EA202000021A1 - Препарат цистеиновой протеиназы пшеницы тритикаина-альфа, полученной в растворимой форме, и способ получения препарата - Google Patents
Препарат цистеиновой протеиназы пшеницы тритикаина-альфа, полученной в растворимой форме, и способ получения препаратаInfo
- Publication number
- EA202000021A1 EA202000021A1 EA202000021A EA202000021A EA202000021A1 EA 202000021 A1 EA202000021 A1 EA 202000021A1 EA 202000021 A EA202000021 A EA 202000021A EA 202000021 A EA202000021 A EA 202000021A EA 202000021 A1 EA202000021 A1 EA 202000021A1
- Authority
- EA
- Eurasian Patent Office
- Prior art keywords
- wheat
- alpha
- triticaine
- soluble form
- preparation
- Prior art date
Links
- 241000209140 Triticum Species 0.000 title abstract 5
- 235000021307 Triticum Nutrition 0.000 title abstract 5
- 238000002360 preparation method Methods 0.000 title abstract 5
- 102000005927 Cysteine Proteases Human genes 0.000 title abstract 3
- 108010005843 Cysteine Proteases Proteins 0.000 title abstract 3
- 238000000034 method Methods 0.000 title abstract 3
- 244000098338 Triticum aestivum Species 0.000 abstract 2
- 239000012634 fragment Substances 0.000 abstract 2
- 102000004169 proteins and genes Human genes 0.000 abstract 2
- 108090000623 proteins and genes Proteins 0.000 abstract 2
- 238000000746 purification Methods 0.000 abstract 2
- 102000004190 Enzymes Human genes 0.000 abstract 1
- 108090000790 Enzymes Proteins 0.000 abstract 1
- 241000588724 Escherichia coli Species 0.000 abstract 1
- 241000235058 Komagataella pastoris Species 0.000 abstract 1
- 101800000268 Leader protease Proteins 0.000 abstract 1
- 108091005804 Peptidases Proteins 0.000 abstract 1
- 239000004365 Protease Substances 0.000 abstract 1
- 102000007056 Recombinant Fusion Proteins Human genes 0.000 abstract 1
- 108010008281 Recombinant Fusion Proteins Proteins 0.000 abstract 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 abstract 1
- 240000004808 Saccharomyces cerevisiae Species 0.000 abstract 1
- 230000001580 bacterial effect Effects 0.000 abstract 1
- 238000004587 chromatography analysis Methods 0.000 abstract 1
- 238000010367 cloning Methods 0.000 abstract 1
- 238000010276 construction Methods 0.000 abstract 1
- 239000003814 drug Substances 0.000 abstract 1
- 230000005714 functional activity Effects 0.000 abstract 1
- 238000002955 isolation Methods 0.000 abstract 1
- 238000004519 manufacturing process Methods 0.000 abstract 1
- 239000013612 plasmid Substances 0.000 abstract 1
- 235000019419 proteases Nutrition 0.000 abstract 1
- 235000018102 proteins Nutrition 0.000 abstract 1
- 230000001225 therapeutic effect Effects 0.000 abstract 1
Classifications
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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
- 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
-
- 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
- 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
- C12N15/79—Vectors or expression systems specially adapted for eukaryotic hosts
- C12N15/80—Vectors or expression systems specially adapted for eukaryotic hosts for fungi
- C12N15/81—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts
- C12N15/815—Vectors or expression systems specially adapted for eukaryotic hosts for fungi for yeasts for yeasts other than Saccharomyces
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Y—ENZYMES
- C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
- C12Y304/22—Cysteine endopeptidases (3.4.22)
- C12Y304/22002—Papain (3.4.22.2)
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Chemical & Material Sciences (AREA)
- Biomedical Technology (AREA)
- Wood Science & Technology (AREA)
- Zoology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- Molecular Biology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Plant Pathology (AREA)
- Biophysics (AREA)
- Physics & Mathematics (AREA)
- Medicinal Chemistry (AREA)
- Mycology (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
- Peptides Or Proteins (AREA)
Abstract
