JPH0699239B2 - Single crystal manufacturing method - Google Patents
Single crystal manufacturing methodInfo
- Publication number
- JPH0699239B2 JPH0699239B2 JP26063086A JP26063086A JPH0699239B2 JP H0699239 B2 JPH0699239 B2 JP H0699239B2 JP 26063086 A JP26063086 A JP 26063086A JP 26063086 A JP26063086 A JP 26063086A JP H0699239 B2 JPH0699239 B2 JP H0699239B2
- Authority
- JP
- Japan
- Prior art keywords
- single crystal
- crystal
- protein
- temperature
- nucleic acid
- 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.)
- Expired - Lifetime
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C30—CRYSTAL GROWTH
- C30B—SINGLE-CRYSTAL GROWTH; UNIDIRECTIONAL SOLIDIFICATION OF EUTECTIC MATERIAL OR UNIDIRECTIONAL DEMIXING OF EUTECTOID MATERIAL; REFINING BY ZONE-MELTING OF MATERIAL; PRODUCTION OF A HOMOGENEOUS POLYCRYSTALLINE MATERIAL WITH DEFINED STRUCTURE; SINGLE CRYSTALS OR HOMOGENEOUS POLYCRYSTALLINE MATERIAL WITH DEFINED STRUCTURE; AFTER-TREATMENT OF SINGLE CRYSTALS OR A HOMOGENEOUS POLYCRYSTALLINE MATERIAL WITH DEFINED STRUCTURE; APPARATUS THEREFOR
- C30B7/00—Single-crystal growth from solutions using solvents which are liquid at normal temperature, e.g. aqueous solutions
Landscapes
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Crystallography & Structural Chemistry (AREA)
- Materials Engineering (AREA)
- Metallurgy (AREA)
- Organic Chemistry (AREA)
- Crystals, And After-Treatments Of Crystals (AREA)
Description
【発明の詳細な説明】 〔発明の概要〕 本発明は、タンパク質等の生体高分子物質の単結晶製造
において、結晶核の数を制御することが困難である点を
解決するため、飽和点近傍の条件で、短時間に温度上昇
または下降させることにより不定形沈殿を作り、これに
より結晶核が形成された後、温度を元に復帰させて不定
形沈殿を消失せしめ、すでに生成された結晶核の成長の
みを行わせるようにしたものである。DETAILED DESCRIPTION OF THE INVENTION [Outline of the Invention] The present invention solves the problem that it is difficult to control the number of crystal nuclei in the production of single crystals of biopolymers such as proteins. Under the conditions of, the amorphous precipitate is created by raising or lowering the temperature in a short time, and after the crystal nucleus is formed by this, the temperature is returned to the original state to eliminate the amorphous precipitate, and the crystal nucleus already generated. It is designed to allow only the growth of.
本発明は、生体高分子物質の単結晶製造方法に関する。 The present invention relates to a method for producing a single crystal of a biopolymer material.
生体高分子単結晶は、タンパク質工学（プロテインエン
ジニアリング）による新しい機能をもつ分子の設計にお
いて重要である。また、高度に配列されたこれらの高分
子は、光電変換等に関する素材として利用することが可
能である。Biopolymer single crystals are important in the design of molecules with new functions by protein engineering. Further, these highly aligned polymers can be used as materials for photoelectric conversion and the like.
従来、タンパク質等の結晶の製造に際しては、タンパク
質等の水溶液に硫酸アンモニウム等の沈殿剤を添加し、
わずかに不定形（アモルファス）のタンパク質沈殿が生
ずる条件とした後、これを静置して結晶化を行わせてい
る。Conventionally, when producing crystals of protein or the like, a precipitating agent such as ammonium sulfate is added to an aqueous solution of protein or the like,
After the conditions under which slightly amorphous (amorphous) protein precipitation occurs, this is left to stand for crystallization.
