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Patent Searching and Data


Title:
PROCESS FOR PRODUCTION OF PROTEASE, PROTEASE SOLUTION, AND SOLUTION OF PRO-FORM OF PROTEASE
Document Type and Number:
WIPO Patent Application WO/2010/021375
Kind Code:
A1
Abstract:
Disclosed is a process for producing a protease.  The process comprises the following steps: a pro-form expression step of introducing DNA encoding a pro-form of the protease into a host cell to induce the expression of the pro-form in the form of an inclusion body; a modification step of modifying the pro-form in the form of an inclusion body to produce a modified pro-form; a refolding step of refolding the modified pro-form in a solution containing a calcium ion and having a pH value of 5 or higher; and a maturation step of maturing the refolded pro-form under alkaline conditions to produce the protease in an active form.  According to the process for producing the protease, the self-degradation of the protease during the production process can be prevented, and a matured form produced from the pro-form can be collected with high efficiency.

Inventors:
KANAYA, Shigenori (1-1 Yamadaoka, Suita-sh, Osaka 71, 〒5650871, JP)
金谷 茂則 (〒71 大阪府吹田市山田丘1番1号 国立大学法人大阪大学内 Osaka, 〒5650871, JP)
TANAKA, Shun-ichi (1-1 Yamadaoka, Suita-sh, Osaka 71, 〒5650871, JP)
田中 俊一 (〒71 大阪府吹田市山田丘1番1号 国立大学法人大阪大学内 Osaka, 〒5650871, JP)
TAKANO, Kazufumi (1-1 Yamadaoka, Suita-sh, Osaka 71, 〒5650871, JP)
Application Number:
JP2009/064629
Publication Date:
February 25, 2010
Filing Date:
August 21, 2009
Export Citation:
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Assignee:
OSAKA UNIVERSITY (1-1 Yamadaoka, Suita-shi Osaka, 71, 〒5650871, JP)
国立大学法人大阪大学 (〒71 大阪府吹田市山田丘1番1号 Osaka, 〒5650871, JP)
KANAYA, Shigenori (1-1 Yamadaoka, Suita-sh, Osaka 71, 〒5650871, JP)
金谷 茂則 (〒71 大阪府吹田市山田丘1番1号 国立大学法人大阪大学内 Osaka, 〒5650871, JP)
TANAKA, Shun-ichi (1-1 Yamadaoka, Suita-sh, Osaka 71, 〒5650871, JP)
田中 俊一 (〒71 大阪府吹田市山田丘1番1号 国立大学法人大阪大学内 Osaka, 〒5650871, JP)
International Classes:
C12N9/52; C11D3/386; C12N15/09
Foreign References:
JPH05260967A1993-10-12
JP2004516845A2004-06-10
Other References:
AKANNAN Y. ET AL.: 'Active subtilisin-like protease from a hyperthermophilic archaeon in a form with a putative prosequence.' APPL. ENVIRON. MICROBIOL. vol. 67, no. 6, 2001, pages 2445 - 2452
PULIDO M. ET AL.: 'Ca2+-dependent maturation of subtilisin from a hyperthermophilic archaeon, Thermococcus kodakaraensis: the propeptide is a potent inhibitor of the mature domain but is not required for its folding.' APPL. ENVIRON. MICROBIOL. vol. 72, no. 6, 2006, pages 4154 - 4162
Attorney, Agent or Firm:
IWATANI, Ryo (KEIHAN Dojima Bldg. 3F, 1-31 Dojima 2-chome, Kita-ku, Osaka-sh, Osaka 03, 〒5300003, JP)
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