccagagcaggaggcgctgttcctggaagaaaccgaacgcgagacagcgaagctgtaccgtattttcgatcaatcccccgaggaagcacgccgtaagggcgacggcacttggatctcctactaccagcgatttctcaaacgcggccaccgccttcgtctggcctggcgccgtctcaaaaaacgcctccgcaagtaa。

[0050] Polypeptide sequence (SEQ ID NO:13) of α-1,3-fucosyltransferase derived from Azospirillum oleiclasticum:

[0051] MIDPRTAAALQEFLANPGLDPSLLDLFLLFGPSYGQRQTRAPLTVAFHDFWPEFDKRENFFTEILSHRFAVTVVEDGCDLAIVSVFGTRHREMRARRSLYFTGENRRPPLDAFDMAVSFDRIDDPRHFRLPLYVMHAFEHMREGATAHFCQPLLPPVPPTRDEFAARKFCAFLYKNPHGTRRNDFFQALHARRHVDSVGWHLNNTGSVVKMGWLPKIRVFAKYRFAFAFENSSHPGYLTEKILDVFQAGAVPLYWGDPDVTRDVVPGSFIDVSRFADDEAAVAAILALDDDYDAYRRCRGVSPFVGAPDFHFDAYRLAEFIESRL。

[0052] Gene sequence (SEQ ID NO:14) of α-1,3-fucosyltransferase derived from Azospirillum oleiclasticum:

[0053] atgatcgacccacgcaccgctgctgctttgcaggaatttctggccaaccctggcttggacccgtcactcctcgacctttttttgctgtttggcccttcctacggtcagcgccagacccgagcaccattgaccgtggcctttcacgacttctggcctgagttcgataagcgcgaaaatttttttaccgaaatcctgtcgcaccgattcgccgtgactgtggtagaagatggctgcgaccttgcaattgtctctgtcttcggcacccgtcatcgcgagatgcgcgcccgccgctccctctacttcacaggcgagaatcgccgtcccccgctggatgctttcgatatggcggtctccttcgaccgcatcgacgacccacgccacttccgtcttccgctttacgtgatgcacgcctttgagcacatgcgtgagggcgcgactgcgcacttctgccagccgctccttccaccggtgcctccaactcgtgacgagttcgcagcgcgcaagttctgcgcgttcctgtacaagaatccccatggcactcgccgcaatgatttttttcaggctcttcacgcccgacgccacgtcgatagcgtcggctggcacctgaacaatacgggctcggtcgtcaaaatgggatggctgccgaaaattcgcgtgttcgcgaaatatcgtttcgcgtttgcgtttgagaactccagccacccaggctacctcaccgagaaaatcctggacgttttccaggctggcgccgtccccctctactggggcgaccccgatgtaactcgagatgtggtccctggcagcttcatcgatgtgtcccgcttcgctgacgacgaggcggcggtagcggcaatccttgctctcgatgacgattacgacgcgtatcgccgttgccgaggtgtttccccgttcgtgggtgcccccgacttccacttcgatgcgtatcgactggccgaatttatcgaatctcgtctctaa。

[0054] Polypeptide sequence (SEQ ID NO:15) of α-1,3-fucosyltransferase derived from Roseomonas genomospecies:

[0055] MIDRRTSAFLADFLSKPGGDPDRLDRFLLHGPYRARRGGRPRLKLAFHDFWPEFDKGTNFFIEILSSRFDLSVVEDDSDLAIVSVFGGRHREARSRRSLFFTGENVRPPLDGFDMAVSFDRIDDPRHFRLPLYVMHAYEHMREGAVPHFCSPVLPPVPPTRAEFAERKFCAFLYKNPNGERRNRFFPALDGRRRVDSVGWHLNNTGNVVKMGWLAKIRVFERYRFAFAFENASHPGYLTEKILDVFQAGAVPLYWGDPDLTREVAAGSFIDVSRFATDEEAVEHILAVDDDHDAYCAHRAVPPFLGTEEFHFDAYRLADWIESRL.

[0056] Gene sequence (SEQ ID NO:16) derived from Roseomonas genomospecies:

[0057] atgattgatcgtcgcacgtccgcatttttggctgactttctttcaaaacccggcggcgatccagaccgcttggaccgcttcttgcttcacggtccataccgtgctcgccgtggcggccgaccccgtctgaaactcgctttccacgatttctggcccgaattcgataagggtactaatttttttatcgagattctttcatcacgcttcgacttgtccgttgtggaagatgattccgatctcgccatcgtgtctgttttcggcggccgacaccgcgaggctcgttcccgccgttccctgttttttaccggtgagaatgttcgaccacctctggacggtttcgacatggcggtctcttttgatcgcattgatgatccccgtcattttcgtcttccgctgtacgtcatgcatgcatacgagcatatgcgtgaaggtgctgtaccacacttctgctcacccgtgctccctccggtgcccccgacacgcgcggagtttgccgagcgtaaattctgtgcgttcttgtacaagaacccaaacggcgagcgccgtaatcgtttcttccctgcgctcgacggtcgtcgtcgcgttgactcggtgggttggcatctcaataatacgggcaatgtcgttaagatgggctggctggcgaagatccgcgtgttcgaacgttatcgtttcgcgtttgccttcgaaaacgcctcccacccgggatacctgactgaaaagattctggacgtcttccaggctggagcagtcccgctgtattggggtgacccggacttgacacgcgaggtggctgccggttcgttcatcgacgtttcccgcttcgcaaccgatgaagaggctgtcgaacatatcctcgcggtggatgacgatcacgacgcgtactgcgctcaccgcgcggttcccccgtttctcggcactgaggaattccatttcgatgcttatcgtctggccgactggatcgaatcacgcctttaa。

[0058] Polypeptide sequence (SEQ ID NO: 17) of α-1,3-fucosyltransferase derived from Azospirillum picis:

[0059] MIDARTSAFLADFFAGGRRNPLELERFLLTGPDGPRRGAKPPLKLAFHDFWPEFDSRRNFFTALLATRFAVRVVEDDSDLAIVSVFGGRHREMRSARSLYFTGENRRPPLDSFDMAVSFDRLDDPRHFRLPLYVVHAHDHFRETATPLFCQPVLPPVLPSRREFQDRGFCAFLYKNPHAERRNAFFAMLDARRRVDAVGWHLNNTGSVVRTGWLPKIKVFQRYRFAFAFENSSHPGYLTEKILDAFQAGTVPLYWGDPELRREVAAGSFIDVSAFPDDEAACSHILALDEDYDAWCAVRGVPPFLGTEDFHFDVYRLVEFIEARL。

[0060] Gene sequence (SEQ ID NO: 18) of α-1,3-fucosyltransferase derived from Azospirillum picis:

[0061] atgattgacgctcgaacatcggccttcttggcagatttcttcgcaggtggccgtcgcaacccactcgagctggagcgctttttgctcaccggaccagacggaccccgccgcggtgccaagcctccgcttaagttggcgttccacgatttctggccagaattcgattcccgtcgaaacttctttaccgctctgctcgcaacgcgctttgccgtgcgagtcgtcgaggatgattcggacctggcgatcgttagcgttttcggcggtcgccaccgagaaatgcgctcggctcgctcgctctacttcaccggtgaaaaccgccgcccacccctcgacagcttcgatatggcagtttccttcgaccgactcgatgatccacgccactttcgattgcctctttacgtggtccacgcccacgatcacttccgcgaaaccgcaaccccgttgttctgtcaaccggtgttgccaccagtgctgccctcacgccgagaatttcaagatcgcggattttgtgcttttttgtacaaaaatcctcacgctgagcgtcgcaacgctttctttgccatgctggacgcgcgtcgtcgcgtggatgccgtaggttggcacttgaacaacacgggctcagtggttcgcactggatggttgccaaaaatcaaggtcttccaacgctaccgttttgcattcgccttcgagaattcctcccatccaggctatctgacagaaaagattttggatgcattccaggcgggcaccgtgcctctgtactggggtgacccagagctccgccgagaagtagccgccggttcgtttatcgacgttagcgcattcccagatgacgaagcagcatgcagccatatcttggcgctggatgaggattatgatgcatggtgcgccgttcgaggcgtcccacctttcctgggcacagaggattttcacttcgatgtctatcgcttggtggaattcatcgaagcccgcttgtaa。

[0062] Polypeptide sequence (SEQ ID NO:19) of α-1,3-fucosyltransferase derived from Helicobacter sp.:

[0063] MFQPLLDAFIESTHLDETTHKPPLNIALANWWPLKNSEKKGFRDFILHFILKQRYKIILHSNPNEPSDLVFGNPLEQARKILSYQNTKRVFYTGENEAPNFNLFDYAIGFDELDFNDRYLRMPLYYAYLHYKAMLVNDTTSPYKLKTLYTLKKPSHKFKENHPHLCALIHNESDPLKRGFASFVASNANAPIRNAFYDALNAIEPVASGGSVKNTLGYKVKNKNEFLSQYKFNLCFENSQGYGYVTEKILDAYFSHTIPIYWGSPSVAKDFNPKSFVNVHDFKNFDEAIDYIKYLHTHQNAYLDMLYENPLNTIDGKAGFYQDLSFEKILDFFKNILENDTIYHCNDAHYSALHRDLNEPLVSVDGLRVNYDDLRVNYDDLRVNYDDLRVNYDDLRVNYDDLRVNYDDLRVNYDDLRVNYDDLRVNYERLLSKATPLLELSQNTSFKIYRKAYQKSLPLLRAIRRWVKK。

[0064] Gene sequence (SEQ ID NO:20) of α-1,3-fucosyltransferase derived from Helicobacter sp.:

[0065]

[0066] Polypeptide sequence (SEQ ID NO:21) of α-1,3-fucosyltransferase derived from Antarcticibacterium arcticum:

[0067] MKNIKLWFTDFYEGFDLADNPLSRILEKNYSIILTKEEPDYLIYSCYGNEFLNYNCIRIYYTGENLTPDFNLCDYAIGFDRLTFRDRYLRYPNYAFFEDQFEQLIKPPNFDKKILSEKKYFCNFIYSNPHAHPARDHFFHKLNSYKEVTSPGTHLKNSNLFVGERFAADWMFSKLEFQSQCKFTIAFENTSSPGYTTEKILHAFISNTIPIYWGDPEVTKDFNPKAFINCHDFENFEAVIERIKEIDGSDELYLSILNEPPFVNNQIPKNLERNKLTSFLQHMFDQEIEVAGRRNHYGTSLKYEKDLKSMVALSQQYKERNKLKRFLDKLGCL。

[0068] Gene sequence (SEQ ID NO:22) of α-1,3-fucosyltransferase derived from Antarcticibacterium arcticum:

[0069]

[0070] Polypeptide sequence (SEQ ID NO:23) of α-1,3-fucosyltransferase derived from Bacteroides congonensis:

[0071] MKKIKVKFVDFFDGFNITANEFFDILKLRYDVEICDVPDYVIYSGFGYEHLKYDCIRIFFTGECQTPDFNECDYAIGFDRLKFGDRYARIPLYNMMQYKSEYKSLLNRKSIISDDIKGRDFCSFVVSNCFADDIRAVFYEKLSQYKHVASGGRYKNNIGGSVKDKKAFLSKYKFNIAFENCSHDGYATEKIMEAFAAGVVPIYYGDPRIAEDFNPKAFVNAHDFSSFDAMIERIKEIDSNDELYLSMLNEPIIQCDADVAELSDFLYSIFDQPLSSVKRRSHSQPAKGMEAMKLRHVFFETKIYKYYRKGMNQFARLRKGTALSSKRTK.

