Fujiyama Lab. ICBiotech. Osaka University  
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Research Topics

Our main interests are development of bioreactors for the production of therapeutic proteins and industrially useful bioresources using plant, insect and microorganism.

1. Production and glycoengineering of useful recombinant glycoproteins in plant cells, insect cells, microorganism and mammalian cells.

Recently, Human pharmaceutical proteins have been produced in lots of bioreactors (Mammalian cells, plants, insects, microorganisms, and so on). Almost all pharmaceutical proteins are glycosylated, and the glycan moiety contributes to the stability and bioactivity. Although plants, insects, and yeasts have advantages of the economical productivity as bioreactors comparing with mammalian cells, the recombinant pharmaceutical proteins actually have host-specific glycans, not human-type. The difference of the glycosylation pattern often causes reduction of the bioactivity and half-life in blood stream, and also shows immunogenicity in human. To solve these problems, we have to humanize such host-specific glycan structures. Our aim is to produce human oriented recombinant pharmaceutical proteins with basic glyco-science and glyco-engineering.

*click on the link to go to PubMed etc. if it is available.


(Arabidopsis thaliana, BY-2 cells, Nicotiana tabacum, Nicotiana benthamiana and MM2d cells)

1) Structural analysis of N- and O-linked oligosaccharides from several plant species

1.1) O-linked oligosaccharide analysis of rice alchol-extracted proteins

Kilcoyne, Shah, Gerlach, Bhavanandan, Nagaraj, Smith, Fujiyama, Sommer, Costello, Olszewski, Joshi. J. Plant Physiol. 166, 219-232, 2009

1.2) N-linked oligosaccharide analysis of Arabidopsis thaliana MM2-d cells

Fujiyama, Tanaka, Terashima, Seki. Plant Biotechnol. 24, 255-259, 2007

1.3) Analysis of sialylated oligosaccharides in Arabidopsis thaliana

Shah, Fujiyama, Flynn, Joshi. Nature Biotechol. 21, 1470-1, 2003

1.4) N-linked oligosaccharide analysis of tobacco BY2 cells

Misaki, Kimura, Fujiyama, Seki. Biosci. Biotechnol. Biochem. 65, 2482-2488, 2001

Palacpac, Kimura, Fujiyama, Yoshida, Seki. Biosci. Biotechnol. Biochem. 63, 35-39, 1999

2) Structural analysis of N-linked oligosaccharides from heterologously expressed pharmaceutical and industrially useful glycoproteins in plant cells.

2.1) HIV-1 vaccine (cholera toxin B subunit-membrane proximal (ectodomain) region of gp41 envelop protein, CTB-MPR649-684) in Nicotiana benthamiana

Matoba, Kajiura, Cherni, Doran, Bomsel, Fujiyama and Mor. Plant Biotechnol J. 7, 129-45, 2009

2.2) Edible vaccine against house dust mite (HDM) allergy (Der p 1 allergen protein) in rice

Yang, Kajiura, Suzuki, Hirose, Fujiyama and Takaiwa. Biochem Biophys Res Commun. 365, 334-9, 2008

2.3) Dengue vaccine (virus envelop protein, D2EIII) in Nicotiana benthamiana

Saejung, Fujiyama, Takasaki, Ito, Hori, Malasit, Watanabe, Kurane, Seki. Vaccine. 25, 6646-54, 2007

2.4) Poliovirus vaccine (poliovirus peptide, PVP) in Nicotiana tabacum

Fujiyama, Saejung, Yanagihara, Nakado, Misaki, Honda, Watanabe, Seki. J Biosci Bioeng. 101, 398-402, 2006

2.5) Mouse monoclonal antibody in BY2 cells

Fujiyama, Misaki, Katsura, Tanaka, Furukawa, Omasa, Seki. J Biosci Bioeng. 101, 212-8, 2006

2.6) Human lactoferrin in rice

Nandi, Yalda, Lu, Nikolov, Misaki, Fujiyama and Huang. Transfenic Res. 14, 237-49, 2005

