Abstract
A silkworm-baculovirus system is particularly effective for producing recombinant proteins, including glycoproteins. However, N-glycan structures in silkworm differ from those in mammals. Glycoproteins in silkworm are secreted as pauci-mannose type N-glycans without sialic acid or galactose residues. Sialic acid on N-glycans plays important roles in protein functions. Therefore, we developed pathways for galactosylation and sialylation in silkworm. Sialylated N-glycans on proteins were successfully produced in silkworm by co-expressing galactosyltransferase and sialyltransferase and providing an external supply of a sialylation-related substrate. α2,3/α2,6 Sialylation to N-glycans was controlled by changing the type of sialyltransferase expressed in silkworm. Furthermore, the co-expression of N-acetylglucosaminyltransferase II facilitated the formation of additional di-sialylated N-glycan structures. Our results provide new information on the control of N-glycosylation in silkworm.
Keywords:
Baculovirus; Expression; N-glycan; Sialylation; Silkworm.
Copyright © 2018 The Society for Biotechnology, Japan. Published by Elsevier B.V. All rights reserved.
MeSH terms
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Animals
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Baculoviridae / genetics*
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Baculoviridae / metabolism
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Bombyx / genetics*
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Bombyx / metabolism
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Cells, Cultured
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Cloning, Molecular
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Galactose / metabolism
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Galactosyltransferases / genetics
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Galactosyltransferases / metabolism
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Gene Expression
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Genetic Vectors* / genetics
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Genetic Vectors* / metabolism
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Glycoproteins / biosynthesis*
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Glycoproteins / isolation & purification
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Glycoproteins / metabolism
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Glycosylation
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Humans
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Mannose / metabolism
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N-Acetylglucosaminyltransferases / genetics
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N-Acetylglucosaminyltransferases / metabolism
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Polysaccharides / metabolism*
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Protein Engineering / methods*
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Protein Processing, Post-Translational / genetics
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Recombinant Proteins* / genetics
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Recombinant Proteins* / isolation & purification
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Recombinant Proteins* / metabolism
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Sialyltransferases / genetics
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Sialyltransferases / metabolism
Substances
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Glycoproteins
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Polysaccharides
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Recombinant Proteins
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Galactosyltransferases
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N-Acetylglucosaminyltransferases
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beta-1,3-galactosyl-0-glycosyl-glycoprotein beta-1,3-N-acetylglucosaminyltransferase
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N-acetyllactosaminide alpha-1,3-galactosyltransferase
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Sialyltransferases
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N-acetyllactosaminide alpha-2,3-sialyltransferase
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Mannose
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Galactose