[0072] Gene sequence (SEQ ID NO:24) of α-1,3-fucosyltransferase derived from Bacteroides congonensis:

[0073] atgaagaaaatcaaggtgaagttcgttgacttctttgatggcttcaacattactgcgaatgagttttttgacatcctcaaactgcgctacgatgtcgaaatctgtgatgtcccagattacgtcatttattctggctttggctacgaacacctcaagtacgactgcatccgcatcttcttcaccggtgaatgccaaaccccagactttaacgaatgtgattacgccattggatttgatcgcctcaaatttggtgatcgctacgcccgtatcccactctacaacatgatgcaatacaagagcgaatacaagtccttgctcaatcgtaaatcaatcatttctgatgatatcaaaggccgtgatttctgttcctttgttgtgtccaattgtttcgcggacgatatccgagctgtgttctacgaaaagttgtcgcaatataagcatgtggcctccggcggccgttacaaaaacaacattggcggctccgtgaaggataagaaggcctttctgtcaaaatacaagttcaacatcgcatttgaaaactgctctcatgatggttatgccaccgaaaaaatcatggaggcttttgccgctggcgtggtgccaatctactacggagacccccgcatcgcggaagattttaatccaaaggcgttcgttaacgcacatgacttctcctcattcgacgcaatgattgaacgcatcaaagagatcgattctaatgacgagttgtacctgtctatgctgaacgaaccaatcatccagtgcgatgccgatgtcgccgagctgtcggacttcctctactccattttcgatcagcctctgtcgtctgtgaaacgccgctcacattcccagcctgctaaaggcatggaggccatgaaactccgtcacgttttctttgaaaccaagatctataaatactaccgcaagggaatgaaccaatttgcgcgtctgcgtaagggtacggctttgagctcaaagcgaaccaagtaa。

[0074] Polypeptide sequence (SEQ ID NO:25) of α-1,3-fucosyltransferase derived from Azospirillum sp. BE72:

[0075] MIDPRTSDFLAEYLASANKDPAVLDRFLLHGPERGGRGARPRLKIAFFDFWPEFDPSANFFVDILSSRFDVSVVDNDCDLAIVSVFGTRHREARTARGLFFTGENVRPPLDGVDISVSFDRIDDPRHYRLPLYVMHAWDHRREGATPHFCHPVLPPVPPTREEAAKRKFCAFLYKNPNCARRNDFFRMLCARRHVDSVGWLLNNTGSVVKMGWLPKIRVFSRYRFAFAFENASYPGYLTEKILDAFQAGTVPLYWGDPGVLRDVAAGSFIDMSRYSSDEEAIDAILAVDDDYDTYRRYRSTAPFLGTEDFYFDAFRLAEWIESRL。

[0076] Gene sequence (SEQ ID NO:26) of α-1,3-fucosyltransferase derived from Azospirillum sp. BE72:

[0077] atgatcgaccctcgcacctccgacttcttggcagaatacctggcgtctgctaataaagatccagccgtcctcgatcgcttcctcttgcacggacccgaacgcggtggtcgcggtgcacgcccccgtcttaaaatcgcctttttcgacttctggccagaattcgatccgtcggcgaatttttttgtcgatatcctttcgtcgcgattcgacgtgtcagttgtcgataatgactgcgaccttgcaattgtttcggttttcggcacacgtcaccgcgaggcccgtactgcgcgtggccttttcttcaccggcgaaaacgtccgccccccactggatggcgttgacatttcggtctcgttcgaccgcatcgacgatccacgccactaccgccttccgctctacgtgatgcatgcctgggaccaccgtcgtgaaggcgctactccacacttctgtcacccagtcttgccgccggtgccaccaacacgtgaagaggcagcaaaacgtaagttttgtgcgttcctctacaaaaaccccaactgcgcgcgccgcaatgatttttttcgtatgttgtgcgcacgccgccacgttgattccgtgggctggctgttgaacaacaccggctctgttgtcaagatgggttggctgccaaagatccgagtgttctcccgttaccgctttgcgttcgcgttcgaaaacgcctcctacccgggatacttgacggagaagattttggacgcttttcaagcgggcaccgttccgctctactggggtgatcctggtgtccttcgcgatgtcgcggctggctcgtttattgatatgtctcgttactctagcgacgaagaggccattgatgctatccttgcagttgatgatgattatgatacctatcgccgataccgctccactgcaccttttttgggcaccgaagacttctatttcgatgctttccgccttgcggaatggatcgagtcccgtctctaa。

[0078] The polypeptide sequence (SEQ ID NO:27) of α-1,3-fucosyltransferase derived from Azospirillum sp. 412522:

[0079] MIDRRTSEFLADHLARADGNPADLDRFLLHGPERGGRGARPRLKIAFFDFWPEFDPSANFFVDILSARFDLSVVDNDCDLAIVSVFGTRHREARTARALFFTGENVRPPLDGVDMSVSFDRIDDPRHYRLPLYVMHAWDHRREGATPHFCQPVLPPVPPTREEAAKRKFCAFLYKNPNCERRNDFFRMLCARRHVESVGWLLNNTGSVVKMGWLPKIRVFSRYRFAFAFENASHPGYLTEKILDAFQAGAVPLYWGDPGVLRDVAAGSFIDVSRYSSDEEAIDAILAVDDDYDTYRRYRGTAPFLGTEDFYFDAFRLAEWIESRL.

[0080] The gene sequence (SEQ ID NO:28) of α-1,3-fucosyltransferase derived from Azospirillum sp. 412522:

[0081] atgatcgaccgacgtacatccgagttcctcgccgatcacttggcccgcgcggatggcaaccctgccgatttggatcgttttctgctgcatggcccggaacgcggcggacgcggcgcgcgcccacgcctgaaaatcgcgttcttcgatttctggcctgagttcgatccatctgctaactttttcgtcgatattctctctgcccgattcgacttgtccgtggttgataatgattgtgatctggcaattgtttccgttttcggcacccgccatcgcgaagcacgtaccgcgcgcgcacttttctttaccggcgagaacgtccgcccaccactcgacggcgtggatatgtctgtgtccttcgatcgcatcgacgatccgcgccactatcgcctgccgctctacgtgatgcatgcgtgggatcaccgccgtgagggcgctactcctcacttctgtcagccagtgttgccacctgtgccccccacccgcgaagaagcggcaaaacgtaagttctgtgccttcttgtataagaatccgaattgcgaacgccgtaacgatttcttccgtatgttgtgtgctcgccgtcacgttgagagcgtgggttggctgctcaacaacactggtagcgtagttaagatgggatggctgccgaagatccgcgtcttctcccgctaccgtttcgccttcgccttcgagaatgcttcccacccgggttacctgaccgagaagattcttgatgcctttcaggcgggtgccgtcccactttactggggcgacccaggagttttgcgcgatgttgccgcgggatcattcatcgacgtgtcccgctacagctccgatgaagaagcaattgatgcaattctcgctgttgatgatgattatgatacctaccgccgatatcgcggcaccgccccgttcctgggcacagaagatttttatttcgacgcattccgtctcgcagagtggatcgaaagccgcctgtaa。

[0082] Polypeptide sequence (SEQ ID NO:29) of α-1,3-fucosyltransferase derived from Azospirillum endophyticum:

[0083] MIDQRTSDFLSEFLASPDRDPSVLDRFLLQGPELGRRGAKPRLKIAFFDFWPEFDPAANLFIDILSARFELSVVDNDCDLAIVSVFGTRHREARTARALFFTGENVRPPLDGVDMSVSFDRIDDPRHYRLPLYVMHAHDHVREGATRHFCQSVLPPVPPTREEAANRKFCAFLYKNPNCERRNAFFRMLCARRHVDSVGWLLNNTGSVVKMGWLPKIRVFSRYRFAFAFENAAYPGYLTEKILDAFQAGVVPLYWGDPGVLRDVAAGSFIDVSRYSSDKEACEAILAIDDDYDSYRRYRSTAPFLGTEDFHFDAFRLAEWIESRL。

[0084] Gene sequence (SEQ ID NO:30) of α-1,3-fucosyltransferase derived from Azospirillum endophyticum:

[0085] atgattgatcaacgaacctcggactttcttagcgaatttctggcctctcctgatcgcgacccgagcgtcttggatcgcttcctcctgcagggtccggaactgggccgccgcggagccaagccgcgtttgaaaatcgctttcttcgacttctggccggaatttgatccagctgcgaatctctttattgatatcttgtccgcgcgattcgaactctccgtggtggataatgattgcgatctggctatcgtatccgttttcggtacccgacaccgcgaagctcgtaccgctcgtgccttgttctttacgggagaaaatgtccgtccgcctctggacggcgttgacatgtcagtgtccttcgaccgaattgatgaccctcgccattaccgcttgccgctctacgtgatgcatgctcatgaccacgttcgtgagggtgcgacccgccatttctgtcagtccgttctgcctccagttcctcccacgcgcgaggaggcagctaaccgaaagttctgcgcattcctttacaaaaacccgaattgcgaacgccgtaatgcattcttccgaatgctgtgcgctcgacgtcacgtggactcggtaggctggctgttgaacaacacaggttccgtcgttaagatgggctggctccctaagatccgtgtgttctctcgctaccgttttgcatttgctttcgagaacgctgcgtatccaggctacttgaccgaaaaaattcttgatgctttccaggccggcgtcgtcccactgtactggggcgatcccggagtcctccgtgatgtcgcggccggttcctttattgatgtgtcccgctactcctcagacaaagaggcttgtgaggctattctggcgatcgatgacgactatgactcctaccgccgttaccgcagcacagcaccgtttctcggcaccgaagatttccacttcgacgccttccgccttgccgaatggattgagtcccgcctctaa。

[0086] Polypeptide sequence (SEQ ID NO:31) of α-1,3-fucosyltransferase derived from Phycisphaerae bacterium:

[0087] MRSIRIAFRGFWPGFKPEEFWQWRPYLTTRYRFVVDEIAPELVICSVFPDPRPVPRGAVRVFYSGEWVEPDMDQWDWAITFCHLDHPRHLREPLWVPHLHRTGLGWEGLIRAAGLPDVPVPERFCAFIYSNEVPRRNAVFDLLSRYKRVDAPGRCRNNMPPIGPDPTAKLAFIARYRFVFAFENRSAPGYATEKLVDPLLARTVPLYWGDPLIGETFDRGSFLDRGACASDEEFVERIVALDRDDAAYRSVRSRPCLLGNAVPPAHADGASLAWWDRVLAGAGELRSDDAVFADQRDPGGPARRAHDRAHGR.

[0088] Gene sequence (SEQ ID NO:32) of α-1,3-fucosyltransferase derived from Phycisphaerae bacterium:

[0089] atgcgctctatccgcattgcattccgcggcttctggccaggctttaagccggaagaattctggcagtggcgtccttacctgaccactcgataccgcttcgtggtggacgaaattgcaccagaactggtgatctgttccgtgtttcctgatccgcgtccagttccacgtggtgcggttcgcgtgttctattcgggcgaatgggtcgaacctgatatggatcagtgggactgggcaatcactttctgccacctcgaccaccctcgccacctccgtgaaccattgtgggtgccgcacctgcatcgcaccggtttgggctgggagggtttgatccgcgccgccggtctgccggatgtccccgtgccagagcgtttctgtgcctttatttactctaacgaggtcccacgtcgcaacgccgtctttgatctcctgtcccgatataagcgcgtcgatgccccgggccgttgccgtaataatatgcccccaatcggccctgatccgactgctaaactggcctttatcgctcgctatcgcttcgtattcgccttcgaaaaccgatccgcgcccggctacgccaccgaaaagttggtggaccccttgcttgcacgcaccgtgccactttattggggtgacccgctcatcggcgagaccttcgaccgcggttccttccttgatcgcggcgcatgcgcaagcgatgaagagttcgtcgaacgtatcgtggctctcgaccgcgacgatgccgcctaccgctccgtgcgttcacgtccctgcctgcttggaaatgcagtgccaccggcgcacgctgacggcgcctccctggcgtggtgggatcgcgtgttggcgggcgctggcgaactccgatcggatgatgcggtcttcgcggatcagcgcgacccaggtggccccgcacgacgtgcccacgaccgagcacacggtcgctaa。

[0090] Polypeptide sequence (SEQ ID NO:33) of α-1,3-fucosyltransferase derived from Helicobacter acinonychis:

[0091] MFQPLLDAYTDSAYSDASDHKPPLNIAIANGWGGTKGFEASVLYFILSWRYKIALHENPNKPADLVFSNPLGQARKILSYQNTKRVFYTGENEAPNFNLFDYAIGFDGLDFKERYLRMPLYYASLHDKAQSVNDTTAPYTLKSDSLYALKKPAHCFKEDHPHLCSVVKGESDPFKRGFASFVASNANAPIRNAFCEALNSVEPVTGGGAVKNTLGYNVTNKSEFLSQYKFNLCFENAQGYGYVTEKIIDAYFSHTIPIYWGSPSVAQDFNPKSFVNVHDFKDFDGAIDYIRYLHTHENAYLDMLYENPLNVIDGKACFYQDLSFKKILDFFKTILENDTIYHNNPFVFDRDLHEPLVSIDNLRADLLLLKDNYDGLKTDYDGLKTDYDGLKTDYDGLKTDYDGLKTDYDGLKTDYDHLFKSALPLLELSQTTSFKIYHKIYQKTLPLLCMARKLVKK。

[0092] Gene sequence (SEQ ID NO:34) of α-1,3-fucosyltransferase derived from Helicobacter acinonychis:

[0093]

[0094] Polypeptide sequence (SEQ ID NO:35) of α-1,3-fucosyltransferase derived from Helicobacter melei:

