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WO2002102845A1 - Procede de production d'un polypeptide fonctionnel provenant de fibroine de soie, et son utilisation - Google Patents

Procede de production d'un polypeptide fonctionnel provenant de fibroine de soie, et son utilisation Download PDF

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Publication number
WO2002102845A1
WO2002102845A1 PCT/JP2002/005663 JP0205663W WO02102845A1 WO 2002102845 A1 WO2002102845 A1 WO 2002102845A1 JP 0205663 W JP0205663 W JP 0205663W WO 02102845 A1 WO02102845 A1 WO 02102845A1
Authority
WO
WIPO (PCT)
Prior art keywords
silk
aqueous solution
chain
chain polypeptide
cocoon
Prior art date
Legal status (The legal status 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 status listed.)
Ceased
Application number
PCT/JP2002/005663
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English (en)
Japanese (ja)
Inventor
Kozo Tsubouchi
Hiroo Yamada
Yoko Takasu
Hiroshi Nakao
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
National Institute of Agrobiological Sciences
Kowa Co Ltd
Original Assignee
National Institute of Agrobiological Sciences
Kowa Co Ltd
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 National Institute of Agrobiological Sciences, Kowa Co Ltd filed Critical National Institute of Agrobiological Sciences
Priority to JP2003506317A priority Critical patent/JPWO2002102845A1/ja
Publication of WO2002102845A1 publication Critical patent/WO2002102845A1/fr
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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Classifications

