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.
Saito
K
K
Department of Biotechnology, Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan.
Yamazaki
H
H
Ohnishi
Y
Y
Fujimoto
S
S
Takahashi
E
E
Horinouchi
S
S
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