National Institute of Advanced Industrial Science and Technology

Daiichi Sankyo Company, Limited

Kyowa Hakko Kirin Co., Limited

Eisai R&d Management Co., Ltd.

Kirin Beer Kabushiki Kaisha

Tokyo Metropolitan Organization for Medical Research

Neo-morgan Laboratory Incorporated

Yoshifumi Jigami

Yasunori Chiba

Kazuo Kobayashi

Kimihisa Ichikawa

Koichi Nonaka

Kenichi Nakayama

Yoriko Takeuchi, Legal Representative

Mariko Umemura

Takeshi Suzuki

Kosuke Kuroda

Makoto Takeuchi

Kappei Tsukahara

Mamiko Tsuchiya

Hitoshi Sakuraba

Toshihiro Komeda

Shigemi Sugioka

Aruto Yoshida

Hiroko Abe

Hiroshi Manya

Tamao Endo

Yoshie Takahashi

Michiyo Okamoto

Yoshinori Kitagawa

Nagako Kawashima

Yuki Takaoka

Akiko Itadani

Takehiko Yoko-O

Morihisa Fujita

Minako Ono

Ken-ichi Nakayama

This invention provides a means for high-level secretory production of a protein, and, in particular, a protein having a complicated structure such as an antibody, in a host cell such as a yeast cell. This invention provides a method for high-level secretory production of a foreign protein with the use of a transformed host cell having one or more types of chaperone protein genes and via suppression of O sugar chain inherent to a host cell such as a yeast cell.

Method for high-level secretory production of protein

The present invention enables screening for compounds that inhibit the transport of GPI-anchored proteins to fungal cell walls, using a simple assay for transacylation to GlcN-PI using membrane fraction expressing GWT1 protein. New antifungal agents can be created that inhibit the synthesis of fungal cell walls and also inhibit adhesion to host cells by inhibiting the transport of GPI-anchored proteins to fungal cell walls.

Method of screening for compounds that inhibit the enzymatic activity of GWT1 gene product

This invention provides a means for enabling high-level secretory production of proteins, in particular proteins having complicated structures such as antibodies, in host cells such as yeast cells. The invention also provides transformed yeast cells having the activated HAC1 gene and the RRBP1 gene and a method for enabling high-level secretory production of foreign proteins using such transformed host cells by inhibiting O-sugar chain formation indigenous to host cells such as yeast cells.

This invention is to provide a process for producing a glycoprotein comprising a mammalian type sugar chain, characterized in that the process comprises introducing an &#x3b1;-1,2-mannosidase gene into a methylotrophic yeast having a mutation of a sugar chain biosynthesizing enzyme gene, so that the &#x3b1;-1,2-mannosidase gene is expressed under the control of a potent promoter in the yeast; culturing in a medium the methylotrophic yeast cells with a heterologous gene transferred thereinto; and obtaining the glycoprotein comprising a mammalian type sugar chain from the culture. Using the newly created methylotrophic yeast having a sugar chain mutation, a neutral sugar chain identical with a high mannose type sugar chain produced by mammalian cells such as human cells, or a glycoprotein comprising such a neutral sugar chain, can be produced in a large amount at a high purity. By introducing a mammalian type sugar chain biosynthesizing gene into the above-described mutant, a mammalian type sugar chain, such as a hybrid or complex, or a protein comprising a mammalian type sugar chain can be efficiently produced.

Methylotrophic yeast producing mammalian type sugar chain

A method for breeding yeast having thermotolerance or recovering growth activity and a method for breeding yeast which produces beta-glucan efficiently as well as an yeast obtained by such methods for breeding are presented by a method for breeding yeast having thermotolerance or recovering growth activity including a step for controlling proofreading function of DNA polymerase in a loss-of-function mutant of yeast (for example, a step for including mutant pol3 gene or mutant cdc6&#x2212; gene in a gene-disruptant.

Breeding method for yeast, yeast and a production method for glycoprotein or beta-glucan

This invention provides proteins and genes thereof involved in the GPI lipid remodeling process and, thereby and constructing a system for screening for useful substances such as anticancer agents and a system for detecting abnormalities in the GPI lipid remodeling process.

Lipid remodeling of GPI-anchored proteins

A process for producing a lysosomal enzyme having a mannose-6-phosphate-containing acidic sugar chain, wherein the process comprising: culturing in a medium yeast cells obtained by introducing a lysosomal enzyme gene into a sugar chain biosynthetic enzyme gene mutant strain of yeast, collecting a lysosomal enzyme having a phosphate-containing sugar chain from the culture, and then treating the enzyme with &#x3b1;-mannosidase; and pharmaceutical compositions for treatment of human lysosomal enzyme deficiencies produced by the process. The genetic engineering technique using the yeast according to the present invention allows large-amount and high-purity production of a glycoprotein having a phosphate-containing acidic sugar chain which can serve as a labeling marker for transporting into lysosomes in cells of mammals such as human. The glycoprotein having a phosphate-containing acidic sugar chain according to the invention may be utilized as a drug effective in treatment of human lysosomal enzyme deficiencies, etc.

Glycoprotein and process for producing the same

The present invention relates to an enzyme having novel N-acetylglucosaminyltransferase (OMGnT) activity, a polynucleotide encoding the enzyme, a recombinant polynucleotide comprising the polynucleotide, a host cell comprising the recombinant polynucleotide, a method for producing an enzyme protein having OMGnT activity by culturing the host cell in a medium, and a method for modifying a sugar chain structure with the enzyme and a carbohydrate having a sugar chain structure modified by the method. This enzyme enables the production of complex carbohydrates, which could not be formed with conventional glycosyltransferases.

N-acetylglucosaminlytransferase and polynucleotide encoding the same

The present invention relates to a gene for an enzyme involving in the synthesis of GDP-L-fucose. Particularly, the present invention relates to a GDP-4-keto-6-deoxy-D-mannose-3, 5-epimerase-4-reductase gene derived from, and a process for producing GDP-L-fucose using the gene. An enzyme encoded by the gene is (a) a protein comprising an amino acid sequence represented by SEQ ID NO: 1; or (b) a protein comprising an amino acid sequence derived from the amino acid sequence of SEQ ID NO: 1 by deletion, substitution, addition or insertion of one or several amino acid residues, and having GDP-4-keto-6-deoxy-D-mannose-3, 5-epimerase-4-reductase activity. The present invention enables efficient mass production of GDP-L-fucose which is essential in performing addition of fucose, which has a very important function in sugar chains.

Gdp-4-keto-6-deoxy-d-mannose-3,5-epimerase-4-reductase gene derived from arabidopsis thaliana

Growing community of inventors

Ibaraki, Japan

Yoshifumi Jigami