Extracellular production and characterization of two streptomyces L-asparaginases.
L-Asparaginase (ASNase) has proved its use in medical and food industries. Sequence-based screening showed the thermophilic Streptomyces strain Streptomyces thermoluteus subsp. fuscus NBRC 14270 (14270 ASNase) to positive against predicted ASNase primary sequences. The 14270 ASNase gene and four L-asparaginase genes from Streptomyces coelicolor, Streptomyces avermitilis, and Streptomyces griseus (SGR ASNase) were expressed in Streptomyces lividans using a hyperexpression vector: pTONA5a. Among those genes, only 14270 ASNase and SGR ASNase were successful for overexpression and detected in culture supernatants without an artificial signal peptide. Comparison of the two Streptomyces enzymes described above demonstrated that 14270 ASNase was superior to SGR ASNase in terms of optimum temperature, thermal stability, and pH stability.
Hatanaka
Tadashi
T
Okayama Prefectural Technology Center for Agriculture, Forestry and Fisheries, Research Institute for Biological Sciences (RIBS), Okayama, 7549-1 Kibichuo-cho, Kaga-gun, Okayama 716-1241, Japan. hatanaka@bio-ribs.com
Usuki
Hirokazu
H
Arima
Jiro
J
Uesugi
Yoshiko
Y
Yamamoto
Yukihiro
Y
Kumagai
Yuya
Y
Yamasato
Akihiro
A
Mukaihara
Takafumi
T
D000595:Amino Acid Sequence; D001215:Asparaginase / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D001216:Asparagine / Q000378:metabolism; D003001:Cloning, Molecular; D005798:Genes, Bacterial; D007527:Isoenzymes / Q000096:biosynthesis* / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D008969:Molecular Sequence Data; D010957:Plasmids / Q000235:genetics / Q000378:metabolism; D048372:Streptomyces coelicolor / Q000201:enzymology / Q000235:genetics; D013305:Streptomyces griseus / Q000201:enzymology / Q000235:genetics; D048170:Streptomyces lividans / Q000201:enzymology / Q000235:genetics; D013696:Temperature
D007527:Isoenzymes; D001216:Asparagine; D001215:Asparaginase