Изобретение относится к области молекулярной биологии, препаративной биохимии, биотехнологии, биофармакологии, а именно к созданию рекомбинантных белков семейства цистеиновых протеиназ пшеницы (Triticum aestivum) в растворимой форме и препараты белка тритикаина-альфа, состоящие из фрагмента тритикаина-альфа пшеницы и способов их получения. Изобретение может быть использовано в исследовательских целях для изучения функционирования папаин-подобных цистеиновых протеиназ, а также в медицине для разработки ферментных терапевтических препаратов, и представляет собой способ получения в растворимой форме рекомбинантных функционально активных вариантов цистеиновой протеазы пшеницы Triticum aestivum, включающий конструирование плазмидных ДНК для клонирования в экспрессионных системах E.coli и P.pastoris. Путем трансформации клеток E.coli штамма Rosetta garni В (DE3) и клеток P.pastoris штамма GS115 плазмидными ДНК pET15-6HIS-Triticain--GM, pET15-Triticain--GM-6HIS и pPIC9-Triticain--GM соответственно, получены штаммы - продуценты усеченной формы тритикаина-альфа пшеницы, с последующим культивированием клеток-хозяев, выделением экспрессирующегося белка и очисткой хроматографическими методами. Изобретение обеспечивает возможность получения вариантов биологически активного фрагмента протеазы пшеницы в растворимой форме в бактериальной и дрожжевой экспрессионных системах, а также получение препарата тритикаина-альфа с высоким и стабильным выходом, уровнем очистки и функциональной активности.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| RU2017122806A RU2676322C1 (ru) | 2017-06-28 | 2017-06-28 | Препарат цистеиновой протеиназы пшеницы тритикаина-альфа, полученной в растворимой форме, и способ получения препарата |
| PCT/RU2018/050071 WO2019004878A1 (ru) | 2017-06-28 | 2018-06-28 | Препарат цистеиновой протеиназы пшеницы тритикаина-альфа, полученной в растворимой форме, и способ получения препарата |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| EA202000021A1 true EA202000021A1 (ru) | 2020-04-22 |
| EA038578B1 EA038578B1 (ru) | 2021-09-17 |
Family
ID=64741789
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| EA202000021A EA038578B1 (ru) | 2017-06-28 | 2018-06-28 | Препарат цистеиновой протеиназы пшеницы тритикаина-альфа, полученной в растворимой форме, и способ получения препарата |
Country Status (4)
| Country | Link |
|---|---|
| US (1) | US11447780B2 (ru) |
| EA (1) | EA038578B1 (ru) |
| RU (1) | RU2676322C1 (ru) |
| WO (1) | WO2019004878A1 (ru) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110791534B (zh) * | 2019-11-27 | 2022-12-30 | 安徽师范大学 | 一种提高水溶性藻青素外源合成产量的方法 |
| RU2740319C1 (ru) * | 2020-03-19 | 2021-01-13 | Общество с ограниченной ответственностью "Альфа-Тритикаин" | Способ повышения активности тритикаина-альфа |
| CN117143257B (zh) * | 2023-10-31 | 2024-02-09 | 深圳市帝迈生物技术有限公司 | Trim28-krab-znf10二元复合物、制备方法及用于前列腺癌筛查的试剂盒 |
Family Cites Families (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8148105B2 (en) * | 2007-03-16 | 2012-04-03 | The Board Of Trustees Of The Leland Stanford Junior University | Scaleable manufacturing process for cysteine endoprotease B, isoform 2 |
| RU2603054C2 (ru) * | 2015-02-17 | 2016-11-20 | Федеральное государственное бюджетное образовательное учреждение высшего образования Первый Московский государственный медицинский университет имени И.М. Сеченова Министерства здравоохранения Российской Федерации (ФГБОУ ВО Первый МГМУ им. И.М. Сеченова Минздрава России) | Способ получения белков семейства цистеиновых протеаз пшеницы (triticum aestivum) и препарат белка тритикаин-альфа, полученный этим способом |
-
2017
- 2017-06-28 RU RU2017122806A patent/RU2676322C1/ru active
-
2018
- 2018-06-28 US US16/626,206 patent/US11447780B2/en active Active
- 2018-06-28 EA EA202000021A patent/EA038578B1/ru unknown
- 2018-06-28 WO PCT/RU2018/050071 patent/WO2019004878A1/ru active Application Filing
Also Published As
| Publication number | Publication date |
|---|---|
| EA038578B1 (ru) | 2021-09-17 |
| US11447780B2 (en) | 2022-09-20 |
| US20220162619A1 (en) | 2022-05-26 |
| RU2676322C1 (ru) | 2018-12-27 |
| WO2019004878A1 (ru) | 2019-01-03 |
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