WO2011083735A1 - Matériau composite utilisant des cristaux de protéine et son procédé de production - Google Patents

Matériau composite utilisant des cristaux de protéine et son procédé de production Download PDF

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Publication number
WO2011083735A1
WO2011083735A1 PCT/JP2010/073782 JP2010073782W WO2011083735A1 WO 2011083735 A1 WO2011083735 A1 WO 2011083735A1 JP 2010073782 W JP2010073782 W JP 2010073782W WO 2011083735 A1 WO2011083735 A1 WO 2011083735A1
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WIPO (PCT)
Prior art keywords
composite material
protein
metal
crystal
item
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PCT/JP2010/073782
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English (en)
Japanese (ja)
Inventor
隆史 上野
聡 安部
進 北川
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国立大学法人京都大学
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Application filed by 国立大学法人京都大学 filed Critical 国立大学法人京都大学
Priority to JP2011548973A priority Critical patent/JPWO2011083735A1/ja
Publication of WO2011083735A1 publication Critical patent/WO2011083735A1/fr

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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
    • 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
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • 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)
    • C12N9/24—Hydrolases (3) acting on glycosyl compounds (3.2)
    • C12N9/2402—Hydrolases (3) acting on glycosyl compounds (3.2) hydrolysing O- and S- glycosyl compounds (3.2.1)
    • C12N9/2462—Lysozyme (3.2.1.17)
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P41/00—Processes using enzymes or microorganisms to separate optical isomers from a racemic mixture