    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/96—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution
    • A61K8/98—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin
    • A61K8/987—Cosmetics or similar toiletry preparations characterised by the composition containing materials, or derivatives thereof of undetermined constitution of animal origin of species other than mammals or birds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K8/00—Cosmetics or similar toiletry preparations
    • A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
    • A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
    • A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P17/00—Drugs for dermatological disorders
    • A61P17/02—Drugs for dermatological disorders for treating wounds, ulcers, burns, scars, keloids, or the like
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
    • A61Q19/00—Preparations for care of the skin
    • 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
    • C07K14/43504—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates
    • C07K14/43563—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects
    • C07K14/43586—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from invertebrates from insects from silkworms
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates to a method for producing silk fiber mouth-in L-chain polypeptide having an excellent cell growth promoting action and its application to the fields of medicine and cosmetics.
  • silk fiber mouth-in has been studied as a wound covering material in the form of a powder or a film, as it has an effect of promoting the healing of skin tissue that has been lost due to wounds, burns, etc. Nos. 2 5 4 1 6 4, 8-1 9 8 7 0 7, 9 1 9 12 9 10, 11 1 10 4 2 2 8). In addition, silk fiber mouth-in has been applied as a cosmetic because it has such an effect.
  • An object of the present invention is to provide a naturally derived component excellent in the ability to regenerate defective skin tissue, that is, a cell growth promoting effect, and to provide the use thereof. Disclosure of the invention
  • the inventors succeeded in isolating a silk fibroin H chain and a low molecular weight silk fibrous in L chain different from its degradation product. Further, they have found that the silk-fiber mouth-in L chain has an excellent skin fibroblast proliferation promoting action, and is useful as a wound healing promoter, a cell culture bed, and a cosmetic. Reached.
  • the present invention provides a method for scouring a cocoon layer or a cocoon thread of a raw cocoon, a dried cocoon or a cooked cocoon or a cocoon thread, a raw silk, a silk fabric, or a remnant thereof, and dissolving the obtained fiber-in-fiber in a neutral salt aqueous solution.
  • An object of the present invention is to provide a method for producing a silk-fiber mouth-in L-chain polypeptide characterized by fractional precipitation.
  • the present invention also provides a cell growth promoter comprising a silk-fiber opening in L chain polypeptide as an active ingredient.
  • the present invention also provides a wound healing promoting agent containing a silk-fiber opening in L chain polypeptide as an active ingredient.
  • the present invention also provides a cell culture bed containing the silk-fiber opening in L chain polypeptide.
  • the present invention also provides a cosmetic containing the silk-fiber in L chain polypeptide.
  • the present invention also provides use of silk fibrous in-L chain polypeptide for producing a cell growth promoter or a wound healing promoter.
  • the present invention provides a method for promoting wound healing, which comprises administering a silk-fiber opening in L chain polypeptide.
  • FIG. 1 is a view showing an electrophoresis (SDS-PAGE) image of each precipitate.
  • Figure 2 is a place where dissolved when the L i SCN was dissolved Fuibu opening in with C a C 1 2 It is a figure which shows the electrophoresis image of the fibroin of the case.
  • Silkworms secrete liquid silk into the silk gland cavity of the body and are called liquid silk.
  • Liquid silk consists of silk fiber mouth and silk sericin, and liquid silk fiber mouth has a molecular weight of about 370,000 (Tasi ro Yutaka and Otsuki Eiichi, Jounal of Cell Biology, Vol. 46, PI (1970) ).
  • Silk fiber mouth with a molecular weight of about 370,000 is divided into molecular weights of about 350,000 and 25,000 by reduction.
  • silk fiber mouth with a molecular weight of about 250,000 is called silk fiber mouth in L chain
  • a silk fiber mouth having a molecular weight of about 350,000 is called a silk fiber mouth heavy chain.
  • Silkworms spin liquid silk during cocoon production to make cocoons (consisting of cocoons and pupae). It is known that cocoon silk has silk fibrous mouth in the center and silk sericin around it, and the abundance ratio is 70-80 (fibrous mouth in): 20-30 (sericin).
  • Raw silk is made by collecting several to several ten cocoons (reeling). The process of removing silk sericin from cocoon, raw silk or raw weave is called scouring, and the fiber after scouring is called silk thread or silk fiber mouth. The scouring does not always completely remove silk sericin. In 5-minute kneading and 7-minute kneading, half or about 70% of sericin may be removed.
  • silk thread is used because it has undergone a scouring process.
  • Raw weave refers to unrefined silk fabric.
  • silk refers to fiber mouth-in alone, sericin alone, or coexistence of fiber mouth-in and sericin at the same time.
  • silk fiber mouth and fibroin and silk sericin and sericin have the same meaning.
  • Silk fibroin refers to silk fibroin film, silk fiber mouth powder, silk fiber mouth, etc. The silk fiber mouth is obtained by scouring cocoon thread, raw silk, raw woven fabric, or their residual yarn. Say fiber.
