Characterization of a gene cluster for glycogen biosynthesis and a heterotetrameric ADP-glucose pyrophosphorylase from Bacillus stearothermophilus.
A chromosomal region of Bacillus stearothermophilus TRBE14 which contains genes for glycogen synthesis was cloned and sequenced. This region includes five open reading frames (glgBCDAP). It has already been demonstrated that glgB encodes branching enzyme (EC 2.4.1.18 [H. Takata et al., Appl. Environ. Microbiol. 60:3096-3104, 1994]). The putative GlgC (387 amino acids [aa]) and GlgD (343 aa) proteins are homologous to bacterial ADP-glucose pyrophosphorylase (AGP [EC 2.7.7.27]): the sequences share 42 to 70% and 20 to 30% identities with AGP, respectively. Purification of GlgC and GlgD indicated that AGP is an alpha2beta2-type heterotetrameric enzyme consisting of these two proteins. AGP did not seem to be an allosteric enzyme, although the activities of most bacterial AGPs are known to be allosterically controlled. GlgC protein had AGP activity without GlgD protein, but its activity was lower than that of the heterotetrameric enzyme. The GlgA (485 aa) and GlgP (798 aa) proteins were shown to be glycogen synthase (EC 2.4.1.21) and glycogen phosphorylase (EC 2.4.1.1), respectively. We constructed plasmids harboring these five genes (glgBCDAP) and assayed glycogen production by a strain carrying each of the derivative plasmids on which the genes were mutated one by one. Glycogen metabolism in B. stearothermophilus is discussed on the basis of these results.
Takata
H
H
Biochemical Research Laboratory, Ezaki Glico Co., Ltd., Nishiyodogawa-ku, Osaka, Japan. biochem-lab@glico.co.jp
Takaha
T
T
Okada
S
S
Takagi
M
M
Imanaka
T
T
D015061:1,4-alpha-Glucan Branching Enzyme / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D002876:Chromosomes, Bacterial; D004269:DNA, Bacterial; D004926:Escherichia coli; D005798:Genes, Bacterial; D001411:Geobacillus stearothermophilus / Q000201:enzymology / Q000235:genetics*; D005936:Glucans / Q000096:biosynthesis; D051599:Glucose-1-Phosphate Adenylyltransferase; D006003:Glycogen / Q000096:biosynthesis*; D006006:Glycogen Synthase / Q000737:chemistry / Q000378:metabolism; D008969:Molecular Sequence Data; D005810:Multigene Family; D009713:Nucleotidyltransferases / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism; D006005:Phosphorylases / Q000737:chemistry / Q000378:metabolism; D017386:Sequence Homology, Amino Acid
D004269:DNA, Bacterial; D005936:Glucans; D006003:Glycogen; D006005:Phosphorylases; D006006:Glycogen Synthase; D015061:1,4-alpha-Glucan Branching Enzyme; D009713:Nucleotidyltransferases; D051599:Glucose-1-Phosphate Adenylyltransferase