title: Targeted glycoengineering extends the protein N-glycosylation pathway in the silkworm silk gland.
abstract: The silkworm silk glands are powerful secretory organs that can produce and secrete proteins at high levels. As such, it has been suggested that the biosynthetic and secretory power of the silk gland can be harnessed to produce and secrete recombinant proteins in tight or loose association with silk fibers. However, the utility of the silkworm platform is constrained by the fact that it has a relatively primitive protein N-glycosylation pathway, which produces relatively simple insect-type, rather than mammalian-type N-glycans. In this study, we demonstrate for the first time that the silk gland protein N-glycosylation pathway can be glycoengineered. We accomplished this by using a dual piggyBac vector encoding two distinct mammalian glycosyltransferases under the transcriptional control of a posterior silk gland (PSG)-specific promoter. Both mammalian transgenes were expressed and each mammalian N-glycan processing activity was induced in transformed silkworm PSGs. In addition, the transgenic animals produced endogenous glycoproteins containing significant proportions of mammalian-type, terminally galactosylated N-glycans, while the parental animals produced none. This demonstration of the ability to glycoengineer the silkworm extends its potential utility as a recombinant protein production platform.
authors.affiliation: Department of Molecular Biology, University of Wyoming, Laramie, WY 82071, USA.
authors.forename: Hideaki
authors.initials: H
authors.lastname: Mabashi-Asazuma
authors.affiliation: Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Bong-Hee
authors.initials: BH
authors.lastname: Sohn
authors.affiliation: Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Young-Soo
authors.initials: YS
authors.lastname: Kim
authors.affiliation: Institute of Biological Chemistry, Academia Sinica 128, Nankang, Taipei 115, Taiwan.
authors.forename: Chu-Wei
authors.initials: CW
authors.lastname: Kuo
authors.affiliation: Institute of Biological Chemistry, Academia Sinica 128, Nankang, Taipei 115, Taiwan.
authors.forename: Kay-Hooi
authors.initials: KH
authors.lastname: Khoo
authors.affiliation: Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Cheryl A
authors.initials: CA
authors.lastname: Kucharski
authors.affiliation: Eck Institute for Global Health, Department of Biological Sciences, University of Notre Dame, Notre Dame, IN 46556, USA.
authors.forename: Malcolm J
authors.initials: MJ
authors.lastname: Fraser
authors.affiliation: Department of Molecular Biology, University of Wyoming, Laramie, WY 82071, USA. Electronic address: DLJarvis@uwyo.edu.
authors.forename: Donald L
authors.initials: DL
authors.lastname: Jarvis
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D012831:Bombyx / Q000201:enzymology / Q000235:genetics*; D005088:Exocrine Glands / Q000378:metabolism*; D005260:Female; D005822:Genetic Vectors; D006023:Glycoproteins / Q000096:biosynthesis* / Q000235:genetics; D006031:Glycosylation; D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism; D007814:Larva / Q000201:enzymology; D008297:Male; D011134:Polysaccharides / Q000378:metabolism; D015202:Protein Engineering; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D047011:Silk
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides; D011994:Recombinant Proteins; D047011:Silk; D016695:Glycosyltransferases
keywords: Glycoengineering; Glycosylation; Transgenic silkworm; piggyBac