Extracellular production of a sphingomyelinase from Streptomyces griseocarneus using Streptomyces lividans.
The structural gene for sphingomyelinase (SMase) from Streptomyces griseocarneus, was introduced into Streptomyces lividans using a shuttle vector, pUC702, for Escherichia coli/S. lividans. High-level secretory production of SMase was achieved using the promoter, signal sequence and terminator regions of phospholipase D from Streptoverticillium cinnamoneum. The transformant constitutively expressed a high specific activity of SMase extracellularly during batch culture. Maximum SMase activity (555 ± 114 U/mg protein) was with 1.75 M MgCl(2) which was about 50-fold more than that with 10 mM MgCl(2).
Department of Symbiotic Systems Science and Technology, Graduate School of Symbiotic Systems Science and Technology, Fukushima University, Kanayagawa, Fukushima, Japan. sugimori@sss.fukushima-u.ac.jp
Daisuke
D
Sugimori
Yu
Y
Tomita
Yusaku
Y
Matsumoto
Chiaki
C
Ogino
D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors / Q000235:genetics; D013108:Sphingomyelin Phosphodiesterase / Q000235:genetics / Q000378:metabolism*; D013302:Streptomyces / Q000235:genetics*; D048170:Streptomyces lividans / Q000235:genetics* / Q000378:metabolism*
D013108:Sphingomyelin Phosphodiesterase