Production of trehalose synthase from a basidiomycete, Grifola frondosa, in Escherichia coli.
The genomic DNA and cDNA for a gene encoding a novel trehalose synthase (TSase) catalyzing trehalose synthesis from alpha-D-glucose 1-phosphate and D-glucose were cloned from a basidiomycete, Grifola frondosa. Nucleotide sequencing showed that the 732-amino-acid TSase-encoding region was separated by eight introns. Consistent with the novelty of TSase, there were no homologous proteins registered in the data-bases. Recombinant TSase with a histidine tag at the NH2-terminal end, produced in Escherichia coli, showed enzyme activity similar to that purified from the original G. frondosa strain. Incubation of alpha-D-glucose 1-phosphate and D-glucose in the presence of recombinant TSase generated trehalose, in agreement with the enzymatic property of TSase that the equilibrium lay far in the direction of trehalose synthesis.
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan.
K
K
Saito
H
H
Yamazaki
Y
Y
Ohnishi
S
S
Fujimoto
E
E
Takahashi
S
S
Horinouchi
D000595:Amino Acid Sequence; D001483:Base Sequence; D001487:Basidiomycota / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development; D003001:Cloning, Molecular; D018076:DNA, Complementary; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics*; D005947:Glucose / Q000378:metabolism; D005964:Glucosyltransferases / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis; D017422:Sequence Analysis, DNA; D014199:Trehalose / Q000096:biosynthesis
D018076:DNA, Complementary; D011994:Recombinant Proteins; D014199:Trehalose; D005964:Glucosyltransferases; C099692:trehalose synthase; D005947:Glucose