Cloning and characterization of a gene encoding trehalose phosphorylase (TP) from Pleurotus sajor-caju

Complementary DNA for a gene encoding trehalose phosphorylase (TP) that reversibly catalyzes trehalose synthesis and degradation from α-glucose-1-phosphate (α-Glc-1-P) and glucose was cloned from Pleurotus sajor-caju. The cDNA of P. sajor-caju TP (designated PsTP, GenBank Accession No. AF149777) enc...

Full description

Saved in:
Bibliographic Details
Published in:Protein expression and purification 2003-08, Vol.30 (2), p.194-202
Main Authors: Han, Sang-Eun, Kwon, Hawk-Bin, Lee, Seung-Bum, Yi, Bu-Young, Murayama, Ikuo, Kitamoto, Yutaka, Byun, Myung-Ok
Format: Article
Language:English
Subjects:
Amino Acid Sequence
Cloning, Molecular
Complementation of yeast Δtps1, Δtps2 mutant
Enzyme Stability
Gene Expression Regulation, Enzymologic
Genetic Complementation Test
Glucosyltransferases - chemistry
Glucosyltransferases - genetics
Glucosyltransferases - isolation & purification
Glucosyltransferases - metabolism
Hydrogen-Ion Concentration
Kinetics
Molecular Sequence Data
Pleurotus - enzymology
Pleurotus - genetics
Pleurotus sajor-caju
Saccharomyces cerevisiae
Sequence Alignment
Temperature
Trehalose
Trehalose - biosynthesis
Trehalose - chemistry
Trehalose - metabolism
Trehalose phosphorylase
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Complementary DNA for a gene encoding trehalose phosphorylase (TP) that reversibly catalyzes trehalose synthesis and degradation from α-glucose-1-phosphate (α-Glc-1-P) and glucose was cloned from Pleurotus sajor-caju. The cDNA of P. sajor-caju TP (designated PsTP, GenBank Accession No. AF149777) encodes a polypeptide of 751 amino acids with a deduced molecular mass of 83.7 kDa. The PsTP gene is expressed in mycelia, pilei, and stipes of fruiting bodies. Trehalose phosphorylase PsTP was purified from PsTP-transformed Escherichia coli. The enzyme catalyzes both the phosphorolysis of trehalose to produce α-Glc-1-P and glucose, and the synthesis of trehalose. The apparent K m values for trehalose and Pi in phosphorolytic reaction at pH 7.0 were 74.8 and 5.4 mM, respectively. The PsTP gene complemented Saccharomyces cerevisiae Δ tps1, Δ tps2 double-mutant cells, allowing their growth on glucose medium. Furthermore, yeast transformed with PsTP produced 2–2.5-fold more trehalose than non-transformants or cells transformed with empty vector only.
ISSN:1046-5928
1096-0279
DOI:10.1016/S1046-5928(03)00104-9