WO2010058707A1 - アフィニティタグが結合した融合コラゲナーゼおよびその製造法 - Google Patents

アフィニティタグが結合した融合コラゲナーゼおよびその製造法 Download PDF

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Publication number
WO2010058707A1
WO2010058707A1 PCT/JP2009/068968 JP2009068968W WO2010058707A1 WO 2010058707 A1 WO2010058707 A1 WO 2010058707A1 JP 2009068968 W JP2009068968 W JP 2009068968W WO 2010058707 A1 WO2010058707 A1 WO 2010058707A1
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WO
WIPO (PCT)
Prior art keywords
collagenase
amino acid
fusion
acid sequence
seq
Prior art date
Application number
PCT/JP2009/068968
Other languages
English (en)
French (fr)
Japanese (ja)
Inventor
崇恵 福島
健吾 横山
弘一郎 村島
昌史 後藤
洋平 山形
Original Assignee
明治製菓株式会社
国立大学法人東北大学
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by 明治製菓株式会社, 国立大学法人東北大学 filed Critical 明治製菓株式会社
Priority to EP09827488.9A priority Critical patent/EP2363461B1/en
Priority to US13/129,969 priority patent/US20110294192A1/en
Priority to CN2009801457257A priority patent/CN102216454B/zh
Priority to NZ593239A priority patent/NZ593239A/xx
Priority to DK09827488T priority patent/DK2363461T3/en
Priority to KR1020117012485A priority patent/KR101700974B1/ko
Priority to ES09827488T priority patent/ES2531402T3/es
Priority to AU2009318554A priority patent/AU2009318554B2/en
Priority to JP2010539201A priority patent/JP5698536B2/ja
Priority to CA2744286A priority patent/CA2744286C/en
Publication of WO2010058707A1 publication Critical patent/WO2010058707A1/ja

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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/52—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from bacteria or Archaea
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/78—Connective tissue peptides, e.g. collagen, elastin, laminin, fibronectin, vitronectin or cold insoluble globulin [CIG]
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K19/00—Hybrid peptides, i.e. peptides covalently bound to nucleic acids, or non-covalently bound protein-protein complexes
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
    • C12N15/62—DNA sequences coding for fusion proteins
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
    • C12N15/09—Recombinant DNA-technology
    • C12N15/63—Introduction of foreign genetic material using vectors; Vectors; Use of hosts therefor; Regulation of expression
    • C12N15/70—Vectors or expression systems specially adapted for E. coli
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K2319/00—Fusion polypeptide
    • C07K2319/20—Fusion polypeptide containing a tag with affinity for a non-protein ligand
    • C07K2319/21—Fusion polypeptide containing a tag with affinity for a non-protein ligand containing a His-tag

Definitions

  • a fusion collagenase having a collagen binding site is selectively collected by purifying a culture obtained by culturing the host cell according to ⁇ 15> or ⁇ 16> by affinity chromatography corresponding to an affinity tag.
  • a method for producing a fusion collagenase characterized by the above.
  • a method of removing a collagenase in which a part or all of CBD is degraded from a culture solution obtained by culturing a host cell expressing the fusion collagenase, and selectively recovering a single fusion collagenase having CBD. Provided. According to the present invention, it has become possible to efficiently produce a fusion collagenase having CBD.
  • DNA encoding an affinity tag can be amplified by PCR using a genomic DNA, cDNA, plasmid or the like containing a gene encoding the affinity tag as a template.
  • DNA encoding a histidine tag it can be amplified by PCR using a commercially available vector pET-24a (+) as a template.

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  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Molecular Biology (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • General Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Medicinal Chemistry (AREA)
  • Microbiology (AREA)
  • Biophysics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Plant Pathology (AREA)
  • Physics & Mathematics (AREA)
  • Toxicology (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)
  • Preparation Of Compounds By Using Micro-Organisms (AREA)
  • Peptides Or Proteins (AREA)
PCT/JP2009/068968 2008-11-19 2009-11-06 アフィニティタグが結合した融合コラゲナーゼおよびその製造法 WO2010058707A1 (ja)

Priority Applications (10)