  • the conventional method for producing silk thread is to start with raw cocoons produced by sericulture farmers. Untreated cocoons) are dried cocoons, boiled cocoons, and reeled into raw silk to make raw weave. Next, the raw silk or raw weave is refined to obtain silk yarn or silk fabric. Debris generated in these processes is called residual yarn. Dry cocoons are prepared by gradually lowering the raw cocoons from a temperature of 115 to 12 O: to a temperature of about 80 ° C in 5 to 6 hours. Boiled cocoons are treated with steam and hot water at 100 to 105 ° C for about 10 minutes.
  • silk fiber mouths can also be obtained from liquid silk.
  • a gel-like content liquid silk
  • a gel-like content liquid silk
  • this method requires dissection of the silkworm, removal of the silk gland from the body of the silkworm, and removal of the silk fiber mouth from the silk gland cavity.
  • the maximum amount of silk-fiber mouth obtained from one silkworm is about 0.4 g.Since it is easy to contain impurities such as silk fluid and silk gland cells, and it takes time to obtain silk fibroin, It is not an industrial production method.
  • an industrially advantageous raw cocoon, a cocoon layer or a cocoon thread of a dried or boiled cocoon, or a raw silk, a silk fabric, or a residual thread thereof is used as a material. Since silk fibroin is easily decomposed during storage, it is most desirable to use fresh raw cocoons as a raw material.
  • Raw cocoons, dry cocoons, raw silk, silk threads, and their remaining threads are preferably stored at a temperature of room temperature or less to block light.
  • these raw materials are scoured to remove sericin.
  • the scouring means include urea aqueous solution treatment, alkali aqueous solution treatment, enzyme aqueous solution treatment, and high-pressure hot water treatment, and urea aqueous solution treatment is particularly preferable because it is mild. In any treatment, scouring is easier and faster as the raw material is separated.
  • the treatment with the urea aqueous solution is, for example, more than 30% ((weight (g) volume (not more than mlj, simply shown as%)) or more from the viewpoint of preventing decomposition of silk fiber mouth, reaction efficiency and industrial operability.
  • Urea Mercaptoethanol or the like may be added to the aqueous solution.
  • the treatment in this case means that the raw material is immersed in an aqueous urea solution, and stirring may be performed during this.
  • the treatment with an alkaline aqueous solution is carried out, for example, with an alkaline aqueous solution having a pH of 10 to 12 by appropriately changing the conditions at the boiling temperature under atmospheric pressure and in the range of 2 to 60 minutes. If the pH is less than 10, scouring is not sufficient, and if the pH is more than 12, silk fibroin is rapidly decomposed and control becomes difficult.
  • the preferred pH is from 10.5 to 11.5.
  • Examples of the alkaline aqueous solution to be used include an aqueous solution of an alkaline sodium salt such as sodium carbonate, sodium hydrogen carbonate, sodium silicate, sodium metasilicate, sodium phosphate, and sodium hydroxide.
  • the liquid is particularly preferable because it has an appropriate buffer effect.
  • the pH, temperature, time, etc. during the refining can be controlled by appropriately changing the processing conditions. For example, temperatures above the boiling temperature (eg, 110 ° and 120 °) When treating with C), adjust the pH toward neutral or shorten the time. When processing at a temperature lower than the boiling temperature, change the conditions appropriately, such as increasing the pH and lengthening the time. Furthermore, when using other sodium salts of sodium carbonate, it is needless to say that conditions are appropriately changed in accordance with sodium carbonate.
  • the treatment in this case means that the raw material is immersed in an aqueous alkaline solution, and stirring may be performed during this.
  • Enzyme aqueous scouring is a method in which evening protein degrading enzyme is applied to the scouring of raw silk and cocoons.
  • papain was often used, but in recent years Alcalase (Koshindo Chemical Industry) has been used.
  • Pretreatment is required for refining the enzyme aqueous solution with alcalase, and the pretreatment is performed at pH 9 to 10, preferably at pH 9.0 to 9.6. This is performed within 60 minutes, preferably within 10 minutes.
  • alcalase is added and scouring is performed at 50 to 60 ° C.
  • the scouring time is preferably within 60 minutes, particularly preferably within 20 minutes. In this case, it is preferable to separate the cocoon layer as much as possible. Also, stirring may be performed during this time.
  • Silk scouring is usually performed at temperatures near the boiling point of water at atmospheric pressure. But,
  • Refining at a high temperature of 10 o ° c or more (also called high-pressure hot water treatment or high-pressure refining) is also possible.
  • the solubility of silk sericin is greatly affected by the temperature of the water, for example, by soaking it in hot water for 30 to 100 minutes at 110 ° C, 20 to 60 minutes at 120 ° C, and 10 to 30 minutes at 130 ° C. Scouring can be performed without using a dissolving agent such as an alkali, and L chains that do not decompose can be obtained.
  • the fibroin fiber obtained by scouring is dissolved in a neutral salt aqueous solution and subjected to fractional precipitation.
  • the neutral salt examples include lithium thiocyanate, calcium chloride, lithium bromide, sodium thiocyanate, and the like.
  • concentration of the neutral salt varies depending on the type of the neutral salt, a saturated aqueous solution or a concentration of 50% saturation or more is preferable for any neutral salt. Particularly, a concentration of 80% saturated aqueous solution or more is preferable. In the case of lithium thiocyanate, the saturated aqueous solution is about 80%.
  • the pH of these aqueous neutral salt solutions is 5-8.
  • acetone-alcohol particularly ethanol
  • ethanol for crystallizing the amorphous silk fibroin.