Fujiyama, Sakai, Misaki, Yanagishita, Honda, Anzai and Seki. Biosci Biotechnol Biochem. 68, 2565-70, 2004

3) Glycoengineering of plant cells

3.1) Cloning and characterization of plant glycan modifying enzymes

Characterization of AtALG3

Kajiura, Seki, Fujiyama. Glycobiology. 20, 736-51, 2010


Characterization of NtGnTI

Dohi, Isoyama-Tanaka, Tokuda, Fujiyama. J Biosci Bioeng. 109, 388-91, 2010


Characterization of AtMANI

Kajiura, Koiwa, Nakazawa, Okazawa, Kobayashi, Seki, Fujiyama. Glycobiology. 20, 235-47, 2010


Analysis of CMP-sialic acid transporter-like proteins in rice

Takashima, Seino, Nakano, Fujiyama, Tsujimoto, Ishida, Hashimoto. Phytochemistry. 70, 1973-81, 2009


Cloning and Characterization AtCMPKDO-S

Misaki, Kajiura, Fujii, Fujiyama, Seki. J Biosci Bioeng. 108, 527-9, 2009


Cloning and gene expression analysis of tomato endo-beta-N-acetylglucosaminidase

Nakamura, Inoue, Maeda, Nakano, Hosoi, Fujiyama, Kimura. Biosci Biotechnol Biochem.73, 461-64, 2009


Characterization of Almond alfa-mannosidase

Misaki, Fujiyama, Yokoyama, Ido, Miyauchi, Yoshida, Seki. J Biosci Bioeng. 96, 187-92, 2003


Identification of Arabidopsis thaliana alfa-mannosidase

Fujiyama, Kira, Iizuka, Kimura, Seki. J Biosci Bioeng. 92, 401-4, 2001


3.2) Introduction of mammalian glycan modifying enzyme

Expression of human UDP-galactose transporter (hUGT1) in Nicotiana tabacum

Khalil, Kajiura, Fujiyama, Koike, Ishida, Tanaka. J Biosci Bioeng. 109, 159-69, 2010


Mouse antibody production in BY2 cells expressing human β1,4-galactosyltransferase

Fujiyama, Furukawa, Katsura, Misaki, Omasa, Seki. Biochem Biophys Res Commun. 358, 85-91, 2007


Expression of human CMP-N-acetylneuraminic acid synthetase and CMP-sialic acid transporter in BY2 cells

Misaki, Fujiyama, Seki. Biochem Biophys Res Commun. 339, 1184-9, 2006


Glycan analysis of suspension medium from human β1,4-galactosyltransferase expressing BY2 cells

Misaki, Kimura, Palacpac, Yoshida, Fujiyama, Seki. Glycobiology. 13, 199-205, 2003


Stable expression of human β1,4-galactosyltransferase in BY2 cells

Palacpac, Yoshida, Sakai, Kimura, Fujiyama, Yoshida, Seki. Proc Natl Acad Sci USA. 96, 4692-7, 1999


3.3) Functional glycobiology in plant

Salt tolerance of Arabidopsis thaliana requires maturation of N-glycosylated proteins in the Golgi apparatus

Kang, Frank, Kang, Kajiura, Vikram, Ueda, Kim, Bahk, Triplett, Fujiyama, Lee, Schaewen, Koiwa. Proc Natl Acad Sci USA. 105, 5933-8, 2008


Salt-adaptaion induces the glycan structures of N-glycans

Kimura, Watanabe, Kimura, Maeda, Murata, Fujiyama. Biosci Biotechnol Biochem. 72, 514-22, 2008


4) Engineering of the cellular localization of glycan modifying enzymes

ER localization of heterologously expressed mouse antibody in BY-2 cells

Fujiyama, Misaki, Sakai, Omasa, Seki. J Biosci Bioeng. 107, 165-72

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Insect cells

(Sf9 cells and Bombix mori)