[0095] MFQPLLDVFAQSTHLPQAPQKKPLKLGLKWYGDGVEDFKSCCFYHVLSSHYDISFTDKNTCDLFLRGFLFHDKDLIECAKLRMMFMGENARIDFNLYDFGMGFDDLSFHDRYLRVPLYYLSLWMFFKLAMGPNSPFQIDESIQRAFYALSPDKSPDHPYSENAHHGTITLNTSFSDNYPHLHALASDQSDPTKRAFASFVASNPKAPVRNAFYQLLNAYRPIGGGGGVFNTIGGPVANKWEFLSQYKFNLCFENSRGYGYTTEKIIDAYFAHTIPIYWGNPAVAKDFNPKSFVNVHDFKDFDEAIDFIVYLDTHPNAYLEMLHTHPLNSIEGQPRFYQDLSFEKILGFLQKALDCQEIYHEHSLYPAHDDPMFLYSGKQLCKFLAKKILKRLGKLSS。

[0096] Gene sequence (SEQ ID NO:36) of α-1,3-fucosyltransferase derived from Helicobacter melei:

[0097]

[0098] Polypeptide sequence (SEQ ID NO: 37) of α-1,3-fucosyltransferase derived from Helicobacter bizzozeronii:

[0099] MFQPLLDVFAHSTHLPQAPQKKPLKLGLKWYGDCEDFKGWCFYHILNSHYDISFTDKNTCDLFLRGFLFDDKELIECTKLRMMFMGENARIDFNLYDFGMGFDDLNFHDRYLRVPLYYQSFYWFLHIITNASNSPFKLDMAQETLHSPLILHIPHASTSTKISFFATYPHLNALAREQKNPLERDFASFVASNWEAPMRNASYQKLNDYRPVAGGGRVFNTTGKPVSNKHEFLSQYKFNLCFENSLGMGYTTEKIVDAYFAHTIPIYWGNPLVHLDFNPKSFVNVHDFDNLDEALDFVRYLDTHDNAYLEMLHAHPLNTSEGKPRFCHDLSFKVILDFLINAIESPHIYHEQINVSSKSLYTPDRTSLLELFSGREHLHMAFKKLRAKTRSMLKTAAKTLRIIH。

[0100] Gene sequence (SEQ ID NO: 38) of α-1,3-fucosyltransferase derived from Helicobacter bizzozeronii:

[0101]

[0102] Polypeptide sequence (SEQ ID NO:39) of α-1,3-fucosyltransferase derived from Helicobacter bizzozeronii CIII-1:

[0103] MSPSSLANLRKFFERFFMFQPLLDAFIEATKLPPPPPPLKKPLNLGLQWYHGFDDFRGWFFYEILQHQYAISFDEKMTYDCLFGVYPRTLEGLYQALDNPNKRLVFVGENERIDFNIYDFAMGFDHLEFGDRYLRVPLYYQSLYQFLHAITHASHAPFKLNASDPLSLENKTPFSATYPHLDALAREQKNPLERKFSSFVASNWEAPMRNASYQKLNDYRPVAGGGRVFNTIGKPVSNKHEFLSQYKFNLCFENSLGMGYTTEKIVDAYFAHTIPIYWGNPLVHLDFNPKSFVNVHDFDNLDEALDFVRYLDTHDNAYLEMLHAHPLNTSEGKPRFCHDLSFKVILDFLINAIESPHIYHEQINVSSKSLYTPDRTSLLELFSGREHLHMAFKKLRAKTRSMLKTAAKTLRIIH。

[0104] Gene sequence (SEQ ID NO:40) of α-1,3-fucosyltransferase derived from Helicobacter bizzozeronii CIII-1:

[0105]

[0106] Polypeptide sequence of α-1,3-fucosyltransferase derived from Helicobacter cynogastricus (SEQ ID NO: 41):

[0107] MFAPLLEAFIDSSRLPPPPPNKKPLVLWIRQGENEADFKDWLFYKILGQRYNLSRAPNDYVCYLGGHHRSLEELGRILKIPTKRIAYIGENERIDFNVYDFGIGFDDLEFNDRYLRVPLYYQAIWWCSEIAITCANSPFKQAKLENLAFGPSKNSTPIDFRTTYPQIDSLVREQSDPFKREFASFVASNINAPVRNALYQIFNTYKPVAGGGGVFNTIGSLVQNKREFLSGYKFNLCPENSQGLGYTTEKIVDAYFAHTIPIYWGNPQVAKDFNPKSFVNVHDFEDFQEALDFVRYLDTHKNAYLDMLYSHPLNIYGNKHRFYKDLSFSKILSFLQNALECPYIYHECTNLINLLYQEQNNEKQEAPKELTGRQHLAIAFNKLLQKVRRVLT。

[0108] Gene sequence of α-1,3-fucosyltransferase derived from Helicobacter cynogastricus (SEQ ID NO: 42):

[0109]

[0110] Polypeptide sequence (SEQ ID NO: 43) of α-1,3-fucosyltransferase derived from Helicobacter ailurogastricus in fasting state:

[0111] MFAPLLRVFIESSQLKNAPVKKPIAIGVANWFAGDTLGNFKSWFFYHILATQHDITLTTSPKDPCALVFGASFREEGYHFFRSILDFPQKRIGHTGENERIDFNVYDFGMGFDEIEFNDRYLRVPLYYLNLWQMHANLLKHTNTPFHLETTNPTKADFQKVHPQIDSLAREQSDPLKRGFASFVASNPNAPIRNAFYQELNTYKPVGGGVFNTIGHLVQNKHEFISGYKFNLCFENSIGLGYTTEKIIDAYFAHTIPIYWGNPEVAKDFNPKSFVNVHDFKDFKEALDFIRYLDTHDNAYLDMLHAHPLNTYEGKPRFYQDLSFSKILNFLQNAIECPHLYHEHTSFNHTGTSLKHAFLSKLTQIKAKFSRAQDTR。

[0112] Gene sequence (SEQ ID NO: 44) of α-1,3-fucosyltransferase derived from Helicobacter ailurogastricus in fasting state:

[0113]

[0114] Polypeptide sequence (SEQ ID NO: 45) of α-1,3-fucosyltransferase derived from Candidate division KSB1 bacterium:

[0115] MNIRIDILCHNRFNKADSIIVRLLKKHYDVTIDRENPDYVFIQSHRFHDTGNYQSDPIRIYWQVEANVPDFNLFDYAILRYDDIRYDDRCYFGDFGWWHHEFLANYDQAETRRRKELLTKKKFCNFIYSHSCKERDQFFHLLNNYKRIDSFGKHLNNMGDLPPENQRYASGWFPSSIELKRPYRFSIAFENVYYPGYTTEKILTSFLAGTIPIYWGNPRISQEFNSKAFINCHEYESFDAVCERVIEIDNDTTMIQQMVKAPSMTVEQHEKLQYALAKHEPDLESFLCRIIDQPIAKAHRRTLNRFSNKIYAFRNDPWYKFGRMGKRGKMIFLIRRILIRIFTSKKS。

[0116] Gene sequence (SEQ ID NO: 46) of α-1,3-fucosyltransferase derived from Candidate division KSB1 bacterium:

[0117]

[0118] Polypeptide sequence (SEQ ID NO:47) of an α-1,3-fucosyltransferase derived from a Lachnospiraceae bacterium:

[0119] MKKIRIAFTDFWGVFDPNDNFIMDALRKNFEVEISDKDPEFVFCSIFGRRHLKYDCAKIFYTGENIDPDFNLVDYALGFPEIDYYDRYLRLPHYLLYPRACKLALSKPSMSDEELLNRKFCNYVISNALSSPERGIMIDKLERYKPLASGGRYHNNVGGPVADKIDFSRGYKFSIAFENSGSRGYTTEKIMESFASQTIPIYWGNPDIAKEFNPDSFINCHDFANFDEVVDFVKKIDSDDTAYLNMVKAPMIRDDSLAAKCLDEDYLSDFLFKICSQDPTAAIRRNRVYIGKHYEDEAKLHEKLDRVLRLPRRAVRGMKNRIKRVE。

[0120] Gene sequence (SEQ ID NO:48) derived from a Lachnospiraceae bacterium:

[0121] atgaaaaaaatccgcatcgcttttaccgatttttggggtgtgtttgatcccaatgataactttatcatggacgcgctgcgtaagaatttcgaagtggagatctcagataaagacccggaattcgttttctgttccatcttcggccgacgccatctcaagtacgattgtgccaaaatcttctacacaggagaaaacatcgatccagactttaacttggtggattacgcacttggctttcccgaaattgactactacgaccgttacctccgactccctcactacctcctgtacccccgcgcatgtaaactggcgctctccaagccatctatgtctgatgaagagctgcttaaccgtaaattctgcaactacgtgatctccaatgcgttgtcatctccggagcgaggaattatgatcgacaaactcgaacgctacaagccattggcctctggtggtcgctaccacaacaatgtaggcggccctgtggctgacaagatcgatttctcgcgcggatacaagttcagcattgcgtttgaaaattccggcagccgtggttacaccacggagaaaattatggaatcgttcgcttcgcagaccatccccatctactggggaaaccccgatatcgctaaagaattcaacccggattcctttatcaactgccacgatttcgcgaatttcgacgaagtggttgactttgtgaagaagatcgattcagacgacaccgcatacctgaacatggtcaaagcccctatgattcgcgacgactccctggcagcaaagtgcctggatgaagattacctgtcggactttctcttcaaaatttgctcccaagaccccaccgccgccatccgtcgtaaccgcgtgtacatcggcaagcattacgaggatgaagccaagcttcacgaaaagctggaccgcgtcctgcgcctccctcgtcgcgccgtacgcggcatgaagaatcgcatcaaacgcgttgaataa。

[0122] Polypeptide sequence (SEQ ID NO:49) of α-1,3-fucosyltransferase derived from Bacteroidota bacterium:

[0123] MKIPKIAVQAKSPINSKLILKFMEIERISFELKDNPLDADYIFLFPLPFEEMLEWCHKYKNKILIFIGAESYIPDFNLFDYTFSYDKVEFGDRHMQIQFQALLSYYLPQKLLVGKDLLKKKENFCNFIYSNPKAHPNRDLFFHTLNKYKKVDSLGAHLKNVNLNIGDRKLDNYFQESIKQKESYKFSISFENALHKGYNTEKIISSMEAYTIPIYWGDSEIGEFYNSRSFINCHNYQSFNEVINFIIEIDNDDDLFLEILNQPWRTKEQEIKFVDQKKSFINRLNHIFSQPYPETFRKPMGTYNDFYSKKISTYNKTSSVLLKLWRNIHHQAHLVKKALKK。

[0124] Polypeptide sequence (SEQ ID NO:50) of α-1,3-fucosyltransferase derived from Bacteroidota bacterium:

[0125]

[0126] Polypeptide sequence (SEQ ID NO: 51) of α-1,3-fucosyltransferase derived from Wenyingzhuangia fucanilytica:

[0127] MKTIYIDFLGFDLSLNKEDNWIVSLLSRSYHVKISKDAPYVFVGSFDPYSYRTEITDKISIYIPGEAIFPDFNFFDYALGFDEFNYDDRYFRWLPLGNTMARGKFVNNIQKPFGRKFCNFIYGNPDAHPNRDLLFHMLNKYKKVDSLGAHLKNTNIDIEPRNGNWYQGSIDIKSEYKFSLSLENSLMKGYTTEKIISSFQAKSIPIYWGNPNVTKEIDPDGFINCHDYESFDHVVEKVKEIDNDKTKYLKMLQSAKSLFFEETFHENQNKKLLLFFENIFDKEFKDAKRKPVGYWTSRHLDTLLIKPEKKQSLTSRLKFWK。

[0128] Gene sequence (SEQ ID NO: 52) of α-1,3-fucosyltransferase derived from Wenyingzhuangia fucanilytica:

[0129] atgaagactatctatattgattttttgggcttcgatctctccctcaacaaggaggacaactggatcgtttccctgctttctcgatcctaccatgtgaaaatctccaaggacgcaccatatgttttcgtaggttccttcgacccgtactcttaccgcaccgagattaccgacaaaatcagcatctacatcccgggtgaagcgatctttccggacttcaatttcttcgactacgcccttggatttgatgaatttaattacgacgatcgatattttcgctggttgccgctcggtaacaccatggctcgtggaaagtttgtgaacaacatccaaaagccctttggacgcaaattctgcaattttatttacggaaatcctgatgcacacccgaaccgcgatcttctcttccatatgcttaacaaatacaaaaaggtggactctttgggcgcgcatctgaaaaatacaaacatcgacatcgagccacgcaatggcaattggtaccaaggttcaattgacatcaaatctgagtacaaattctccctttctcttgagaactccctcatgaagggttacaccaccgaaaagattatctcgtcatttcaggcgaagtcgatcccgatttactggggcaacccaaacgtcacaaaggaaatcgacccggacggctttattaactgtcatgactacgagtcgttcgaccatgttgttgagaaagtaaaagagattgacaacgataagactaagtacctcaagatgctccaatccgccaaatccctgtttttcgaggaaacgttccatgaaaaccagaacaaaaagcttttgctgttctttgagaatattttcgacaaggaattcaaggatgccaaacgcaagcccgtcggttactggacaagccgccacttggatacactccttattaagccagaaaagaagcagtcattgacctcccgtctcaagttctggaaataa。

