Ajinomoto Co., Ltd.

Noriko Tsurui

Hiroshi Itaya

Yoshimi Kikuchi

Naoko Yamada

Yumi Ito

Yoshihiko Matsuda

Teruhisa Mannen

Takahiro Nonaka

Yukari Kashima

A novel technique for improving secretory production of a heterologous protein by coryneform bacteria is described, as well as a method for secretory production of a heterologous protein. A coryneform bacterium is described that has been modified to have a specific mutation so as to harbor a phoS gene, and when cultured, is able to to produce a heterologous protein by secretory production.

Method for secretory production of protein

A method of purifying a fusion protein, including: (1) adjusting the pH of an aqueous phase, containing a fusion protein which is a fusion of a protein having a self-assembly capability and a tamet protein, and which aqueous phase has a first pH, to a second pH, to obtain a remaining aqueous phase and a solid fraction containing an amount of the fusion protein; (2) separating the solid fraction from the remaining aqueous phase, to obtain a separated solid fraction; and (3) dissolving the separated solid fraction in a solution having a pH of 12 or lower but higher than the second pH by at least 0.1 pH units, wherein the protein having a self-assembly capability is a cell surface protein.

Method for producing protein by precipitation

A novel technique for improving secretory production of a heterologous protein by coryneform bacteria is described, and thereby a method for secretory production of a heterologous protein is provided. A coryneform bacterium is cultured so that it secretes a heterologous protein, the bacterium having a genetic construct which includes a promoter sequence that functions in the coryneform bacterium, a nucleic acid sequence coding for a signal peptide that functions in the coryneform bacterium, which is ligated downstream from the promoter sequence, and a nucleic acid sequence coding for a fusion protein having an amino acid sequence that includes Gln-Glu-Thr and the heterologous protein, which is ligated downstream from the nucleic acid sequence coding for the signal peptide.

Growing community of inventors

Kanagawa, Japan

Noriko Tsurui