しかして、かかるタンパク質等の結晶製造に当たり、大
型で格子欠陥の少ない結晶を得ることが要求される。こ
こにおいて、良質の結晶を得るために、ゆっくりと不定
形沈殿を生じさせることが重要であり、半透膜を介して
沈殿剤を加える等の方法が取られていた。Therefore, in producing crystals of such proteins, it is required to obtain crystals of large size and few lattice defects. Here, in order to obtain high-quality crystals, it is important to slowly cause amorphous precipitation, and methods such as adding a precipitant through a semipermeable membrane have been taken.
しかるに、かかる方法では、結晶核の形成と結晶の成長
をそれぞれ独立に調整することができないため、核の数
を制御することが困難であり、また結晶成長速度を制御
することも難しかった。However, in such a method, the formation of crystal nuclei and the growth of crystals cannot be adjusted independently, so that it is difficult to control the number of nuclei and it is also difficult to control the crystal growth rate.
本発明は、従って、かかる従来技術の問題点を解消し、
より大きな単結晶を生成することのできる方法を提供す
ることを目的とする。The present invention thus solves the problems of the prior art,
An object is to provide a method capable of producing a larger single crystal.
本発明によれば即ち液体溶媒中においてタンパク質また
は核酸の単結晶を製造する方法が提供されるのであっ
て、この方法は、タンパク質または核酸の液体溶媒中の
溶液を一時的に昇温もしくは降温することにより前記タ
ンパク質または核酸の結晶核を形成させ、その後前記溶
液の温度を所定の温度に変えることによって不定形沈殿
物が析出しない条件として、すでに形成された結晶核を
おだやかに成長させることを特徴とする。According to the present invention, that is, there is provided a method for producing a single crystal of a protein or nucleic acid in a liquid solvent, which method temporarily raises or lowers the temperature of a solution of the protein or nucleic acid in the liquid solvent. Thereby forming crystal nuclei of the protein or nucleic acid, and then changing the temperature of the solution to a predetermined temperature to gently grow the already formed crystal nuclei under the condition that an amorphous precipitate does not precipitate. And
本発明では、不定形沈殿物に比べて結晶の溶解度が低い
ことに着目し、一旦不定形沈殿を作り、結晶核を形成さ
せた後、結晶核のみを残し、不定形沈殿を消失させるこ
とによって、その後の新たな核形成を抑えるとともに、
核の成長をゆるやかに行わせることにより欠陥の少ない
結晶を生成させるものである。In the present invention, focusing on the fact that the solubility of crystals is lower than that of amorphous precipitates, once the amorphous precipitates are formed, and after the crystal nuclei are formed, only the crystal nuclei are left and the amorphous precipitates are eliminated. , While suppressing new nucleation after that,
By gradually growing the nuclei, crystals with few defects are generated.
本発明で用いることのできるタンパク質の例としては、
ミオグロビン、リボヌクレアーゼＳなどがある。また、
核酸の例としては、転移RNAなどがある。Examples of proteins that can be used in the present invention include:
Examples include myoglobin and ribonuclease S. Also,
Examples of nucleic acids include transfer RNA.
本発明の方法においては、核形成の条件を制御できるた
め、核の数を制限することができ、より大きな単結晶の
製造を可能とする。また、結晶の成長条件を独立に調整
することが可能であるため、双晶等の形成が抑えられ、
良質の単結晶を得ることができる。In the method of the present invention, since the conditions for nucleation can be controlled, the number of nuclei can be limited, and a larger single crystal can be produced. In addition, since it is possible to independently adjust the crystal growth conditions, formation of twins and the like can be suppressed,
A high quality single crystal can be obtained.
以下に、実施例を挙げ、本発明をさらに説明する。 Hereinafter, the present invention will be further described with reference to examples.
