Novel cyclodextrin glucanotransferase, process for producing the same and process for producing cyclodextrin by using this enzyme

• US 20030194796A1
• Filed: 04/14/2003
• Published: 10/16/2003
• Est. Priority Date: 05/23/2000
• Status: Active Grant

• Current Assignee: Nihon Shokuhin Kako Co., Ltd.
• Original Assignee: Nihon Shokuhin Kako Co., Ltd.
• Inventors: Yamamoto, Takeshi, Sone, Hironobu, Yamamoto, Mikio, Ide, Takahiro, Watanabe, Yoshimi, Unno, Takehiro, Takada, Masayasu
• Granted Patent: US 6,924,136 B2
• Time in Patent Office: Days
• US Class Current: 435/193
• CPC Class Codes: C12N 9/1074 Cyclomaltodextrin glucanotransferase (2.4.1.19)
• CPC Class Codes: C12P 19/18 produced by the action of a glycosyl transferase, e.g. alpha-, beta- or gamma-cyclodextrins

Abstract

CGTase having the following enzymatic chemical properties: (1) Function and substrate specificity: funtioning on starch, dextrin, amylopectin, and amylose to produce primarily γ-CD, with the quantities of β- and α-CD produced being smaller than the quantity of γ-CD produced; (2) Optimum pH: 10.5-11.0; (3) Optimum temperature: About 60° C.; (4) Stable pH: 6-11; and (5) Temperature stability: With a 15 minute-treatment at 50° C., residual activity of not less than 90 percent is exhibited. A method of manufacturing cyclodextrin glucanotransferase in which a microorganism belonging to the species Bacillus clarkii (for example, FERM BP-7156) and having the ability to produce cyclodextrin glucanotransferase is cultured, cyclodextrin glucanotransferase is produced in the cultured product, and the cyclodextrin glucanotransferase that has been produced is collected. A method of manufacturing CD in which CGTase produced by the species Bacillus clarkii is reacted with a solution comprising starch or the like to produce principally γ-cyclodextrin. γ-Cyclodextrin glucanotransferase (CGTase) capable of predominantly producing γ-CD, a microorganism having the ability to produce γ-CGTase, and a method of manufacturing γ-CD using γ-CGTase are provided.

Claims

1. Cyclodextrin glucanotransferase having the enzymatic chemical properties listed below:

(1) Function and substrate specificity;

functioning on starch, dextrin, amylopectin, and amylose to produce primarily γ

-cyclodextrin, with the quantities of β

- and α

-cyclodextrin produced being smaller than the quantity of γ

-cyclodextrin produced;

(2) Optimum pH;

10.5-11.0;

(3) Optimum temperature;

About 60°

C.;

(4) Stable pH;

6-11;

(5) Temperature stability;

With a 15 minute-treatment at 50°

C., residual activity of not less than 90 percent is exhibited.

5. The manufacturing method according to claim 4, wherein the cyclodextrin glucanotransferase is the cyclodextrin glucanotransferase according to claim 1.

2. A method of manufacturing cyclodextrin glucanotransferase characterized in that a microorganism belonging to the species Bacillus clarkii and having the ability to produce cyclodextrin glucanotransferase is cultured, cyclodextrin glucanotransferase is produced in the cultured product, and the cyclodextrin glucanotransferase that has been produced is collected.

3. The manufacturing method according to claim 2, wherein the microorganism belonging to the species Bacillus clarkii and having the ability to produce cyclodextrin glucanotransferase is Bacillus clarkii strain 7364 (FERM BP-7156).

4. A method of manufacturing cyclodextrin characterized in that cyclodextrin glucanotransferase produced by the species Bacillus clarkii is reacted with a solution comprising at least one member selected from among the group consisting of starch, dextrin, amylopectin, and amylose to produce principally γ

-cyclodextrin.

6. The manufacturing method according to claim 4 or 5, characterized in that the γ

-cyclodextrin produced is separated from other cyclodextrins.

7. Bacillus clarkii strain 7364 (FERM BP-7156) being the species Bacillus clarkii having the ability to produce cyclodextrin glucanotransferase.