[0130] Polypeptide sequence of α-1,3-fucosyltransferase derived from Phycisphaerales bacterium (SEQ ID NO: 53):

[0131] MKNTIKIMVSDRRPTSGILNCVYEILEKDYNVVECADADYFISGSWEPLKNARLIQDRISIYLNTEATAPDFNLFDYAVGFDDLHFGDRYLKYLPAFMGAVGPEYMTVSDPLARRFCNFIYSNPISHPNRELVFHKLSRNYKQVDSLGSILKNVSFDISPRNGDWYPGNVEAKSRYKFSVAFENAYYPGYTSEKILTSYQARSIPIYWGNPDIHKIANPDSFINCHDYDNFDQVVEKVRELDTNDEKYLTMLNADKGAMGTDEFFAENHTRFTSFFRNIFDQPLEKARRRPVGAAVDGHRNTLILINNLGYSDVYENQAKELIKKNKPKEAYDKISKAIELRPGFPDYYFIRANILYSMKDFESSMQSLSVAIQIDPSNTAYTKLMSTLQQQIFSP。

[0132] Gene sequence of α-1,3-fucosyltransferase derived from Phycisphaerales bacterium (SEQ ID NO: 54):

[0133]

[0134] Polypeptide sequence (SEQ ID NO:55) of α-1,3-fucosyltransferase derived from Succinivibrio dextrinosolvens:

[0135] MKKINIKFASGTNSDFIQEILSYLKLRFDVTIDERNFDYFFCNELIYRNKDSFGELFKLPPRVIRFFWGGEAVYPDLNLFDYACCYDNFDSTRILKIPNIYIRDFNWLKKEKYHELFTYSKNASEILRQKTKFCNFIYGNPHSHPIRDSLFFKLSEYKKVDSLGSHLNNIKIVNSRSDTDWLKKSIDLKSPYKFSIAAENAFFKGYVSEKLMTSMLANTIPIYFGTNDVVKEFNPKSFINVNDFSSLDDLLEKVKQIDNDDDLYCEMMSEPWITQKQCEETVSGYNKFLEQFLSIFDLPLDIAVRRPMGCWTDFIYPNFYKDLNSTANQIHYSFKDLLFSIKNYPSLYKQICIFGIKFKLRKRPIILKK。

[0136] Gene sequence (SEQ ID NO:56) of α-1,3-fucosyltransferase derived from Succinivibrio dextrinosolvens:

[0137]

[0138] Polypeptide sequence of α-1,3-fucosyltransferase from Candidatus Akkermansia timonensis (SEQ ID NO:57):

[0139] MFLPSVPIDCLWGICIIISSILNNIRMKVKVGFIEGAEEWQINFFLRRFQKIDGSVLEFEIDKERADFLIAFPWLYMSSRENYLQFLEESRGKIVIMDVLGEALAPNLNLFDYHIGFDAPDDDGRLLCMSYLFDLRMKLKDLHTMNPDDALCGKDGFCNYIYSHGLGHPYRIQLFSELSAYKKVDAIGKHLNNTPCLIPREAEDWLAGSVLLKKPYKFSIACENSWYRRYTMEKIITSFLACTVPVYWGNPLVEEEYNPKAFINCHRYSSLKEVVAEIKRIDEDEALWKAMMAEPRRLPWQIEREQEKKDKFNAELMKIFTSPVEHVRKRGDGLWMNNYRNFFTDKMTMKKEDDRKKLKSALKEWNKRLFPRF。

[0140] Gene sequence of α-1,3-fucosyltransferase from Candidatus Akkermansia timonensis (SEQ ID NO:58):

[0141]

[0142] Polypeptide sequence (SEQ ID NO: 59) of an α-1,3-fucosyltransferase derived from Akkermansia sp.:

[0143] MKNCKISFVRNTPSWLIENVISGLSDIEGIKFHHLKEGGDYLFATPFMYSSFVSFSDFLDKAPSAITIMHCVGEAIMPDLNIFDYYIGFYPEACGDRIIQRLYERELRRKISECKKRPAVQILKEKKAFCNFIYSNGKGHPMRGQLFDLLSHYRKVDAPGRYKHNIDILTSRHNKDWLSESIDLKNPYKFSISAENAWFPGYTSEKLLTSMLADTLPIYWGNPEISNEFNPDSFIHVDHYSNLNELLEFIIRVDLDDELWCRIMDEPWRLKKQIEEDDRIFEHYVKRYGNIFKQPIEKAFRKGNGYFEDMYRNQMKAGMGLRLLNENISSNCMKFIKNMPRFFRKK。

[0144] Gene sequence (SEQ ID NO: 60) of an α-1,3-fucosyltransferase derived from Akkermansia sp.:

[0145]

[0146] Polypeptide sequence (SEQ ID NO: 61) of an α-1,3-fucosyltransferase derived from Roseburia sp.:

[0147] MKKKIKIYMLHRCVYDADSLYRYLHLHSCENLEYEFVWNPSDPEYLIATELIWEDISSWKEFVKLYRKAKVHIYHAGECIAPDLNLFDYAICFDAHLNCDDRICRTPARLLFEDMIFDFVNSLDSEEKAHKELQKKTGFCNFMYSNKSGHANRTKIFHKIDQYKHVDSLGKYLNNKSVPMSMRIKRQKGWRNVIRESIDIKSNYKFSIAFENASYAGYTSEKILSSLEAHTVPIYWGNPFIAEELNEEAFINCHKYRNFDEVLERIKEIDNDDRIWCHMVSQPWLTQEQEKREREETEKYYRFMNHIFMQPVERAGRKGEGYWPEKYQRFFLQNRNVLLILKEYIFVKAKSMIKKVMEKLGLR。

[0148] Gene sequence (SEQ ID NO: 62) of an α-1,3-fucosyltransferase derived from Roseburia sp.:

[0149]

[0150] Polypeptide sequence (SEQ ID NO:63) of an α-1,3-fucosyltransferase derived from a Bacteroidales bacterium:

[0151] MIKICCLHKNVSDIPSMLRFLLLDPSFAGKLVWDEEAPDILIASEWIYYKSALFRKFRDLYGKSGIKVSFMGEAIEPDFNVFDYCVGFSDGFKDNPAFLRLPSPYHMFSEFLKEKENPIKTPDEARAELERKSGFCSFLYSNPKANPVRDQLFYEISAYKKVDSLGRHLNNVSIPGTGYIGHKGEGVLIKANYKFSISSENSCFRGYTSEKLLTSLEAHSLPIYWGNPNVADDINPECFIDAGKMSGMDELISVIERFDNDDALWCEMVSKPWMTPSQESAHKIRTEAFLQGMRRLLTGELPPIMARGYHVDMYRDHFLSGEFFLDSRKEKLRNRLSQLSLSRHPQG。

[0152] Gene sequence (SEQ ID NO:64) of an α-1,3-fucosyltransferase derived from a Bacteroidales bacterium:

[0153]

[0154] Polypeptide sequence (SEQ ID NO: 65) of α-1,3-fucosyltransferase derived from Treponema socranskii:

[0155] MKQIRVFMIHNKIHSVDDFLSYLNIKDLSSNYEFIWDENNPDYLISSEQIYFFKKSFSLFKKLYSKKRINIFMGGEYMIPDFNVFDYAVGYHDDLKCNDRYAQLLTPDIFFNRGFIKRDAFKKISKDEAYEILKTKTGFCDFLYSNSRAHPNRDFFFHLLSKYKKVDSLGAWLNNVGRKGTGYMGHEGDIIPIKAPYKFSIAFENCYGKGYTSEKILTSLQAHCIPIYWGDPDIDRIVNSKAFVNCNEFNSFEEVIEKIKQIDQDDALWCEMASQPWQNEENIAYAKERNEYYYNFWINIFEQDYEKCHRVACGTHPENYIEWVKSAYPIVDGRLKTIMIRKMKTLLRIIRTK。

[0156] Gene sequence (SEQ ID NO: 66) of α-1,3-fucosyltransferase derived from Treponema socranskii:

[0157]

[0158] Polypeptide sequence (SEQ ID NO: 67) of α-1,3-fucosyltransferase derived from Blautia sp.:

[0159] MKQIRILPLINNDYNIEWLYQLFLLDKEKYEIVVDYEDPQYVLVVISHVIYRNALLYKQLQNIYSEDRIIIFLGDEAISPDMNLFDYAITYDDELFMRDRICRRPTVLWMRGGDKICKKNHLEYAEAEQKYDTRKFCSFIYSNAKANTHRASIFYGLNSYQNVDALGKYLHNCDIEITRNEKNWFELSIEQKENYRFSIAAENSFFRGYTSEKIISSFLAHSIPIYWGNPNIEKEFNPKAFINCHNYKDLNEVIEKVKTINEDKALWSYIVTQPWQTDEQISLQNEQINNYRSFTDHIFAQDIKDAKRIYQGTAVWNYIDFWMNKTVGM。

[0160] Gene sequence (SEQ ID NO: 68) of α-1,3-fucosyltransferase derived from Blautia sp.:

[0161] atgaaacaaatccgtatccttcctctgattaataacgattataacatcgaatggttgtaccaactgtttctcttggacaaggaaaagtacgaaatcgtcgtggattacgaggacccacagtacgtgctcgtagtcattagccacgtcatctaccgcaacgctttgttgtacaagcaactgcagaacatctattccgaagaccgtattattatttttctgggcgatgaagcaatctcacccgacatgaaccttttcgactacgctatcacctacgatgatgaactttttatgcgagaccgtatctgccgccgcccaaccgtgctctggatgcgcggcggagataaaatttgcaagaagaaccatctggagtacgccgaagccgagcagaagtacgacacgcgaaagttttgttctttcatctacagcaatgcgaaagccaatacccaccgcgcaagcatcttttatggcctgaactcataccaaaacgtggatgcactgggcaaatatttgcataactgcgatatcgaaatcacacgtaacgaaaagaattggttcgaattgagcattgaacaaaaggaaaattaccgattttctattgccgcggaaaactcttttttccgcggttacacgtccgagaagatcatctcgtcctttctggctcacagcatccctatttactggggcaacccaaacatcgaaaaggaattcaaccctaaggccttcatcaattgtcacaactataaagaccttaacgaggtcatcgaaaaagtcaagacaatcaatgaggacaaagctctctggtcttacattgttacgcagccctggcaaactgatgaacaaatttccctgcagaacgagcagatcaacaactatcgatccttcaccgatcacatcttcgcacaggatatcaaggatgctaagcgaatctaccagggtactgccgtctggaactatatcgacttctggatgaataagacggtcggcatgtaa。

[0162] The polypeptide sequence (SEQ ID NO:69) of an α-1,3-fucosyltransferase derived from a Lachnospiraceae bacterium:

[0163] MEAYKRKIVYLHPIYLNGDKNKVVEKIKDEFRLQNNESCNVEFEWSMEKPEYIIATELIYVDRKIAKLFFDLVEKNPNAILIFIGYEFVSPDMNIFDYALSFDRNCKMQDRIVKVPTIYHFAGYMLETKNQMSYEEALKEYEKRKFCNFIYSNSNAHPVRDQIFYALERYKRVDSLGGHLNNMGNTITRYEKDWGYQSINMKAQYRFSISAGNAIGSGNTDEKLLTSFQAHSIPIFWGDPDVEDEFNPDAFINVGKFSTLEDLVFEVSRIDQDPHLWSKMVSAPWQTPEQEREKIKEEERYTEFWLNVFCQDISKAHRAPQGTFTSVYQKVYYGMLKKCVGRSGIIEQLRERCHHLKIRRR。

[0164] The gene sequence (SEQ ID NO:70) of an α-1,3-fucosyltransferase derived from a Lachnospiraceae bacterium:

[0165]

[0166] Polypeptide sequence of GDP-D-mannose-4,6-dehydratase from Escherichia coli (E. coli) (SEQ ID NO:71):

[0167] MSKVALITGVTGQDGSYLAEFLLEKGYEVHGIKRRASSFNTERVDHIYQDPHTCNPKFHLHYGDLSDTSNLTRILREVQPDEVYNLGAMSHVAVSFESPEYTADVDAMGTLRLLEAIRFLGLEKKTRFYQASTSELYGLVQEIPQKETTPFYPRSPYAVAKLYAYWITVNYRESYGMYACNGILFNHESPRRGETFVTRKITRAIANIAQGLESCLYLGNMDSLRDWGHAKDYVKMQWMMLQQEQPEDFVIATGVQYSVRQFVEMAAAQLGIKLRFEGTGVEEKGIVVSVTGHDAPGVKPGDVIIAVDPRYFRPAEVETLLGDPTKAHEKLGWKPEITLREMVSEMVANDLEAAKKHSLLKSHGYDVAIALES。