ミオグロビン1gを30mlの燐酸ナトリウム緩衝液に溶解す
る。別に、20℃で調製した飽和硫酸アンモニウム水溶液
70mlを加え、混合して均一化する。得られた液を、密閉
できる容器に入れ、37℃で６時間放置して不定形沈殿を
生じさせた後、20℃に降温してそのまま静置する。２日
後に小さな結晶数個が観察され、２週間後には1mm程度
の単結晶が得られた。1 g of myoglobin is dissolved in 30 ml of sodium phosphate buffer. Separately, saturated ammonium sulfate aqueous solution prepared at 20 ° C
Add 70 ml, mix and homogenize. The obtained solution is placed in a container which can be sealed and left at 37 ° C. for 6 hours to cause an amorphous precipitate, then the temperature is lowered to 20 ° C. and the mixture is allowed to stand as it is. Several small crystals were observed after 2 days, and a single crystal of about 1 mm was obtained after 2 weeks.
なお、上昇（ないし下降）させる温度および時間を制御
することにより核の形成数が制御可能である。The number of nuclei formed can be controlled by controlling the temperature and time for raising (or lowering).
本発明によれば、結晶核の数を制限できるため、同量の
出発材料からより大きな単結晶を得ることができる。ま
た、結晶がゆっくり成長するため、良質の単結晶を得る
ことができる。According to the present invention, since the number of crystal nuclei can be limited, a larger single crystal can be obtained from the same amount of starting material. Further, since the crystal grows slowly, a good quality single crystal can be obtained.
───────────────────────────────────────────────────── フロントページの続き (72)発明者 端谷 隆文 神奈川県川崎市中原区上小田中1015番地 富士通株式会社内 (72)発明者 河又 聡子 神奈川県川崎市中原区上小田中1015番地 富士通株式会社内 ─────────────────────────────────────────────────── ─── Continuation of the front page (72) Takafumi Hataya, Takafumi Hataya, 1015 Kamiodanaka, Nakahara-ku, Kawasaki-shi, Kanagawa, Fujitsu Limited Within
Claims (1)
の単結晶を製造するに当たり、タンパク質または核酸の
液体溶媒中の溶液を一時的に昇温もしくは降温すること
により前記タンパク質または核酸の結晶核を形成させ、
その後前記溶液の温度を所定の温度に変えることによっ
て不定形沈澱物が析出しない条件として、すでに形成さ
れた結晶核をおだやかに成長させることを特徴とする、
単結晶製造方法。1. When producing a single crystal of a protein or nucleic acid in a liquid solvent, a crystal nucleus of the protein or nucleic acid is formed by temporarily raising or lowering the temperature of a solution of the protein or nucleic acid in the liquid solvent. ,
Thereafter, by changing the temperature of the solution to a predetermined temperature, as a condition that the amorphous precipitate does not precipitate, the already formed crystal nuclei are gently grown,
Single crystal manufacturing method.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP26063086A JPH0699239B2 (en) | 1986-11-04 | 1986-11-04 | Single crystal manufacturing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP26063086A JPH0699239B2 (en) | 1986-11-04 | 1986-11-04 | Single crystal manufacturing method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| JPS63117999A JPS63117999A (en) | 1988-05-21 |
| JPH0699239B2 true JPH0699239B2 (en) | 1994-12-07 |
Family
ID=17350588
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| JP26063086A Expired - Lifetime JPH0699239B2 (en) | 1986-11-04 | 1986-11-04 | Single crystal manufacturing method |
Country Status (1)
| Country | Link |
|---|---|
| JP (1) | JPH0699239B2 (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPH0745359B2 (en) * | 1987-12-25 | 1995-05-17 | 東レ株式会社 | Method for producing organic compound crystal |
| JP5929480B2 (en) * | 2012-02-01 | 2016-06-08 | 三菱レイヨン株式会社 | Method for purifying methacrylic acid |
-
1986
- 1986-11-04 JP JP26063086A patent/JPH0699239B2/en not_active Expired - Lifetime
Also Published As
| Publication number | Publication date |
|---|---|
| JPS63117999A (en) | 1988-05-21 |
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