Cloning and characterization of the glycogen branching enzyme gene existing in tandem with the glycogen debranching enzyme from Pectobacterium chrysanthemi PY35.
The glycogen branching enzyme gene (glgB) from Pectobacterium chrysanthemi PY35 was cloned, sequenced, and expressed in Escherichia coli. The glgB gene consisted of an open reading frame of 2196bp encoding a protein of 731 amino acids (calculated molecular weight of 83,859Da). The glgB gene is upstream of glgX and the ORF starts the ATG initiation codon and ends with the TGA stop codon at 2bp upstream of glgX. The enzyme was 43-69% sequence identical with other glycogen branching enzymes. The enzyme is the most similar to GlgB of E. coli and contained the four regions conserved among the alpha-amylase family. The glycogen branching enzyme (GlgB) was purified and the molecular weight of the enzyme was estimated to be 84kDa by SDS-PAGE. The glycogen branching enzyme was optimally active at pH 7 and 30 degrees C.
Lim
Woo Jin
WJ
Division of Applied Life Science, Gyeongsang National University, Chinju 660-701, Republic of Korea.
Park
Sang Ryeol
SR
Kim
Min Keun
MK
An
Chang Long
CL
Yun
Ho Jong
HJ
Hong
Su Young
SY
Kim
Eun Ju
EJ
Shin
Eun Chule
EC
Lee
Shin Woo
SW
Lim
Yong Pyo
YP
Yun
Han Dae
HD
D015061:1,4-alpha-Glucan Branching Enzyme / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D000595:Amino Acid Sequence; D001483:Base Sequence; D003001:Cloning, Molecular; D017124:Conserved Sequence; D004269:DNA, Bacterial / Q000235:genetics; D004755:Enterobacteriaceae / Q000201:enzymology* / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005798:Genes, Bacterial; D006004:Glycogen Debranching Enzyme System / Q000235:genetics*; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010802:Phylogeny; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D006004:Glycogen Debranching Enzyme System; D011994:Recombinant Proteins; D015061:1,4-alpha-Glucan Branching Enzyme