[0168] Gene sequence of GDP-D-mannose-4,6-dehydratase from Escherichia coli (E. coli) (SEQ ID NO:72):

[0169]

[0170] Polypeptide sequence (SEQ ID NO: 73) of GDP-L-fucose synthase derived from Escherichia coli (E. coli) MG1655:

[0171] MSKQRVFIAGHRGMVGSAIRRQLEQRGDVELVLRTRDELNLLDSRAVHDFFASERIDQVYLAAAKVGGIVANNTYPADFIYQNMMIESNIIHAAHQNDVNKLLFLGSSCIYPKLAKQPMAESELLQGTLEPTNEPYAIAKIAGIKLCESYNRQYGRDYRSVMPTNLYGPHDNFHPSNSHVIPALLRRFHEATAQNAPDVVVWGSGTPMREFLHVDDMAAASIHVMELAHEVWLENTQPMLSHINVGTGVDCTIRELAQTIAKVVGYKGRVVFDASKPDGTPRKLLDVTRLHQLGWYHEISLEAGLASTYQWFLENQDRFRG。

[0172] Gene sequence (SEQ ID NO: 74) of GDP-L-fucose synthase derived from Escherichia coli (E. coli) MG16555:

[0173] atgagtaaacaacgagtttttattgctggtcatcgcgggatggtcggttccgccatcaggcggcagctcgaacagcgcggtgatgtggaactggtattacgcacccgcgacgagctgaacctgctggacagccgcgccgtgcatgatttctttgccagcgaacgtattgaccaggtctatctggcggcggcgaaagtgggcggcattgttgccaacaacacctatccggcggatttcatctaccagaacatgatgattgagagcaacatcattcacgccgcgcatcagaacgacgtgaacaaactgctgtttctcggatcgtcctgcatctacccgaaactggcaaaacagccgatggcagaaagcgagttgttgcagggcacgctggagccgactaacgagccttatgctattgccaaaatcgccgggatcaaactgtgcgaatcatacaaccgccagtacggacgcgattaccgctcagtcatgccgaccaacctgtacgggccacacgacaacttccacccgagtaattcgcatgtgatcccagcattgctgcgtcgcttccacgaggcgacggcacagaatgcgccggacgtggtggtatggggcagcggtacaccgatgcgcgaatttctgcacgtcgatgatatggcggcggcgagcattcatgtcatggagctggcgcatgaagtctggctggagaacacccagccgatgttgtcgcacattaacgtcggcacgggcgttgactgcactatccgcgagctggcgcaaaccatcgccaaagtggtgggttacaaaggccgggtggtttttgatgccagcaaaccggatggcacgccgcgcaaactgctggatgtgacgcgcctgcatcagcttggctggtatcacgaaatctcactggaagcggggcttgccagcacttaccagtggttccttgagaatcaagaccgctttcgggggtaa。

[0174] Promoter Ptuf (SEQ ID NO:75) derived from Corynebacterium glutamicum ATCC 13032:

[0175] TTGCTTTTCGACGCCCCACCCCGCGCGTTTTAGCGTGTCAGTAGGCGCGTAGGGTAAGTGGGGTAGCGGCTTGTTAGATATCTTGAAATCGGCTTTCAACAGCATTGATTTCGATGTATTTAGCTGGCCGTTACCCTGCGAATGTCCACAGGGTAGCTGGTAGTTTGAAAATCAACGCCGTTGCCCTTAGGATTCAGTAACTGGCACATTTTGTAATGCGCTAGATCTGTGTGCTCAGTCTTCCAGGCTGCTTATCACAGTGAAAGCAAAACCAATTCGTGGCTGCGAAAGTCGTAGCCACCACGAAGTCCAGGAGGACATACA。

[0176] Promoter Psod (SEQ ID NO:76) derived from Corynebacterium glutamicum ATCC 13032:

[0177] tagctgccaattattccgggcttgtgacccgctacccgataaataggtcggctgaaaaatttcgttgcaatatcaacaaaaaggcctatcattgggaggtgtcgc accaagtacttttgcgaagcgccatctgacggattttcaaaagatgtatatgctcggtgcggaaacctacgaaaggattttttaccc。

[0178] Polypeptide sequence of lactose permease LacY (SEQ ID NO:77) derived from Escherichia coli:

[0179] MYYLKNTNFWMFGLFFFFYFFIMGAYFPFFPIWLHDINHISKSDTGIIFAAISLFSLLFQPLFGLLSDKLGLRKYLLWIITGMLVMFAPFFIFIFGPLLQYNILVGSIVGGIYLGFCFNAGAPAVEAFIEKVSRRSNFEFGRARMFGCVGWALCASIVGIMFTINNQFVFWLGSGCALILAVLLFFAKTDAPSSATVANAVGANHSAFSLKLALELFRQPKLWFLSLYVIGVSCTYDVFDQQFANFFTSFFATGEQGTRVFGYVTTMGELLNASIMFFAPLIINRIGGKNALLLAGTIMSVRIIGSSFATSALEVVILKTLHMFEVPFLLVGCFKYITSQFEVRFSATIYLVCFCFFKQLAMIFMSVLAGNMYESIGFQGAYLVLGLVALGFTLISVFTLSGPGPLSLLRRQVNEVA。

[0180] Gene sequence (SEQ ID NO:78) of lactose permease LacY derived from Escherichia coli:

[0181]

[0182] Promoter Pcg2195 (SEQ ID NO:79) derived from Corynebacterium glutamicum ATCC 13032:

[0183] gagtatcgaaaaatttccacgtcaagttaactgcgttaataaaggtggagaataagttgtttccaagatcaattcaaggaaagttgcattttcgcaggtcagtgttaccccctaagactacccctttccattgcatacaaaggaaatacatatagacttttgggcattagattacctcgataaaagtttagggaatctaaattcattgatcaagacttgctgtcgcctagctctaattcacttgagcccggctgctaaaggtcaagatcattgaatgcactacttgctagcagtcatctgaaaaaacgacgttggttcgtagtcgctggaaatttaataattcctccgtccccttcaactagggggtggaaacccgactatttccgaaggactattctc。

[0184] Polypeptide sequence of α-1,2-fucosyltransferase HpfutC derived from Helicobacter pylori (SEQ ID NO:80):

[0185] MAFKVVQICGGLGNQMFQYAFAKSLQKHSNTPVLLDITSFDWSDRKMQLELFPIDLPYASAKEIAIAKMQHLPKLVRDALKCMGFDRVSQEIVFEYEPKLLKPSRLTYFFGYFQDPRYFDAISPLIKQTFTLPPPPENNKNNNKKEEEYQCKLSLILAAKNSVFVHIRRGDYVGIGCQLGIDYQKKALEYMAKRVPNMELFVFCEDLEFTQNLDLGYPFMDMTTRDKEEEAYWDMLLMQSCQHGIIANSTYSWWAAYLIENPEKIIIGPKHWLFGHENILCKEWVKIESHFEVKSQKYNA。

[0186] Gene sequence (SEQ ID NO: 81) of α-1,2-fucosyltransferase HpfutC derived from Helicobacter pylori:

[0187] atggccttcaaggttgtgcagatttgcggtggactcggcaatcagatgttccagtatgcattcgcgaaatcgttgcaaaaacattctaacaccccagtgctgctggacatcacctcatttgattggtctgatcgcaagatgcagctggagctgtttccgattgatcttccctatgcgtccgccaaggagatcgcaattgctaaaatgcaacacctacccaaattggtccgtgatgcattgaagtgcatgggtttcgaccgcgtatcgcaggaaatcgtgtttgaatatgagcctaagctgctgaagccatctcggctcacctacttcttcgggtatttccaagatccacgctacttcgatgccatttcccctctcatcaagcagaccttcacgctacctccgccgccagaaaataataaaaacaacaacaagaaggaagaggaataccagtgcaagctgtccttgattcttgctgcgaagaatagcgtctttgttcacatcaggcgtggcgactacgtcggtatcggctgccagttgggaatcgactaccaaaaaaaggcgctggagtacatggctaaacgagttcccaacatggaactctttgtgttttgcgaagacctggagttcacccaaaacctcgaccttggctacccattcatggacatgaccacacgcgacaaagaagaagaggcttactgggatatgctgctcatgcagagctgtcagcacggcatcatagccaactccacttattcctggtgggcagcatacctcattgaaaaccctgagaagatcatcattggcccaaagcactggttgtttggtcatgagaacatcctgtgcaaagagtgggtgaaaattgagtcccacttcgaagtcaagtcacaaaagtacaacgcctaa。

[0188] Polypeptide sequence (SEQ ID NO:82) of DdahC derived from Campylobacter jejuni:

[0189] MMQKDSKIYIAGHSGLVGSAILNELKQQGYKNLVFKTHFELDLTNQKAVADFFEREKPEYVILAAAKAGGILANNTYRADFIYQNLMIECNVIHNAYLHKVKKLLFIASTTVYPKNATLPTSEEQMLSGDLEYTNKPYAIAKISGLMLCESYNLQYNTNFIAITPTNLYGNNDKFDLEKSHVLPGILRKMHLAKLLNEKRYEDLLNDLKFDSIEEAKNYLKKFGVDKDNVEIWGSGKPTREFLHSQDLANACLFIMNNIDFKDLKSDNIEIINTHLNIGPHKNITIKELAELIKNIVGFKGKLVFNLNRPDGAMQKFTDCSKIHSLGWKHKIELEDGIKMMYKWYLKEQNIRQ.

[0190] Gene sequence (SEQ ID NO:83) of DdahC derived from Campylobacter jejuni:

[0191]

[0192] The polypeptide sequence (SEQ ID NO:84) of OsGME derived from rice (Oryza sativa):

[0193] MGSSEKNGTAYGEYTYAELEREQYWPSEKLRISITGAGGFIGSHIARRLKSEGHYIIASDWKKNEHMTEDMFCHEFHLVDLRVMDNCLKVTNGVDHVFNLAADMGGMGFIQSNHSVIMYNNTMISFNMLEAARINGVKRFFYASSACIYPEFKQLETNVSLKESDAWPAEPQDAYGLEKLATEELCKHYTKDFGIECRVGRFHNIYGPFGTWKGGREKAPAAFCRKAQTSTDRFEMWGDGLQTRSFTFIDECVEGVLRLTKSDFREPVNIGSDEMVSMNEMAEIILSFEDRELPIHHIPGPEGVRGRNSDNTLIKEKLGWAPTMKLKDGLRFTYFWIKEQIEKEKTQGVDIAGYGSSKVVSTQAPVQLGSLRAADGKE。

[0194] The gene sequence (SEQ ID NO:85) of OsGME derived from rice (Oryza sativa):

[0195]

[0196] In a seventh aspect, the present invention provides a fucosyllactose, which is prepared by the method described in the sixth aspect.

[0197] In an eighth aspect, the present invention provides the use of the fucosyllactose described in the seventh aspect in the preparation of drugs, health products or food additives.

[0198] Compared with the prior art, the present invention has the following beneficial effects:

[0199] By optimizing the codons of the encoding gene of fucosyltransferase, the present invention can obtain a heterologously expressed fucosyltransferase, which effectively catalyzes the reaction and improves the yields of 3-fucosyllactose and 2'-fucosyllactose. Detailed implementation manners

[0200] To further illustrate the technical means and effects adopted by the present invention, the present invention will be further described below with reference to examples. It can be understood that the specific implementation manners described herein are only used to explain the present invention, rather than limiting the present invention.

[0201] For those not specifying specific techniques or conditions in the examples, the techniques or conditions described in the literature in this field or according to the product specifications are followed. For reagents or instruments without indicating the manufacturer, they are all conventional products that can be obtained through regular channels.

[0202] Example 1

[0203] Cultivation of Corynebacterium glutamicum and detection of products.

[0204] Two different culture media were used to culture the strain, namely the seed medium LBHI and the fermentation medium FM20.

[0205] The components of the seed medium LBHI are as follows: yeast powder 10 g / L, peptone 5 g / L, NaCl 10 g / L, brain heart infusion (BHI) 18.5 g / L.

[0206] The components of the seed medium LBHI+K15 (also known as LBHI+kan) are as follows: LBHI medium, kanamycin 15 mg / L.

[0207] The components of the LBHI+sucrose medium are as follows: LBHI medium, sucrose 10 g / L.

