Digestion by serine proteases enhances salt tolerance of glutaminase in the marine bacterium Micrococcus luteus K-3.
Salt-tolerant glutaminase (Micrococcus glutaminase, with an apparent molecular mass of 48.3 kDa, intact glutaminase) from the marine bacterium Micrococcus luteus K-3 was digested using protease derived from M. luteus K-3. The digestion products were a large fragment (apparent molecular mass of 38.5 kDa, the glutaminase fragment) and small fragments (apparent molecular mass of 8 kDa). The digestion was inhibited by phenylmethanesulfonyl fluoride (PMSF). Digestion of intact glutaminase by serine proteases including trypsin, elastase, lysyl endopeptidase, and arginylendopeptidase also produced the glutaminase fragment. The N-terminus of the glutaminase fragment was the same as that of intact glutaminase. The N-termini of two small fragments were Ala394 and Ala396, respectively. The enzymological and kinetic properties of the glutaminase fragment were almost the same as those of intact glutaminase except for salt-tolerant behavior. The glutaminase fragment was a higher salt-tolerant enzyme than the intact glutaminase, suggesting that Micrococcus glutaminase is digested in the C-terminal region by serine protease from M. luteus K-3 to confer salt tolerance on glutaminase.
Yoshimune
Kazuaki
K
Department of Applied Chemistry, Faculty of Engineering, Oita University, Dannoharu, Oita 870-1192, Japan.
Yamashita
Ryoko
R
Masuo
Naohisa
N
Wakayama
Mamoru
M
Moriguchi
Mitsuaki
M
D000595:Amino Acid Sequence; D005972:Glutaminase / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D016982:Micrococcus luteus / Q000201:enzymology* / Q000235:genetics / Q000378:metabolism; D008969:Molecular Sequence Data; D008970:Molecular Weight; D010446:Peptide Fragments / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D012697:Serine Endopeptidases / Q000378:metabolism*; D012965:Sodium Chloride / Q000378:metabolism
D010446:Peptide Fragments; D011994:Recombinant Proteins; D012965:Sodium Chloride; D012697:Serine Endopeptidases; D005972:Glutaminase