Application Number Priority Date Filing Date Title
EP09827488.9A EP2363461B1 (en) 2008-11-19 2009-11-06 Fusion collagenase to which affinity tag is attached, and method for producing same
US13/129,969 US20110294192A1 (en) 2008-11-19 2009-11-06 Fusion collagenase in which affinity tag is linked and method for producing the same
CN2009801457257A CN102216454B (zh) 2008-11-19 2009-11-06 结合了亲和标记物的融合胶原酶及其制造法
NZ593239A NZ593239A (en) 2008-11-19 2009-11-06 Fusion Collagenase in which affinity TAG is linked and method for producing the same
DK09827488T DK2363461T3 (en) 2008-11-19 2009-11-06 Fusion collagenase associated with affinity tags, and process for its preparation
KR1020117012485A KR101700974B1 (ko) 2008-11-19 2009-11-06 친화성 태그가 결합된 융합 콜라게나제 및 그의 제조방법
ES09827488T ES2531402T3 (es) 2008-11-19 2009-11-06 Colagenasa de fusión a la que se une una marca de afinidad, y método para producir la misma
AU2009318554A AU2009318554B2 (en) 2008-11-19 2009-11-06 Fusion collagenase to which affinity tag is attached, and method for producing same
JP2010539201A JP5698536B2 (ja) 2008-11-19 2009-11-06 アフィニティタグが結合した融合コラゲナーゼおよびその製造法
CA2744286A CA2744286C (en) 2008-11-19 2009-11-06 Fusion collagenase in which affinity tag is linked and method for producing the same

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2008-295922 2008-11-19
JP2008295922 2008-11-19

Publications (1)

Publication Number Publication Date
WO2010058707A1 true WO2010058707A1 (ja) 2010-05-27

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Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2009/068968 WO2010058707A1 (ja) 2008-11-19 2009-11-06 アフィニティタグが結合した融合コラゲナーゼおよびその製造法

Country Status (11)

Country Link
US (1) US20110294192A1 (zh)
EP (1) EP2363461B1 (zh)
JP (1) JP5698536B2 (zh)
KR (1) KR101700974B1 (zh)
CN (1) CN102216454B (zh)
AU (1) AU2009318554B2 (zh)
CA (1) CA2744286C (zh)
DK (1) DK2363461T3 (zh)
ES (1) ES2531402T3 (zh)
NZ (1) NZ593239A (zh)
WO (1) WO2010058707A1 (zh)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2012124338A1 (ja) * 2011-03-16 2012-09-20 国立大学法人東北大学 生体組織分析用プローブ及びその利用法
WO2014119334A1 (ja) * 2013-02-01 2014-08-07 国立大学法人東北大学 生体組織からの細胞の分離方法
JP2015506176A (ja) * 2012-01-12 2015-03-02 オーキシリウム インターナショナル ホールディングス，インコーポレイテッド クロストリジウム・ヒストリチクム（ｃｌｏｓｔｒｉｄｉｕｍｈｉｓｔｏｌｙｔｉｃｕｍ）酵素およびその使用のための方法
WO2019187691A1 (ja) * 2018-03-26 2019-10-03 株式会社カネカ コラゲナーゼ活性を有するポリペプチド及びその製造方法
US11220666B2 (en) * 2017-09-13 2022-01-11 MB PHARMA s.r.o. Bacterial strain clostridium histolyticum and methods of use thereof

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
PT107276B (pt) * 2013-11-07 2018-06-07 Univ Aveiro Método para produção de colagenase recombinante para digestão de colagénios
CN115969318A (zh) 2017-03-01 2023-04-18 恩多风投有限公司 用于评估和治疗橘皮组织的装置和方法
KR102642381B1 (ko) 2017-03-28 2024-02-28 엔도 벤쳐즈 리미티드 개선된 콜라게나제 생성 방법
CN113025599B (zh) * 2021-04-02 2023-09-12 重庆科润生物医药研发有限公司 一种重组溶组织梭菌i型胶原酶及其制备方法和应用
CN114836461B (zh) * 2022-05-31 2024-03-29 华南理工大学 表达胶原蛋白酶的重组质粒、酵母菌株及其发酵培养基和发酵培养方法

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WO2008124166A2 (en) * 2007-04-09 2008-10-16 The Board Of Trustees Of The University Of Arkansas Fusion proteins of collagen-binding domain and parathyroid hormone

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JP2003284553A (ja) * 2002-03-28 2003-10-07 Seikagaku Kogyo Co Ltd 生理活性タンパク質
WO2008124166A2 (en) * 2007-04-09 2008-10-16 The Board Of Trustees Of The University Of Arkansas Fusion proteins of collagen-binding domain and parathyroid hormone

