Cyclodextrin glucanotransferase and its method of manufacture

• US 6,924,136 B2
• Filed: 05/23/2001
• Issued: 08/02/2005
• Est. Priority Date: 05/23/2000
• Status: Expired due to Term

• Current Assignee: Nihon Shokuhin Kako Co., Ltd.
• Original Assignee: Nihon Shokuhin Kako Co., Ltd.
• Inventors: Yamamoto, Takeshi, Ide, Takahiro, Watanabe, Yoshimi, Takada, Masayasu, Sone, Hironobu, Yamamoto, Mikio, Unno, Takehiro
• Primary Examiner(s): Gitomer, Ralph
• Application Number:
  US10/296,129
• Publication Number:
  US 20030194796A1
• Time in Patent Office: 1,532 Days
• Field of Search: 435/193, 435/71.2, 435/97, 435/183, 435/252.31, 435/832, 536/46, 536/103, 536/124
• 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 enzymatically acts on starch, dextrin, amylopectin, and amylose to produce primarily γ-CD, with quantities of β- and α-CD produced being smaller than the quantity of γ-CD produced, has an optimum pH of 10.5-11.0, an optimum temperature of about 60° C., a stable pH of 6-11, and temperature stability, exhibits residual activity of not less than 90 percent with a 15 minute-treatment at 50° C., is produced by culturing a Bacillus clarkii (for example, FERM B-7156), reacts with starch or the like to produce principally γ-cyclodextrin.

Claims

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

a) function and substrate specificity;

enzymatically acting 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;

b) optimum pH;

10.5-11.0;

c) optimum temperature;

about 60°

C.;

d) stable pH;

6-11;

e) temperature stability;

with a 15 minute-treatment at 50°

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

2. A method of manufacturing cyclodextrin glucanotransferase wherein a microorganism belonging to the species Bacillus clarkii that produces cyclodextrin glucanotransferase is cultured to produce cyclodextrin glucanotransferase, 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 ability to produce cyclodextrin glucanotransferase is Bacillus clarkii strain 7364 (FERM BP-7156).

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

-cyclodextrin.

5. The manufacturing method according to claim 4, wherein the cyclodextrin glucanotransferase having the enzymatic chemical properties listed below:

a) function and substrate specificity;

enzymatically acting 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;

b) optimum pH;

10.5-11.0;

c) optimum temperature;

about 60°

C.;

d) stable pH;

6-11;

e) temperature stability;

with a 15 minute-treatment at 50°

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

6. The manufacturing method according to claim 4, wherein 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.