Recombinant thermostable enzyme which releases trehalose from non-reducing saccharide
AssigneeKabushiki Kaisha Hayashibara Seibutsu Kagaku Kenkyujo
InventorsHitoshi Mitsuzumi, Michio Kubota, Toshiyuki Sugimoto
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Abstract
Disclosed is a recombinant thermostable enzyme which has a molecular weight of about 54,000-64,000 daltons and a pI of about 5.6-6.6, and releases trehalose from non-reducing saccharides having a trehalose structure as an end unit and a degree of glucose polymerization of at least 3. The enzyme has a satisfactorily-high thermostability, i.e. it is not substantially inactivated even when incubated in an aqueous solution (pH 7.0) at 85° C. for 60 min, and this facilitates the production of trehalose on an industrial scale and in a satisfactorily-high yield.
Claims17
An isolated DNA molecule encoding a thermostable enzyme having the following physico-chemical properties:
- (1) Action
- Releasing trehalose from non-reducing saccharides having a trehalose structure as an end unit and a degree of glucose polymerization of at least 3, but not substantially acting on maltooligosaccharides having a degree of glucose polymerization of at least 3;
- (2) Molecular weight
- About 54,000-64,000 daltons on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE);
- and
- (3) Thermostability
- Substantially not inactivated even when incubated in an aqueous solution (pH 7.0) at 85° C. for 60 min,
- wherein said DNA molecule has a base sequence selected from the group consisting of SEQ ID NO:2 and a sequence similar to SEQ ID NO:2 where one or more bases in SEQ ID NO:2 is replaced with a different base without altering the corresponding amino acid sequence of SEQ ID NO:1.
An isolated recombinant DNA molecule comprising a nucleotide sequence encoding a thermostable enzyme having the following physico-chemical properties:
- (1) Action
- Releasing trehalose from non-reducing saccharides having a trehalose structure as an end unit and a degree of glucose polymerization of at least 3, but not substantially acting on maltooligosaccharides having a degree of glucose polymerization of at least 3;
- (2) Molecular weight
- About 54,000-64,000 daltons on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE);
- and
- (3) Thermostability
- Substantially not inactivated even when incubated in an aqueous solution (pH 7.0) at 85° C. for 60 min, and
- wherein said encoded thermostable enzyme has the amino acid sequence of SEQ ID NO:1, or a variant thereof which has the same physico-chemical properties as the thermostable enzyme of SEQ ID NO:1.
Dependencies are inferred from the claim text.