  • the concentration of alcohol or acetone is low, the H chain selectively precipitates, and when the concentration is high, the L chain selectively precipitates.Therefore, the L chain is selectively precipitated by adjusting the concentration of these solvents. It can be done.
  • ethanol is added to a calcium chloride aqueous solution for precipitation
  • the concentration is less than 82.4%
  • mainly H chains are precipitated
  • concentration exceeds 82.4% mainly L chains are precipitated.
  • the silk-fiber-mouth L chain polypeptide thus obtained exhibits an excellent cell growth promoting action. Therefore, various cell culture additives (especially additives for animal cells including humans), cell culture beds, and wound healing promotion. It is useful as an agent or cosmetic (hereinafter, also referred to as an external preparation).
  • an agent or cosmetic hereinafter, also referred to as an external preparation.
  • silk L chain When used as a wound healing promoter, a cell culture bed, or a cosmetic, it is preferable to use silk L chain as an external preparation applied to a wound site or skin in the form of a hide mouth gel, film, powder, or the like.
  • a silk-fiber mouth-in L chain aqueous solution may be poured on a flat plate and dried.
  • this film is crushed, or the aqueous solution of silk fibroin L chain is sprayed in the air, or the aqueous solution of silk fiber mouth L chain is stirred or the precipitate obtained by adding alcohol to the aqueous solution of silk fiber mouth L chain is washed with water. It may be dried, crushed.
  • the silk-fiber mouth L chain can be used in the form of a usual external preparation, for example, an ointment, a cream, a cataplasm and the like.
  • a silk-fiber mouth L chain may be blended with a usual ointment base, a cream base, a cataplasm and the like.
  • the amount of the silk-fiber mouth L chain to be added to these external preparations varies depending on the dosage form and the base, but is 0.01 to 30 (weight / weight)%, particularly 0.1 to: L 0 (weight). / Wt)% is preferred.
  • Example 1 (Function of promoting cell growth when coated on cell culture vessel)
  • the cocoon layer of the day-crossed hybrid (13 7 x 14 6) was separated into 5 or 6 cocoons and separated well. Then, the mixture was treated with 30 times the volume (VZW) of a 5 M aqueous urea solution at 100 ° (:, 5 minutes) with good stirring to dissolve and remove the sericin. It was dried and used as a five-mouthed mouth (raw material).
  • the precipitates of these H and L chains were dissolved in 9M Li SCN, and dialyzed four times with 50 volumes of water for 30 minutes. Further, the amounts of these proteins were measured by the UV method, and diluted with water to adjust to 0.025% and 0.025%, respectively.
  • the absorbance at 275 mn of the protein solution was measured, and the protein concentration at an absorbance of 1.0 was calculated as 1 mg / mL.
  • the cells used were frozen human dermal fibroblasts (adult-derived) purchased from Kurabo Industries.
  • a low blood blue medium for human skin fibroblast proliferation purchased from Kurabo Industries was used.
  • the growth rate of the H chain and the L chain of the Petri dish coated with the in-mouth polypeptide was superior to that of the control.
  • Example 2 Cell growth promoting function when added to cell culture medium
  • the experimental method is the same as Example 1 except for the following.
  • the experiment for adding the aqueous solution of the in-chain H chain and L chain to the medium was performed as follows. Ma Put 2niL of medium into a Ruchiwell plate (6 holes, Falcon), inoculate about 100,000 cells, and replace the medium with the aqueous solution (25 ⁇ gZmL) of the in-chain H chain and L chain one day later. The culture was continued for another 4 days. Table 2 shows the number of viable cells when the fibrous opening in polypeptide was added to the control group in which the number of viable cells when the fibrous opening in polypeptide was not added was 100%.
  • Light chain (25 / mL) 11 is 1.8 99%
  • the growth rate was better than that of the control when the fibrous mouth in polypeptide was added to the medium.
  • a neutral salt solution is used as a solvent for silk protein.
  • a saturated aqueous solution of Li SCN lithium thiocyanate
  • CaCl 2 calcium chloride
  • C aC 1 2 is dissolved well silk protein, it takes a heating tends to reduce the molecular weight of the silk protein.
  • the molecular weight of the H chain is more likely to be reduced by heat, acid, alkali, light, etc. than the L chain. Therefore, as in Example 1, by dissolving the silk protein below 80 C, the recovery of H chains with C a C 1 2 can be.
  • CaCl 2 is used, part of the H chain is degraded, but L chain is hardly degraded.
  • the Fuibu port in showing the C a C 1 2 (85) and L i SCN electrophoresis image of Fuibu port in the lysates was dissolved in (room temperature) in Fig. In Fig. 2, CaCl 2 No H chain can be found in the dissolved fiber mouth.
  • H chain When dissolving the Fuiburoi down with CaC 1 2, when performing a dissolution temperature of about 85 ° C, H chain mostly decompose, most of the L chain remain without being decomposed. Thus, L chains are more stable than H chains and are easier to handle.
  • Table 3 shows the results of preparing 0.1% and 0.5% aqueous solutions of the H chain and L chain obtained in Example 1, respectively, and observing the state of the solution at room temperature.
  • the H chain aqueous solution started gelling after 1 day at a concentration of 0.5% and completely gelled after 7 days. Even at 0.1%, the viscosity increased as a sign of gelation after 1 day, a loose gel was formed after 7 days, and completely gelled after 14 days.
  • the L chain aqueous solution was completely liquid until 7 days even at a concentration of 0.5%, and the viscosity slightly increased after 14 days. It was stable for more than 14 days at a concentration of 0.1%.
  • the silk fiber mouth L chain of the present invention has an excellent cell growth promoting action and high stability, and is therefore useful as a medicine for promoting wound healing and a cosmetic for skin care.