Definitions

  • the present invention relates to a composite material using protein crystals and a method for producing the same.
  • Metal activity varies greatly depending on the carrier on which the metal is supported.
  • the metal carrier inorganic materials such as silica, titania, zirconia, zeolite and activated carbon are often used.
  • Non-Patent Document 1 discloses that Pd and Pt are supported after virus particles are crosslinked with glutaraldehyde.
  • Non-Patent Document 2 discloses that Ag or Au is supported on tetragonal lysozyme.
  • proteins have many functional groups that can coordinate with metals such as SH, COOH, and NH 2 , so that metal ions can be coordinated, but retain metal atoms in their pores. Protein crystals are not known.
  • An object of the present invention is to provide a composite material in which a metal is supported in pores of a protein crystal.
  • the present invention provides the following composite material and method for producing the same.
  • Item 1 A composite material having a metal in the pores of a protein crystal.
  • the metal is Ti, V, Cr, Mn, Fe, Co, Ni, Zn, Ga, Ge, Zr, Nb, Mo, Tc, Ru, Rh, Pd, Ag, Cd, In, Sn, Hf, Ta, W.
  • the metal is at least one selected from the group consisting of Ru, Rh, Pd, Ag, Cd, In, Sn, Hf, Ta, W, Re, Os, Ir, Pt, Au, Hg, Tl, and Pb.
  • Item 4. The composite material according to Item 1.
  • Item 4. Item 2. The composite material according to Item 1, wherein the metal is a metal exhibiting at least one type of ferromagnetism selected from the group consisting of Fe, Co, Ni, and Gd.
  • the composite material according to Item 1, wherein the crystal is selected from the group consisting of tetragonal, orthorhombic, triclinic, monoclinic, hexagonal, trigonal and cubic.
  • the protein is selected from the group consisting of thermolysin, elastase, esterase, lipase, nitrilase, hydantoinase, protease, asparaginase, urease, insulin, somatotin, glucose isomerase, xylanase, concanavalin A, ribonuclease, myoglobin, ferritin and lysozyme.
  • the composite material according to 1. Item 7.
  • the protein is selected from the group consisting of thermolysin, elastase, esterase, lipase, nitrilase, hydantoinase, protease, asparaginase, urease, insulin, somatotin, glucose isomerase, xylanase, concanavalin A, ribonuclease, myoglobin, ferritin and lysozyme.
  • the composite material according to 1. Item 8.
  • Item 2. The composite material according to Item 1, wherein the protein is lysozyme.
  • Item 9 A method for producing a composite material having a metal in pores of a protein crystal, comprising impregnating a protein crystal with a solution of a metal compound, washing the protein, and reducing metal ions bound to the protein.
  • a composite material in which metal is filled in the pores of a protein crystal can be obtained.
  • the inside of these pores is an asymmetric environment with functional groups such as protein amino groups, carboxyl groups, imidazole groups, thiol groups, and a specific configuration. Suitable as a carrier.
  • the enzyme can be stabilized and a composite material with metal can be provided while maintaining the activity of the enzyme.
  • a metal exhibiting ferromagnetism such as Fe, Co, Ni, Gd is used as the metal, a composite material having magnetism can be obtained.
  • FIG. 4 shows that high magnetic anisotropy (Hc) is obtained by increasing the Co content.
  • A: CoPt / tetragonal crystal HEWL Co: Pt 1: 41.
  • B: CoPt / orthogonal crystal HEWL Co: Pt 1: 5.
  • a transmission electron microscope (TEM) image of a composite material synthesized in the presence of Co ions, Pt ions, and cross-linked lysozyme crystals and a histogram of fine particle diameter are shown.
  • the protein used in the present invention is not particularly limited as long as it can be crystallized, and includes any protein.
  • proteins include enzymes, proteins that form biological structures (collagen, keratin, etc.), hormones, receptors, muscle constituent proteins (actin, myosin, etc.), antibodies, nutritional proteins (egg, soybean, milk, etc.) Protein), albumin and the like.
  • the enzyme include hydrolase, isomerase, lyase, ligase, transferase, and oxidoreductase, and hydrolase is preferable.
  • hydrolases examples include thermolysin, elastase, esterase, lipase, nitrilase, hydantoinase, protease, asparaginase, urease, lysozyme and the like, and lysozyme is preferred.
  • protein crystals examples include tetragonal, orthorhombic, triclinic, monoclinic, hexagonal, trigonal and cubic, with orthorhombic and tetragonal being preferred.
  • the size of the pores is 0.1 to 500 nm, preferably 0.5 to 50 nm.
  • a crystal having a larger pore size is preferable because it can carry a large amount of metal.
  • the protein crystal may support the metal as it is, but it is preferable to stabilize the structure by crosslinking the protein with a crosslinking agent.
  • the protein cross-linking agents cross-linking agents having an aldehyde group such as glutaraldehyde 2 or more, formaldehyde, or an isocyanate, isothiocyanate, maleimide, OH such as succinimide, SH, and side chain functional groups of the protein, such as NH 2 reaction
  • a cross-linking agent having two or more groups capable of forming the same is used in an amount of 0.1 to 100,000 parts by weight, preferably 1 to 5000 parts by weight, and more preferably 10 to 200 parts by weight with respect to 100 parts by weight of protein crystals.
  • metals Ti, V, Cr, Mn, Fe, Co, Ni, Zn, Ga, Ge, Zr, Nb, Mo, Tc, Ru, Rh, Pd, Ag, Cd, In, Sn, Hf, Ta, W, Re, Os, Ir, Pt, Au, Hg, Tl, Pb and the like can be mentioned, and one or more of these metals can be used.
  • Metals are water, alcohol (methanol, ethanol, isopropanol, etc.), acetone in the form of metal compounds such as acetate, nitrate, fluoride, chloride, bromide, iodide, sulfate, perchlorate, phosphate, etc. It is dissolved in a polar solvent such as ketone, DMF, DMSO.
  • a composite material having a metal in the pores of the protein crystal can be produced by impregnating a protein crystal in the metal compound solution, washing, and then reducing the metal ion bound to the protein.
  • a metal exhibiting ferromagnetism such as Fe, Co, Ni, or Gd is used alone or in combination with another metal, a composite material having magnetism can be obtained.
  • the metal is supported in an amount of 0.1 to 1,000,000 parts by weight, preferably 1 to 1000 parts by weight, more preferably 10 to 100 parts by weight, per 100 parts by weight of protein crystals.
  • the metal can be bonded to both the surface of the crystal and the pores, but the surface area is overwhelmingly large in the pores, so that it is substantially supported in the pores.
  • the protein crystal is impregnated with the solvent of the metal compound, and the metal ion is captured in the protein side chain functional group (NH 2 , COOH, imidazole, SH, etc.) in the pores of the protein.
  • protein side chain functional group NH 2 , COOH, imidazole, SH, etc.
  • metal ions those having weak bonds with these side chain functional groups are removed in the subsequent washing process, but those having strong bonds with side chain functional groups are retained in the pores of the protein crystal.
  • metals softer large transition metal ions (Ru, Rh, Pd, Ag, Cd, In, Sn, Hf, Ta, W, Re, Os, Ir, Pt, Au, Hg, Tl, Pb, etc.) are fine. Strongly held in the hole.
  • These metals and metals (Ti, V, Cr, Mn, Fe, Co, Ni, Zn) that are slightly weakly bonded to proteins in the pores are removed by washing depending on the type of protein or crystal structure.
  • the held metal ions are reduced by a reducing agent, and a metal is deposited in the pores.
  • the reducing agent include metal hydrides such as NaBH 4 , hypophosphite, sulfite, ascorbic acid, light, methanol, hydrazine, electrochemical reduction, and the like, and NaBH 4 is preferable.
  • CO: Pt addition is the concentration ratio of CoCl 2 and K 2 PtCl 4 in which crystals are immersed.

