PROCESS FOR PRODUCING PROTEIN CAPABLE OF FORMING INCLUSION BODY

• Main Authors: SUENAGA, KIYOTAKA, HIRASHIMA, MASAKI, NAKATAKE, HIROSHI, MAEDA, HIROAKI, META, AKIHIRO
• Format: Patent
• Language: English; French; German
• Published: 02.05.2012
• Subjects: BEER; BIOCHEMISTRY; CHEMISTRY; COMPOSITIONS THEREOF; CULTURE MEDIA; ENZYMOLOGY; FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIREDCHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERSFROM A RACEMIC MIXTURE; METALLURGY; MICROBIOLOGY; MICROORGANISMS OR ENZYMES; MUTATION OR GENETIC ENGINEERING; ORGANIC CHEMISTRY; PEPTIDES; PROPAGATING, PRESERVING OR MAINTAINING MICROORGANISMS; SPIRITS; VINEGAR; WINE

A process for preparing an inclusion body-forming protein is provided. A nucleic acid fragment consisting of a nucleotide sequence coding for a modified alkaline phosphatase signal peptide (modified APSP) where leucine at the 13th position in the amino acid sequence shown in SEQ ID NO: 1 is substituted with proline and/or alanine at the 21st position is substituted with the other amino acid, downstream of which nucleotide sequence is bound a nucleotide sequence of a gene of a protein of interest, and a process for preparing an inclusion body-forming protein consisting of the following steps: (1) preparing an expression vector in which a nucleic acid fragment is incorporated consisting of a nucleotide sequence coding for a modified signal peptide downstream of which is bound a nucleotide sequence of a gene of a protein of interest, (2) preparing a host cell transformed with the expression vector that produces an inclusion body-forming protein, and (3) purifying the inclusion body-forming protein from culture obtained by culturing the host cell that produces the inclusion body-forming protein. The above other amino acid is selected from the group consisting of aspartic acid, glutamic acid, lysine, histidine, phenylalanine and tyrosine.