Kirin Beer Kabushiki Kaisha

National Institute of Advanced Industrial Science and Technology

Toshihiro Komeda

Nobuo Kato

Yasuyoshi Sakai

Kazuo Kobayashi

Akihiro Iwamatsu

Yasunori Chiba

Keiji Kondo

Yoshifumi Jigami

Yukio Tamai

Hisako Suda

Yoshinori Kitagawa

Nagako Kawashima

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

This invention relates to a promoter for dihydroxyacetone synthase gene from ; an expression cassette or vector comprising the promoter and a heterologous gene; a transformant transformed with this vector; and a method for producing an expression product of the heterologous gene which comprises culturing the transformant to express the heterologous gene.

Candida boidini dihydroxyacetone synthase promoter

The present invention relates to a DNA comprising the nucleotide sequence shown in SEQ ID NO:1, or a DNA for enhancing promoter activity which comprises a nucleotide sequence having deletions, substitutions, additions or insertions of one or more nucleotides in the nucleotide sequence shown in SEQ ID NO:1; to a mutant promoter comprising one or more fragments of the DNA; to a recombinant expression vector comprising the mutant promoter together with a heterologous gene; to a transformant prepared by transforming a host cell with the vector; to a process for preparing an expression product, comprising culturing the transformant in a medium, and recovering the expression product of a heterologous gene from the obtained culture; and to a method for enhancing a promoter activity, characterized in that one or more fragments of the above-defined DNA are located at at least one position preceding, following or within a selective promoter in a forward or reverse direction.

DNA sequences enhancing promoter activity

Promoter and terminator sequences of formate dehydrogenase gene of

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Kanagawa, Japan

Toshihiro Komeda