Method of heat-stabilizing α-glucan phosphorylase(GP)

Details Method of heat-stabilizing α-glucan phosphorylase(GP) Method of heat-stabilizing α-glucan phosphorylase(GP)

2004-06-15

2009-08-04

Saidha, Tekchand (Department: 1652)

Chemistry: molecular biology and microbiology

Enzyme , proenzyme; compositions thereof; process for...

Transferase other than ribonuclease

C536S023200

Reexamination Certificate

active

07569377

ABSTRACT:
An α-glucan phosphorylase having improved thermostability, which obtained by modifying natural α-glucan phosphorylase, and a method for producing this α-glucan phosphorylase having improved thermostability are provided. The natural α-glucan phosphorylase is derived from a plant, this α-glucan phosphorylase having improved thermostability has an amino acid residue which is different from that of the natural α-glucan phosphorylase in at least one position selected from the group consisting of a position corresponding to position4in a motif sequence1L or1H, a position corresponding to position4in a motif sequence2, and a position corresponding to position7in a motif sequence3L or3H, and wherein the enzyme activity of α-glucan phosphorylase having improved thermostability at 37° C., after heating in a 20 mM citrate buffer (pH 6.7) at 60° C. for 10 minutes, is 20% or more of the enzyme activity of the α-glucan phosphorylase having improved thermostability at 37° C., before heating.

REFERENCES:
patent: 10-14580 (1998-01-01), None
Sequence search alignment between Accession No. Q9LKJ3 (2000) and Applicants' SEQ ID No. 2.
International Search Report for corresponding Application No. PCT/JP2004/008362 mailed Aug. 17, 2004.
Shin, H.J. et al.; Formation of a αD-glucose-1-phosphate by thermophilic α-1, 4-D-glucan phosphorylase., Journal of Industrial Microbiology, vol. 24, pp. 89 to 93, 2000.
Takaha, T. et al., Structure and Properties ofThermus aquaticusα-Glucan Phosphorylase Express inEscherichia coli.J.Appl.Glycosci., vol. 48, No. 1, pp. 71 to 78, 2000.
Nakano, K. et al., The complete amino acid sequence of potato alpha-glucan phosphorylase., J.Biol.Chem., vol. 261, pp. 8230 to 8236, 1986.
Mori, H. et al., Potato tuber type H phosphory lase isozyme., Molecular cloning, nucleotide sequence, and expression of a full-length cDNA inEscherichia coli.J.Biol.Chem., vol. 266, pp. 18446 to 18453, 1991.

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