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Abstract

L'invention concerne des promoteurs de prolifération cellulaire, des promoteurs de cicatrisation de plaie, des supports de culture cellulaire, et des cosmétiques, contenant, en tant que principe actif, un polypeptide à chaîne L de fibroïne de soie. L'invention concerne un procédé de production de ce polypeptide à chaîne L. En raison de ses excellentes propriétés de promotion de prolifération cellulaire et de sa haute fiabilité, le polypeptide à chaîne L de fibroïne de soie est utile en tant que médicament, notamment en tant que promoteur de cicatrisation de plaie, et que cosmétique pour soin de la peau.
PCT/JP2002/005663 2001-06-14 2002-06-07 Procede de production d'un polypeptide fonctionnel provenant de fibroine de soie, et son utilisation Ceased WO2002102845A1 (fr)

Priority Applications (1)

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JP2003506317A JPWO2002102845A1 (ja) 2001-06-14 2002-06-07 絹フィブロイン由来機能性ポリペプチドの製造法及びその利用

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JP2001-180169 2001-06-14
JP2001180169 2001-06-14

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Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005510268A (ja) * 2001-10-25 2005-04-21 ユニヴァーシティー オブ コネティカット 生物活性材料、生物活性材料の製法およびその使用方法
CN103290085A (zh) * 2013-05-13 2013-09-11 湖州新天丝生物技术有限公司 一种蚕丝蛋白粉及其制备方法
JP2014221011A (ja) * 2013-05-13 2014-11-27 独立行政法人物質・材料研究機構 細胞培養用支持体の製造方法、細胞培養用支持体および細胞培養方法
JP2015100326A (ja) * 2013-11-27 2015-06-04 国立研究開発法人農業生物資源研究所 カイコのセリシン１変異系統
WO2015190292A1 (fr) * 2014-06-12 2015-12-17 株式会社アーダン Pommade contenant de la fibroïne hydrolysée et son procédé de production
WO2019054506A1 (fr) * 2017-09-15 2019-03-21 Ｓｐｉｂｅｒ株式会社 Procédé de fabrication de fibrilles
KR102295015B1 (ko) * 2020-03-27 2021-08-27 주식회사 바이오스탠다드 수용화 상태로 장기간 안정성을 가지는 실크 피브로인 단백질 수용액