[0208] The components of the fermentation medium FM20 are as follows: glucose 30 g / L, lactose 20 g / L, peptone 4 g / L, yeast extract 2 g / L, (NH4)2SO4 10 g / L, urea 5 g / L, KH2PO4 1 g / L, K2HPO4 1 g / L, MgSO4 0.25 g / L, MOPS 42 g / L, CaC12 10 mg / L, Biotin 0.2 mg / L, Protocatechuic acid 0.03 mg / L, FeSO4·7H2O 10 mg / L, MnSO4·H2O 10 mg / L, ZnSO4·7H2O 1 mg / L, CuSO4 0.2 mg / L, NiCl2·6H2O 0.02 mg / L, pH 7.0).

[0209] The seed medium is sterilized by autoclaving (121 °C, 20 min), and the fermentation medium is sterilized by filtration (0.22 μm filter membrane). When necessary, the medium is made selective by adding antibiotics (e.g., kanamycin). When preparing solid plates, 2% agar is added additionally to the medium.

[0210] When the strain is fermented, the colonies on the plate are picked into the seed medium for overnight culture. The strain after overnight culture is inoculated into a 96-well plate containing the fermentation medium at an inoculation amount of 5%. After culturing the plate at 30 °C, 900 rpm, and a humidity of 80% for 48 h, 80 °C water bath is used for 15 min for the whole culture, then centrifuged and the supernatant is taken to measure the product titer of the whole culture, or the plate is centrifuged and the supernatant is taken to measure the product titer in the supernatant after culture.

[0211] Detection of product titer: The determination is analyzed by HPLC-RID of Agilent. By analyzing the refractive index change of the mobile phase when the sample passes through, the concentration of the substance is measured. An Agilent hilic-z 4 μm, 4.6 * 250 mm column and mobile phase are used to separate all sugars in an isocratic flow rate mode. The mobile phase contains 700 mL of acetonitrile, 300 mL of ultrapure water, 0.23 g of ammonium acetate, 1.5 mL of 30% ammonia water, the flow rate is 0.8 mL / min, the injection volume is 3 μL, the column temperature is 35 °C, and the differential detector temperature is 35 °C.

[0212] Genetic modification of Corynebacterium glutamicum: Using the sacB modification method, specifically, using a plasmid based on pK18mobsacB, through homologous recombination, using the sucrase encoded by the sacB gene of Bacillus subtilis as a counter-selection marker. This enzyme catalyzes the hydrolysis of sucrose and the synthesis of high-molecular-weight fructose polymers called levans. When the sacB gene is expressed in Corynebacterium glutamicum, the strain cannot grow in a medium containing sucrose. The genetic modification operation is carried out with reference to (Lothar Eggeling and Michael Bott, Handbook of Corynebacterium glutamicum, 1st Edition, 2005).

[0213] Kanamycin resistance and sacB gene counter-selection method for recombinant bacteria: Electrotransform the foreign plasmid into the strain and spread it on an LBHI + kan plate. Pick clones from the LBHI + kan plate and culture them overnight in antibiotic-free LBHI. Spread the overnight culture on an LBHI + sucrose plate and culture until clones form. Dot the clones grown on the LBHI + sucrose plate onto an antibiotic-free LBHI plate and an LBHI + kan plate. Use primers to perform PCR confirmation on the clones that do not grow on the LBHI + kan plate but grow on the antibiotic-free LBHI plate.

[0214] Example 2

[0215] Construct the recombinant Corynebacterium glutamicum strain manBC-gmdwcaG.

[0216] Using the Corynebacterium glutamicum ATCC 13032 genome as a template, homologous arms manB-Up and manB-Down were obtained by PCR amplification using primer pairs manB-U-F / manB-U-R and manB-D-F / manB-D-R, respectively (the PCR system used 2×Phanta Max Master Mix (Dye Plus), Vazyme); the plasmid pK18mobsacB was digested with restriction enzymes EcoRI and XbaI to obtain a linearized fragment line-pK18mobsacB; the above three fragments: manB-Up, manB-Down, and line-pK18mobsacB were seamlessly assembled using a recombinant cloning kit (ClonExpress Multis One Step Cloning Kit, Vazyme, catalog number: C113-02), and the reaction system and reaction conditions were all referred to the kit instructions. After seamless assembly, it was transformed into Trans1 T1 competent cells to obtain the recombinant plasmid pK18mobsacB-manB. The verified plasmid was electrotransformed into Corynebacterium glutamicum ATCC13032, and the manB promoter -10 region sequence "TAGGAT" was replaced with "TATAAT" and the manB promoter -35 region sequence "TTCGGA" was replaced with "TTGGCA" by successive selection with kanamycin resistance and the sacB gene to obtain the strain manB with modified manB promoter -10 and -35 regions.

[0217] Using the Corynebacterium glutamicum ATCC 13032 genome as a template, homologous arms manC-Up and manC-Down were obtained by PCR amplification using primer pairs manC-U-F / manC-U-R and manC-D-F / manC-D-R, respectively (the PCR system used 2×Phanta Max Master Mix (Dye Plus), Vazyme); the plasmid pK18mobsacB was digested with restriction enzymes EcoRI and XbaI to obtain the linearized fragment line-pK18mobsacB; the above three fragments, manC-Up, manC-Down, and line-pK18mobsacB, were seamlessly assembled using a recombinant cloning kit (ClonExpress Multis One Step Cloning Kit, Vazyme, catalog number: C113-02), and the reaction system and reaction conditions were all referred to the kit instructions. After seamless assembly, Trans1 T1 competent cells were transformed to obtain the recombinant plasmid pK18mobsacB-manC. The verified plasmid was electrotransformed into the previously obtained Corynebacterium glutamicum recombinant strain manB, and the manC promoter -10 region sequence "TAAAGT" was replaced with "TATAAT" by successive selection using kanamycin resistance and the sacB gene counter-selection to obtain the strain manBC with the modified manC promoter -10 region.

[0218] Using the Corynebacterium glutamicum ATCC 13032 genome as a template, PCR amplifications were performed using primer pairs gmdwcaG-U-F / gmdwcaG-U-R, gmdwcaG-D-F / gmdwcaG-D-R, and Ptuf-F / Ptuf-R to obtain the upstream homologous arms gmdwcaG-Up, gmdwcaG-Down, and the promoter Ptuf (the PCR system used 2×Phanta Max Master Mix, (DyePlus), Vazyme); using the Escherichia coli MG1655 genome as a template, PCR amplification was performed using the primer pair gmd-F / wcaG-R to obtain the gene fragment gmdwcaG (the PCR system used 2×Phanta Max Master Mix, (Dye Plus), Vazyme); the plasmid pK18mobsacB was digested with the restriction enzymes EcoRI and XbaI to obtain the linearized fragment line-pK18mobsacB; the above 5 fragments: gmdwcaG-Up, gmdwcaG-Down, Ptuf, gmdwcaG, line-pK18mobsacB were seamlessly assembled using a recombinant cloning kit (ClonExpress Multis One Step Cloning Kit, Vazyme, product number: C113-02), and the reaction system and reaction conditions were all referred to the kit instructions. After seamless assembly, Trans1T1 competent cells were transformed to obtain the recombinant plasmid pK18mobsacB-gmdwcaG. The verified plasmid was electrotransformed into the previously obtained Corynebacterium glutamicum recombinant strain manBC, and the basic strain manBC-gmdwcaG into which the exogenous gmd and wcaG were introduced was further obtained by successive selection using kanamycin resistance and the sacB gene. The polypeptide sequence of GDP-D-mannose-4,6-dehydratase Gmd is SEQ ID NO:71, and the gene sequence is SEQ ID NO:72. The polypeptide sequence of GDP-L-fucose synthase WcaG is SEQ ID NO:73, and the gene sequence is SEQ ID NO:74. The gene sequence of the promoter Ptuf is SEQ ID NO:75, and the primer sequences are shown in Table 1.

[0219] Table 1

[0220] Sequence number Primer name Primer sequence SEQ ID NO:86 manB-U-F aaacagctatgacatgattacgaattccgtgccgattgcggaaagttc SEQ ID NO:87 manB-U-R tatacccctgccaaaggatccaatgccaatagcaaaccttatgattcgtcctg SEQ ID NO:88 manB-D-F ctattggcattggatcctttggcaggggtataattgcaagcgttattttgttccc SEQ ID NO:89 manB-D-R gtaaaacgacggccagtgccaagcttgtttgccgcatccacagcaac SEQ ID NO:90 manC-U-F ggggatcctctagagtcgacctgcagatgtgcgaaacgtcaccctg SEQ ID NO:91 manC-U-R tagattcattatacatttgcctgaaatgagttgaattcac SEQ ID NO:92 manC-D-F aactcatttcaggcaaatgtataatgaatctacattaattttctgaaatggatttgc SEQ ID NO:93 manC-D-R gtaaaacgacggccagtgccaagctttgggtgtccagaatgctgttg SEQ ID NO:94 gmdwcaG-U-F cgagctcggtacccggggatcctctagatcttccttagcgatatcaccagg SEQ ID NO:95 gmdwcaG-U-R gggtggggcgtcgaaaagcaaatggcacacagctacgcag SEQ ID NO:96 gmdwcaG-D-F caagaccgctttcgggggtaactgaactcctcaacgttatggc SEQ ID NO:97 gmdwcaG-D-R gtaaaacgacggccagtgccaagcttgataacggcgaggaaagccc SEQ ID NO:98 Ptuf-F ttgcttttcgacgccccac SEQ ID NO:99 Ptuf-R gatgagagcgacttttgacattgtatgtcctcctggacttcgtg SEQ ID NO:100 gmd-F atgtcaaaagtcgctctcatcac SEQ ID NO:101 wcaG-R ttacccccgaaagcggtcttg

[0221] Example 3

[0222] Construction of an α-1,3-fucosyltransferase-lactose permease expression plasmid.

[0223] Using the Corynebacterium glutamicum ATCC 13032 genome as a template, the promoter Psod (SEQ ID NO:76) was obtained by PCR amplification using the primer pair Psod-F / Psod-R. The PCR system used 2×Phanta Max Master Mix, (DyePlus), Vazyme; using the Escherichia coli MG1655 genome as a template, the lactose permease gene fragment LacY was obtained by PCR amplification using the primer pair LacY-F / LacY-R (the PCR system used 2×Phanta Max Master Mix, (DyePlus), Vazyme); the plasmid pJC1 was digested with the restriction enzymes BamHI and SalI to obtain the linearized fragment line-pJC1; the above three fragments: Psod, LacY, and line-pJC1 were seamlessly assembled using a recombinant cloning kit (ClonExpress Multis OneStep Cloning Kit, Vazyme, product number: C113-02). The reaction system and reaction conditions were all carried out according to the kit instructions. After seamless assembly, Trans1 T1 competent cells were transformed to obtain the recombinant plasmid pJC1-Psod_LacY. The polypeptide sequence of LacY is SEQ ID NO:77, and the gene sequence is SEQ ID NO:78.

[0224] By mining databases for α-1,3-fucosyltransferase, polypeptide sequences were obtained, including those from Candidatus Akkermansia timonensis (SEQ ID NO:1), Akkermansia sp. (SEQ ID NO:3), Akkermansia sp. (SEQ ID NO:5), Akkermansia sp. (SEQ ID NO:7), Akkermansiaceaebacterium (SEQ ID NO:9), Akkermansia sp. (SEQ ID NO:11), Azospirillum oleiclasticum (SEQ ID NO:13), Roseomonas genomospecies (SEQ ID NO:15), Azospirillum picis (SEQ ID NO:17), Helicobacter sp. (SEQ ID NO:19), Antarcticibacterium arcticum (SEQ ID NO:21), Bacteroides congonensis (SEQ ID NO:23), Azospirillum sp. BE72 (SEQ ID NO:25), and Azospirillum sp. 412522The polypeptide sequence (SEQ ID NO: 27) of (412522), the polypeptide sequence (SEQ ID NO: 29) from Azospirillum endophyticum, the polypeptide sequence (SEQ ID NO: 31) from Phycisphaerae bacterium, the polypeptide sequence (SEQ ID NO: 33) from Helicobacter acinonychis, the polypeptide sequence (SEQ ID NO: 35) from Helicobacter mehlei, the polypeptide sequence (SEQ ID NO: 37) from Helicobacter bizzozeronii, the polypeptide sequence (SEQ ID NO: 39) from Helicobacter bizzozeronii CIII-1, the polypeptide sequence (SEQ ID NO: 41) from Helicobacter cynogastricus, the polypeptide sequence (SEQ ID NO: 43) from Helicobacter ailurogastricus, the polypeptide sequence (SEQ ID NO: 45) from Candidate division KSB1 bacterium, the polypeptide sequence (SEQ ID NO: 47) from Lachnospiraceae bacterium, the polypeptide sequence (SEQ ID NO: 49) from Bacteroidotabacterium, the polypeptide sequence (SEQ ID NO: 51) from Wenyingzhuangia fucanilytica, the polypeptide sequence (SEQ ID NO: 53) from Phycisphaerales bacterium, the polypeptide sequence (SEQ ID NO: 55) from Succinivibrio dextrinosolvens, the polypeptide sequence (SEQ ID NO: 57) from Candidatus Akkermansia timonensis, the polypeptide sequence (SEQ ID NO: 59) from Akkermansia sp., the polypeptide sequence from Roseburia sp.) The polypeptide sequences (SEQ ID NO: 61), the polypeptide sequence from Bacteroidales bacterium (SEQ ID NO: 63), the polypeptide sequence from Treponema socranskii (SEQ ID NO: 65), the polypeptide sequence from Blautia sp. (SEQ ID NO: 67), the polypeptide sequence from Lachnospiraceae bacterium (SEQ ID NO: 69). Without changing the amino acid sequence of the polypeptide, the gene sequences encoding the above polypeptides were replaced with codons preferred by Glutamicibacter. After codon optimization, their gene sequences are SEQ ID NO: 2, SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 8, SEQ ID NO: 10, SEQ ID NO: 12, SEQ ID NO: 14, SEQ ID NO: 16, SEQ ID NO: 18, SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26, SEQ ID NO: 28, SEQ ID NO: 30, SEQ ID NO: 32, SEQ ID NO: 34, SEQ ID NO: 36, SEQ ID NO: 38, SEQ ID NO: 40, SEQ ID NO: 42, SEQ ID NO: 44, SEQ ID NO: 46, SEQ ID NO: 48, SEQ ID NO: 50, SEQ ID NO: 52, SEQ ID NO: 54, SEQ ID NO: 56, SEQ ID NO: 58, SEQ ID NO: 60, SEQ ID NO: 62, SEQ ID NO: 64, SEQ ID NO: 66, SEQ ID NO: 68, SEQ ID NO: 70.