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  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Zoology (AREA)
  • Engineering & Computer Science (AREA)
  • Wood Science & Technology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Peptides Or Proteins (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Abstract

L'invention porte sur un matériau composite, un métal étant contenu dans un pore de cristal de protéine ; et sur son procédé de production.
PCT/JP2010/073782 2010-01-05 2010-12-28 Matériau composite utilisant des cristaux de protéine et son procédé de production WO2011083735A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
JP2011548973A JPWO2011083735A1 (ja) 2010-01-05 2010-12-28 タンパク質結晶を用いた複合材料及びその製造方法

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2010-000366 2010-01-05
JP2010000366 2010-01-05

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WO2011083735A1 true WO2011083735A1 (fr) 2011-07-14

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2018519838A (ja) * 2015-07-15 2018-07-26 ザイムトロニクス エルエルシーＺｙｍｔｒｏｎｉｘ， Ｌｌｃ 自動バイオナノ触媒製造

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008088059A (ja) * 2006-09-07 2008-04-17 Univ Nagoya 改変アポフェリチンの製造方法とその利用

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2008088059A (ja) * 2006-09-07 2008-04-17 Univ Nagoya 改変アポフェリチンの製造方法とその利用

Non-Patent Citations (11)

* Cited by examiner, † Cited by third party
Title
HIROYASU TABE ET AL.: "Kinzoku Ion ni yoru Tanpakushitsu Kessho no Kinoka", CSJ: THE CHEMICAL SOCIETY OF JAPAN KOEN YOKOSHU, vol. 90, no. 3, March 2010 (2010-03-01), pages 631 *
SATOSHI ABE ET AL.: "Metal Accumulation on the Surface of Porous Protein Crystal", CSJ: THE CHEMICAL SOCIETY OF JAPAN KOEN YOKOSHU, vol. 89, no. 2, 2009, pages 1259 *
SATOSHI ABE ET AL.: "Takosei Tanpakushitsu Kessho eno Kinzoku Ion Shuseki o Riyo shita Co-Pt Bimetal Ryushi no Sakusei to Kinoka", SYMPOSIUM ON COORDINATION CHEMISTRY OF JAPAN KOEN YOSHISHU, vol. 60, September 2010 (2010-09-01), pages 157 *
SATOSHI ABE ET AL.: "Takosei Tanpakushitsu Kessho o Riyo shita Muki Zairyo Gosei", POLYMER PREPRINTS, vol. 58, no. 2, 2009, JAPAN, pages 5245 *
SATOSHI ABE ET AL.: "Tanpakushitsu Kessho Saiko Kukannai deno Kinzoku Zairyo no Gosei to Kinoka", CSJ: THE CHEMICAL SOCIETY OF JAPAN KOEN YOKOSHU, vol. 90, no. 3, March 2010 (2010-03-01), pages 642 *
SATOSHI ABE ET AL.: "Tanpakushitsu Kessho Takosei Kukan o Riyo shita Kinzoku Ion no Shuseki", POLYMER PREPRINTS, vol. 57, no. 2, 2008, JAPAN, pages 4862 *
TAKAFUMI UENO: "Design of Protein Scaffolds for Chemical Reactions Catalyzed by Metal Complexes and Nanoparticles", BULL. JPN. SOC. COORD. CHEM., vol. 51, 2008, pages 20 - 30 *
TAKASHI UENO ET AL.: "Tanpakushitsu Kessho Kukan o Riyo shita Kinzoku Sakutai Shuseki", SYMPOSIUM ON COORDINATION CHEMISTRY OF JAPAN KOEN YOSHISHU, vol. 58, 2008, pages 118 *
UENO, T. ET AL.: "Crystal structure based design of functional metal/protein hybrids.", J. INORG. BIOCHEM., vol. 101, no. 11-12, 2007, pages 1667 - 1675, XP022382121, DOI: doi:10.1016/j.jinorgbio.2007.06.025 *
UENO, T. ET AL.: "Elucidation of metal-ion accumulation induced by hydrogen bonds on protein surfaces by using porous lysozyme crystals containing RhIII ions as the model surfaces.", CHEM. EUR. J., vol. 16, no. 9, March 2010 (2010-03-01), pages 2730 - 2740 *
UENO, T. ET AL.: "Process of accumulation of metal ions on the interior surface of apo- ferritin: crystal structure of a series of apo- ferritins containing variable quantities of Pd(II) ions.", J. AM. CHEM. SOC., vol. 131, no. 14, 2009, pages 5094 - 5100 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JP2018519838A (ja) * 2015-07-15 2018-07-26 ザイムトロニクス エルエルシーＺｙｍｔｒｏｎｉｘ， Ｌｌｃ 自動バイオナノ触媒製造

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