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TAMAI E ET AL.: "High-level expression of his- tagged clostridial collagenase in Clostridium perfringens.", APPL MICROBIOL BIOTECHNOL., vol. 80, no. 4, September 2008 (2008-09-01), pages 627 - 35, XP019623735 *
YOSHIHARA K ET AL.: "Cloning and nucleotide sequence analysis of the colH gene from Clostridium histolyticum encoding a collagenase and a gelatinase.", J BACTERIOL., vol. 176, no. 21, 1994, pages 6489 - 96, XP008150460 *
YOSHITOMO OKA: "Tokushu/Saibo Baiyo Saibo Baiyo wa dokomade Kano ka, 2) Suito Saibo", BYOTAI SEIRI, vol. 6, no. 11, 1987, pages 852 - 856, XP008150462 *

Cited By (16)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP5997131B2 (ja) * 2011-03-16 2016-09-28 国立大学法人東北大学 生体組織分析用プローブ及びその利用法
CN103562719A (zh) * 2011-03-16 2014-02-05 国立大学法人东北大学 生物体组织分析用探针及其使用方法
JPWO2012124338A1 (ja) * 2011-03-16 2014-07-17 国立大学法人東北大学 生体組織分析用プローブ及びその利用法
KR101921353B1 (ko) * 2011-03-16 2018-11-23 도호쿠 다이가쿠 생체 조직 분석용 프로브 및 그 이용법
WO2012124338A1 (ja) * 2011-03-16 2012-09-20 国立大学法人東北大学 生体組織分析用プローブ及びその利用法
US9354240B2 (en) 2011-03-16 2016-05-31 Tohoku University Probe for analyzing biological tissue and method for utilizing same
JP2018075011A (ja) * 2012-01-12 2018-05-17 オーキシリウム インターナショナル ホールディングス，インコーポレイテッド クロストリジウム・ヒストリチクム（ｃｌｏｓｔｒｉｄｉｕｍ ｈｉｓｔｏｌｙｔｉｃｕｍ）酵素およびその使用のための方法
JP2015506176A (ja) * 2012-01-12 2015-03-02 オーキシリウム インターナショナル ホールディングス，インコーポレイテッド クロストリジウム・ヒストリチクム（ｃｌｏｓｔｒｉｄｉｕｍｈｉｓｔｏｌｙｔｉｃｕｍ）酵素およびその使用のための方法
JP2019103505A (ja) * 2012-01-12 2019-06-27 オーキシリウム インターナショナル ホールディングス，インコーポレイテッド クロストリジウム・ヒストリチクム（ｃｌｏｓｔｒｉｄｉｕｍ ｈｉｓｔｏｌｙｔｉｃｕｍ）酵素およびその使用のための方法
JP2020188810A (ja) * 2012-01-12 2020-11-26 オーキシリウム インターナショナル ホールディングス，インコーポレイテッド クロストリジウム・ヒストリチクム（ｃｌｏｓｔｒｉｄｉｕｍ ｈｉｓｔｏｌｙｔｉｃｕｍ）酵素およびその使用のための方法
JPWO2014119334A1 (ja) * 2013-02-01 2017-01-26 国立大学法人東北大学 生体組織からの細胞の分離方法
WO2014119334A1 (ja) * 2013-02-01 2014-08-07 国立大学法人東北大学 生体組織からの細胞の分離方法
JP2018198613A (ja) * 2013-02-01 2018-12-20 国立大学法人東北大学 生体組織からの細胞の分離方法
US10870833B2 (en) 2013-02-01 2020-12-22 Tohoku University Method for separating cell from biological tissue
US11220666B2 (en) * 2017-09-13 2022-01-11 MB PHARMA s.r.o. Bacterial strain clostridium histolyticum and methods of use thereof
WO2019187691A1 (ja) * 2018-03-26 2019-10-03 株式会社カネカ コラゲナーゼ活性を有するポリペプチド及びその製造方法

Also Published As

Publication number Publication date
CN102216454A (zh) 2011-10-12
CA2744286C (en) 2018-07-31
KR20110106286A (ko) 2011-09-28
AU2009318554A1 (en) 2011-06-23
NZ593239A (en) 2013-01-25
KR101700974B1 (ko) 2017-01-31
JP5698536B2 (ja) 2015-04-08
JPWO2010058707A1 (ja) 2012-04-19
CN102216454B (zh) 2013-05-29
EP2363461B1 (en) 2014-12-24
ES2531402T3 (es) 2015-03-13
AU2009318554B2 (en) 2016-06-09
EP2363461A1 (en) 2011-09-07
EP2363461A4 (en) 2012-08-01
CA2744286A1 (en) 2010-05-27
US20110294192A1 (en) 2011-12-01
DK2363461T3 (en) 2015-03-02

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