1) Structural analysis of N-linked oligosaccharides from insect cells

N-linked glycan structures of mouse interferon-β produced in Bobbyx mori larvae

Misaki, Nagaya, Fujiyama, Yahagihara, Honda, Seki. Biochem Biophys Res Commun. 311, 979-86, 2003

2) Cloning and characterization of insect glycan modifying enzymes

Cloning and characterization of BmFDL

Nomura, Ikeda, Ishiyama, Mita, Tamura, Okada, Fujiyama, Usami. J Biosci Bioeng. 110, 386-91, 2010


Cloning and characterization of BmGlcNAcase1, 2

Okada, Ishiyama, Sezutsu, Usami, Tamura, Mita, Fujiyama, Seki. Biosci Biotechnol Biochem. 71, 1626-35, 2007


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(Schizosaccharomyces pombe, Pichia species and E. coli)

1) Introduction of eukaryotic glycan modifying enzymes into E. coli and yeast

2) Efficient production of proteins from human, plant and insect cells

Cloning and characterization of BmGlcNAcase1, 2

Okada, Ishiyama, Sezutsu, Usami, Tamura, Mita, Fujiyama, Seki. Biosci Biotechnol Biochem. 71, 1626-35, 2007


2.1) Honey bee β1,3-GalT production in E. coli

Kimura, Sakamura, Ushijima, Hama, Kajiura, Fujiyama, Okihara, Hashimoto, Sugimoto, Yamada. Biosci Biotechnol Biochem. 71, 1111-4, 2007


2.2) Dengue vaccine (virus envelop protein, D2EIII) production in E. coli

Saejung, Puttikhunt, Prommool, Sojikul, Tanaka, Fujiyama, Malasit, Seki. J Biosci Bioeng. 102, 333-9, 2006


2.3) Human GnT-1 production in E. coli

Fujiyama, Ido, Misaki, Moran, Yanagihara, Honda, Nishimura, Yoshida, Seki. J Biosci Bioeng. 92, 569-74, 2001


2.4) Human ER Mannosidase production in E. coli

Fujiyama, Sakuradani, Moran, Yoshida, Seki. J Biosci Bioeng. 91, 419-21, 2001

Moran, Sakuradani, Jun, Fujiyama, Seki, Yoshida. 86, 277-83, 1998


2.5) Human β-GalT1 production in E. coli

Shibatani, Fujiyama, Nishiguchi, Seki, Maekawa. J Biosci Bioeng. 91, 85-71, 2001


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Mammalian cells

(CHO;Chinese Hamster Ovary) cells, COS7 (African Green Monkey SV40-transf’d kidney fibroblast cell line) cells, HEK (Human embryonic kidney) 293 cells and RAW (Mouse leukaemic monocyte macrophage cell line) cells)

1) Structural analysis of N-linked oligosaccharides of the recombinant proteins produced in mammalian cells

Kim, Tokunaga, Ozaki, Ishibashi, Honda, Kajiura, Fujiyama, Asano, Kumagai, Omasa, Ohtake. Appl. Microbiol. Biotechnol. 85, 535-42, 2010


2) Development of drug delivery system using plant-produced biopharmaceuticals

3) Functional analysis of recycling endosome

Misaki, Morimatsu, Uemura, Waguri, Miyoshi, Taniguchi, Matsuda, Taguchi. J. Cell. Biol. 191, 23-9, 2010


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2. Exploration of novel effective microorganisms for production of useful bioresources.

1) Yeast species isolated from mangrove in Thailand

Isolation of new yeast species

Kaewwichian, Yongmanitchai, Srisuk, Fujiyama, Limtong. FEMS Yeast Res. 10, 214-20, 2010

Limtong, Yongmanitchai, Kawasaki, Fujiyama. FEMS Yeast Res. 9, 504-10, 2009


One of the strains isolated in Thailand shows highly lipid production like linoleic acid (C17H31COOH, 18:2 (n-6)) and stearic acid (C17H35COOH).


2) Root nodule bacteria isolated from Iriomote island

We explore the possibilities for industrial application.


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