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04300897A (ja) * 1991-03-28 1992-10-23 Kiyoichi Matsumoto 絹フィブロイン溶液
JPH07173192A (ja) * 1993-12-16 1995-07-11 Kiyoichi Matsumoto 絹フィブロインの新規溶媒
JPH11180999A (ja) * 1997-12-25 1999-07-06 T Hasegawa Co Ltd 精製絹フィブロイン溶液、その製造法及びそれを含有する飲食品
JP2001122894A (ja) * 1999-10-23 2001-05-08 Rep Korea ゲル濾過法による高純度シルクペプチドの製造方法
JP2001163899A (ja) * 1999-12-09 2001-06-19 Natl Inst Of Sericultural & Entomological Science 機能性絹フィブロインの製造方法とその利用

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH04300897A (ja) * 1991-03-28 1992-10-23 Kiyoichi Matsumoto 絹フィブロイン溶液
JPH07173192A (ja) * 1993-12-16 1995-07-11 Kiyoichi Matsumoto 絹フィブロインの新規溶媒
JPH11180999A (ja) * 1997-12-25 1999-07-06 T Hasegawa Co Ltd 精製絹フィブロイン溶液、その製造法及びそれを含有する飲食品
JP2001122894A (ja) * 1999-10-23 2001-05-08 Rep Korea ゲル濾過法による高純度シルクペプチドの製造方法
JP2001163899A (ja) * 1999-12-09 2001-06-19 Natl Inst Of Sericultural & Entomological Science 機能性絹フィブロインの製造方法とその利用

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
OYAMA F. ET AL.: "Studies on immunological properties of fibroin heavy and light chains", J. BIOCHEM., vol. 96, no. 6, 1984, TOKYO, pages 1689 - 1694, XP002954485 *
YAMAGUCHI K. ET AL.: "Primary structure of the silk fibroin light chain determined by cDNA sequencing and peptide analysis", J. MOL. BIOL., vol. 210, no. 1, 1989, pages 127 - 139, XP002954484 *

Cited By (10)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2005510268A (ja) * 2001-10-25 2005-04-21 ユニヴァーシティー オブ コネティカット 生物活性材料、生物活性材料の製法およびその使用方法
CN103290085A (zh) * 2013-05-13 2013-09-11 湖州新天丝生物技术有限公司 一种蚕丝蛋白粉及其制备方法
JP2014221011A (ja) * 2013-05-13 2014-11-27 独立行政法人物質・材料研究機構 細胞培養用支持体の製造方法、細胞培養用支持体および細胞培養方法
JP2015100326A (ja) * 2013-11-27 2015-06-04 国立研究開発法人農業生物資源研究所 カイコのセリシン１変異系統
WO2015190292A1 (fr) * 2014-06-12 2015-12-17 株式会社アーダン Pommade contenant de la fibroïne hydrolysée et son procédé de production
JP2016000710A (ja) * 2014-06-12 2016-01-07 株式会社アーダン 加水分解フィブロインを含む軟膏及びその製造方法
WO2019054506A1 (fr) * 2017-09-15 2019-03-21 Ｓｐｉｂｅｒ株式会社 Procédé de fabrication de fibrilles
JPWO2019054506A1 (ja) * 2017-09-15 2020-10-22 Ｓｐｉｂｅｒ株式会社 フィブリルの製造方法
JP7303550B2 (ja) 2017-09-15 2023-07-05 Ｓｐｉｂｅｒ株式会社 フィブリルの製造方法
KR102295015B1 (ko) * 2020-03-27 2021-08-27 주식회사 바이오스탠다드 수용화 상태로 장기간 안정성을 가지는 실크 피브로인 단백질 수용액

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