[0225] Add the promoter Pcg2195 sequence (SEQ ID NO: 79) in front of all α-1,3-fucosyltransferase gene sequences for gene synthesis. The gene synthesis vector uses the pJC1-Psod_LacY plasmid, and the gene synthesis sequence is located after the restriction enzyme site BamHI. The recombinant plasmids are named pJC1-α-1,3-FucT1-LacY, pJC1-α-1,3-FucT2-LacY, pJC1-α-1,3-FucT3-LacY, pJC1-α-1,3-FucT4-LacY, pJC1-α-1,3-FucT5-LacY, pJC1-α-1,3-FucT6-LacY, pJC1-α-1,3-FucT7-LacY, pJC1-α-1,3-FucT8-LacY, pJC1-α-1,3-FucT9-LacY, pJC1-α-1,3-FucT10-LacY, pJC1-α-1,3-FucT11-LacY, pJC1-α-1,3-FucT12-LacY, pJC1-α-1,3-FucT13-LacY, pJC1-α-1,3-FucT14-LacY, pJC1-α-1,3-FucT15-LacY, pJC1-α-1,3-FucT16-LacY, pJC1-α-1,3-FucT17-LacY, pJC1-α-1,3-FucT18-LacY, pJC1-α-1,3-FucT19-LacY, pJC1-α-1,3-FucT20-LacY, pJC1-α-1,3-FucT21-LacY, pJC1-α-1,3-FucT22-LacY, pJC1-α-1,3-FucT23-LacY, pJC1-α-1,3-FucT24-LacY, pJC1-α-1,3-FucT25-LacY, pJC1-α-1,3-FucT26-LacY, pJC1-α-1,3-FucT27-LacY, pJC1-α-1,3-FucT28-LacY, pJC1-α-1,3-FucT29-LacY, pJC1-α-1,3-FucT30-LacY, pJC1-α-1,3-FucT31-LacY, pJC1-α-1,3-FucT32-LacY, pJC1-α-1,3-FucT33-LacY, pJC1-α-1,3-FucT34-LacY, pJC1-α-1,3-FucT35-LacY (the recombinant plasmids correspond to the aforementioned α-1,3-fucosyltransferases and their gene sequences in the written order. Taking pJC1-α-1,3-FucT1-LacY as an example, the α-1,The 3-fucosyltransferase sequence is SEQ ID NO: 1, and the corresponding gene sequence is SEQ ID NO: 2. Taking pJC1-α-1,3-FucT2-LacY as an example, the α-1,3-fucosyltransferase sequence corresponding to pJC1-α-1,3-FucT1-LacY is SEQ ID NO: 3, and the corresponding gene sequence is SEQ ID NO: 4. The same applies to others. The primer sequences are shown in Table 2.,

[0226] Table 2

[0227]

[0228] Example 4

[0229] Construct a recombinant Corynebacterium glutamicum strain to synthesize 3-fucosyllactose.

[0230] The recombinant plasmids pJC1-α-1,3-FucT1-LacY, pJC1-α-1,3-FucT2-LacY, pJC1-α-1,3-FucT3-LacY, pJC1-α-1,3-FucT4-LacY, pJC1-α-1,3-FucT5-LacY, pJC1-α-1,3-FucT6-LacY, pJC1-α-1,3-FucT7-LacY, pJC1-α-1,3-FucT8-LacY, pJC1-α-1,3-FucT9-LacY, pJC1-α-1,3-FucT10-LacY, pJC1-α-1,3-FucT11-LacY, pJC1-α-1,3-FucT12-LacY, pJC1-α-1,3-FucT13-LacY, pJC1-α-1,3-FucT14-LacY, pJC1-α-1,3-FucT15-LacY, pJC1-α-1,3-FucT16-LacY, pJC1-α-1,3-FucT17-LacY, pJC1-α-1,3-FucT18-LacY, pJC1-α-1,3-FucT19-LacY, pJC1-α-1,3-FucT20-LacY, pJC1-α-1,3-FucT21-LacY, pJC1-α-1,3-FucT22-LacY, pJC1-α-1,3-FucT23-LacY, pJC1-α-1,3-FucT24-LacY, pJC1-α-1,3-FucT25-LacY, pJC1-α-1,3-FucT26-LacY, pJC1-α-1,3-FucT27-LacY, pJC1-α-1,3-FucT28-LacY, pJC1-α-1,3-FucT29-LacY, pJC1-α-1,3-FucT30-LacY, pJC1-α-1,3-FucT31-LacY, pJC1-α-1,3-FucT32-LacY, pJC1-α-1,3-FucT33-LacY, pJC1-α-1,3-FucT34-LacY, pJC1-α-1,3-FucT35-LacY obtained by gene synthesis of the above Example 3 were respectively electrotransformed into the aforementioned obtained basic strain manBC-gmdwcaG to obtain the Corynebacterium glutamicum recombinant strains Cg2FL-α-1,3-FucT1, Cg2FL-α-1,3-FucT2, Cg2FL-α-1,3-FucT3, Cg2FL-α-1,3-FucT4, Cg2FL-α-1,3-FucT5, Cg2FL-α-1,3-FucT6, Cg2FL-α-1,3-FucT7, Cg2FL-α-1,3-FucT8, Cg2FL-α-1,3-FucT9, Cg2FL-α-1,3-FucT10, Cg2FL-α-1,3-FucT11, Cg2FL-α-1,3-FucT12, Cg2FL-α-1,3-FucT13, Cg2FL-α-1,3-FucT14, Cg2FL-α-1,3-FucT15, Cg2FL-α-1,3-FucT16, Cg2FL-α-1,3-FucT17, Cg2FL-α-1,3-FucT18, Cg2FL-α-1,3-FucT19, Cg2FL-α-1,3-FucT20, Cg2FL-α-1,3-FucT21, Cg2FL-α-1,3-FucT22, Cg2FL-α-1,3-FucT23, Cg2FL-α-1,3-FucT24, Cg2FL-α-1,3-FucT25, Cg2FL-α-1,3-FucT26, Cg2FL-α-1,3-FucT27, Cg2FL-α-1,3-FucT28, Cg2FL-α-1,3-FucT29, Cg2FL-α-1,3-FucT30, Cg2FL-α-1,3-FucT31, Cg2FL-α-1,3-FucT32, Cg2FL-α-1,3-FucT33, Cg2FL-α-1,3-FucT34, Cg2FL-α-1,3-FucT35, the recombinant strains, the aforementioned recombinant plasmids and the corresponding α-1,3-FucT contained are as shown in Table 3 below.,

[0231] Table 3

[0232]

[0233]

[0234]

[0235] The active α-1,3-fucosyltransferase was screened out.,

[0236] Table 4

[0237]

[0238]

[0239] Example 5

[0240] Sequence analysis of α-1,3-fucosyltransferase.,

[0241] The active α-1,3-fucosyltransferase obtained from the test of Example 4 was classified as follows according to the yield of the corresponding recombinant strain: 1) α-1,3-fucosyltransferases (high-activity transferases) corresponding to recombinant strains with a 3-FL yield higher than 1 g / L, including α-1,3-FucT1, α-1,3-FucT2, α-1,3-FucT3, α-1,3-FucT4, α-1,3-FucT5, α-1,3-FucT6, α-1,3-FucT7, α-1,3-FucT8, α-1,3-FucT9; 2) α-1,3-fucosyltransferases (low-activity transferases) corresponding to recombinant strains with a 3-FL yield lower than 1 g / L, including α-1,3-FucT10, α-1,3-FucT11, α-1,3-FucT12, α-1,3-FucT13, α-1,3-FucT14, α-1,3-FucT15, α-1,3-FucT16. By comparing the sequences of low-activity and high-activity α-1,3-fucosyltransferases, it can be found that both low-activity and high-activity α-1,3-fucosyltransferases have the amino acid sequence of the conserved GDP-fucose binding domain Y[T / L / V / A][S / T]EK, where X can be any different amino acid residue (Table 5). Having the GDP-fucose binding domain is a necessary condition for the catalytic function of α-1,3-fucosyltransferase, but there are significant differences in the activities of transferases with this domain, and other domains also have an important impact on protein activity. Further comparative analysis reveals that: in addition to the conserved GDP-fucose binding domain, the high-activity fucosyltransferase also has a conserved motif GXHLNNT, while this motif is not conserved in the low-activity fucosyltransferase (Table 5). This indicates that this domain is crucial for the activity of α-1,3-fucosyltransferase.

[0242] Table 5

[0243]

[0244] Example 6

[0245] Application of the transferase in a strain with a new pathway for 3-fucosyllactose synthesis.

[0246] The patent with the application number CN 202410437877.0 (incorporated herein in its entirety by reference) discloses a new pathway for the synthesis of 2'-fucosyllactose, which allows the synthesis of GDP-L-fucose in cells through a new pathway different from the traditional de novo synthesis pathway of GDP-L-fucose, and then the synthesis of 2'-fucosyllactose from GDP-L-fucose and lactose. The new pathway for the synthesis of GDP-L-fucose includes: the synthesis of GDP-4-keto-6-deoxy-D-mannose by catalyzing GDP-D-mannose with GDP-D-mannose-4,6-dehydratase (GMD), and then the synthesis of GDP-D-rhamnose by catalyzing GDP-4-keto-6-deoxy-D-mannose with a reductase capable of catalyzing the conversion of GDP-4-keto-6-deoxy-D-mannose to GDP-D-rhamnose, and then the synthesis of GDP-L-fucose by catalyzing GDP-D-rhamnose with GDP-D-rhamnose-3,5-epimerase (GRE). Among them, a recombinant Corynebacterium glutamicum strain Cg2FL-11 was constructed for the production of 2'-fucosyllactose. The construction method of Cg2FL-11 is as follows:

[0247] The gene sequences of the α-1,2-fucosyltransferase HpfutC of Helicobacter pylori (SEQ ID NO: 80, and the amino acid sequence of this enzyme is SEQ ID NO: 81) and the lactose permease LacY gene sequence of Escherichia coli (SEQ ID NO: 78, and the amino acid sequence of this enzyme is SEQ ID NO: 77) were synthesized. The gene synthesis vector used the pUC57 plasmid, and the recombinant plasmids were named pUC57-HpfutC and pUC57-LacY respectively.

[0248] Using the genome of Corynebacterium glutamicum ATCC 13032 as a template, homologous arms HpfutC-lacY-Up, HpfutC-lacY-Down, promoter Psod, and Pcg2195 were obtained by PCR amplification using primer pairs HpfutC-lacY-U-F / HpfutC-lacY-U-R, HpfutC-lacY-D-F / HpfutC-lacY-D-R, Psod-F-1 / Psod-R-1, and Pcg2195-F-1 / Pcg2195-R-1, respectively (the PCR system used 2×Phanta Max Master Mix, (Dye Plus), Vazyme); using pUC57-LacY as a template, the lactose permease LacY gene fragment was obtained by PCR amplification using primer pair LacY-F / LacY-R (the PCR system used 2×Phanta Max Master Mix, (Dye Plus), Vazyme); using pUC57-HpfutC as a template, the α-1,2-fucosyltransferase HpfutC gene fragment was obtained by PCR amplification using primer pair HpfutC-F / HpfutC-R; the plasmid pK18mobsacB was digested with restriction enzymes EcoR I and Xba I to obtain the linearized fragment line-pK18mobsacB. The above 7 fragments: HpfutC-lacY-Up, Pcg2195, HpfutC, Psod, LacY, HpfutC-lacY-Down, and line-pK18mobsacB were seamlessly assembled using a recombinant cloning kit (ClonExpress Multis One Step Cloning Kit, Vazyme, product number: C113-02). The reaction system and reaction conditions were all referred to the kit instructions. After seamless assembly, Trans1 T1 competent cells were transformed to obtain the recombinant plasmid pK18mobsacB-HpfutC-lacY. The verified plasmid was electrotransformed into Corynebacterium glutamicum ATCC 13032, and kanamycin resistance and sacB gene counterselection were used successively to introduce the genes encoding HpfutC and LacY at the poxB locus of the Corynebacterium glutamicum ATCC 13032 genome. To enable the strain to have sufficient fucosyltransferase activity, the α-1,2-fucosyltransferase HpfutC gene sequence was inserted at the cg0554 locus and tnp2b locus of the Corynebacterium glutamicum genome in a similar manner.On this basis, the -10 region sequence "TAGGAT" of the phosphomannomutase ManB promoter and the -10 region sequence "TAAAGT" of the mannose-1-phosphate guanylyltransferase ManC promoter were replaced with "TATAAT" by the same sacB gene counter-selection method to increase the supply of the 2'-FL precursor GDP-L-fucose. Thus, the basic strain HpfutC-LacY-ManB-ManC was obtained, and the primers are shown in Table 6.

[0249] Table 6

[0250]

[0251]

[0252] The reductase DdahC derived from Campylobacter jejuni was synthesized, with its amino acid sequence being SEQ ID NO:82, its gene sequence being SEQ ID NO:83, the gene synthesis vector being pUC57, and the recombinant plasmid being named pUC57-11. The GDP-mannose-3,5-epimerase OsGRE derived from Oryza sativa was synthesized, with its amino acid sequence being SEQ ID NO:84, its gene sequence being SEQ ID NO:85, the gene synthesis vector being pRSFDuet, and the recombinant plasmid being named pRSFDuet-OsGRE.

[0253] Using the genome of Corynebacterium glutamicum ATCC 13032 as a template, the promoters Pgap, Pcg2195, and Psod were obtained by PCR amplification using the primer pairs Pgap-F-2 / Pgap-R-2, Pcg2195-F-2 / Pcg2195-R-2, and Psod-F-2 / Psod-R-2 in Table 7, respectively (the PCR system used 2×Phanta Max Master Mix, (Dye Plus), Vazyme). Using the genome of Escherichia coli MG1655 as a template, the gene fragment encoding GDP-D-mannose-4,6-dehydratase EcGMD was obtained by PCR amplification using the primer pair EcGMD-F and EcGMD-R shown in Table 7. Using pRSFDuet-OsGRE as a template, the gene fragment encoding GDP-D-rhamnose-3,5-epimerase OsGRE was obtained by PCR amplification using the primer pair OsGME-F and OsGME-R shown in Table 7. Using pUC57-11 as a template, the gene fragment encoding DdahC was obtained by PCR amplification using the corresponding primer pair 11-F-1 / 11-R-1 shown in Table 7 (the PCR system used 2×Phanta Max Master Mix, (Dye Plus), Vazyme); the plasmid pJC1 was digested with the restriction enzymes BamH I and SalI to obtain the linearized fragment line-pJC1. The fragments Pgap, Pcg2195, Psod, EcGMD, OsGME, line-pJC1, and the DdahC gene fragment were assembled seamlessly using a recombinant cloning kit (ClonExpress Multis One Step Cloning Kit, Vazyme, product number: C113-02). The reaction system and reaction conditions were all referred to the kit instructions. After seamless assembly, Trans1 T1 competent cells were transformed to obtain the recombinant plasmid: pJC1-Pgap_EcGMD-Pcg2195_11-Psod_OsGME, which contains the following elements: Pgap, Pcg2195, Psod, EcGMD, OsGME, line-pJC1, and DdahC.

[0254] Table 7

[0255]

[0256]

[0257] The above recombinant plasmid was electrotransformed into the basic strain HpfutC-LacY-ManB-ManC to obtain the recombinant Corynebacterium glutamicum strain Cg2FL-11.

[0258] The recombinant Corynebacterium glutamicum strain Cg2FL-11 was cultured and analyzed, and its 2'-FL yield could reach 78 mg / L. The generated 2'-FL was analyzed by LC-MS and the molecular weight was found to be 511 [M+Na]. + , and both the retention time and the molecular weight were consistent with the standard product, thus confirming that the aforementioned metabolic pathway can achieve the synthesis of 2'-FL.

[0259] Furthermore, a control recombinant Corynebacterium glutamicum strain without GDP-D-rhamnose-3,5-epimerase was also constructed. Using the aforementioned pUC57-11 as a template, the primer pair 11-F-2 / 11-R-2 in Table 7 above was used for PCR amplification to obtain the gene of DdahC (the PCR system used 2×Phanta Max Master Mix, (Dye Plus), Vazyme). Using the recombinant cloning kit (ClonExpress Multis One Step Cloning Kit, Vazyme, product number: C113-02), the gene of DdahC was seamlessly assembled with the aforementioned obtained Pgap, Pcg2195, EcGMD, and line-pJC1 gene fragments. The reaction system and reaction conditions were all referred to the kit instructions. After seamless assembly, it was transformed into Trans1 T1 competent cells to obtain the recombinant plasmid pJC1-Pgap_EcGMD-Pcg2195_11, which was electrotransformed into the aforementioned obtained basic strain HpfutC-LacY-ManB-ManC to obtain the control recombinant strain Cg2FL-26.

[0260] The recombinant Corynebacterium glutamicum strain Cg2FL-26 was cultured and analyzed, and it did not show the ability to produce 2'-FL, indicating that Ddahc is a monofunctional protein and only has reducing activity but no isomerization activity towards GDP-4-keto-6-deoxy-D-mannose. At this time, GME is necessary for the production of 2'-FL.

[0261] In the recombinant strain Cg2FL-11 constructed above, the sacB counter-selection method was used to knockout lacY and HpfutC in this strain first. Then, the expression plasmids of pJC1-α-1,3-FucT1-LacY, pJC1-α-1,3-FucT2-LacY, pJC1-α-1,3-FucT3-LacY, pJC1-α-1,3-FucT4-LacY, pJC1-α-1,3-FucT5-LacY, pJC1-α-1,3-FucT6-LacY, pJC1-α-1,3-FucT7-LacY, pJC1-α-1,3-FucT8-LacY, pJC1-α-1,3-FucT9-LacY, pJC1-α-1,3-FucT10-LacY, pJC1-α-1,3-FucT11-LacY, pJC1-α-1,3-FucT12-LacY, pJC1-α-1,3-FucT13-LacY, pJC1-α-1,3-FucT14-LacY, pJC1-α-1,3-FucT15-LacY, pJC1-α-1,3-FucT16 constructed in the foregoing embodiments were electrotransformed respectively to obtain the modified strains NCg3FL-α-1,3-FucT1, NCg3FL-α-1,3-FucT2, NCg3FL-α-1,3-FucT3, NCg3FL-α-1,3-FucT4, NCg3FL-α-1,3-FucT5, NCg3FL-α-1,3-FucT6, NCg3FL-α-1,3-FucT7, NCg3FL-α-1,3-FucT8, NCg3FL-α-1,3-FucT9, NCg3FL-α-1,3-FucT10, NCg3FL-α-1,3-FucT11, NCg3FL-α-1,3-FucT12, NCg3FL-α-1,3-FucT13, NCg3FL-α-1,3-FucT14, NCg3FL-α-1,3-FucT15, NCg3FL-α-1,3-FucT16. Cultivation and supernatant product determination were carried out in the manner described in Example 1. The relative changes in the product titers of 3-fucosyllactose detected are shown in Table 8 below. The relative changes in the product titers were determined in the following manner: taking the 3-FL product titer of NCg3FL-Fuct1 as 100%, dividing the 3-FL product titers of the remaining strains by the product titer of NCg3FL-Fuct1, and then multiplying by 100%.

[0262] Table 8

[0263]

[0264]

[0265] It can be seen from the tabular results that after culturing the transformed strains NCg3FL-α-1,3-FucT1, NCg3FL-α-1,3-FucT2, NCg3FL-α-1,3-FucT3, NCg3FL-α-1,3-FucT4, NCg3FL-α-1,3-FucT5, NCg3FL-α-1,3-FucT6, NCg3FL-α-1,3-FucT7, NCg3FL-α-1,3-FucT8 and NCg3FL-α-1,3-FucT9, the yield of 3-fucosyllactose is effectively increased.

[0266] In summary, by codon-optimizing the encoding gene of fucosyltransferase, the present invention can obtain a heterologously expressed fucosyltransferase, and the fucosyltransferase effectively catalyzes the reaction and increases the yields of 3-fucosyllactose and 2'-fucosyllactose.

[0267] The applicant declares that the present invention uses the above embodiments to illustrate the detailed method of the present invention, but the present invention is not limited to the above detailed method, that is, it does not mean that the present invention must rely on the above detailed method to be implemented. Those skilled in the art should understand that any improvement to the present invention, the equivalent replacement of each raw material of the product of the present invention, the addition of auxiliary components, the selection of specific methods, etc. all fall within the protection scope and the disclosure scope of the present invention.

Claims

1. A fucosyltransferase-encoding gene, characterized in that, The nucleic acid sequence of the fucosyltransferase-encoding gene includes any one or a combination of at least two of the following sequences: (1) Any one or a combination of at least two of the sequences shown in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16 or SEQ ID NO:18; (2) A nucleic acid sequence encoding the amino acid sequence of SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 or SEQ ID NO:17; (3) A nucleic acid sequence encoding a fucosyltransferase activity obtained by substituting, deleting or adding one or more nucleotides to the sequence shown in SEQ ID NO:2, SEQ ID NO:4, SEQ ID NO:6, SEQ ID NO:8, SEQ ID NO:10, SEQ ID NO:12, SEQ ID NO:14, SEQ ID NO:16 or SEQ ID NO:18; (4) A nucleic acid sequence having at least 80% sequence homology with the nucleic acid sequence described in (1), (2) or (3) and having the same or similar function.

2. The fucosyltransferase encoded by the fucosyltransferase-encoding gene according to claim 1, characterized in that, The amino acid sequence of the fucosyltransferase includes any one or a combination of at least two of the following sequences: (1) Any one or a combination of at least two of the sequences shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 or SEQ ID NO:17; (2) An amino acid sequence having fucosyltransferase activity formed by substituting, deleting or adding one or more amino acids to the amino acid sequence shown in SEQ ID NO:1, SEQ ID NO:3, SEQ ID NO:5, SEQ ID NO:7, SEQ ID NO:9, SEQ ID NO:11, SEQ ID NO:13, SEQ ID NO:15 or SEQ ID NO:17; (3) An amino acid sequence having at least 80% sequence homology with the amino acid sequence described in (1) or (2) and having the same or similar function.

3. A genetically engineered strain that simultaneously expresses fucosyltransferase and lactose permease, characterized in that, The genetically engineered strain contains the fucosyltransferase-encoding gene described in claim 1; Preferably, the genetically engineered strain includes any one of Corynebacterium glutamicum, Escherichia coli, Bacillus subtilis or Saccharomyces cerevisiae.

4. A method for preparing the genetically engineered strain according to claim 3, characterized in that, The preparation method includes: transducing a recombinant vector containing the fucosyltransferase-encoding gene described in claim 1 into a Corynebacterium glutamicum strain, thereby obtaining the genetically engineered strain.

5. Use of the fucosyltransferase-encoding gene described in claim 1 in the preparation of fucosyllactose.

6. The application according to claim 5, wherein The fucosyllactose includes 3-fucosyllactose and / or 2'-fucosyllactose.

7. A method for preparing fucosyllactose, characterized in that, The method includes: overexpressing the fucosyltransferase-encoding gene described in claim 1 to obtain the fucosyllactose.

8. The method for preparing fucosyllactose according to claim 7, wherein The method for overexpressing the fucosyltransferase-encoding gene described in claim 1 includes introducing the fucosyltransferase-encoding gene described in claim 1 into a free expression vector of a host cell or integrating the fucosyltransferase-encoding gene described in claim 1 into the chromosome of the host cell.

9. A fucosyllactose, characterized in that, The fucosyllactose is prepared by the method described in claim 7 or 8.

10. Use of the fucosyllactose described in claim 9 in the preparation of a drug, a health product, or a food additive.

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